Sample records for human tph2 transcripts

  1. Tryptophan hydroxylase 2 (TPH2) in a neuronal cell line: modulation by cell differentiation and NRSF/rest activity.

    PubMed

    Gentile, Maria Teresa; Nawa, Yukino; Lunardi, Gianluigi; Florio, Tullio; Matsui, Hiroaki; Colucci-D'Amato, Luca

    2012-12-01

    Serotonin (5-HT) is a neurotransmitter involved in many aspects of the neuronal function. The synthesis of 5-HT is initiated by the hydroxylation of tryptophan, catalyzed by tryptophan hydroxylase (TPH). Two isoforms of TPH (TPH1 and TPH2) have been identified, with TPH2 almost exclusively expressed in the brain. Following TPH2 discovery, it was reported that polymorphisms of both gene and non-coding regions are associated with a spectrum of psychiatric disorders. Thus, insights into the mechanisms that specifically regulate TPH2 expression and its modulation by exogenous stimuli may represent a new therapeutic approach to modify serotonergic neurotransmission. To this aim, a CNS-originated cell line expressing TPH2 endogenously represents a valid model system. In this study, we report that TPH2 transcript and protein are modulated by neuronal differentiation in the cell line A1 mes-c-myc (A1). Moreover, we show luciferase activity driven by the human TPH2 promoter region and demonstrate that upon mutation of the NRSF/REST responsive element, the promoter activity strongly increases with cell differentiation. Our data suggest that A1 cells could represent a model system, allowing an insight into the mechanisms of regulation of TPH2 and to identify novel therapeutic targets in the development of drugs for the management of psychiatric disorders. © 2012 The Authors Journal of Neurochemistry © 2012 International Society for Neurochemistry.

  2. Association of regulatory TPH2 polymorphisms with higher reduction in depressive symptoms in children and adolescents treated with fluoxetine.

    PubMed

    Gassó, Patricia; Rodríguez, Natalia; Boloc, Daniel; Blázquez, Ana; Torres, Teresa; Gortat, Ana; Plana, Maria Teresa; Lafuente, Amalia; Mas, Sergi; Arnaiz, Joan Albert; Lázaro, Luisa

    2017-07-03

    Genetic variability related to the brain serotonergic system has a significant impact on both the susceptibility to psychiatric disorders, such as major depressive disorder (MDD), and the response to antidepressant drugs, such as fluoxetine. TPH2 is one of the most important serotonergic candidate genes in selective serotonin reuptake inhibitors (SSRIs) pharmacogenetic studies. The aim of the present study was to evaluate the influence of regulatory polymorphisms that are specifically located in human TPH2 transcription factor binding sites (TFBSs), and therefore could be functional by altering gene expression, on clinical improvement in children and adolescents treated with fluoxetine. The selection of SNPs was also based on their linkage disequilibrium with TPH2 rs4570625, a genetic variant with questionable functionality, which was previously associated with clinical response in our pediatric population. A total of 83 children and adolescents were clinically evaluated 12weeks after initiating antidepressant treatment with fluoxetine for the first time. Clinical improvement was assessed by reductions in depressive symptoms measured using the Children's Depression Inventory (CDI) scale. The polymorphisms rs11179002, rs60032326 and rs34517220 were, for the first time in the literature, significantly associated with higher clinical improvement. The strongest association was found for rs34517220. In particular, minor allele homozygotes showed higher score reductions on the CDI scale compared with the major allele carriers. Interestingly, this polymorphism is located in a human TPH2 TFBS for two relevant transcription factors in the serotoninergic neurons, Foxa1 and Foxa2, which together with the high level of significance found for this SNP, could indicate that rs34517220 is in fact the crucial functional genetic variant related to the fluoxetine response. These results provide new evidence for the role of regulatory genetic variants that could modulate human TPH2 expression in the SSRI antidepressant response. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Functional Constituents of a Local Serotonergic System, Intrinsic to the Human Coronary Artery Smooth Muscle Cells

    PubMed Central

    Baskar, Kannan; Sur, Swastika; Selvaraj, Vithyalakashmi; Agrawal, Devendra K.

    2015-01-01

    Human coronary artery smooth muscle cells (HCASMCs) play an important role in the pathogenesis of coronary atherosclerosis and coronary artery diseases (CAD). Serotonin is a mediator known to produce vascular smooth muscle cell (VSMC) mitogenesis and contribute to coronary atherosclerosis. We hypothesize that the human coronary artery smooth muscle cell possesses certain functional constituents of the serotonergic system such as: tryptophan hydroxylase and serotonin transporter. Our aim was to examine the presence of functional tryptophan hydroxylase-1 (TPH1) and serotonin transporter (SERT) in HCASMCs. The mRNA transcripts by qPCR and protein expression by Western blot of TPH1 and SERT were examined. The specificity and accuracy of the primers were verified using DNA gel electrophoresis and sequencing of qPCR products. The functionality of SERT was examined using a fluorescence dye-based serotonin transporter assay. The enzymatic activity of TPH was evaluated using UPLC. The HCASMCs expressed both mRNA transcripts and protein of SERT and TPH. The qPCR showed a single melt curve peak for both transcripts and in sequence analysis the amplicons were aligned with the respective genes. SERT and TPH enzymatic activity was present in the HCASMCs. Taken together, both TPH and SERT are functionally expressed in HCASMCs. These findings are novel and represent an initial step in examining the clinical relevance of the serotonergic system in HCASMCs and its role in the pathogenesis of coronary atherosclerosis and CAD. PMID:25861735

  4. Stimulation of 5-HT2C Receptors Improves Cognitive Deficits Induced by Human Tryptophan Hydroxylase 2 Loss of Function Mutation

    PubMed Central

    Del'Guidice, Thomas; Lemay, Francis; Lemasson, Morgane; Levasseur-Moreau, Jean; Manta, Stella; Etievant, Adeline; Escoffier, Guy; Doré, François Y; Roman, François S; Beaulieu, Jean-Martin

    2014-01-01

    Polymorphisms in the gene encoding the serotonin synthesis enzyme Tph2 have been identified in mental illnesses, including bipolar disorder, major depression, autism, schizophrenia, and ADHD. Deficits in cognitive flexibility and perseverative behaviors are shared common symptoms in these disorders. However, little is known about the impact of Tph2 gene variants on cognition. Mice expressing a human TPH2 variant (Tph2-KI) were used to investigate cognitive consequences of TPH2 loss of function and pharmacological treatments. We applied a recently developed behavioral assay, the automated H-maze, to study cognitive functions in Tph2-KI mice. This assay involves the consecutive discovery of three different rules: a delayed alternation task, a non-alternation task, and a delayed reversal task. Possible contribution of locomotion, reward, and sensory perception were also investigated. The expression of loss-of-function mutant Tph2 in mice was associated with impairments in reversal learning and cognitive flexibility, accompanied by perseverative behaviors similar to those observed in human clinical studies. Pharmacological restoration of 5-HT synthesis with 5-hydroxytryptophan or treatment with the 5-HT2C receptor agonist CP809.101 reduced cognitive deficits in Tph2-KI mice and abolished perseveration. In contrast, treatment with the psychostimulant methylphenidate exacerbated cognitive deficits in mutant mice. Results from this study suggest a contribution of TPH2 in the regulation of cognition. Furthermore, identification of a role for a 5-HT2 receptor agonist as a cognition-enhancing agent in mutant mice suggests a potential avenue to explore for the personalized treatment of cognitive symptoms in humans with reduced 5-HT synthesis and TPH2 polymorphisms. PMID:24196946

  5. Neuronal Tryptophan Hydroxylase Expression in BALB/cJ and C57Bl/6J Mice

    PubMed Central

    Bach, Helene; Arango, Victoria; Huang, Yung-Yu; Leong, Sharlene; Mann, J. John; Underwood, Mark D.

    2014-01-01

    BALB/c is an inbred stress-sensitive mouse strain exhibiting low brain serotonin (5-HT) content and a 5-HT biosynthetic enzyme tryptophan hydroxylase (Tph2) variant reported to have lower catalytic activity compared to other inbred base strains. To evaluate other mechanisms that may explain low 5-HT, we compared BALB/cJ mice and a control inbred strain C57Bl/6J mice, for expression of Tph2 mRNA, TPH2 protein and regional levels of 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA). Tph2 mRNA and TPH2 protein in brainstem dorsal raphe nuclei (DRN) was assayed by in situ hybridization and immunocytochemistry respectively. 5-HT and 5-HIAA were determined by high pressure liquid chromatography (HPLC). BALB/cJ mice had 20% less Tph2 mRNA and 28% fewer TPH2 immunolabeled neurons than C57Bl/6J mice (t = -2.59, p = 0.02). The largest difference in Tph2 transcript expression was in rostral DRN (t = 2.731, p = 0.008). 5-HT was 15% lower in the midbrain of BALB/cJ compared to C57Bl/6J mice (p < 0.05). The behavioral differences in BALB/cJ mice relative to the C57Bl/6J strain may be due in part, to fewer 5-HT neurons and lower Tph2 gene expression resulting in less 5-HT neurotransmission. Future studies quantifying expression per neuron are needed to determine whether less expression is explained by fewer neurons or also less expression per neuron, or both. PMID:21740442

  6. Neuronal tryptophan hydroxylase expression in BALB/cJ and C57Bl/6J mice.

    PubMed

    Bach, Helene; Arango, Victoria; Huang, Yung-Yu; Leong, Sharlene; Mann, J John; Underwood, Mark D

    2011-09-01

    BALB/c is an inbred stress-sensitive mouse strain exhibiting low brain serotonin (5-HT) content and a 5-HT biosynthetic enzyme tryptophan hydroxylase (Tph2) variant reported to have lower catalytic activity compared with other inbred base strains. To evaluate other mechanisms that may explain low 5-HT, we compared BALB/cJ mice and a control inbred strain C57Bl/6J mice, for expression of Tph2 mRNA, TPH2 protein and regional levels of 5-HT and its metabolite 5-hydroxyindoleacetic acid. Tph2 mRNA and TPH2 protein in brainstem dorsal raphe nuclei was assayed by in situ hybridization and immunocytochemistry respectively. 5-HT and 5-hydroxyindoleacetic acid were determined by HPLC. BALB/cJ mice had 20% less Tph2 mRNA and 28% fewer TPH2 immunolabeled neurons than C57Bl/6J mice (t = -2.59, p = 0.02). The largest difference in Tph2 transcript expression was in rostral dorsal raphe nuclei (t = 2.731, p = 0.008). 5-HT was 15% lower in the midbrain and 18% lower in the cerebral cortex of BALB/cJ compared with C57Bl/6J mice (p < 0.05). The behavioral differences in BALB/cJ mice relative to the C57Bl/6J strain may be due in part, to fewer 5-HT neurons and lower Tph2 gene expression resulting in less 5-HT neurotransmission. Future studies quantifying expression per neuron are needed to determine whether less expression is explained by fewer neurons or also less expression per neuron, or both. © 2011 The Authors. Journal of Neurochemistry © 2011 International Society for Neurochemistry.

  7. Differential tissue distribution of tryptophan hydroxylase isoforms 1 and 2 as revealed with monospecific antibodies.

    PubMed

    Sakowski, Stacey A; Geddes, Timothy J; Thomas, David M; Levi, Edi; Hatfield, James S; Kuhn, Donald M

    2006-04-26

    Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the synthesis of the neurotransmitter serotonin. Once thought to be a single-gene product, TPH is now known to exist in two isoforms-TPH1 is found in the pineal and gut, and TPH2 is selectively expressed in brain. Heretofore, probes used for localization of TPH protein or mRNA could not distinguish between the TPH isoforms because of extensive homology shared by them at the nucleotide and amino acid level. We have produced monospecific polyclonal antibodies against TPH1 and TPH2 using peptide antigens from nonoverlapping sequences in the respective proteins. These antibodies allow the differentiation of TPH1 and TPH2 upon immunoblotting, immunoprecipitation, and immunocytochemical staining of tissue sections from brain and gut. TPH1 and TPH2 antibodies do not cross-react with either tyrosine hydroxylase or phenylalanine hydroxylase. Analysis of mouse tissues confirms that TPH1 is the predominant form expressed in pineal gland and in P815 mastocytoma cells with a molecular weight of 51 kDa. TPH2 is the predominant enzyme form expressed in brain extracts from mesencephalic tegmentum, striatum, and hippocampus with a molecular weight of 56 kDa. Antibody specificity against TPH1 and TPH2 is retained across mouse, rat, rabbit, primate, and human tissues. Antibodies that distinguish between the isoforms of TPH will allow studies of the differential regulation of their expression in brain and periphery.

  8. TPH2 polymorphisms and expression in Prader-Willi syndrome subjects with differing genetic subtypes.

    PubMed

    Henkhaus, Rebecca S; Bittel, Douglas C; Butler, Merlin G

    2010-09-01

    Prader-Willi syndrome (PWS) is a genetic imprinting disease that causes developmental and behavioral disturbances resulting from loss of expression of genes from the paternal chromosome 15q11-q13 region. In about 70% of subjects, this portion of the paternal chromosome is deleted, while 25% have two copies of the maternal chromosome 15, or uniparental maternal disomy (UPD; the remaining subjects have imprinting center defects. There are several documented physical and behavioral differences between the two major PWS genetic subtypes (deletion and UPD) indicating the genetic subtype plays a role in clinical presentation. Serotonin is known to be disturbed in PWS and affects both eating behavior and compulsion, which are reported to be abnormal in PWS. We investigated the tryptophan hydroxylase gene (TPH2), the rate-limiting enzyme in the production of brain serotonin, by analyzing three different TPH2 gene polymorphisms, transcript expression, and correlation with PWS genetic subtype. DNA and RNA from lymphoblastoid cell lines derived from 12 PWS and 12 comparison subjects were used for the determination of genetic subtype, TPH2 polymorphisms and quantitative RT-PCR analysis. A similar frequency of TPH2 polymorphisms was seen in the PWS and comparison subjects with PWS deletion subjects showing increased expression with one or more TPH2 polymorphism. Both PWS deletion and PWS UPD subjects had significantly lower TPH2 expression than control subjects and PWS deletion subjects had significantly lower TPH2 expression compared with PWS UPD subjects. PWS subjects with 15q11-q13 deletions had lower TPH2 expression compared with PWS UPD or control subjects, requiring replication and further studies to identify the cause including identification of disturbed gene interactions resulting from the deletion process.

  9. Polymorphism of the Tryptophan Hydroxylase 2 (TPH2) Gene Is Associated with Chimpanzee Neuroticism

    PubMed Central

    Morimura, Naruki; Udono, Toshifumi; Hayasaka, Ikuo; Humle, Tatyana; Murayama, Yuichi; Ito, Shin'ichi; Inoue-Murayama, Miho

    2011-01-01

    In the brain, serotonin production is controlled by tryptophan hydroxylase 2 (TPH2), a genotype. Previous studies found that mutations on the TPH2 locus in humans were associated with depression and studies of mice and studies of rhesus macaques have shown that the TPH2 locus was involved with aggressive behavior. We previously reported a functional single nucleotide polymorphism (SNP) in the form of an amino acid substitution, Q468R, in the chimpanzee TPH2 gene coding region. In the present study we tested whether this SNP was associated with neuroticism in captive and wild-born chimpanzees living in Japan and Guinea, respectively. Even after correcting for multiple tests (Bonferroni p = 0.05/6 = 0.008), Q468R was significantly related to higher neuroticism (β = 0.372, p = 0.005). This study is the first to identify a genotype linked to a personality trait in chimpanzees. In light of the prior studies on humans, mice, and rhesus macaques, these findings suggest that the relationship between neuroticism and TPH2 has deep phylogenetic roots. PMID:21765945

  10. Intraspinal serotonergic neurons consist of two, temporally distinct populations in developing zebrafish

    PubMed Central

    Montgomery, Jacob E.; Wiggin, Timothy D.; Rivera-Perez, Luis M.; Lillesaar, Christina; Masino, Mark A.

    2015-01-01

    Zebrafish intraspinal serotonergic neuron (ISN) morphology and distribution have been examined in detail at different ages; however, some aspects of the development of these cells remain unclear. Although antibodies to serotonin (5-HT) have detected ISNs in the ventral spinal cord of embryos, larvae, and adults, the only tryptophan hydroxylase (tph) transcript that has been described in the spinal cord is tph1a. Paradoxically, spinal tph1a is expressed transiently in embryos, which brings the source of 5-HT in the ISNs of larvae and adults into question. Because the pet1 and tph2 promoters drive transgene expression in the spinal cord, we hypothesized that tph2 is expressed in spinal cords of zebrafish larvae. We confirmed this hypothesis through in situ hybridization. Next, we used 5-HT antibody labeling and transgenic markers of tph2-expressing neurons to identify a transient population of ISNs in embryos that was distinct from ISNs that appeared later in development. The existence of separate ISN populations may not have been recognized previously due to their shared location in the ventral spinal cord. Finally, we used transgenic markers and immunohistochemical labeling to identify the transient ISN population as GABAergic Kolmer-Agduhr double-prime (KA″) neurons. Altogether, this study revealed a novel developmental paradigm in which KA″ neurons are transiently serotonergic before the appearance of a stable population of tph2-expressing ISNs. PMID:26437856

  11. A Functional Tph2 C1473G Polymorphism Causes an Anxiety Phenotype via Compensatory Changes in the Serotonergic System

    PubMed Central

    Berger, Stefan M; Weber, Tillmann; Perreau-Lenz, Stephanie; Vogt, Miriam A; Gartside, Sarah E; Maser-Gluth, Christiane; Lanfumey, Laurence; Gass, Peter; Spanagel, Rainer; Bartsch, Dusan

    2012-01-01

    The association of single-nucleotide polymorphisms (SNPs) in the human tryptophan hydroxylase 2 (TPH2) gene with anxiety traits and depression has been inconclusive. Observed inconsistencies might result from the fact that TPH2 polymorphisms have been studied in a genetically heterogeneous human population. A defined genetic background, control over environmental factors, and the ability to analyze the molecular and neurochemical consequences of introduced genetic alterations constitute major advantages of investigating SNPs in inbred laboratory mouse strains. To investigate the behavioral and neurochemical consequences of a functional C1473G SNP in the mouse Tph2 gene, we generated congenic C57BL/6N mice homozygous for the Tph2 1473G allele. The Arg447 substitution in the TPH2 enzyme resulted in a significant reduction of the brain serotonin (5-HT) in vivo synthesis rate. Despite decreased 5-HT synthesis, we could detect neither a reduction of brain region-specific 5-HT concentrations nor changes in baseline and stress-induced 5-HT release using a microdialysis approach. However, using a [35S]GTP-γ-S binding assay and 5-HT1A receptor autoradiography, a functional desensitization of 5-HT1A autoreceptors could be identified. Furthermore, behavioral analysis revealed a distinct anxiety phenotype in homozygous Tph2 1473G mice, which could be reversed with chronic escitalopram treatment. Alterations in depressive-like behavior could not be detected under baseline conditions or after chronic mild stress. These findings provide evidence for an involvement of functional Tph2 polymorphisms in anxiety-related behaviors, which are likely not caused directly by alterations in 5-HT content or release but are rather due to compensatory changes during development involving functional desensitization of 5-HT1A autoreceptors. PMID:22491354

  12. Increased ethanol consumption despite taste aversion in mice with a human tryptophan hydroxylase 2 loss of function mutation.

    PubMed

    Lemay, Francis; Doré, François Y; Beaulieu, Jean-Martin

    2015-11-16

    Polymorphisms in the gene encoding the brain serotonin synthesis enzyme Tph2 have been identified in mental illnesses, with co-morbidity of substance use disorder. However, little is known about the impact of Tph2 gene variants on addiction. Mice expressing a human Tph2 loss of function variant were used to investigate consequences of aversive conditions on ethanol intake. Mice were familiarized either with ethanol or a solution containing both ethanol and the bittering agent quinine. Effect of familiarization to ethanol or an ethanol-quinine solution was then evaluated using a two-bottles preference test in Tph2-KI and control littermates. Mice from both genotypes displayed similar levels of ethanol consumption and quinine avoidance when habituated to ethanol alone. In contrast, addition of quinine to ethanol during the familiarization period resulted in a reduction of avoidance for the quinine-ethanol solution only in mutant mice. These results indicate that loss of function mutation in Tph2 results in greater motivation for ethanol consumption under aversive conditions and may confer enhanced sensitivity to alcohol use disorder. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  13. Generation of a Tph2 Conditional Knockout Mouse Line for Time- and Tissue-Specific Depletion of Brain Serotonin

    PubMed Central

    Migliarini, Sara; Pacini, Giulia; Pasqualetti, Massimo

    2015-01-01

    Serotonin has been gaining increasing attention during the last two decades due to the dual function of this monoamine as key regulator during critical developmental events and as neurotransmitter. Importantly, unbalanced serotonergic levels during critical temporal phases might contribute to the onset of neuropsychiatric disorders, such as schizophrenia and autism. Despite increasing evidences from both animal models and human genetic studies have underpinned the importance of serotonin homeostasis maintenance during central nervous system development and adulthood, the precise role of this molecule in time-specific activities is only beginning to be elucidated. Serotonin synthesis is a 2-step process, the first step of which is mediated by the rate-limiting activity of Tph enzymes, belonging to the family of aromatic amino acid hydroxylases and existing in two isoforms, Tph1 and Tph2, responsible for the production of peripheral and brain serotonin, respectively. In the present study, we generated and validated a conditional knockout mouse line, Tph2 flox/flox, in which brain serotonin can be effectively ablated with time specificity. We demonstrated that the Cre-mediated excision of the third exon of Tph2 gene results in the production of a Tph2 null allele in which we observed the near-complete loss of brain serotonin, as well as the growth defects and perinatal lethality observed in serotonin conventional knockouts. We also revealed that in mice harbouring the Tph2 null allele, but not in wild-types, two distinct Tph2 mRNA isoforms are present, namely Tph2Δ3 and Tph2Δ3Δ4, with the latter showing an in-frame deletion of amino acids 84–178 and coding a protein that could potentially retain non-negligible enzymatic activity. As we could not detect Tph1 expression in the raphe, we made the hypothesis that the Tph2Δ3Δ4 isoform can be at the origin of the residual, sub-threshold amount of serotonin detected in the brain of Tph2 null/null mice. Finally, we set up a tamoxifen administration protocol that allows an efficient, time-specific inactivation of brain serotonin synthesis. On the whole, we generated a suitable genetic tool to investigate how serotonin depletion impacts on time-specific events during central nervous system development and adulthood life. PMID:26291320

  14. Blunted epidermal L-tryptophan metabolism in vitiligo affects immune response and ROS scavenging by Fenton chemistry, part 1: Epidermal H2O2/ONOO(-)-mediated stress abrogates tryptophan hydroxylase and dopa decarboxylase activities, leading to low serotonin and melatonin levels.

    PubMed

    Schallreuter, Karin U; Salem, Mohamed A E L; Gibbons, Nick C J; Martinez, Aurora; Slominski, Radomir; Lüdemann, Jürgen; Rokos, Hartmut

    2012-06-01

    Vitiligo is characterized by a progressive loss of inherited skin color. The cause of the disease is still unknown. To date, there is accumulating in vivo and in vitro evidence for massive oxidative stress via hydrogen peroxide (H(2)O(2)) and peroxynitrite (ONOO(-)) in the skin of affected individuals. Autoimmune etiology is the favored theory. Since depletion of the essential amino acid L-tryptophan (Trp) affects immune response mechanisms, we here looked at epidermal Trp metabolism via tryptophan hydroxylase (TPH) with its downstream cascade, including serotonin and melatonin. Our in situ immunofluorescence and Western blot data reveal significantly lower TPH1 expression in patients with vitiligo. Expression is also low in melanocytes and keratinocytes under in vitro conditions. Although in vivo Fourier transform-Raman spectroscopy proves the presence of 5-hydroxytryptophan, epidermal TPH activity is completely absent. Regulation of TPH via microphthalmia-associated transcription factor and L-type calcium channels is severely affected. Moreover, dopa decarboxylase (DDC) expression is significantly lower, in association with decreased serotonin and melatonin levels. Computer simulation supports H(2)O(2)/ONOO(-)-mediated oxidation/nitration of TPH1 and DDC, affecting, in turn, enzyme functionality. Taken together, our data point to depletion of epidermal Trp by Fenton chemistry and exclude melatonin as a relevant contributor to epidermal redox balance and immune response in vitiligo.

  15. Determination of a risk management primer at petroleum-contaminated sites: developing new human health risk assessment strategy.

    PubMed

    Park, In-Sun; Park, Jae-Woo

    2011-01-30

    Total petroleum hydrocarbon (TPH) is an important environmental contaminant that is toxic to human and environmental receptors. However, human health risk assessment for petroleum, oil, and lubricant (POL)-contaminated sites is especially challenging because TPH is not a single compound, but rather a mixture of numerous substances. To address this concern, this study recommends a new human health risk assessment strategy for POL-contaminated sites. The strategy is based on a newly modified TPH fractionation method and includes an improved analytical protocol. The proposed TPH fractionation method is composed of ten fractions (e.g., aliphatic and aromatic EC8-10, EC10-12, EC12-16, EC16-22 and EC22-40). Physicochemical properties and toxicity values of each fraction were newly defined in this study. The stepwise ultrasonication-based analytical process was established to measure TPH fractions. Analytical results were compared with those from the TPH Criteria Working Group (TPHCWG) Direct Method. Better analytical efficiencies in TPH, aliphatic, and aromatic fractions were achieved when contaminated soil samples were analyzed with the new analytical protocol. Finally, a human health risk assessment was performed based on the developed tiered risk assessment framework. Results showed that a detailed quantitative risk assessment should be conducted to determine scientifically and economically appropriate cleanup target levels, although the phase II process is useful for determining the potency of human health risks posed by POL-contamination. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Identification of a Novel Allosteric Inhibitory Site on Tryptophan Hydroxylase 1 Enabling Unprecedented Selectivity Over all Related Hydroxylases

    PubMed Central

    Petrassi, Mike; Barber, Rob; Be, Celine; Beach, Sarah; Cox, Brian; D’Souza, Anne-Marie; Duggan, Nick; Hussey, Martin; Fox, Roy; Hunt, Peter; Jarai, Gabor; Kosaka, Takatoshi; Oakley, Paul; Patel, Viral; Press, Neil; Rowlands, David; Scheufler, Clemens; Schmidt, Oliver; Srinivas, Honnappa; Turner, Mary; Turner, Rob; Westwick, John; Wolfreys, Alison; Pathan, Nuzhat; Watson, Simon; Thomas, Matthew

    2017-01-01

    Pulmonary arterial hypertension (PAH) has demonstrated multi-serotonin receptor dependent pathologies, characterized by increased tone (5-HT1B receptor) and complex lesions (SERT, 5-HT1B, 5-HT2B receptors) of the pulmonary vasculature together with right ventricular hypertrophy, ischemia and fibrosis (5-HT2B receptor). Selective inhibitors of individual signaling elements – SERT, 5-HT2A, 5HT2B, and combined 5-HT2A/B receptors, have all been tested clinically and failed. Thus, inhibition of tryptophan hydroxylase 1 (TPH1), the rate limiting step in 5-HT synthesis, has been suggested as a more broad, and thereby more effective, mode of 5-HT inhibition. However, selectivity over non-pathogenic enzyme family members, TPH2, phenylalanine hydroxylase, and tyrosine hydroxylase has hampered therapeutic development. Here we describe the site/sequence, biochemical, and biophysical characterization of a novel allosteric site on TPH1 through which selectivity over TPH2 and related aromatic amino acid hydroxylases is achieved. We demonstrate the mechanism of action by which novel compounds selectively inhibit TPH1 using surface plasma resonance and enzyme competition assays with both tryptophan ligand and BH4 co-factor. We demonstrate 15-fold greater potency within a human carcinoid cell line versus the most potent known TPH1/2 non-specific inhibitor. Lastly, we detail a novel canine in vivo system utilized to determine effective biologic inhibition of newly synthesized 5-HT. These findings are the first to demonstrate TPH1-selective inhibition and may pave the way to a truly effective means to reduce pathologic 5-HT and thereby treat complex remodeling diseases such as PAH. PMID:28529483

  17. Evaluation of gastrointestinal solubilization of petroleum hydrocarbon residues in soil using an in vitro physiologically based model.

    PubMed

    Holman, Hoi-Ying N; Goth-Goldstein, Regine; Aston, David; Yun, Mao; Kengsoontra, Jenny

    2002-03-15

    Petroleum hydrocarbon residues in weathered soils may pose risks to humans through the ingestion pathway. To understand the factors controlling their gastrointestinal (GI) absorption, a newly developed experimental extraction protocol was used to model the GI solubility of total petroleum hydrocarbon (TPH) residues in highly weathered soils from different sites. The GI solubility of TPH residues was significantly higher for soil contaminated with diesel than with crude oil. Compared to the solubility of TPH residues during fasted state,the solubility of TPH residues during fat digestion was much greater. Diesel solubility increased from an average of 8% during the "gallbladder empty" phase of fasting (and less than 0.2% during the otherfasting phase) to an average of 16% during fat digestion. For crude oil, the solubility increased from an average of 1.2% during the gallbladder empty phase of fasting (and undetectable during the other fasting phase) to an average of 4.5% during fat digestion. Increasing the concentration of bile salts also increased GI solubility. GI solubility was reduced by soil organic carbon but enhanced by the TPH content.

  18. Genetic depletion of brain 5HT reveals a common molecular pathway mediating compulsivity and impulsivity.

    PubMed

    Angoa-Pérez, Mariana; Kane, Michael J; Briggs, Denise I; Sykes, Catherine E; Shah, Mrudang M; Francescutti, Dina M; Rosenberg, David R; Thomas, David M; Kuhn, Donald M

    2012-06-01

    Neuropsychiatric disorders characterized by behavioral disinhibition, including disorders of compulsivity (e.g. obsessive-compulsive disorder; OCD) and impulse-control (e.g. impulsive aggression), are severe, highly prevalent and chronically disabling. Treatment options for these diseases are extremely limited. The pathophysiological bases of disorders of behavioral disinhibition are poorly understood but it has been suggested that serotonin dysfunction may play a role. Mice lacking the gene encoding brain tryptophan hydroxylase 2 (Tph2-/-), the initial and rate-limiting enzyme in the synthesis of serotonin, were tested in numerous behavioral assays that are well known for their utility in modeling human neuropsychiatric diseases. Mice lacking Tph2 (and brain 5HT) show intense compulsive and impulsive behaviors to include extreme aggression. The impulsivity is motor in form and not cognitive because Tph2-/- mice show normal acquisition and reversal learning on a spatial learning task. Restoration of 5HT levels by treatment of Tph2-/- mice with its immediate precursor 5-hydroxytryptophan attenuated compulsive and impulsive-aggressive behaviors. Surprisingly, in Tph2-/- mice, the lack of 5HT was not associated with anxiety-like behaviors. The results indicate that 5HT mediates behavioral disinhibition in the mammalian brain independent of anxiogenesis. © 2012 The Authors. Journal of Neurochemistry © 2012 International Society for Neurochemistry.

  19. Vitamin D hormone regulates serotonin synthesis. Part 1: relevance for autism.

    PubMed

    Patrick, Rhonda P; Ames, Bruce N

    2014-06-01

    Serotonin and vitamin D have been proposed to play a role in autism; however, no causal mechanism has been established. Here, we present evidence that vitamin D hormone (calcitriol) activates the transcription of the serotonin-synthesizing gene tryptophan hydroxylase 2 (TPH2) in the brain at a vitamin D response element (VDRE) and represses the transcription of TPH1 in tissues outside the blood-brain barrier at a distinct VDRE. The proposed mechanism explains 4 major characteristics associated with autism: the low concentrations of serotonin in the brain and its elevated concentrations in tissues outside the blood-brain barrier; the low concentrations of the vitamin D hormone precursor 25-hydroxyvitamin D [25(OH)D3]; the high male prevalence of autism; and the presence of maternal antibodies against fetal brain tissue. Two peptide hormones, oxytocin and vasopressin, are also associated with autism and genes encoding the oxytocin-neurophysin I preproprotein, the oxytocin receptor, and the arginine vasopressin receptor contain VDREs for activation. Supplementation with vitamin D and tryptophan is a practical and affordable solution to help prevent autism and possibly ameliorate some symptoms of the disorder. © FASEB.

  20. Methylation of the tryptophan hydroxylase‑2 gene is associated with mRNA expression in patients with major depression with suicide attempts.

    PubMed

    Zhang, Yuqi; Chang, Zaohuo; Chen, Jionghua; Ling, Yang; Liu, Xiaowei; Feng, Zhang; Chen, Caixia; Xia, Minghua; Zhao, Xingfu; Ying, Wang; Qing, Xu; Li, Guilin; Zhang, Changsong

    2015-08-01

    Tryptophan hydroxylase-2 (TPH2) contributes to alterations in the function of neuronal serotonin (5-HT), which are associated with various psychopathologies, including major depressive disorder (MDD) or suicidal behavior. The methylation of a single CpG site in the promoter region of TPH2 affects gene expression. Suicide and MDD are strongly associated and genetic factors are at least partially responsible for the variability in suicide risk. The aim of the present study was to investigate whether variations in TPH2 methylation in peripheral blood samples may predispose patients with MDD to suicide attempts. TPH2 mRNA expression levels differed significantly between 50 patients with MDD who had attempted suicide (MDD + suicide group) and 75 control patients with MDD (MDD group); TPH2 expression levels were significantly decreased (P=0.0005) in the patients who had attempted suicide. Furthermore, the frequency of TPH2 methylation was 36.0% in the MDD + suicide group, while it was 13.0% in the MDD group. The results of the present study demonstrated that methylation in the promoter region of TPH2 significantly affected the mRNA expression levels of TPH2, thus suggesting that methylation of the TPH2 promoter may silence TPH2 mRNA expression in MDD patients with or without suicidal behavior. In addition, there was a significant correlation between the methylation status of the TPH2 promoter and depression, hopelessness and cognitive impairment in the MDD + suicide group. In conclusion, the present study demonstrated that TPH2 expression was regulated by DNA methylation of the TPH2 promoter region in patients with MDD.

  1. Seasonal postembryonic maturation of the diurnal rhythm of serotonin in the chicken pineal gland.

    PubMed

    Piesiewicz, Aneta; Kedzierska, Urszula; Turkowska, Elzbieta; Adamska, Iwona; Majewski, Pawel M

    2015-02-01

    Previously, we have demonstrated the postembryonic development of chicken (Gallus gallus domesticus L.) pineal gland functions expressed as changes in melatonin (MEL) biosynthesis. Pineal concentrations of MEL and its precursor serotonin (5-HT) were shown to increase between the 2nd and 16th day of life. We also found that levels of the mRNAs encoding the enzymes participating in the final two steps of MEL biosynthesis from 5-HT: arylalkylamine-N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT), as well as their enzymatic activities, were raised during postembryonic development. Moreover, the manner of these changes was season-of-hatch dependent, even in animals kept under constant laboratory conditions (L:D 12:12). The most pronounced changes were seen in the concentrations of 5-HT and MEL, as well as in Aanat mRNA level and its enzymatic activity. The high daily variability in 5-HT content suggested that season- and age-dependent changes in the activity of the chicken pineal gland might rely on the availability of 5-HT, i.e. it may be limited by changes in pineal tryptophan (TRP) and/or 5-hydroxytryptophan (5-HTP) levels as well as by the activity of tryptophan hydroxylase (TPH) and aromatic l-amino acid decarboxylase (AADC): two enzymes participating in the conversion of TRP to 5-HT. The present study was undertaken with the following objectives: (1) to examine whether the pineal concentration of the 5-HT precursors TRP and 5-HTP exhibit age- and season-related changes; (2) to look for season-related differences in the transcription of the Tph1 and Ddc genes encoding enzymes TPH and AADC; (3) to identify the step(s) in postembryonic development in which these season-related variations in pineal gland function are most pronounced. Male Hy-line chickens hatched in the summer or winter, from eggs laid by hens held in L:D 16:8 conditions were kept from the day of hatch in L:D 12:12 conditions. At the age of 2 or 9 days, animals were sacrificed every 2 or 4 h over a 24-h period and their pineal glands were isolated under dim red light and processed for the measurement of (i) the pineal content of TRP, 5-HTP and 5-HT, and (ii) the level of Tph1 and Ddc mRNAs. Circadian rhythmicity of all the measured parameters was evaluated by the cosinor method. The pineal levels of TRP and 5-HT as well as the Tph1 and Ddc transcripts changed during postembryonic development in a season-related way. Whereas, the 5-HTP concentration did not vary between animals from both age groups, regardless of the season. Circadian rhythmicity of all the measured parameters was dependent on both the age and the season of hatch, and was greatest in older animals in the summer. These findings indicated that the efficiency of season-related MEL biosynthesis, reported previously, is limited by 5-HT availability and this limitation depends on the transcription of both the Tph1 and Ddc genes. Moreover, Ddc mRNA level in 9-d-old birds changed rhythmically, even though this gene is generally considered to be arrhythmic.

  2. Gene Therapy by Targeted Adenovirus-mediated Knockdown of Pulmonary Endothelial Tph1 Attenuates Hypoxia-induced Pulmonary Hypertension

    PubMed Central

    Morecroft, Ian; White, Katie; Caruso, Paola; Nilsen, Margaret; Loughlin, Lynn; Alba, Raul; Reynolds, Paul N; Danilov, Sergei M; Baker, Andrew H; MacLean, Margaret R

    2012-01-01

    Serotonin is produced by pulmonary arterial endothelial cells (PAEC) via tryptophan hydroxylase-1 (Tph1). Pathologically, serotonin acts on underlying pulmonary arterial cells, contributing to vascular remodeling associated with pulmonary arterial hypertension (PAH). The effects of hypoxia on PAEC-Tph1 activity are unknown. We investigated the potential of a gene therapy approach to PAH using selective inhibition of PAEC-Tph1 in vivo in a hypoxic model of PAH. We exposed cultured bovine pulmonary arterial smooth muscle cells (bPASMCs) to conditioned media from human PAECs (hPAECs) before and after hypoxic exposure. Serotonin levels were increased in hypoxic PAEC media. Conditioned media evoked bPASMC proliferation, which was greater with hypoxic PAEC media, via a serotonin-dependent mechanism. In vivo, adenoviral vectors targeted to PAECs (utilizing bispecific antibody to angiotensin-converting enzyme (ACE) as the selective targeting system) were used to deliver small hairpin Tph1 RNA sequences in rats. Hypoxic rats developed PAH and increased lung Tph1. PAEC-Tph1 expression and development of PAH were attenuated by our PAEC-Tph1 gene knockdown strategy. These results demonstrate that hypoxia induces Tph1 activity and selective knockdown of PAEC-Tph1 attenuates hypoxia-induced PAH in rats. Further investigation of pulmonary endothelial-specific Tph1 inhibition via gene interventions is warranted. PMID:22525513

  3. Targeting brain serotonin synthesis: insights into neurodevelopmental disorders with long-term outcomes related to negative emotionality, aggression and antisocial behaviour.

    PubMed

    Lesch, Klaus-Peter; Araragi, Naozumi; Waider, Jonas; van den Hove, Daniel; Gutknecht, Lise

    2012-09-05

    Aggression, which comprises multi-faceted traits ranging from negative emotionality to antisocial behaviour, is influenced by an interaction of biological, psychological and social variables. Failure in social adjustment, aggressiveness and violence represent the most detrimental long-term outcome of neurodevelopmental disorders. With the exception of brain-specific tryptophan hydroxylase-2 (Tph2), which generates serotonin (5-HT) in raphe neurons, the contribution of gene variation to aggression-related behaviour in genetically modified mouse models has been previously appraised (Lesch 2005 Novartis Found Symp. 268, 111-140; Lesch & Merschdorf 2000 Behav. Sci. Law 18, 581-604). Genetic inactivation of Tph2 function in mice led to the identification of phenotypic changes, ranging from growth retardation and late-onset obesity, to enhanced conditioned fear response, increased aggression and depression-like behaviour. This spectrum of consequences, which are amplified by stress-related epigenetic interactions, are attributable to deficient brain 5-HT synthesis during development and adulthood. Human data relating altered TPH2 function to personality traits of negative emotionality and neurodevelopmental disorders characterized by deficits in cognitive control and emotion regulation are based on genetic association and are therefore not as robust as the experimental mouse results. Mouse models in conjunction with approaches focusing on TPH2 variants in humans provide unexpected views of 5-HT's role in brain development and in disorders related to negative emotionality, aggression and antisocial behaviour.

  4. Tph2 gene deletion enhances amphetamine-induced hypermotility: effect of 5-HT restoration and role of striatal noradrenaline release.

    PubMed

    Carli, Mirjana; Kostoula, Chrysaugi; Sacchetti, Giuseppina; Mainolfi, Pierangela; Anastasia, Alessia; Villani, Claudia; Invernizzi, Roberto William

    2015-11-01

    Variants of tryptophan hydroxylase-2 (Tph2), the gene encoding enzyme responsible for the synthesis of brain serotonin (5-HT), have been associated with neuropsychiatric disorders, substance abuse and addiction. This study assessed the effect of Tph2 gene deletion on motor behavior and found that motor activity induced by 2.5 and 5 mg/kg amphetamine was enhanced in Tph2(-/-) mice. Using the in vivo microdialysis technique we found that the ability of amphetamine to stimulate noradrenaline (NA) release in the striatum was reduced by about 50% in Tph2(-/-) mice while the release of dopamine (DA) was not affected. Tph2 deletion did not affect the release of NA and DA in the prefrontal cortex. The role of endogenous 5-HT in enhancing the effect of amphetamine was confirmed showing that treatment with the 5-HT precursor 5-hydroxytryptophan (10 mg/kg) restored tissue and extracellular levels of brain 5-HT and the effects of amphetamine on striatal NA release and motor activity in Tph2(-/-) mice. Treatment with the NA precursor dihydroxyphenylserine (400 mg/kg) was sufficient to restore the effect of amphetamine on striatal NA release and motor activity in Tph2(-/-) mice. These findings indicate that amphetamine-induced hyperactivity is attenuated by endogenous 5-HT through the inhibition of striatal NA release. Tph2(-/-) mice may be a useful preclinical model to assess the role of 5-HT-dependent mechanisms in the action of psychostimulants. Acute sensitivity to the motor effects of amphetamine has been associated to increased risk of psychostimulant abuse. Here, we show that deletion of Tph2, the gene responsible for brain 5-HT synthesis, enhances the motor effect of amphetamine in mice through the inhibition of striatal NA release. This suggests that Tph2(-/-) mice is a useful preclinical model to assess the role of 5-HT-dependent mechanisms in psychostimulants action. Tph2, tryptophan hydroxylase-2. © 2015 International Society for Neurochemistry.

  5. Sex-dependent programming effects of prenatal glucocorticoid treatment on the developing serotonin system and stress-related behaviors in adulthood

    PubMed Central

    Hiroi, Ryoko; Carbone, David L.; Zuloaga, Damian G.; Bimonte-Nelson, Heather A.; Handa, Robert J.

    2016-01-01

    Prenatal stress and overexposure to glucocorticoids (GC) during development may be associated with an increased susceptibility to a number of diseases in adulthood including neuropsychiatric disorders, such as depression and anxiety. In animal models, prenatal overexposure to GC results in hyper-responsiveness to stress in adulthood, and females appear to be more susceptible than males. Here, we tested the hypothesis that overexposure to GC during fetal development has sex-specific programming effects on the brain, resulting in altered behaviors in adulthood. We examined the effects of dexamethasone (DEX; a synthetic GC) during prenatal life on stress-related behaviors in adulthood and on the tryptophan hydroxylase-2 (TpH2) gene expression in the adult dorsal raphe nucleus (DRN). TpH2 is the rate-limiting enzyme for serotonin (5-HT) synthesis and has been implicated in the etiology of human affective disorders. Timed-pregnant rats were treated with DEX from gestational days 18–22. Male and female offspring were sacrificed on the day of birth (postnatal day 0; P0), P7, and in adulthood (P80-84) and brains were examined for changes in TpH2 mRNA expression. Adult animals were also tested for anxiety- and depressive- like behaviors. In adulthood, prenatal DEX increased anxiety- and depressive- like behaviors selectively in females, as measured by decreased time spent in the center of the open field and increased time spent immobile in the forced swim test, respectively. Prenatal DEX increased TpH2 mRNA selectively in the female caudal DRN at P7, whereas it decreased TpH2 mRNA selectively in the female caudal DRN in adulthood. In animals challenged with restraint stress in adulthood, TpH2 mRNA was significantly lower in rostral DRN of prenatal DEX treated females compared to vehicle treated females. These data demonstrated that prenatal overexposure to GC alters the development of TpH2 gene expression and these alterations correlated with lasting behavioral changes found in adult female offspring. PMID:26844389

  6. Theoretical study of singlet oxygen molecule generation via an exciplex with valence-excited thiophene.

    PubMed

    Sumita, Masato; Morihashi, Kenji

    2015-02-05

    Singlet-oxygen [O2((1)Δg)] generation by valence-excited thiophene (TPH) has been investigated using multireference Møller-Plesset second-order perturbation (MRMP2) theory of geometries optimized at the complete active space self-consistent field (CASSCF) theory level. Our results indicate that triplet TPH(1(3)B2) is produced via photoinduced singlet TPH(2(1)A1) because 2(1)A1 TPH shows a large spin-orbit coupling constant with the first triplet excited state (1(3)B2). The relaxed TPH in the 1(3)B2 state can form an exciplex with O2((3)Σg(-)) because this exciplex is energetically more stable than the relaxed TPH. The formation of the TPH(1(3)B2) exciplex with O2((3)Σg(-)) whose total spin multiplicity is triplet (T1 state) increases the likelihood of transition from the T1 state to the singlet ground or first excited singlet state. After the transition, O2((1)Δg) is emitted easily although the favorable product is that from a 2 + 4 cycloaddition reaction.

  7. Genetic moderation of cocaine subjective effects by variation in the TPH1, TPH2, and SLC6A4 serotonin genes.

    PubMed

    Patriquin, Michelle A; Hamon, Sara C; Harding, Mark J; Nielsen, Ellen M; Newton, Thomas F; De La Garza, Richard; Nielsen, David A

    2017-10-01

    This study investigated variants of tryptophan hydroxylase (TPH)1, TPH2, and SLC6A4 in the moderation of the subjective effects of cocaine. Non-treatment-seeking cocaine-dependent individuals (N=66) were intravenously administered saline and cocaine (40 mg) in a randomized order. Participants self-reported subjective effects of cocaine using a visual analog scale starting before administration of saline or cocaine (-15 min) to up to 20 min after infusion. Self-report ratings on the visual analog scale ranged from 0 (no effect) to 100 (greatest effect). Participants were genotyped for the TPH1 rs1799913, TPH2 rs4290270, and SLC6A4 5-HTTLPR variants. Repeated-measures analysis of covariance was used to examine changes in subjective effect scores over time while controlling for population structure. Participants carrying the TPH1 rs1799913 A allele reported greater subjective response to cocaine for 'stimulated' and 'access' relative to the CC genotype group. Those carrying the TPH2 rs4290270 A allele reported higher 'good effect' and lower 'depressed' effect relative to the TT genotype group. Those carrying the SLC6A4 5-HTTLPR S' allele reported greater 'desire' and 'access' compared with the L'L' genotype group. These findings indicate that TPH1, TPH2, and SLC6A4 variants moderate the subjective effects of cocaine in non-treatment-seeking cocaine-dependent participants.

  8. Anoxic biodegradation of petroleum hydrocarbons in saline media using denitrifier biogranules.

    PubMed

    Moussavi, Gholamreza; Shekoohiyan, Sakine; Naddafi, Kazem

    2016-07-01

    The total petroleum hydrocarbons (TPH) biodegradation was examined using biogranules at different initial TPH concentration and contact time under anoxic condition in saline media. The circular compact biogranules having the average diameter between 2 and 3mm were composed of a dense population of Bacillus spp. capable of biodegrading TPH under anoxic condition in saline media were formed in first step of the study. The biogranules could biodegrade over 99% of the TPH at initial concentration up to 2g/L at the contact time of 22h under anoxic condition in saline media. The maximum TPH biodegradation rate of 2.6 gTPH/gbiomass.d could be obtained at initial TPH concentration of 10g/L. Accordingly, the anoxic biogranulation is a possible and promising technique for high-rate biodegradation of petroleum hydrocarbons in saline media. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Pilot scale feasibility study for in-situ chemical oxidation using H2O2 solution conjugated with biodegradation to remediate a diesel contaminated site.

    PubMed

    Kim, Insu; Lee, Minhee

    2012-11-30

    A pilot scale test for a process combining in-situ chemical oxidation using H(2)O(2) solution with biodegradation was performed to remove TPH from a diesel contaminated military site. In batch experiments, when 20% H(2)O(2) solution was used for TPH contaminated soil, TPH removal efficiency was 63.5%. Batch experiments investigating biodegradation by adding indigenous microorganisms in pre-H(2)O(2)-treated soil were also performed, and TPH removal efficiency of biodegradation was 48.5%, showing an improvement of 19.4% by biodegradation even after chemical oxidation. For a pilot scale feasibility test, a site contaminated with diesel (2.5 m × 2.7 m × 1 m) in Korea was selected, and five injection wells and one extraction well were installed in the site. After 0.3 pore volumes of 17.5% H(2)O(2) solution flushing for 15 days, TPH removal efficiency of the site was 51.5%. Seven days after the H(2)O(2) solution flushing was finished, a mixed indigenous microorganism cultured solution (43 L) was injected into the wells two times. After the injection of the cultured solution, the average concentration of TPH in the site decreased to 777 mg/kg, showing that an additional 19.6% of TPH was removed by biodegradation (total TPH removal efficiency: 71.1%). Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Tryptophan hydroxylase 2 aggregates through disulfide cross-linking upon oxidation: Possible link to serotonin deficits and non-motor symptoms in Parkinson's disease

    PubMed Central

    Kuhn, Donald M.; Sykes, Catherine E.; Geddes, Timothy J.; Jaunarajs, Karen L. Eskow; Bishop, Christopher

    2010-01-01

    Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by the loss of dopamine neurons of the nigrostriatal system, resulting in severe motor disturbances. Although much less appreciated, non-motor symptoms are also very common in PD and many can be traced to serotonin neuronal deficits. Tryptophan hydroxylase 2 (TPH2), the rate-limiting enzyme in the serotonin biosynthesis, is a phenotypic marker for serotonin neurons and is known to be extremely labile to oxidation. Therefore, the oxidative processes that prevail in PD could cause TPH2 misfolding and modify 5HT neuronal function much as is seen in dopamine neurons. Oxidation of TPH2 inhibits enzyme activity and leads to the formation of high molecular weight aggregates in a dithiothreitol-reversible manner. Cysteine-scanning mutagenesis shows that as long as a single cysteine residue (out of a total of 13 per monomer) remains in TPH2, it cross-links upon oxidation and only cysteine-less mutants are resistant to this effect. The effects of oxidants on TPH2 catalytic function and cross-linking are also observed in intact TPH2-expressing HEK293 cells. Oxidation shifts TPH2 from the soluble compartment into membrane fractions and large inclusion bodies. Sequential non-reducing/reducing two-dimensional SDS-PAGE and immunoblotting confirmed that TPH2 was one of a small number of cytosolic proteins that form disulfide-bonded aggregates. The propensity of TPH2 to misfold upon oxidation of its cysteine residues is responsible for its catalytic lability and may be related to loss of serotonin neuronal function in PD and the emergence of non-motor (psychiatric) symptoms. PMID:21105877

  11. Serotonergic systems in the balance: CRHR1 and CRHR2 differentially control stress-induced serotonin synthesis.

    PubMed

    Donner, Nina C; Siebler, Philip H; Johnson, Danté T; Villarreal, Marcos D; Mani, Sofia; Matti, Allison J; Lowry, Christopher A

    2016-01-01

    Anxiety and affective disorders are often associated with hypercortisolism and dysfunctional serotonergic systems, including increased expression of TPH2, the gene encoding the rate-limiting enzyme of neuronal serotonin synthesis. We previously reported that chronic glucocorticoid exposure is anxiogenic and increases rat Tph2 mRNA expression, but it was still unclear if this also translates to increased TPH2 protein levels and in vivo activity of the enzyme. Here, we found that adult male rats treated with corticosterone (CORT, 100 μg/ml) via the drinking water for 21 days indeed show increased TPH2 protein expression in the dorsal and ventral part of the dorsal raphe nucleus (DRD, DRV) during the light phase, abolishing the enzyme's diurnal rhythm. In a second study, we systemically blocked the conversion of 5-hydroxytryptophan (5-HTP) to serotonin immediately before rats treated with CORT or vehicle were either exposed to 30 min acoustic startle stress or home cage control conditions. This allowed us to measure 5-HTP accumulation as a direct readout of basal versus stress-induced in vivo TPH2 activity. As expected, basal TPH2 activity was elevated in the DRD, DRV and MnR of CORT-treated rats. In response to stress, a multitude of serotonergic systems reacted with increased TPH2 activity, but the stress-, anxiety-, and learned helplessness-related dorsal and caudal DR (DRD/DRC) displayed stress-induced increases in TPH2 activity only after chronic CORT-treatment. To address the mechanisms underlying this region-specific CORT-dependent sensitization, we stereotaxically implanted CORT-treated rats with cannulae targeting the DR, and pharmacologically blocked either corticotropin-releasing hormone receptor type 1 (CRHR1) or type 2 (CRHR2) 10 min prior to acoustic startle stress. CRHR2 blockade prevented stress-induced increases of TPH2 activity within the DRD/DRC, while blockade of CRHR1 potentiated stress-induced TPH2 activity in the entire DR. Stress-induced TPH2 activity in the DRD/DRC furthermore predicted TPH2 activity in the amygdala and in the caudal pontine reticular nucleus (PnC), while serotonin synthesis in the PnC was strongly correlated with the maximum startle response. Our data demonstrate that chronically elevated glucocorticoids sensitize stress- and anxiety-related serotonergic systems, and for the first time reveal competing roles of CRHR1 and CRHR2 on stress-induced in vivo serotonin synthesis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Serotonergic systems in the balance: CRHR1 and CRHR2 differentially control stress-induced serotonin synthesis

    PubMed Central

    Donner, Nina C.; Siebler, Philip H.; Johnson, Danté T.; Villarreal, Marcos D.; Mani, Sofia; Matti, Allison J.; Lowry, Christopher A.

    2015-01-01

    Anxiety and affective disorders are often associated with hypercortisolism and dysfunctional serotonergic systems, including increased expression of TPH2, the gene encoding the rate-limiting enzyme of neuronal serotonin synthesis. We previously reported that chronic glucocorticoid exposure is anxiogenic and increases rat Tph2 mRNA expression, but it was still unclear if this also translates to increased TPH2 protein levels and in vivo activity of the enzyme. Here, we found that adult male rats treated with corticosterone (CORT, 100 μg/ml) via the drinking water for 21 days indeed show increased TPH2 protein expression in the dorsal and ventral part of the dorsal raphe nucleus (DRD, DRV) during the light phase, abolishing the enzyme’s diurnal rhythm. In a second study, we systemically blocked the conversion of 5-hydroxytryptophan (5-HTP) to serotonin immediately before rats treated with CORT or vehicle were either exposed to 30 min acoustic startle stress or home cage control conditions. This allowed us to measure 5-HTP accumulation as a direct readout of basal versus stress-induced in vivo TPH2 activity. As expected, basal TPH2 activity was elevated in the DRD, DRV and MnR of CORT-treated rats. In response to stress, a multitude of serotonergic systems reacted with increased TPH2 activity, but the stress-, anxiety-, and learned helplessness-related dorsal and caudal DR (DRD/DRC) displayed stress-induced increases in TPH2 activity only after chronic CORT-treatment. To address the mechanisms underlying this region-specific CORT-dependent sensitization, we stereotaxically implanted CORT-treated rats with cannulae targeting the DR, and pharmacologically blocked either corticotropin-releasing hormone receptor type 1 (CRHR1) or type 2 (CRHR2) 10 min prior to acoustic startle stress. CRHR2 blockade prevented stress-induced increases of TPH2 activity within the DRD/DRC, while blockade of CRHR1 potentiated stress-induced TPH2 activity in the entire DR. Stress-induced TPH2 activity in the DRD/DRC furthermore predicted TPH2 activity in the amygdala and in the caudal pontine reticular nucleus (PnC), while serotonin synthesis in the PnC was strongly correlated with the maximum startle response. Our data demonstrate that chronically elevated glucocorticoids sensitize stress- and anxiety-related serotonergic systems, and for the first time reveal competing roles of CRHR1 and CRHR2 on stress-induced in vivo serotonin synthesis. PMID:26454419

  13. Chronic central serotonin depletion attenuates ventilation and body temperature in young but not adult Tph2 knockout rats.

    PubMed

    Kaplan, Kara; Echert, Ashley E; Massat, Ben; Puissant, Madeleine M; Palygin, Oleg; Geurts, Aron M; Hodges, Matthew R

    2016-05-01

    Genetic deletion of brain serotonin (5-HT) neurons in mice leads to ventilatory deficits and increased neonatal mortality during development. However, it is unclear if the loss of the 5-HT neurons or the loss of the neurochemical 5-HT led to the observed physiologic deficits. Herein, we generated a mutant rat model with constitutive central nervous system (CNS) 5-HT depletion by mutation of the tryptophan hydroxylase 2 (Tph2) gene in dark agouti (DA(Tph2-/-)) rats. DA(Tph2-/-) rats lacked TPH immunoreactivity and brain 5-HT but retain dopa decarboxylase-expressing raphe neurons. Mutant rats were also smaller, had relatively high mortality (∼50%), and compared with controls had reduced room air ventilation and body temperatures at specific postnatal ages. In adult rats, breathing at rest and hypoxic and hypercapnic chemoreflexes were unaltered in adult male and female DA(Tph2-/-) rats. Body temperature was also maintained in adult DA(Tph2-/-) rats exposed to 4°C, indicating unaltered ventilatory and/or thermoregulatory control mechanisms. Finally, DA(Tph2-/-) rats treated with the 5-HT precursor 5-hydroxytryptophan (5-HTP) partially restored CNS 5-HT and showed increased ventilation (P < 0.05) at a developmental age when it was otherwise attenuated in the mutants. We conclude that constitutive CNS production of 5-HT is critically important to fundamental homeostatic control systems for breathing and temperature during postnatal development in the rat. Copyright © 2016 the American Physiological Society.

  14. Interaction of brain 5-HT synthesis deficiency, chronic stress and sex differentially impact emotional behavior in Tph2 knockout mice.

    PubMed

    Gutknecht, Lise; Popp, Sandy; Waider, Jonas; Sommerlandt, Frank M J; Göppner, Corinna; Post, Antonia; Reif, Andreas; van den Hove, Daniel; Strekalova, Tatyana; Schmitt, Angelika; Colaςo, Maria B N; Sommer, Claudia; Palme, Rupert; Lesch, Klaus-Peter

    2015-07-01

    While brain serotonin (5-HT) function is implicated in gene-by-environment interaction (GxE) impacting the vulnerability-resilience continuum in neuropsychiatric disorders, it remains elusive how the interplay of altered 5-HT synthesis and environmental stressors is linked to failure in emotion regulation. Here, we investigated the effect of constitutively impaired 5-HT synthesis on behavioral and neuroendocrine responses to unpredictable chronic mild stress (CMS) using a mouse model of brain 5-HT deficiency resulting from targeted inactivation of the tryptophan hydroxylase-2 (Tph2) gene. Locomotor activity and anxiety- and depression-like behavior as well as conditioned fear responses were differentially affected by Tph2 genotype, sex, and CMS. Tph2 null mutants (Tph2(-/-)) displayed increased general metabolism, marginally reduced anxiety- and depression-like behavior but strikingly increased conditioned fear responses. Behavioral modifications were associated with sex-specific hypothalamic-pituitary-adrenocortical (HPA) system alterations as indicated by plasma corticosterone and fecal corticosterone metabolite concentrations. Tph2(-/-) males displayed increased impulsivity and high aggressiveness. Tph2(-/-) females displayed greater emotional reactivity to aversive conditions as reflected by changes in behaviors at baseline including increased freezing and decreased locomotion in novel environments. However, both Tph2(-/-) male and female mice were resilient to CMS-induced hyperlocomotion, while CMS intensified conditioned fear responses in a GxE-dependent manner. Our results indicate that 5-HT mediates behavioral responses to environmental adversity by facilitating the encoding of stress effects leading to increased vulnerability for negative emotionality.

  15. Common variants in the TPH2 promoter confer susceptibility to paranoid schizophrenia.

    PubMed

    Yi, Zhenghui; Zhang, Chen; Lu, Weihong; Song, Lisheng; Liu, Dentang; Xu, Yifeng; Fang, Yiru

    2012-07-01

    Serotonergic system-related genes may be good candidates in investigating the genetic basis of schizophrenia. Our previous study suggested that promoter region of tryptophan hydroxylase 2 gene (TPH2) may confer the susceptibility to paranoid schizophrenia. In this study, we investigated whether common variants within TPH2 promoter may predispose to paranoid schizophrenia in Han Chinese. A total of 509 patients who met DSM-IV criteria for paranoid schizophrenia and 510 matched healthy controls were recruited for this study. Five polymorphisms within TPH2 promoter region were tested. No statistically significant differences were found in allele or genotype frequencies between schizophrenic patients and healthy controls. The frequency of the rs4448731T-rs6582071A-rs7963803A-rs4570625T-rs11178997A haplotype was significantly higher in cases compared to the controls (P = 0.003; OR = 1.49; 95% CI, 1.15-1.95). Our results suggest that the common variants within TPH2 promoter are associated with paranoid schizophrenia in Han Chinese. Further studies in larger samples are warranted to elucidate the role of TPH2 in the etiology of paranoid schizophrenia.

  16. Placental lactogens induce serotonin biosynthesis in a subset of mouse beta cells during pregnancy

    PubMed Central

    Schraenen, A.; Lemaire, K.; de Faudeur, G.; Hendrickx, N.; Granvik, M.; Van Lommel, L.; Mallet, J.; Vodjdani, G.; Gilon, P.; Binart, N.; in’t Veld, P.

    2010-01-01

    Aims/hypothesis Upregulation of the functional beta cell mass is required to match the physiological demands of mother and fetus during pregnancy. This increase is dependent on placental lactogens (PLs) and prolactin receptors, but the mechanisms underlying these events are only partially understood. We studied the mRNA expression profile of mouse islets during pregnancy to gain a better insight into these changes. Methods RNA expression was measured ex vivo via microarrays and quantitative RT-PCR. In vivo observations were extended by in vitro models in which ovine PL was added to cultured mouse islets and MIN6 cells. Results mRNA encoding both isoforms of the rate-limiting enzyme of serotonin biosynthesis, tryptophan hydroxylase (TPH), i.e. Tph1 and Tph2, were strongly induced (fold change 25- to 200-fold) during pregnancy. This induction was mimicked by exposing islets or MIN6 cells to ovine PLs for 24 h and was dependent on janus kinase 2 and signal transducer and activator of transcription 5. Parallel to Tph1 mRNA and protein induction, islet serotonin content increased to a peak level that was 200-fold higher than basal. Interestingly, only a subpopulation of the beta cells was serotonin-positive in vitro and in vivo. The stored serotonin pool in pregnant islets and PL-treated MIN6 cells was rapidly released (turnover once every 2 h). Conclusions/interpretation A very strong lactogen-dependent upregulation of serotonin biosynthesis occurs in a subpopulation of mouse islet beta cells during pregnancy. Since the newly formed serotonin is rapidly released, this lactogen-induced beta cell function may serve local or endocrine tasks, the nature of which remains to be identified. Electronic supplementary material The online version of this article (doi:10.1007/s00125-010-1913-7) contains supplementary material, which is available to authorised users. PMID:20938637

  17. Physical Weight Loading Induces Expression of Tryptophan Hydroxylase 2 in the Brain Stem

    PubMed Central

    Shim, Joon W.; Dodge, Todd R.; Hammond, Max A.; Wallace, Joseph M.; Zhou, Feng C.; Yokota, Hiroki

    2014-01-01

    Sustaining brain serotonin is essential in mental health. Physical activities can attenuate mental problems by enhancing serotonin signaling. However, such activity is not always possible in disabled individuals or patients with dementia. Knee loading, a form of physical activity, has been found to mimic effects of voluntary exercise. Focusing on serotonergic signaling, we addressed a question: Does local mechanical loading to the skeleton elevate expression of tryptophan hydroxylase 2 (tph2) that is a rate-limiting enzyme for brain serotonin? A 5 min knee loading was applied to mice using 1 N force at 5 Hz for 1,500 cycles. A 5-min treadmill running was used as an exercise (positive) control, and a 90-min tail suspension was used as a stress (negative) control. Expression of tph2 was determined 30 min – 2 h in three brain regions ––frontal cortex (FC), ventromedial hypothalamus (VMH), and brain stem (BS). We demonstrated for the first time that knee loading and treadmill exercise upregulated the mRNA level of tph2 in the BS, while tail suspension downregulated it. The protein level of tph2 in the BS was also upregulated by knee loading and downregulated by tail suspension. Furthermore, the downregulation of tph2 mRNA by tail suspension can be partially suppressed by pre-application of knee loading. The expression of tph2 in the FC and VMH was not significantly altered with knee loading. In this study we provided evidence that peripheral mechanical loading can activate central tph2 expression, suggesting that physical cues may mediate tph2-cathalyzed serotonergic signaling in the brain. PMID:24416346

  18. Autoantibodies against aromatic amino acid hydroxylases in patients with autoimmune polyendocrine syndrome type 1 target multiple antigenic determinants and reveal regulatory regions crucial for enzymatic activity.

    PubMed

    Bratland, Eirik; Magitta, Ng'weina Francis; Bøe Wolff, Anette Susanne; Ekern, Trude; Knappskog, Per Morten; Kämpe, Olle; Haavik, Jan; Husebye, Eystein Sverre

    2013-06-01

    Patients with autoimmune polyendocrine syndrome type 1 (APS-1) frequently have autoantibodies directed against the aromatic amino acid hydroxylases tryptophan hydroxylase (TPH) and tyrosine hydroxylase (TH). We aimed to characterize these autoantibodies with regard to their antigenic determinants, their influence on enzymatic activity and their clinical associations. In particular, we wanted to compare autoantibodies against the two different isoforms of TPH, which display different tissue distribution. Using sera from 48 Scandinavian APS-1 patients we identified 36 patients (75%) with antibodies against one or more of these three enzymes. Antibodies against TPH1, but not TPH2, were associated with malabsorption in the whole Scandinavian cohort, while TH antibodies were associated with dental enamel hypoplasia in Norwegian patients. Subsequent experiments with selected patient sera indicated that while the C-terminal domain was the immunodominant part of TPH1, the epitopes of TPH2 and TH were mainly located in the N-terminal regulatory domains. We also identified a TPH1 specific epitope involved in antibody mediated inhibition of enzyme activity, a finding that provides new insight into the enzymatic mechanisms of the aromatic amino acid hydroxylases and knowledge about structural determinants of enzyme autoantigens. In conclusion, TPH1, TPH2 and TH all have unique antigenic properties in spite of their structural similarity. Copyright © 2012 Elsevier GmbH. All rights reserved.

  19. Autoantibodies to human tryptophan hydroxylase and aromatic L-amino acid decarboxylase.

    PubMed

    Dal Pra, Chiara; Chen, Shu; Betterle, Corrado; Zanchetta, Renato; McGrath, Vivienne; Furmaniak, Jadwiga; Rees Smith, Bernard

    2004-03-01

    To assess the prevalence of autoantibodies (Abs) to tryptophan hydroxylase (TPH) and aromatic l-amino acid decarboxylase (AADC) in patients with different autoimmune diseases and to analyse their respective epitopes. TPH and AADC Abs were measured in an immunoprecipitation assay using (35)S-labelled full-length and fragments of TPH and AADC. Patients with different autoimmune adrenal diseases (n=84), non-adrenal autoimmune diseases (n=37), idiopathic vitiligo (n=8) and 56 healthy blood donors were studied. Fourteen of twenty-three (61%) of patients with autoimmune polyglandular syndrome (APS) type I and 1/34 (3%) of patients with isolated Addison's disease (AD) were positive for TPH Abs. None of the patients with APS type II (n=27), coeliac disease (n=10), autoimmune thyroid disease (AITD) (n=11), type 1 diabetes mellitus (DM) (n=16) or idiopathic vitiligo (n=8) was positive for TPH Abs. AADC Abs were detected in 12/23 (52%) patients with APS type I, in 1/29 (3%) patients with APS type II and 1/34 (3%) patients with isolated AD. None of the patients with coeliac disease, type 1 DM, AITD or idiopathic vitiligo was positive for AADC Abs. TPH Abs were found to interact with the C-terminal amino acids (aa) 308-423, central aa 164-205 and N-terminal aa 1-105 of the TPH molecule. AADC Ab binding epitopes were within the C-terminal aa 382-483, the central aa 243-381 and the N-terminal aa 1-167. Our study suggests that TPH Abs and AADC Abs react with several different epitopes and that different epitopes are recognized by different sera. The prevalence of TPH Abs and AADC Abs in patients with APS type I in our study is in agreement with previous reports. TPH Abs and AADC Abs were found very rarely in patients with other forms of autoimmune adrenal disease and were not detected in patients with non-adrenal autoimmune diseases.

  20. Melatonin Synthesis: Acetylserotonin O-Methyltransferase (ASMT) Is Strongly Expressed in a Subpopulation of Pinealocytes in the Male Rat Pineal Gland.

    PubMed

    Rath, Martin F; Coon, Steven L; Amaral, Fernanda G; Weller, Joan L; Møller, Morten; Klein, David C

    2016-05-01

    The rat pineal gland has been extensively used in studies of melatonin synthesis. However, the cellular localization of melatonin synthesis in this species has not been investigated. Here we focus on the localization of melatonin synthesis using immunohistochemical methods to detect the last enzyme in melatonin synthesis, acetylserotonin O-methyltransferase (ASMT), and in situ hybridization techniques to study transcripts encoding ASMT and two other enzymes in melatonin synthesis, tryptophan hydroxylase (TPH)-1 and aralkylamine N-acetyltransferase. In sections of the rat pineal gland, marked cell-to-cell differences were found in ASMT immunostaining intensity and in the abundance of Tph1, Aanat, and Asmt transcripts. ASMT immunoreactivity was localized to the cytoplasm in pinealocytes in the parenchyma of the superficial pineal gland, and immunopositive pinealocytes were also detected in the pineal stalk and in the deep pineal gland. ASMT was found to inconsistently colocalize with S-antigen, a widely used pinealocyte marker; this colocalization was seen in cells throughout the pineal complex and also in displaced pinealocyte-like cells of the medial habenular nucleus. Inconsistent colocalization between ASMT and TPH protein was also detected in the pineal gland. ASMT protein was not detected in extraepithalamic parts of the central nervous system or in peripheral tissues. The findings in this report are of special interest because they provide reason to suspect that melatonin synthesis varies significantly among individual pinealocytes.

  1. Melatonin Synthesis: Acetylserotonin O-Methyltransferase (ASMT) Is Strongly Expressed in a Subpopulation of Pinealocytes in the Male Rat Pineal Gland

    PubMed Central

    Coon, Steven L.; Amaral, Fernanda G.; Weller, Joan L.; Møller, Morten; Klein, David C.

    2016-01-01

    The rat pineal gland has been extensively used in studies of melatonin synthesis. However, the cellular localization of melatonin synthesis in this species has not been investigated. Here we focus on the localization of melatonin synthesis using immunohistochemical methods to detect the last enzyme in melatonin synthesis, acetylserotonin O-methyltransferase (ASMT), and in situ hybridization techniques to study transcripts encoding ASMT and two other enzymes in melatonin synthesis, tryptophan hydroxylase (TPH)-1 and aralkylamine N-acetyltransferase. In sections of the rat pineal gland, marked cell-to-cell differences were found in ASMT immunostaining intensity and in the abundance of Tph1, Aanat, and Asmt transcripts. ASMT immunoreactivity was localized to the cytoplasm in pinealocytes in the parenchyma of the superficial pineal gland, and immunopositive pinealocytes were also detected in the pineal stalk and in the deep pineal gland. ASMT was found to inconsistently colocalize with S-antigen, a widely used pinealocyte marker; this colocalization was seen in cells throughout the pineal complex and also in displaced pinealocyte-like cells of the medial habenular nucleus. Inconsistent colocalization between ASMT and TPH protein was also detected in the pineal gland. ASMT protein was not detected in extraepithalamic parts of the central nervous system or in peripheral tissues. The findings in this report are of special interest because they provide reason to suspect that melatonin synthesis varies significantly among individual pinealocytes. PMID:26950199

  2. The peroxidase-mediated biodegradation of petroleum hydrocarbons in a H2O2-induced SBR using in-situ production of peroxidase: Biodegradation experiments and bacterial identification.

    PubMed

    Shekoohiyan, Sakine; Moussavi, Gholamreza; Naddafi, Kazem

    2016-08-05

    A bacterial peroxidase-mediated oxidizing process was developed for biodegrading total petroleum hydrocarbons (TPH) in a sequencing batch reactor (SBR). Almost complete biodegradation (>99%) of high TPH concentrations (4g/L) was attained in the bioreactor with a low amount (0.6mM) of H2O2 at a reaction time of 22h. A specific TPH biodegradation rate as high as 44.3mgTPH/gbiomass×h was obtained with this process. The reaction times required for complete biodegradation of TPH concentrations of 1, 2, 3, and 4g/L were 21, 22, 28, and 30h, respectively. The catalytic activity of hydrocarbon catalyzing peroxidase was determined to be 1.48U/mL biomass. The biodegradation of TPH in seawater was similar to that in fresh media (no salt). A mixture of bacteria capable of peroxidase synthesis and hydrocarbon biodegradation including Pseudomonas spp. and Bacillus spp. were identified in the bioreactor. The GC/MS analysis of the effluent indicated that all classes of hydrocarbons could be well-degraded in the H2O2-induced SBR. Accordingly, the peroxidase-mediated process is a promising method for efficiently biodegrading concentrated TPH-laden saline wastewater. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. TPH2 gene polymorphisms in the regulatory region are associated with paranoid schizophrenia in Northern Han Chinese.

    PubMed

    Xu, X M; Ding, M; Pang, H; Wang, B J

    2014-03-12

    In the last years, serotonin (5-HT) has been related with the pathophysiology of several psychiatric disorders, including schizophrenia. Thus, genes related to the serotonergic (5-HTergic) system are good candidate genes for schizophrenia. The rate-limiting enzyme of 5-HT synthesis is tryptophan hydroxylase 2 (TPH2). Single nucleotide polymorphisms (SNPs) in the regulatory regions of TPH2 gene may affect gene expression and biosynthesis of 5-HT triggering to various neuropsychiatric disorders related to 5-HT dysfunction. The present study explored the association of SNPs within the TPH2 gene with paranoid schizophrenia in Han Chinese. A total of 164 patients with schizophrenia and 244 healthy controls were genotyped for six TPH2 SNPs (rs4570625, rs11178997, rs11178998, rs41317118, rs17110747, and rs41317114). Significant group differences were observed in the allele and genotype frequencies of rs4570625 and in the frequencies of GTA and TTA haplotypes corresponding to rs4570625-rs11178997-rs11178998. Our findings suggest that common genetic variations of TPH2 are likely to contribute to genetic susceptibility to paranoid schizophrenia in Han Chinese. Further studies in larger samples are needed to replicate this association.

  4. Etiological classification of depression based on the enzymes of tryptophan metabolism.

    PubMed

    Fukuda, Katsuhiko

    2014-12-24

    Viewed in terms of input and output, the mechanisms of depression are still akin to a black box. However, there must be main pivots for diverse types of depression. From recent therapeutic observations, both the serotonin (5-HT) and kynurenine pathways of tryptophan metabolism may be of particular importance to improved understanding of depression. Here, I propose an etiological classification of depression, based on key peripheral and central enzymes of tryptophan metabolism. Endogenous depression is caused by a larger genetic component than reactive depression. Besides enterochromaffin and mast cells, tryptophan hydroxylase 1 (TPH1), primarily expressed in the gastrointestinal tract, is also found in 5-hydroxytryptophan-producing cells (5-HTP cells) in normal intestinal enterocytes, which are thought to essentially shunt 5-HT production in 5-HT-producing cells. Genetic studies have reported an association between TPH1 and depression, or the responsiveness of depression to antidepressive medication. Therefore, it is possible that hypofunctional 5-HTP cells (reflecting TPH1 dysfunction) in the periphery lead to deficient brain 5-HT levels. Additionally,it has been reported that higher TPH2 expression in depressed suicides may reflect a homeostatic response to deficient 5-HT levels. Subsequently, endogenous depression may be caused by TPH1 dysfunction combined with compensatory TPH2 activation. Reactive depression results from life stresses and involves the hypothalamic-pituitary-adrenal axis, with resulting cortisol production inducing tryptophan 2,3-dioxygenase (TDO) activation. In secondary depression, caused by inflammation, infection, or oxidative stress, indoleamine 2,3-dioxygenase (IDO) is activated. In both reactive and secondary depression, the balance between 3-hydroxykynurenine (3-HK) and kynurenic acid may shift towards 3-HK production via kynurenine-3-monooxygenase (KMO) activation. By shifting the equilibrium position of key enzymes of tryptophan metabolism, the classical classification of depression can be reorganized, as below. Peripheral classification of depression by key enzymes: TPH1 dysfunction, TDO activation, IDO activation. Central classification: TPH2 activation, KMO activation. Etiological classification of depression expressed by peripheral (TPH1, TDO, IDO) and central (TPH2, KMO)enzymes of tryptophan metabolism may enable depression to be viewed as a clear box, with the inner components available for inspection and treatment.

  5. Interactions between resin monomers and commercial composite resins with human saliva derived esterases.

    PubMed

    Jaffer, F; Finer, Y; Santerre, J P

    2002-04-01

    Cholesterol esterase (CE) and pseudocholinesterase (PCE) have been reported to degrade commercial and model composite resins containing bisphenylglycidyl dimethacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA) or the latter in combination with urethane modified BisGMA monomer systems. In addition, human saliva has been shown to contain esterase like activities similar to CE and PCE. Hence, it was the aim of the current study to determine to what extent human saliva could degrade two common commercial composite resins (Z250 from 3M Inc. and Spectrum TPH from L.D. Caulk) which contain the above monomer systems. Saliva samples from different volunteers were collected, processed, pooled, and freeze-dried. TEGDMA and BisGMA monomers were incubated with human saliva derived esterase activity (HSDEA) and their respective hydrolysis was monitored using high performance liquid chromatography (HPLC). Both monomers were completely hydrolyzed within 25 h by HSDEA. Photopolymerized composites were incubated with buffer or human saliva (pH 7.0 and 37 C) for 2, 8 and 16 days. The incubation solutions were analyzed using HPLC and mass spectrometry. Surface morphology characterization was carried out using scanning electron microscopy. Upon biodegradation, the Z250 composite yielded higher amounts of BisGMA and TEGDMA related products relative to the TPH composite. However, there were higher amounts of ethoxylated bis-phenol A released from the TPH material. In terms of total mass of products released, human saliva demonstrated a greater ability to degrade Z250. In summary, HSDEA has been shown to contain esterase activities that can readily catalyze the biodegradation of current commercial composite resins.

  6. Differential Patterning of Genes Involved in Serotonin Metabolism and Transport in Extra-embryonic Tissues of the Mouse

    PubMed Central

    Wu, Hsiao-Huei; Choi, Sera; Levitt, Pat

    2016-01-01

    Introduction Serotonin (5-HT) is an important neuromodulator, but recently has been shown to be involved in neurodevelopment. Although previous studies have demonstrated that the placenta is a major source of forebrain 5-HT during early forebrain development, the processes of how 5-HT production, metabolism, and transport from placenta to fetus are regulated are unknown. As an initial step in determining the mechanisms involved, we investigated the expression patterns of genes critical for 5-HT system function in mouse extraembryonic tissues. Methods Mid- through late gestation expression of 5-HT system-related enzymes, Tph1, Ddc, Maoa, and 5-HT transporters, Sert/Slc6a4, Oct3/Slc22a3, Vmat2/Slc18a2, and 5-HT in placenta and yolk sac were examined, with cell type-specific resolution, using multiplex fluorescent in situ hybridization to co-localize transcripts and immunocytochemistry to co-localize the corresponding proteins and neurotransmitter. Results Tph1 and Ddc are found in the syncytiotrophoblast I (SynT-I) and sinusoidal trophoblast giant cells (S-TGC), whereas Maoa is expressed in SynT-I, syncytiotrophoblast II (SynT-II) and S-TGC. Oct3 expression is observed in the SynT-II only, while Vmat2 is mainly expressed in S-TGC. Surprisingly, there were comparatively high expression of Tph1, Ddc, and Maoa in the yolk sac visceral endoderm. Discussion In addition to trophoblast cells, visceral endoderm cells in the yolk sac may contribute to fetal 5-HT production. The findings raise the possibility of a more complex regulation of 5-HT access to the fetus through the differential roles of trophoblasts that surround maternal and fetal blood space and of yolk sac endoderm prior to normal degeneration. PMID:27238716

  7. TBT and TPhT persistence in a sludged soil.

    PubMed

    Marcic, Christophe; Le Hecho, Isabelle; Denaix, Laurence; Lespes, Gaëtane

    2006-12-01

    The persistence of tributyltin (TBT) and triphenyltin (TPhT) in soils was studied, taking into consideration the quantity of sewage sludge, TBT and TPhT concentrations in soil as well as the soil pH. The organotin compounds (OTC) were introduced into the soil via a spiked urban sludge, simulating agricultural practise. OTC speciation was achieved after acidic extraction of soil samples followed by gas chromatography-pulsed flame photometric analysis (GC-PFPD). Leaching tests conducted on a spiked sludge showed that more than 98% of TBT are sorbed on the sludge. TBT persistence in soil appeared to depend on its initial concentration in sludge. Thus, it was more important when concentration is over 1000 microg(Sn) kg(-1) of sludge. More than 50% of the initial TBT added into the soil were still present after 2 months, whatever the experimental conditions. The main degradation product appeared to be dibutyltin. About 90% of TPhT were initially sorbed on sludge, whatever the spiking concentration in sludge was. However, TPhT seemed to be quantitatively exchangeable at the solid/liquid interface, according to the leaching tests. It was also significantly degraded in sludged soil as only about 20% of TPhT remain present after 2 months, the monophenyltin being the main degradation product. pH had a significant positive effect on TBT and particularly TPhT persistence, according to the initial amounts introduced into the soil. Thus, at pH over 7 and triorganotin concentration over 100 microg(Sn) kg(-1), less than 10% of TBT but about 60% of TPhT were degraded. When the sludge was moderately contaminated by triorganotins (typically 50 microg(Sn) kg(-1) in our conditions) the pH had no effect on TBT and TPhT persistence.

  8. Enhancement of Gastric Ulcer Healing and Angiogenesis by Hepatocyte Growth Factor Gene Mediated by Attenuated Salmonella in Rats

    PubMed Central

    2017-01-01

    The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 109 cfu), vehicle (TP, 1 × 109 cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm2, which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm2, respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P < 0.05). The microvessel density in the ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm2, which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site. PMID:28049228

  9. Atmospheric chemical reactions of alternatives of polybrominated diphenyl ethers initiated by OH: A case study on triphenyl phosphate.

    PubMed

    Yu, Qi; Xie, Hong-Bin; Chen, Jingwen

    2016-11-15

    Many studies have been performed to evaluate the environmental risk caused by alternative flame retardants (AFRs) of polybrominated diphenyl ethers due to their ubiquitous occurrence in the environment. However, as an indispensable component of the environmental risk assessment, the information on atmospheric fate of AFRs is limited although some AFRs have been frequently and highly detected in the atmosphere. Here, a combined quantum chemical method and kinetics modeling were used to investigate atmospheric transformation mechanism and kinetics of AFRs initiated by OH in the presence of O2, taking triphenyl phosphate (TPhP) as a case. Results show that the pathway involving initial OH addition to phenyl of TPhP to form TPhP-OH adduct, and subsequent reaction of the TPhP-OH adduct with O2 to finally form phenol phosphate, is the most favorable for the titled reaction. The calculated overall reaction rate constant is 1.6×10(-12)cm(3) molecule(-1)s(-1), translating 7.6days atmospheric lifetime of TPhP. This clarifies that gaseous TPhP has atmospheric persistence. In addition, it was found that ice surface, as a case of ubiquitous water in the atmosphere, has little effect on the kinetics of the rate-determining step in the OH-initiated TPhP reaction. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. AGN-2979, an inhibitor of tryptophan hydroxylase activation, does not affect serotonin synthesis in Flinders Sensitive Line rats, a rat model of depression, but produces a significant effect in Flinders Resistant Line rats

    PubMed Central

    Kanemaru, Kazuya; Nishi, Kyoko; Diksic, Mirko

    2009-01-01

    The neurotransmitter, serotonin, is involved in several brain functions, including both normal, physiological functions, and pathophysiological functions. Alterations in any of the normal parameters of serotonergic neurotransmission can produce several different psychiatric disorders, including major depression. In many instances, brain neurochemical variables are not able to be studied properly in humans, thus making the use of good animal models extremely valuable. One of these animal models is the Flinders Sensitive Line (FSL) of rats, which has face, predictive and constructive validities in relation to human depression. The objective of this study was to quantify the effect of the tryptophan hydroxylase (TPH) activation inhibitor, AGN-2979, on the FSL rats (rats with depression-like behaviour), and compare it to the effect on the Flinders Resistant Line (FRL) of rats used as the control rats. The effect was evaluated by measuring changes in regional serotonin synthesis in the vehicle treated rats (FSL-VEH and FRL-VEH) relative to those measured in the AGN-2979 treated rats (FSL-AGN and FRL-AGN). Regional serotonin synthesis was measured autoradiographically in more than thirty brain regions. The measurements were performed using α-[14C]methyl-L-tryptophan as the tracer. The results indicate that AGN-2979 did not produce a significant reduction of TPH activity in the AGN-2979 group relative to the vehicle group (a reduction would have been observed if there had been an activation of TPH by the experimental set up) in the FSL rats. On the other hand, there was a highly significant reduction of synthesis in the FRL rats treated by AGN-2979, relative to the vehicle group. Together, the results demonstrate that in the FSL rats, AGN-2979 does not affect serotonin synthesis. This suggests that there was no activation of TPH in the FSL rats during the experimental procedure, but such activation did occur in the FRL rats. Because of this finding, it could be hypothesised that TPH in the FSL rats cannot be easily activated. This may contribute to the development of depressive-like symptoms in the FSL rats (“depressed” rats), as they cannot easily modulate their need for elevated amounts of this neurotransmitter, and possibly other neurotransmitters. Further, because these rats represent a very good model of human depression, one can hypothesize that humans who do not have readily activated TPH may be more prone to develop depression. PMID:19463878

  11. Response of the microbial community to seasonal groundwater level fluctuations in petroleum hydrocarbon-contaminated groundwater.

    PubMed

    Zhou, Ai-xia; Zhang, Yu-ling; Dong, Tian-zi; Lin, Xue-yu; Su, Xiao-si

    2015-07-01

    The effects of seasonal groundwater level fluctuations on the contamination characteristics of total petroleum hydrocarbons (TPH) in soils, groundwater, and the microbial community were investigated at a typical petrochemical site in northern China. The measurements of groundwater and soil at different depths showed that significant TPH residue was present in the soil in this study area, especially in the vicinity of the pollution source, where TPH concentrations were up to 2600 mg kg(-1). The TPH concentration in the groundwater fluctuated seasonally, and the maximum variation was 0.8 mg L(-1). The highest TPH concentrations were detected in the silty clay layer and lied in the groundwater level fluctuation zones. The groundwater could reach previously contaminated areas in the soil, leading to higher groundwater TPH concentrations as TPH leaches into the groundwater. The coincident variation of the electron acceptors and TPH concentration with groundwater-table fluctuations affected the microbial communities in groundwater. The microbial community structure was significantly different between the wet and dry seasons. The canonical correspondence analysis (CCA) results showed that in the wet season, TPH, NO3(-), Fe(2+), TMn, S(2-), and HCO3(-) were the major factors correlating the microbial community. A significant increase in abundance of operational taxonomic unit J1 (97% similar to Dechloromonas aromatica sp.) was also observed in wet season conditions, indicating an intense denitrifying activity in the wet season environment. In the dry season, due to weak groundwater level fluctuations and low temperature of groundwater, the microbial activity was weak. But iron and sulfate-reducing were also detected in dry season at this site. As a whole, groundwater-table fluctuations would affect the distribution, transport, and biodegradation of the contaminants. These results may be valuable for the control and remediation of soil and groundwater pollution at this site and in other petrochemical-contaminated areas. Furthermore, they are probably helpful for reducing health risks to the general public from contaminated groundwater.

  12. Intermittent hypoxia promotes recovery of respiratory motor function in spinal cord-injured mice depleted of serotonin in the central nervous system.

    PubMed

    Komnenov, Dragana; Solarewicz, Julia Z; Afzal, Fareeza; Nantwi, Kwaku D; Kuhn, Donald M; Mateika, Jason H

    2016-08-01

    We examined the effect of repeated daily exposure to intermittent hypoxia (IH) on the recovery of respiratory and limb motor function in mice genetically depleted of central nervous system serotonin. Electroencephalography, diaphragm activity, ventilation, core body temperature, and limb mobility were measured in spontaneously breathing wild-type (Tph2(+/+)) and tryptophan hydroxylase 2 knockout (Tph2(-/-)) mice. Following a C2 hemisection, the mice were exposed daily to IH (i.e., twelve 4-min episodes of 10% oxygen interspersed with 4-min normoxic periods followed by a 90-min end-recovery period) or normoxia (i.e., sham protocol, 21% oxygen) for 10 consecutive days. Diaphragm activity recovered to prehemisection levels in the Tph2(+/+) and Tph2(-/-) mice following exposure to IH but not normoxia [Tph2(+/+) 1.3 ± 0.2 (SE) vs. 0.3 ± 0.2; Tph2(-/-) 1.06 ± 0.1 vs. 0.3 ± 0.1, standardized to prehemisection values, P < 0.01]. Likewise, recovery of tidal volume and breathing frequency was evident, although breathing frequency values did not return to prehemisection levels within the time frame of the protocol. Partial recovery of limb motor function was also evident 2 wk after spinal cord hemisection. However, recovery was not dependent on IH or the presence of serotonin in the central nervous system. We conclude that IH promotes recovery of respiratory function but not basic motor tasks. Moreover, we conclude that spontaneous or treatment-induced recovery of respiratory and motor limb function is not dependent on serotonin in the central nervous system in a mouse model of spinal cord injury.

  13. 76 FR 75584 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-02

    ... Holder firm. Each FBW user has an FBW Login ID. The Exchange proposes to charge a Requesting TPH $100 per Login ID per month. There will be a cap of $2,000 per month for any Requesting TPH. Therefore, any TPH that requests access to the FBW Market Access Controls Window for more than 20 login IDs will not be...

  14. Functional Role of Serotonin in Insulin Secretion in a Diet-Induced Insulin-Resistant State

    PubMed Central

    Kim, Kyuho; Oh, Chang-Myung; Ohara-Imaizumi, Mica; Park, Sangkyu; Namkung, Jun; Yadav, Vijay K.; Tamarina, Natalia A.; Roe, Michael W.; Philipson, Louis H.; Karsenty, Gerard; Nagamatsu, Shinya

    2015-01-01

    The physiological role of serotonin, or 5-hydroxytryptamine (5-HT), in pancreatic β-cell function was previously elucidated using a pregnant mouse model. During pregnancy, 5-HT increases β-cell proliferation and glucose-stimulated insulin secretion (GSIS) through the Gαq-coupled 5-HT2b receptor (Htr2b) and the 5-HT3 receptor (Htr3), a ligand-gated cation channel, respectively. However, the role of 5-HT in β-cell function in an insulin-resistant state has yet to be elucidated. Here, we characterized the metabolic phenotypes of β-cell-specific Htr2b−/− (Htr2b βKO), Htr3a−/− (Htr3a knock-out [KO]), and β-cell-specific tryptophan hydroxylase 1 (Tph1)−/− (Tph1 βKO) mice on a high-fat diet (HFD). Htr2b βKO, Htr3a KO, and Tph1 βKO mice exhibited normal glucose tolerance on a standard chow diet. After 6 weeks on an HFD, beginning at 4 weeks of age, both Htr3a KO and Tph1 βKO mice developed glucose intolerance, but Htr2b βKO mice remained normoglycemic. Pancreas perfusion assays revealed defective first-phase insulin secretion in Htr3a KO mice. GSIS was impaired in islets isolated from HFD-fed Htr3a KO and Tph1 βKO mice, and 5-HT treatment improved insulin secretion from Tph1 βKO islets but not from Htr3a KO islets. Tph1 and Htr3a gene expression in pancreatic islets was not affected by an HFD, and immunostaining could not detect 5-HT in pancreatic islets from mice fed an HFD. Taken together, these results demonstrate that basal 5-HT levels in β-cells play a role in GSIS through Htr3, which becomes more evident in a diet-induced insulin-resistant state. PMID:25426873

  15. Evaluation of ethyl lactate as solvent in Fenton oxidation for the remediation of total petroleum hydrocarbon (TPH)-contaminated soil.

    PubMed

    Jalilian Ahmadkalaei, Seyedeh Pegah; Gan, Suyin; Ng, Hoon Kiat; Abdul Talib, Suhaimi

    2017-07-01

    Due to the health and environmental risks posed by the presence of petroleum-contaminated areas around the world, remediation of petroleum-contaminated soil has drawn much attention from researchers. Combining Fenton reaction with a solvent has been proposed as a novel way to remediate contaminated soils. In this study, a green solvent, ethyl lactate (EL), has been used in conjunction with Fenton's reagents for the remediation of diesel-contaminated soil. The main aim of this research is to determine how the addition of EL affects Fenton reaction for the destruction of total petroleum hydrocarbons (TPHs) within the diesel range. Specifically, the effects of different parameters, including liquid phase volume-to-soil weight (L/S) ratio, hydrogen peroxide (H 2 O 2 ) concentration and EL% on the removal efficiency, have been studied in batch experiments. The results showed that an increase in H 2 O 2 resulted in an increase in removal efficiency of TPH from 68.41% at H 2 O 2  = 0.1 M to 90.21% at H 2 O 2  = 2 M. The lowest L/S, i.e. L/S = 1, had the highest TPH removal efficiency of 85.77%. An increase in EL% up to 10% increased the removal efficiency to 96.74% for TPH, and with further increase in EL%, the removal efficiency of TPH decreased to 89.6%. EL with an optimum value of 10% was found to be best for TPH removal in EL-based Fenton reaction. The power law and pseudo-first order equations fitted well to the experimental kinetic data of Fenton reactions.

  16. [Influence of estradiol on tryptophan hydroxylase and 5-hydroxytryptamine content in raphe nuclei of rats under forced swimming stress].

    PubMed

    Yang, Fu-zhong; Wu, Yan; Zhang, Wei-guo; Cai, Yi-yun; Shi, Shen-xun

    2010-07-20

    To investigate the effect of estradiol (E2) on tryptophan hydroxylase (TPH) and 5-hydroxytryptamine (5-HT) content in raphe nuclei of rats under forced swimming stress and explore the role of estrogen and stress in disease mechanism of depression in women. At Week 3 post-ovariectomy, 35 ovariectomized (OVX) female SD rats were randomly divided into 5 groups (n = 7): non-stress group, control group, estradiol (E2) group and fluoxetine (FLX) group and E2 plus FLX group. Animals were administered with different drugs for 2 weeks. At Day 14, animals except those in the non-stress group were subjected to the 15 min forced swimming test (FST). At 2 hours post-FST, all animals including those in the non-stress group were perfused with 4% paraformaldehyde and brains removed for TPH and 5-HT immunofluorescence staining. We compared the content of TPH and 5-HT by observing and calculating the integrated optical density (IOD) of immunofluorescent-positive signals in raphe nuclei. (1) The IOD value of TPH- and 5-HT-positive region in raphe nuclei of rats in the control group was significantly lower than that of the non-stress group (P < 0.01); (2) the IOD value of TPH- and 5-HT-positive region in raphe nuclei of rats in the E2, FLX and E2 plus FLX groups was significantly higher than that in the control group (P < 0.05). Forced swimming stress can decrease the TPH and 5-HT content in raphe nuclei. Such changes can be prevented by a pre-administration of estradiol. Similar results are observed with antidepressant fluoxetine. These effects may underlie the role of estradiol and stress in the disease mechanism of depression in women.

  17. Biodegradation of total petroleum hydrocarbons from acidic sludge produced by re-refinery industries of waste oil using in-vessel composting.

    PubMed

    Asgari, Alireza; Nabizadeh, Ramin; Mahvi, Amir Hossein; Nasseri, Simin; Dehghani, Mohammad Hadi; Nazmara, Shahrokh; Yaghmaeian, Kamyar

    2017-01-01

    In Iran, re-refinery industry has been developed many years ago based on the acid-clay treatment. Acidic sludge with high concentration of total petroleum hydrocarbon (TPH) is the final products of some facilities. In this study removal of TPH by aerated in-vessel composting was investigated. In order to microorganisms seeding and nutrient providing, urban immature compost was added as an amendment to acidic sludge. The ratios of acidic sludge (AS) to compost were, 1:0 (as control), 1:5, 1:8, 1:10, 1:15, 1:20, 1:30, 1:40, 1:50, 1:75 and 1:100 (as dry basis) at a C: N: P ratio of 100:5:1 and 45-65% moisture content for 70 days. The removal efficiency in all reactors was more than 48%. The highest and lowest TPH removal was observed in 1:5 (71.56%) and 1:100 (48.53%) mixing ratios, respectively. The results of the control reactors showed that biological treatment was the main mechanism for TPH removal. Experimental data was fitted second order kinetic model ( R 2  > 0.8006). Degradation of TPH in 1:5 mixing ratio (k 2  = 0.0038 gmg -1 d -1 ; half-life = 3.08d) was nearly three times faster than 1:100 mixing ratio (k 2  = 0.0238; half-life = 8.96d). The results of the control reactors showed that biological treatment was the main mechanism for TPH removal. The results of this study revealed in-vessel composting with immature urban compost as the amendment maybe recommended as an effective method for TPH remediation.

  18. Removal of petroleum hydrocarbons from contaminated groundwater by the combined technique of adsorption onto perlite followed by the O3/H2O2 process.

    PubMed

    Moussavi, Gholamreza; Bagheri, Amir

    2012-09-01

    Groundwater contaminated with petroleum hydrocarbons was treated using a combined system of adsorption onto powdered expanded perlite (PEP) followed by the O3/H2O2 process. The pretreatment investigations indicated a high capacity for PEP to remove petroleum hydrocarbons from the contaminated water. An experimental total petroleum hydrocarbon (TPH) adsorption capacity of 275 mg/g PEP was obtained at the natural pH of water. The experimental data fit best with the Freundlich isotherm model and pseudo-second-order adsorption model. The second phase of the experiment evaluated the performance of the O3/H2O2 process in the removal of residual TPH from pretreated water and compared the results with that of raw water. The O3/H202 process attained a maximum TPH removal rate for the pretreated water after 70 min, when 93% of the residual TPH in the effluent of the adsorption system was removed. Overall, the combination of adsorption onto PEP for 100 min and the subsequent treatment with the O3/H2O2 process for 70min eliminated over 99% of the TPH of highly petroleum-contaminated groundwater, with initial values of 162 mg/L. Therefore, we can conclude that the developed treatment system is an appropriate method of remediation for petroleum-contaminated waters.

  19. 4-Chloro-DL-phenylalanine protects against monocrotaline‑induced pulmonary vascular remodeling and lung inflammation.

    PubMed

    Bai, Yang; Wang, Han-Ming; Liu, Ming; Wang, Yun; Lian, Guo-Chao; Zhang, Xin-Hua; Kang, Jian; Wang, Huai-Liang

    2014-02-01

    The present study was performed to investigate the effects of 4-chloro-DL-phenylalanine (PCPA), a tryptophan hydroxylase (Tph) inhibitor (TphI), on pulmonary vascular remodeling and lung inflammation in monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) in rats. Animal models of PAH were established using Sprague-Dawley (SD) rats by a single intraperitoneal injection of MCT (60 mg/kg). PCPA (50 or 100 mg/kg/day) was administered to the rats with PAH. On day 22, hemodynamic measurements and morphological observations of the lung tissues were performed. The levels of Tph-1 and serotonin transporter (SERT) in the lungs were analyzed by immunohistochemistry and western blot analysis. The expression of matrix metalloproteinase (MMP)-2 and MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 and inflammatory cytokines were assayed by western blot analysis. The activity of MMP-2 and MMP-9 was evaluated by gelatin zymography (GZ). MCT markedly promoted PAH, increased the right ventricular hypertrophy index, pulmonary vascular remodeling, lung inflammation and mortality, which was associated with the increased expression of Tph-1, SERT, MMP-2/-9, TIMP-1/-2 and inflammatory cytokines. PCPA markedly attenuated MCT-induced pulmonary vascular remodeling and lung inflammation, inhibited the expression of Tph-1 and SERT and suppressed the expression of MMP-2/-9, TIMP-1/-2, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and intercellular adhesion molecule-1 (ICAM-1). These findings suggest that the amelioration of MCT-induced pulmonary vascular remodeling and lung inflammation by PCPA is associated with the downregulation of Tph-1, SERT, MMP/TIMP and inflammatory cytokine expression in rats.

  20. An experimental study on the bio-surfactant-assisted remediation of crude oil and salt contaminated soils.

    PubMed

    Zhang, Wen; Li, Jianbing; Huang, Guohe; Song, Weikun; Huang, Yuefei

    2011-01-01

    The effect of bio-surfactant (rhamnolipid) on the remediation of crude oil and salt contaminated soil was investigated in this study. The experimental results indicated that there was a distinct decline of total petroleum hydrocarbon (TPH) concentration within the soil when using rhamnolipid during a remediation period of 30 days, with maximum TPH reduction of 86.97%. The most effective remediation that was observed was with rhamnolipid at a concentration of 2 CMC in soil solution, and a first-order TPH degradation rate constant of 0.0866 d(-1). The results also illustrated that salts in soil had a negative impact on TPH reduction, and the degradation rate was negatively correlated with NaCl concentration in soil solution. The analysis of soil TPH fractions indicated that there was a significant reduction of C13-C30 during the remediation process when using bio-surfactant.

  1. Serotonin and the Brain's Rich Club-Association Between Molecular Genetic Variation on the TPH2 Gene and the Structural Connectome.

    PubMed

    Markett, Sebastian; de Reus, Marcel A; Reuter, Martin; Montag, Christian; Weber, Bernd; Schoene-Bake, Jan-Christoph; van den Heuvel, Martijn P

    2017-03-01

    The rich club comprises a densely mutually connected set of hub regions in the brain, thought to serve as a processing and integration core. We assessed the impact of normal variation of the tryptophane hydroxylase 2 gene's promotor region (TPH2 rs4570625) on structural connectivity of the rich club pathways by means of a candidate gene association design. Tryptophane hydroxylase 2 (TPH2) is a rate-limiting enzyme in the biosynthesis of serotonin and is known to inhibit, in addition to its role as a trans-synaptic messenger, axonal and dendritic growth. The TPH2 T-variant has been associated with reduced mRNA expression and reduced serotonin levels, which may particularly influence the development of macroscale anatomical connectivity. Here, we show larger mean connectivity in the rich club in carriers of the T-variant, suggesting potential effects of upregulation of neural connectivity growth in this central core system. In addition, by edge-removal statistics, we show that the TPH2-associated higher levels of rich club connectivity are of importance for the functioning of the total structural network. The observed association is speculated to result from an effect of serotonin levels on brain development, potentially leading to stronger structural connectivity in heavily interconnected hubs. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. An Air-Stripping Packed Bed Combined with a Biofilm-Type Biological Process for Treating BTEX and Total Petroleum Hydrocarbon Contaminated Groudwater

    NASA Astrophysics Data System (ADS)

    Hong, U.; Park, S.; Lim, J.; Lee, W.; Kwon, S.; Kim, Y.

    2009-12-01

    In this study, we examined the removal efficiency of a volatile compound (e.g. toluene) and a less volatile compound [e.g. total petroleum hydrocarbon (TPH)] using an air stripping packed bed combined with a biofilm-type biological process. We hypothesized that this system might be effective and economical to simultaneously remove both volatile and less volatile compounds. The gas-tight reactor has 5.9-inch-diameter and 48.8-inch-height. A spray nozzle was installed at the top cover to distribute the liquid evenly through reactor. The reactor was filled with polypropylene packing media for the increase of volatilization surface area and the growth of TPH degrading facultative aerobic bacteria on the surface of the packing media. In air stripping experiments, 45.6%, 71.7%, 72.0%, and 75.4% of toluene was removed at air injection rates of 0 L/min, 2.5 L/min, 4 L/min, and 6 L/min, respectively. Through the result, we confirmed that toluene removal efficiency increased by injecting higher amounts of air. TPH removal by stripping was minimal. To remove a less volatile TPH by commercial TPH degrading culture (BIO-ZYME B-52), 15-times diluted culture was circulated through the reactor for 2-3 days to build up a biofilm on the surface of packing media with 1 mg-soluble nitrogen source /L-water per 1 ppm of TPH. Experiments evaluating the degree of TPH biodegradation in this system are carrying out.

  3. Optimizing photo-Fenton like process for the removal of diesel fuel from the aqueous phase

    PubMed Central

    2014-01-01

    Background In recent years, pollution of soil and groundwater caused by fuel leakage from old underground storage tanks, oil extraction process, refineries, fuel distribution terminals, improper disposal and also spills during transferring has been reported. Diesel fuel has created many problems for water resources. The main objectives of this research were focused on assessing the feasibility of using photo-Fenton like method using nano zero-valent iron (nZVI/UV/H2O2) in removing total petroleum hydrocarbons (TPH) and determining the optimal conditions using Taguchi method. Results The influence of different parameters including the initial concentration of TPH (0.1-1 mg/L), H2O2 concentration (5-20 mmole/L), nZVI concentration (10-100 mg/L), pH (3-9), and reaction time (15-120 min) on TPH reduction rate in diesel fuel were investigated. The variance analysis suggests that the optimal conditions for TPH reduction rate from diesel fuel in the aqueous phase are as follows: the initial TPH concentration equals to 0.7 mg/L, nZVI concentration 20 mg/L, H2O2 concentration equals to 5 mmol/L, pH 3, and the reaction time of 60 min and degree of significance for the study parameters are 7.643, 9.33, 13.318, 15.185 and 6.588%, respectively. The predicted removal rate in the optimal conditions was 95.8% and confirmed by data obtained in this study which was between 95-100%. Conclusion In conclusion, photo-Fenton like process using nZVI process may enhance the rate of diesel degradation in polluted water and could be used as a pretreatment step for the biological removal of TPH from diesel fuel in the aqueous phase. PMID:24955242

  4. Metabolomics Approach Reveals Integrated Metabolic Network Associated with Serotonin Deficiency

    PubMed Central

    Weng, Rui; Shen, Sensen; Tian, Yonglu; Burton, Casey; Xu, Xinyuan; Liu, Yi; Chang, Cuilan; Bai, Yu; Liu, Huwei

    2015-01-01

    Serotonin is an important neurotransmitter that broadly participates in various biological processes. While serotonin deficiency has been associated with multiple pathological conditions such as depression, schizophrenia, Alzheimer’s disease and Parkinson’s disease, the serotonin-dependent mechanisms remain poorly understood. This study therefore aimed to identify novel biomarkers and metabolic pathways perturbed by serotonin deficiency using metabolomics approach in order to gain new metabolic insights into the serotonin deficiency-related molecular mechanisms. Serotonin deficiency was achieved through pharmacological inhibition of tryptophan hydroxylase (Tph) using p-chlorophenylalanine (pCPA) or genetic knockout of the neuronal specific Tph2 isoform. This dual approach improved specificity for the serotonin deficiency-associated biomarkers while minimizing nonspecific effects of pCPA treatment or Tph2 knockout (Tph2-/-). Non-targeted metabolic profiling and a targeted pCPA dose-response study identified 21 biomarkers in the pCPA-treated mice while 17 metabolites in the Tph2-/- mice were found to be significantly altered compared with the control mice. These newly identified biomarkers were associated with amino acid, energy, purine, lipid and gut microflora metabolisms. Oxidative stress was also found to be significantly increased in the serotonin deficient mice. These new biomarkers and the overall metabolic pathways may provide new understanding for the serotonin deficiency-associated mechanisms under multiple pathological states. PMID:26154191

  5. Metabolomics Approach Reveals Integrated Metabolic Network Associated with Serotonin Deficiency.

    PubMed

    Weng, Rui; Shen, Sensen; Tian, Yonglu; Burton, Casey; Xu, Xinyuan; Liu, Yi; Chang, Cuilan; Bai, Yu; Liu, Huwei

    2015-07-08

    Serotonin is an important neurotransmitter that broadly participates in various biological processes. While serotonin deficiency has been associated with multiple pathological conditions such as depression, schizophrenia, Alzheimer's disease and Parkinson's disease, the serotonin-dependent mechanisms remain poorly understood. This study therefore aimed to identify novel biomarkers and metabolic pathways perturbed by serotonin deficiency using metabolomics approach in order to gain new metabolic insights into the serotonin deficiency-related molecular mechanisms. Serotonin deficiency was achieved through pharmacological inhibition of tryptophan hydroxylase (Tph) using p-chlorophenylalanine (pCPA) or genetic knockout of the neuronal specific Tph2 isoform. This dual approach improved specificity for the serotonin deficiency-associated biomarkers while minimizing nonspecific effects of pCPA treatment or Tph2 knockout (Tph2-/-). Non-targeted metabolic profiling and a targeted pCPA dose-response study identified 21 biomarkers in the pCPA-treated mice while 17 metabolites in the Tph2-/- mice were found to be significantly altered compared with the control mice. These newly identified biomarkers were associated with amino acid, energy, purine, lipid and gut microflora metabolisms. Oxidative stress was also found to be significantly increased in the serotonin deficient mice. These new biomarkers and the overall metabolic pathways may provide new understanding for the serotonin deficiency-associated mechanisms under multiple pathological states.

  6. Metabolomics Approach Reveals Integrated Metabolic Network Associated with Serotonin Deficiency

    NASA Astrophysics Data System (ADS)

    Weng, Rui; Shen, Sensen; Tian, Yonglu; Burton, Casey; Xu, Xinyuan; Liu, Yi; Chang, Cuilan; Bai, Yu; Liu, Huwei

    2015-07-01

    Serotonin is an important neurotransmitter that broadly participates in various biological processes. While serotonin deficiency has been associated with multiple pathological conditions such as depression, schizophrenia, Alzheimer’s disease and Parkinson’s disease, the serotonin-dependent mechanisms remain poorly understood. This study therefore aimed to identify novel biomarkers and metabolic pathways perturbed by serotonin deficiency using metabolomics approach in order to gain new metabolic insights into the serotonin deficiency-related molecular mechanisms. Serotonin deficiency was achieved through pharmacological inhibition of tryptophan hydroxylase (Tph) using p-chlorophenylalanine (pCPA) or genetic knockout of the neuronal specific Tph2 isoform. This dual approach improved specificity for the serotonin deficiency-associated biomarkers while minimizing nonspecific effects of pCPA treatment or Tph2 knockout (Tph2-/-). Non-targeted metabolic profiling and a targeted pCPA dose-response study identified 21 biomarkers in the pCPA-treated mice while 17 metabolites in the Tph2-/- mice were found to be significantly altered compared with the control mice. These newly identified biomarkers were associated with amino acid, energy, purine, lipid and gut microflora metabolisms. Oxidative stress was also found to be significantly increased in the serotonin deficient mice. These new biomarkers and the overall metabolic pathways may provide new understanding for the serotonin deficiency-associated mechanisms under multiple pathological states.

  7. Gestational Diabetes Mellitus From Inactivation of Prolactin Receptor and MafB in Islet β-Cells.

    PubMed

    Banerjee, Ronadip R; Cyphert, Holly A; Walker, Emily M; Chakravarthy, Harini; Peiris, Heshan; Gu, Xueying; Liu, Yinghua; Conrad, Elizabeth; Goodrich, Lisa; Stein, Roland W; Kim, Seung K

    2016-08-01

    β-Cell proliferation and expansion during pregnancy are crucial for maintaining euglycemia in response to increased metabolic demands placed on the mother. Prolactin and placental lactogen signal through the prolactin receptor (PRLR) and contribute to adaptive β-cell responses in pregnancy; however, the in vivo requirement for PRLR signaling specifically in maternal β-cell adaptations remains unknown. We generated a floxed allele of Prlr, allowing conditional loss of PRLR in β-cells. In this study, we show that loss of PRLR signaling in β-cells results in gestational diabetes mellitus (GDM), reduced β-cell proliferation, and failure to expand β-cell mass during pregnancy. Targeted PRLR loss in maternal β-cells in vivo impaired expression of the transcription factor Foxm1, both G1/S and G2/M cyclins, tryptophan hydroxylase 1 (Tph1), and islet serotonin production, for which synthesis requires Tph1. This conditional system also revealed that PRLR signaling is required for the transient gestational expression of the transcription factor MafB within a subset of β-cells during pregnancy. MafB deletion in maternal β-cells also produced GDM, with inadequate β-cell expansion accompanied by failure to induce PRLR-dependent target genes regulating β-cell proliferation. These results unveil molecular roles for PRLR signaling in orchestrating the physiologic expansion of maternal β-cells during pregnancy. © 2016 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.

  8. Bioremediation of diesel and lubricant oil-contaminated soils using enhanced landfarming system.

    PubMed

    Wang, Sih-Yu; Kuo, Yu-Chia; Hong, Andy; Chang, Yu-Min; Kao, Chih-Ming

    2016-12-01

    Lubricant and diesel oil-polluted sites are difficult to remediate because they have less volatile and biodegradable characteristics. The goal of this research was to evaluate the potential of applying an enhanced landfarming to bioremediate soils polluted by lubricant and diesel. Microcosm study was performed to evaluate the optimal treatment conditions with the addition of different additives (nutrients, addition of activated sludge from oil-refining wastewater facility, compost, TPH-degrading bacteria, and fern chips) to enhance total petroleum hydrocarbon (TPH) removal. To simulate the aerobic landfarming biosystem, air in the microcosm headspace was replaced once a week. Results demonstrate that the additives of activated sludge and compost could result in the increase in soil microbial populations and raise TPH degradation efficiency (up to 83% of TPH removal with 175 days of incubation) with initial (TPH = 4100 mg/kg). The first-order TPH degradation rate reached 0.01 1/d in microcosms with additive of activated sludge (mass ratio of soil to inocula = 50:1). The soil microbial communities were determined by nucleotide sequence analyses and 16S rRNA-based denatured gradient gel electrophoresis. Thirty-four specific TPH-degrading bacteria were detected in microcosm soils. Chromatograph analyses demonstrate that resolved peaks were more biodegradable than unresolved complex mixture. Results indicate that more aggressive remedial measures are required to enhance the TPH biodegradation, which included the increase of (1) microbial population or TPH-degrading bacteria, (2) biodegradable carbon sources, (3) nutrient content, and (4) soil permeability. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Detection of ethanol in alcoholic beverages or vapor phase using fluorescent molecules embedded in a nanofibrous polymer.

    PubMed

    Akamatsu, Masaaki; Mori, Taizo; Okamoto, Ken; Komatsu, Hirokazu; Kumagai, Ken; Shiratori, Seimei; Yamamura, Masaki; Nabeshima, Tatsuya; Sakai, Hideki; Abe, Masahiko; Hill, Jonathan P; Ariga, Katsuhiko

    2015-03-25

    An alcohol sensor was developed using the solid-state fluorescence emission of terphenyl-ol (TPhOH) derivatives. Admixtures of TPhOH and sodium carbonate exhibited bright sky-blue fluorescence in the solid state upon addition of small quantities of ethanol. A series of terphenol derivatives was synthesized, and the effects of solvent polarities and the structures of these π-conjugated systems on their fluorescence were systematically investigated by using fluorescence spectroscopy. In particular, π-extended TPhOHs and TPhOHs containing electron-withdrawing groups exhibited significant solvatochromism, and fluorescence colors varied from blue to red. Detection of ethanol contents in alcohol beverages (detection limit ∼ 5 v/v %) was demonstrated using different TPhOHs revealing the effect of molecular structure on sensing properties. Ethanol contents in alcoholic beverages could be estimated from the intensity of the fluorescence elicited from the TPhOHs. Moreover, when terphenol and Na2CO3 were combined with a water-absorbent polymer, ethanol could be detected at lower concentrations. Detection of ethanol vapor (8 v/v % in air) was also accomplished using a nanofibrous polymer scaffold as the immobilized sensing film.

  10. Association between Tryptophan Hydroxylase 2 Gene Polymorphism and Completed Suicide

    ERIC Educational Resources Information Center

    Fudalej, Sylwia; Ilgen, Mark; Fudalej, Marcin; Kostrzewa, Grazyna; Barry, Kristen; Wojnar, Marcin; Krajewski, Pawel; Blow, Frederic; Ploski, Rafal

    2010-01-01

    The association between suicide and a single nucleotide polymorphism (rs1386483) was examined in the recently identified tryptophan hydroxylase 2 (TPH2) gene. Blood samples of 143 suicide victims and 162 age- and sex-matched controls were examined. The frequency of the TT genotype in the TPH2 polymorphism was higher in suicide victims than in…

  11. Assessing soil and groundwater contamination in a metropolitan redevelopment project.

    PubMed

    Yun, Junki; Lee, Ju Young; Khim, Jeehyeong; Ji, Won Hyun

    2013-08-01

    The purpose of this study was to assess contaminated soil and groundwater for the urban redevelopment of a rapid transit railway and a new mega-shopping area. Contaminated soil and groundwater may interfere with the progress of this project, and residents and shoppers may be exposed to human health risks. The study area has been remediated after application of first remediation technologies. Of the entire area, several sites were still contaminated by waste materials and petroleum. For zinc (Zn) contamination, high Zn concentrations were detected because waste materials were disposed in the entire area. For petroleum contamination, high total petroleum hydrocarbon (TPH) and hydrocarbon degrading microbe concentrations were observed at the depth of 7 m because the underground petroleum storage tank had previously been located at this site. Correlation results suggest that TPH (soil) concentration is still related with TPH (groundwater) concentration. The relationship is taken into account in the Spearman coefficient (α).

  12. Central adiponectin administration reveals new regulatory mechanisms of bone metabolism in mice

    PubMed Central

    Wu, Yuwei; Tu, Qisheng; Valverde, Paloma; Zhang, Jin; Murray, Dana; Dong, Lily Q.; Cheng, Jessica; Jiang, Hua; Rios, Maribel; Morgan, Elise; Tang, Zhihui

    2014-01-01

    Adiponectin (APN), the most abundant adipocyte-secreted adipokine, regulates energy homeostasis and exerts well-characterized insulin-sensitizing properties. The peripheral or central effects of APN regulating bone metabolism are beginning to be explored but are still not clearly understood. In the present study, we found that APN-knockout (APN-KO) mice fed a normal diet exhibited decreased trabecular structure and mineralization and increased bone marrow adiposity compared with wild-type (WT) mice. APN intracerebroventricular infusions decreased uncoupling protein 1 (UCP1) expression in brown adipose tissue, epinephrine and norepinephrine serum levels, and osteoclast numbers, whereas osteoblast osteogenic marker expression and trabecular bone mass increased in APN-KO and WT mice. In addition, centrally administered APN increased hypothalamic tryptophan hydroxylase 2 (TPH2), cocaine- and amphetamine-regulated transcript (CART), and 5-hydroxytryptamine (serotonin) receptor 2C (Htr2C) expressions but decreased hypothalamic cannabinoid receptor-1 expression. Treatment of immortalized mouse neurons with APN demonstrated that APN-mediated effects on TPH2, CART, and Htr2C expression levels were abolished by downregulating adaptor protein containing pleckstrin homology domain, phosphotyrosine domain, and leucine zipper motif (APPL)-1 expression. Pharmacological increase in sympathetic activity stimulated adipogenic differentiation of bone marrow stromal cells (BMSC) and reversed APN-induced expression of the lysine-specific demethylases involved in regulating their commitment to the osteoblastic lineage. In conclusion, we found that APN regulates bone metabolism via central and peripheral mechanisms to decrease sympathetic tone, inhibit osteoclastic differentiation, and promote osteoblastic commitment of BMSC. PMID:24780611

  13. High dose sapropterin dihydrochloride therapy improves monoamine neurotransmitter turnover in murine phenylketonuria (PKU).

    PubMed

    Winn, Shelley R; Scherer, Tanja; Thöny, Beat; Harding, Cary O

    2016-01-01

    Central nervous system (CNS) deficiencies of the monoamine neurotransmitters, dopamine and serotonin, have been implicated in the pathophysiology of neuropsychiatric dysfunction in phenylketonuria (PKU). Increased brain phenylalanine concentration likely competitively inhibits the activities of tyrosine hydroxylase (TH) and tryptophan hydroxylase (TPH), the rate limiting steps in dopamine and serotonin synthesis respectively. Tetrahydrobiopterin (BH4) is a required cofactor for TH and TPH activity. Our hypothesis was that treatment of hyperphenylalaninemic Pah(enu2/enu2) mice, a model of human PKU, with sapropterin dihydrochloride, a synthetic form of BH4, would stimulate TH and TPH activities leading to improved dopamine and serotonin synthesis despite persistently elevated brain phenylalanine. Sapropterin (20, 40, or 100mg/kg body weight in 1% ascorbic acid) was administered daily for 4 days by oral gavage to Pah(enu2/enu2) mice followed by measurement of brain biopterin, phenylalanine, tyrosine, tryptophan and monoamine neurotransmitter content. A significant increase in brain biopterin content was detected only in mice that had received the highest sapropterin dose, 100mg/kg. Blood and brain phenylalanine concentrations were unchanged by sapropterin therapy. Sapropterin therapy also did not alter the absolute amounts of dopamine and serotonin in brain but was associated with increased homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA), dopamine and serotonin metabolites respectively, in both wild type and Pah(enu2/enu2) mice. Oral sapropterin therapy likely does not directly affect central nervous system monoamine synthesis in either wild type or hyperphenylalaninemic mice but may stimulate synaptic neurotransmitter release and subsequent metabolism. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Important role of mucosal serotonin in colonic propulsion and peristaltic reflexes: in vitro analyses in mice lacking tryptophan hydroxylase 1.

    PubMed

    Heredia, Dante J; Gershon, Michael D; Koh, Sang Don; Corrigan, Robert D; Okamoto, Takanubu; Smith, Terence K

    2013-12-01

    Although there is general agreement that mucosal 5-hydroxytryptamine (5-HT) can initiate peristaltic reflexes in the colon, recent studies have differed as to whether or not the role of mucosal 5-HT is critical. We therefore tested the hypothesis that the secretion of 5-HT from mucosal enterochromaffin (EC) cells is essential for the manifestation of murine colonic peristaltic reflexes. To do so, we analysed the mechanisms underlying faecal pellet propulsion in isolated colons of mice lacking tryptophan hydroxylase 1 (Tph1(-/-) mice), which is the rate-limiting enzyme in the biosynthesis of mucosal but not neuronal 5-HT. We used video analysis of faecal pellet propulsion, tension transducers to record colonic migrating motor complexes (CMMCs) and intracellular microelectrodes to record circular muscle activity occurring spontaneously or following intraluminal distension. When compared with control (Tph1(+/+)) mice, Tph1(-/-) animals exhibited: (1) an elongated colon; (2) larger faecal pellets; (3) orthograde propulsion followed by retropulsion (not observed in Tph1(+/+) colon); (4) slower in vitro propulsion of larger faecal pellets (28% of Tph1(+/+)); (5) CMMCs that infrequently propagated in an oral to anal direction because of impaired descending inhibition; (6) reduced CMMCs and inhibitory responses to intraluminal balloon distension; (7) an absence of reflex activity in response to mucosal stimulation. In addition, (8) thin pellets that propagated along the control colon failed to do so in Tph1(-/-) colon; and (9) the 5-HT3 receptor antagonist ondansetron, which reduced CMMCs and blocked their propagation in Tph1(+/+) mice, failed to alter CMMCs in Tph1(-/-) animals. Our observations suggest that mucosal 5-HT is essential for reflexes driven by mucosal stimulation and is also important for normal propagation of CMMCs and propulsion of pellets in the isolated colon.

  15. Early life environmental and pharmacological stressors result in persistent dysregulations of the serotonergic system

    PubMed Central

    Wong, Peiyan; Sze, Ying; Gray, Laura Jane; Chang, Cecilia Chin Roei; Cai, Shiwei; Zhang, Xiaodong

    2015-01-01

    Dysregulations in the brain serotonergic system and exposure to environmental stressors have been implicated in the development of major depressive disorder. Here, we investigate the interactions between the stress and serotonergic systems by characterizing the behavioral and biochemical effects of chronic stress applied during early-life or adulthood in wild type (WT) mice and mice with deficient tryptophan hydroxylase 2 (TPH2) function. We showed that chronic mild stress applied in adulthood did not affect the behaviors and serotonin levels of WT and TPH2 knock-in (KI) mice. Whereas, maternal separation (MS) stress increased anxiety- and depressive-like behaviors of WT mice, with no detectable behavioral changes in TPH2 KI mice. Biochemically, we found that MS WT mice had reduced brain serotonin levels, which was attributed to increased expression of monoamine oxidase A (MAO A). The increased MAO A expression was detected in MS WT mice at 4 weeks old and adulthood. No change in TPH2 expression was detected. To determine whether a pharmacological stressor, dexamethasone (Dex), will result in similar biochemical results obtained from MS, we used an in vitro system, SH-SY5Y cells, and found that Dex treatment resulted in increased MAO A expression levels. We then treated WT mice with Dex for 5 days, either during postnatal days 7–11 or adulthood. Both groups of Dex treated WT mice had reduced basal corticosterone and glucocorticoid receptors expression levels. However, only Dex treatment during PND7–11 resulted in reduced serotonin levels and increased MAO A expression. Just as with MS WT mice, TPH2 expression in PND7–11 Dex-treated WT mice was unaffected. Taken together, our findings suggest that both environmental and pharmacological stressors affect the expression of MAO A, and not TPH2, when applied during the critical postnatal period. This leads to long-lasting perturbations in the serotonergic system, and results in anxiety- and depressive-like behaviors. PMID:25964750

  16. Molecular docking of bacosides with tryptophan hydroxylase: a model to understand the bacosides mechanism.

    PubMed

    Rajathei, David Mary; Preethi, Jayakumar; Singh, Hemant K; Rajan, Koilmani Emmanuvel

    2014-08-01

    Tryptophan hydroxylase (TPH) catalyses l-tryptophan into 5-hydroxy-l-tryptophan, which is the first and rate-limiting step of serotonin (5-HT) biosynthesis. Earlier, we found that TPH2 up-regulated in the hippocampus of postnatal rats after the oral treatment of Bacopa monniera leaf extract containing the active compound bacosides. However, the knowledge about the interactions between bacosides with TPH is limited. In this study, we take advantage of in silico approach to understand the interaction of bacoside-TPH complex using three different docking algorithms such as HexDock, PatchDock and AutoDock. All these three algorithms showed that bacoside A and A3 well fit into the cavity consists of active sites. Further, our analysis revealed that major active compounds bacoside A3 and A interact with different residues of TPH through hydrogen bond. Interestingly, Tyr235, Thr265 and Glu317 are the key residues among them, but none of them are either at tryptophan or BH4 binding region. However, its note worthy to mention that Tyr 235 is a catalytic sensitive residue, Thr265 is present in the flexible loop region and Glu317 is known to interacts with Fe. Interactions with these residues may critically regulate TPH function and thus serotonin synthesis. Our study suggested that the interaction of bacosides (A3/A) with TPH might up-regulate its activity to elevate the biosynthesis of 5-HT, thereby enhances learning and memory formation.

  17. TPH-2 Polymorphisms Interact with Early Life Stress to Influence Response to Treatment with Antidepressant Drugs.

    PubMed

    Xu, Zhi; Reynolds, Gavin P; Yuan, Yonggui; Shi, Yanyan; Pu, Mengjia; Zhang, Zhijun

    2016-11-01

    Variation in genes implicated in monoamine neurotransmission may interact with environmental factors to influence antidepressant response. We aimed to determine how a range of single nucleotide polymorphisms in monoaminergic genes influence this response to treatment and how they interact with childhood trauma and recent life stress in a Chinese sample. An initial study of monoaminergic coding region single nucleotide polymorphisms identified significant associations of TPH2 and HTR1B single nucleotide polymorphisms with treatment response that showed interactions with childhood and recent life stress, respectively (Xu et al., 2012). A total of 47 further single nucleotide polymorphisms in 17 candidate monoaminergic genes were genotyped in 281 Chinese Han patients with major depressive disorder. Response to 6 weeks' antidepressant treatment was determined by change in the 17-item Hamilton Depression Rating Scale score, and previous stressful events were evaluated by the Life Events Scale and Childhood Trauma Questionnaire-Short Form. Three TPH2 single nucleotide polymorphisms (rs11178998, rs7963717, and rs2171363) were significantly associated with antidepressant response in this Chinese sample, as was a haplotype in TPH2 (rs2171363 and rs1487278). One of these, rs2171363, showed a significant interaction with childhood adversity in its association with antidepressant response. These findings provide further evidence that variation in TPH2 is associated with antidepressant response and may also interact with childhood trauma to influence outcome of antidepressant treatment. © The Author 2016. Published by Oxford University Press on behalf of CINP.

  18. TPH-2 Polymorphisms Interact with Early Life Stress to Influence Response to Treatment with Antidepressant Drugs

    PubMed Central

    Reynolds, Gavin P.; Yuan, Yonggui; Shi, Yanyan; Pu, Mengjia; Zhang, Zhijun

    2016-01-01

    Background: Variation in genes implicated in monoamine neurotransmission may interact with environmental factors to influence antidepressant response. We aimed to determine how a range of single nucleotide polymorphisms in monoaminergic genes influence this response to treatment and how they interact with childhood trauma and recent life stress in a Chinese sample. An initial study of monoaminergic coding region single nucleotide polymorphisms identified significant associations of TPH2 and HTR1B single nucleotide polymorphisms with treatment response that showed interactions with childhood and recent life stress, respectively (Xu et al., 2012). Methods: A total of 47 further single nucleotide polymorphisms in 17 candidate monoaminergic genes were genotyped in 281 Chinese Han patients with major depressive disorder. Response to 6 weeks’ antidepressant treatment was determined by change in the 17-item Hamilton Depression Rating Scale score, and previous stressful events were evaluated by the Life Events Scale and Childhood Trauma Questionnaire-Short Form. Results: Three TPH2 single nucleotide polymorphisms (rs11178998, rs7963717, and rs2171363) were significantly associated with antidepressant response in this Chinese sample, as was a haplotype in TPH2 (rs2171363 and rs1487278). One of these, rs2171363, showed a significant interaction with childhood adversity in its association with antidepressant response. Conclusions: These findings provide further evidence that variation in TPH2 is associated with antidepressant response and may also interact with childhood trauma to influence outcome of antidepressant treatment. PMID:27521242

  19. Optimization of combined in-vessel composting process and chemical oxidation for remediation of bottom sludge of crude oil storage tanks.

    PubMed

    Koolivand, Ali; Naddafi, Kazem; Nabizadeh, Ramin; Saeedi, Reza

    2017-07-31

    In this research, removal of petroleum hydrocarbons from oily sludge of crude oil storage tanks was investigated under the optimized conditions of in-vessel composting process and chemical oxidation with H 2 O 2 and Fenton. After determining the optimum conditions, the sludge was pre-treated with the optimum state of the oxidation process. Then, the determined optimum ratios of the sludge to immature compost were composted at a C:N:P ratio of 100:5:1 and moisture content of 55% for a period of 10 weeks. Finally, both pre-treated and composted mixtures were again oxidized with the optimum conditions of the oxidants. Results showed that total petroleum hydrocarbons (TPH) removal of the 1:8 and 1:10 composting reactors which were pre-treated with H 2 O 2 were 88.34% and 90.4%, respectively. In addition, reduction of TPH in 1:8 and 1:10 composting reactors which were pre-treated with Fenton were 83.90% and 84.40%, respectively. Without applying the pre-treatment step, the composting reactors had a removal rate of about 80%. Therefore, pre-treatment of the reactors increased the TPH removal. However, post-oxidation of both pre-treated and composted mixtures reduced only 13-16% of TPH. Based on the results, remarkable overall removal of TPH (about 99%) was achieved by using chemical oxidation and subsequent composting process. The study showed that chemical oxidation with H 2 O 2 followed by in-vessel composting is a viable choice for the remediation of the sludge.

  20. A pharmacological evidence of positive association between mouse intermale aggression and brain serotonin metabolism.

    PubMed

    Kulikov, A V; Osipova, D V; Naumenko, V S; Terenina, E; Mormède, P; Popova, N K

    2012-07-15

    The neurotransmitter serotonin (5-HT) is involved in the regulation of mouse intermale aggression. Previously, it was shown that intensity of mouse intermale aggression was positively associated with activity of the key enzyme of 5-HT synthesis - tryptophan hydroxylase 2 (TPH2) in mouse brain. The aim of the present study was to investigate the effect of pharmacological activation or inhibition of 5-HT synthesis in the brain on intermale aggression in two mouse strains differing in the TPH2 activity: C57BL/6J (B6, high TPH2 activity, high aggressiveness) and CC57BR/Mv (BR, low TPH2 activity, low aggressiveness). Administration of 5-HT precursor L-tryptophan (300 mg/kg, i.p.) to BR mice significantly increased the 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) levels in the midbrain as well as the number of attacks and their duration in the resident-intruder test. And vice versa, administration of TPH2 inhibitor p-chlorophenylalanine (pCPA) (300 mg/kg, i.p., for 3 consecutive days) to B6 mice dramatically reduced the 5-HT and 5-HIAA contents in brain structures and attenuated the frequency and the duration of aggressive attacks. At the same time, L-tryptophan or pCPA did not influence the percentage of aggressive mice and the attack latency reflecting the threshold of aggressive reaction. This result indicated that the intensity of intermale aggression, but not the threshold of aggressive reaction is positively dependent on 5-HT metabolism in mouse brain. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Investigation of ethyl lactate as a green solvent for desorption of total petroleum hydrocarbons (TPH) from contaminated soil.

    PubMed

    Jalilian Ahmadkalaei, Seyedeh Pegah; Gan, Suyin; Ng, Hoon Kiat; Abdul Talib, Suhaimi

    2016-11-01

    Treatment of oil-contaminated soil is a major environmental concern worldwide. The aim of this study is to examine the applicability of a green solvent, ethyl lactate (EL), in desorption of diesel aliphatic fraction within total petroleum hydrocarbons (TPH) in contaminated soil and to determine the associated desorption kinetics. Batch desorption experiments were carried out on artificially contaminated soil at different EL solvent percentages (%). In analysing the diesel range of TPH, TPH was divided into three fractions and the effect of solvent extraction on each fraction was examined. The experimental results demonstrated that EL has a high and fast desorbing power. Pseudo-second order rate equation described the experimental desorption kinetics data well with correlation coefficient values, R 2 , between 0.9219 and 0.9999. The effects of EL percentage, initial contamination level of soil and liquid to solid ratio (L/S (v/w)) on initial desorption rate have also been evaluated. The effective desorption performance of ethyl lactate shows its potential as a removal agent for remediation of TPH-contaminated soil worldwide.

  2. Tryptophan hydroxylase type 2 variants modulate severity and outcome of addictive behaviors in Parkinson's disease.

    PubMed

    Cilia, Roberto; Benfante, Roberta; Asselta, Rosanna; Marabini, Laura; Cereda, Emanuele; Siri, Chiara; Pezzoli, Gianni; Goldwurm, Stefano; Fornasari, Diego

    2016-08-01

    Impulse control disorders and compulsive medication intake may occur in a minority of patients with Parkinson's disease (PD). We hypothesize that genetic polymorphisms associated with addiction in the general population may increase the risk for addictive behaviors also in PD. Sixteen polymorphisms in candidate genes belonging to five neurotransmitter systems (dopaminergic, catecholaminergic, serotonergic, glutamatergic, opioidergic) and the BDNF were screened in 154 PD patients with addictive behaviors and 288 PD control subjects. Multivariate analysis investigated clinical and genetic predictors of outcome (remission vs. persistence/relapse) after 1 year and at the last follow-up (5.1 ± 2.5 years). Addictive behaviors were associated with tryptophan hydroxylase type 2 (TPH2) and dopamine transporter gene variants. A subsequent analysis within the group of cases showed a robust association between TPH2 genotype and the severity of addictive behaviors, which survived Bonferroni correction for multiple testing. At multivariate analysis, TPH2 genotype resulted the strongest predictor of no remission at the last follow-up (OR[95%CI], 7.4[3.27-16.78] and 13.2[3.89-44.98] in heterozygous and homozygous carriers, respectively, p < 0.001). The extent of medication dose reduction was not a predictor. TPH2 haplotype analysis confirmed the association with more severe symptoms and lower remission rates in the short- and the long-term (p < 0.005 for all analyses). The serotonergic system is likely to be involved in the pathophysiology of addictive behaviors in PD, modulating the severity of symptoms and the rate of remission at follow-up. If confirmed in larger independent cohorts, TPH2 genotype may become a useful biomarker for the identification of at-risk individuals. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. 78 FR 28666 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-15

    ... enable the TPH to meet its existing obligations to customers. In addition, such procedures must address the TPH's existing relationships with other broker-dealers and third parties. The business continuity... business continuity plan if such TPH has public customers. If the TPH does not have public customers, the...

  4. The use of sensory perception indicators for improving the characterization and modelling of total petroleum hydrocarbon (TPH) grade in soils.

    PubMed

    Roxo, Sónia; de Almeida, José António; Matias, Filipa Vieira; Mata-Lima, Herlander; Barbosa, Sofia

    2016-03-01

    This paper proposes a multistep approach for creating a 3D stochastic model of total petroleum hydrocarbon (TPH) grade in potentially polluted soils of a deactivated oil storage site by using chemical analysis results as primary or hard data and classes of sensory perception variables as secondary or soft data. First, the statistical relationship between the sensory perception variables (e.g. colour, odour and oil-water reaction) and TPH grade is analysed, after which the sensory perception variable exhibiting the highest correlation is selected (oil-water reaction in this case study). The probabilities of cells belonging to classes of oil-water reaction are then estimated for the entire soil volume using indicator kriging. Next, local histograms of TPH grade for each grid cell are computed, combining the probabilities of belonging to a specific sensory perception indicator class and conditional to the simulated values of TPH grade. Finally, simulated images of TPH grade are generated by using the P-field simulation algorithm, utilising the local histograms of TPH grade for each grid cell. The set of simulated TPH values allows several calculations to be performed, such as average values, local uncertainties and the probability of the TPH grade of the soil exceeding a specific threshold value.

  5. Estrogen receptor beta regulates the expression of tryptophan-hydroxylase 2 mRNA within serotonergic neurons of the rat dorsal raphe nuclei

    PubMed Central

    Donner, Nina C; Handa, Robert J

    2009-01-01

    Dysfunctions of the brain serotonin (5-HT) system are often associated with affective disorders, such as depression. The raphe nuclei target the limbic system and most forebrain areas and constitute the main source of 5-HT in the brain. All 5-HT neurons express tryptophan hydroxylase-2 (TPH2), the brain specific, rate-limiting enzyme for 5-HT synthesis. ERbeta agonists have been shown to attenuate anxiety-and despair-like behaviors in rodent models. Therefore, we tested the hypothesis that ERbeta may contribute to the regulation of gene expression in 5-HT neurons of the dorsal raphe nuclei (DRN) by examining the effects of systemic and local application of the selective ERbeta agonist diarylpropionitrile (DPN) on tph2 mRNA expression. Ovariectomized (OVX) female rats were injected subcutaneously (s.c.) with DPN or vehicle once daily for 8 days. In situ hybridization revealed that systemic DPN-treatment elevated basal tph2 mRNA expression in the caudal and mid-dorsal DRN. Behavioral testing of all animals in the open field (OF) and on the elevated plus maze (EPM) on days 6 and 7 of treatment confirmed the anxiolytic nature of ERbeta activation. Another cohort of female OVX rats was stereotaxically implanted bilaterally with hormone-containing wax pellets flanking the DRN. Pellets contained either 17-beta-estradiol (E), DPN, or no hormone. Both DPN and E significantly enhanced tph2 mRNA expression in the mid-dorsal DRN. DPN also increased tph2 mRNA in the caudal DRN. DPN- and E-treated rats displayed a more active stress-coping behavior in the forced-swim test (FST). No behavioral differences were found in the OF or on the EPM. These data indicate that ERbeta acts at the level of the rat DRN to modulate tph2 mRNA expression and thereby influence 5-HT synthesis in DRN subregions. Our results also suggest that local activation of ERbeta neurons in the DRN may be sufficient to decrease despair-like behavior, but not anxiolytic behaviors. PMID:19559077

  6. Optimization of the anaerobic treatment of a waste stream from an enhanced oil recovery process.

    PubMed

    Alimahmoodi, Mahmood; Mulligan, Catherine N

    2011-01-01

    The aim of this work was to optimize the anaerobic treatment of a waste stream from an enhanced oil recovery (EOR) process. The treatment of a simulated waste water containing about 150 mg chemical oxygen demand (COD)/L of total petroleum hydrocarbons (TPH) and the saturation level of CO2 was evaluated. A two-step anaerobic system was undertaken in the mesophilic temperature range (30-40°C). The method of evolutionary operation EVOP factorial design was used to optimize pH, temperature and organic loading rate with the target parameters of CO2 reduction and CH4 production in the first reactor and TPH removal in the second reactor. The results showed 98% methanogenic removal of CO2 and CH4 yield of 0.38 L/gCOD in the first reactor and 83% TPH removal in the second reactor. In addition to enhancing CO2 and TPH removal and CH4 production, application of this method showed the degree of importance of the operational variables and their interactive effects for the two reactors in series. Copyright © 2010 Elsevier Ltd. All rights reserved.

  7. Optimized conditions for phytoremediation of diesel by Scirpus grossus in horizontal subsurface flow constructed wetlands (HSFCWs) using response surface methodology.

    PubMed

    Al-Baldawi, Israa Abdul Wahab; Sheikh Abdullah, Siti Rozaimah; Abu Hasan, Hassimi; Suja, Fatihah; Anuar, Nurina; Mushrifah, Idris

    2014-07-01

    This study investigated the optimum conditions for total petroleum hydrocarbon (TPH) removal from diesel-contaminated water using phytoremediation treatment with Scirpus grossus. In addition, TPH removal from sand was adopted as a second response. The optimum conditions for maximum TPH removal were determined through a Box-Behnken Design. Three operational variables, i.e. diesel concentration (0.1, 0.175, 0.25% Vdiesel/Vwater), aeration rate (0, 1 and 2 L/min) and retention time (14, 43 and 72 days), were investigated by setting TPH removal and diesel concentration as the maximum, retention time within the given range, and aeration rate as the minimum. The optimum conditions were found to be a diesel concentration of 0.25% (Vdiesel/Vwater), a retention time of 63 days and no aeration with an estimated maximum TPH removal from water and sand of 76.3 and 56.5%, respectively. From a validation test of the optimum conditions, it was found that the maximum TPH removal from contaminated water and sand was 72.5 and 59%, respectively, which was a 5 and 4.4% deviation from the values given by the Box-Behnken Design, providing evidence that S. grossus is a Malaysian native plant that can be used to remediate wastewater containing hydrocarbons. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Important role of mucosal serotonin in colonic propulsion and peristaltic reflexes: in vitro analyses in mice lacking tryptophan hydroxylase 1

    PubMed Central

    Heredia, Dante J; Gershon, Michael D; Koh, Sang Don; Corrigan, Robert D; Okamoto, Takanubu; Smith, Terence K

    2013-01-01

    Although there is general agreement that mucosal 5-hydroxytryptamine (5-HT) can initiate peristaltic reflexes in the colon, recent studies have differed as to whether or not the role of mucosal 5-HT is critical. We therefore tested the hypothesis that the secretion of 5-HT from mucosal enterochromaffin (EC) cells is essential for the manifestation of murine colonic peristaltic reflexes. To do so, we analysed the mechanisms underlying faecal pellet propulsion in isolated colons of mice lacking tryptophan hydroxylase 1 (Tph1−/− mice), which is the rate-limiting enzyme in the biosynthesis of mucosal but not neuronal 5-HT. We used video analysis of faecal pellet propulsion, tension transducers to record colonic migrating motor complexes (CMMCs) and intracellular microelectrodes to record circular muscle activity occurring spontaneously or following intraluminal distension. When compared with control (Tph1+/+) mice, Tph1−/− animals exhibited: (1) an elongated colon; (2) larger faecal pellets; (3) orthograde propulsion followed by retropulsion (not observed in Tph1+/+ colon); (4) slower in vitro propulsion of larger faecal pellets (28% of Tph1+/+); (5) CMMCs that infrequently propagated in an oral to anal direction because of impaired descending inhibition; (6) reduced CMMCs and inhibitory responses to intraluminal balloon distension; (7) an absence of reflex activity in response to mucosal stimulation. In addition, (8) thin pellets that propagated along the control colon failed to do so in Tph1−/− colon; and (9) the 5-HT3 receptor antagonist ondansetron, which reduced CMMCs and blocked their propagation in Tph1+/+ mice, failed to alter CMMCs in Tph1−/− animals. Our observations suggest that mucosal 5-HT is essential for reflexes driven by mucosal stimulation and is also important for normal propagation of CMMCs and propulsion of pellets in the isolated colon. PMID:24127620

  9. Central adiponectin administration reveals new regulatory mechanisms of bone metabolism in mice.

    PubMed

    Wu, Yuwei; Tu, Qisheng; Valverde, Paloma; Zhang, Jin; Murray, Dana; Dong, Lily Q; Cheng, Jessica; Jiang, Hua; Rios, Maribel; Morgan, Elise; Tang, Zhihui; Chen, Jake

    2014-06-15

    Adiponectin (APN), the most abundant adipocyte-secreted adipokine, regulates energy homeostasis and exerts well-characterized insulin-sensitizing properties. The peripheral or central effects of APN regulating bone metabolism are beginning to be explored but are still not clearly understood. In the present study, we found that APN-knockout (APN-KO) mice fed a normal diet exhibited decreased trabecular structure and mineralization and increased bone marrow adiposity compared with wild-type (WT) mice. APN intracerebroventricular infusions decreased uncoupling protein 1 (UCP1) expression in brown adipose tissue, epinephrine and norepinephrine serum levels, and osteoclast numbers, whereas osteoblast osteogenic marker expression and trabecular bone mass increased in APN-KO and WT mice. In addition, centrally administered APN increased hypothalamic tryptophan hydroxylase 2 (TPH2), cocaine- and amphetamine-regulated transcript (CART), and 5-hydroxytryptamine (serotonin) receptor 2C (Htr2C) expressions but decreased hypothalamic cannabinoid receptor-1 expression. Treatment of immortalized mouse neurons with APN demonstrated that APN-mediated effects on TPH2, CART, and Htr2C expression levels were abolished by downregulating adaptor protein containing pleckstrin homology domain, phosphotyrosine domain, and leucine zipper motif (APPL)-1 expression. Pharmacological increase in sympathetic activity stimulated adipogenic differentiation of bone marrow stromal cells (BMSC) and reversed APN-induced expression of the lysine-specific demethylases involved in regulating their commitment to the osteoblastic lineage. In conclusion, we found that APN regulates bone metabolism via central and peripheral mechanisms to decrease sympathetic tone, inhibit osteoclastic differentiation, and promote osteoblastic commitment of BMSC. Copyright © 2014 the American Physiological Society.

  10. TPH2 G/T polymorphism is associated with hyperphagia, IQ, and internalizing problems in Prader-Willi syndrome

    PubMed Central

    Dykens, Elisabeth M.; Roof, Elizabeth; Bittel, Douglas; Butler, Merlin G.

    2010-01-01

    Background Prader-Willi syndrome (PWS) is a genetic, neurodevelopmental disorder characterized by intellectual disabilities, growth hormone dysregulation, hyperphagia, increased risks of morbid obesity, compulsive behaviors, and irritability. As aberrant serotonergic functioning is strongly implicated in PWS, we examined associations between the PWS phenotype and polymorphisms in tryptophan hydroxylase 2 (TPH2), the rate-limiting enzyme in the biosynthesis of serotonin in the brain. Methods 92 individuals with PWS aged 4 to 50 years (M = 21.97) were genotyped for the TPH2 G703-T polymorphism. IQ testing was conducted in offspring, and parents completed questionnaires that tapped their child’s compulsivity, hyperphagia, and other behavior problems. Results As expected, the frequency of G/T or T/T polymorphisms in participants with PWS (39%) was similar to rates found in the general population (38%). Compared to those with a homozygous (G/G) genotype, individuals with a T allele had significantly higher hyperphagic behavior, drive, and severity scores, and they also had a younger age of onset of hyperphagia. Those with a T allele also had higher IQ scores than their counterparts. Females with a T allele had significantly higher internalizing symptoms, primarily anxiety and depression, than all others. Conclusions TPH2 G/T polymorphisms, and presumed loss of enzyme function, were associated with specific aspects of the PWS phenotype. Aberrant serotonergic functioning is strongly implicated in hyperphagia in PWS, and females with TPH2 T alleles may be at higher risk for affective or mood disorders. Findings hold promise for examining other serotonin-altering genes in PWS, and for future serotonin-altering treatment trials. PMID:21418060

  11. TPH2 G/T polymorphism is associated with hyperphagia, IQ, and internalizing problems in Prader-Willi syndrome.

    PubMed

    Dykens, Elisabeth M; Roof, Elizabeth; Bittel, Douglas; Butler, Merlin G

    2011-05-01

    Prader-Willi syndrome (PWS) is a genetic, neurodevelopmental disorder characterized by intellectual disabilities, growth hormone dysregulation, hyperphagia, increased risks of morbid obesity, compulsive behaviors, and irritability. As aberrant serotonergic functioning is strongly implicated in PWS, we examined associations between the PWS phenotype and polymorphisms in tryptophan hydroxylase 2 (TPH2), the rate-limiting enzyme in the biosynthesis of serotonin in the brain. Ninety-two individuals with PWS aged 4 to 50 years (M = 21.97) were genotyped for the TPH2 G703-T polymorphism. IQ testing was conducted in offspring, and parents completed questionnaires that tapped their child's compulsivity, hyperphagia, and other behavior problems. As expected, the frequency of G/T or T/T polymorphisms in participants with PWS (39%) was similar to rates found in the general population (38%). Compared to those with a homozygous (G/G) genotype, individuals with a T allele had significantly higher hyperphagic behavior, drive, and severity scores, and they also had a younger age of onset of hyperphagia. Those with a T allele also had higher IQ scores than their counterparts. Females with a T allele had significantly higher internalizing symptoms, primarily anxiety and depression, than all others. TPH2 G/T polymorphisms, and presumed loss of enzyme function, were associated with specific aspects of the PWS phenotype. Aberrant serotonergic functioning is strongly implicated in hyperphagia in PWS, and females with TPH2 T alleles may be at higher risk for affective or mood disorders. Findings hold promise for examining other serotonin-altering genes in PWS, and for future serotonin-altering treatment trials. © 2011 The Authors. Journal of Child Psychology and Psychiatry © 2011 Association for Child and Adolescent Mental Health.

  12. Association of TPH1 with suicidal behaviour and psychiatric disorders in the Chinese population

    PubMed Central

    Liu, X; Li, H; Qin, W; He, G; Li, D; Shen, Y; Shen, J; Gu, N; Feng, G; He, L

    2006-01-01

    Tryptophan hydroxylase (TPH), the rate limiting enzyme in serotonin biosynthesis, is one of the most important regulating factors in the serotonergic system. Recently, polymorphisms of the TPH gene have been identified as being associated with suicide, but the evidence is inconsistent. To investigate the role in suicide of one of the isoforms, TPH1, we examined the association of five single nucleotide polymorphisms (SNPs) in the promoter region and in intron 7 of the TPH1 gene based on a sample from the Chinese population of 810 subjects, of whom 329 had made no suicide attempts (NSA), 297 had made suicide attempts (SA), and 184 were healthy subjects (HS). In this study, we observed statistically significant differences between NSA and HS subjects in allele distributions on one marker, −6526A (p = 0.0329; odds ratio (OR) 1.36; 95% confidence interval (CI) 1.01 to 1.81). No significant difference in genotype distribution or allele frequencies of other polymorphisms was found between the suicide victims and the controls. The overall haplotype frequency was significantly different between cases and healthy controls (p = 0.000024 NSA v HS; p<0.000001, SA v HS; p<0.000001, cases v HS). We found the haplotype TCAAA of −7180/−7065/−6526/218/779 to be strongly associated with suicidal behaviour and psychiatric disorders (p = 0.00243; OR = 1.62; 95% CI 1.17 to 2.24 and p = 0.018; OR = 1.41; 95% CI 1.05 to 1.91), which suggests an association of TPH1 with suicidal behaviour and indicates that TPH1 may play a significant role in the aetiology of psychiatric disorders in the Han Chinese population. PMID:16467214

  13. [The association of polymorphisms in SLC18A1, TPH1 and RELN genes with risk of paranoid schizophrenia].

    PubMed

    Galaktionova, D Iu; Gareeva, A E; Khusnutdinova, E K; Nasedkina, T V

    2014-01-01

    We have developed a biochip for the analysis of polymorphisms in candidate genes for schizophrenia: DISC1, RELN, ZNF804A, PLXNA2, COMT, SLC18A41, CACNA1C, ANK3, TPH1, PLAA and SNAP-25. Using biochip the allele and genotype frequencies in 198 patients with schizophrenia and 192 healthy individuals have been obtained. For SLC18A1 polymorphism rs2270641 A>C, the frequencies of A allele (p = 0.007) and AA genotype (p = 0.002) were lower in patients compared with healthy individuals. A significant association was found between AA genotype (p = 0.036) of the TPH1 polymorphism rs1800532 C>A and schizophrenia. The C allele (p = 0.039) of the RELNpolymorphism rs7341475 C>T were lower in patients with schizophrenia compared with healthy individuals in a tatar population. Genotype AA of the TPH1 polymorphism rs1800532 C>A were more frequent in patients with schizophrenia compared with healthy individuals. Ithas been shown that the C allele (p = 0.0001) and GC (p = = 0.0001) genotype of the PLXNA2 polymorphism rs1327175 G>C are associated with the family history in patients with paranoid schizophrenia. The obtained data suggest that SLC18A1, TPH1 and RELN gene polymorphisms are associated with the risk of paranoid schizophrenia.

  14. Tributyltin and triphenyltin induce spermatogenesis in ovary of female abalone, Haliotis gigantea.

    PubMed

    Horiguchi, T; Kojima, M; Kaya, M; Matsuo, T; Shiraishi, H; Morita, M; Adachi, Y

    2002-01-01

    Two-month flow-through exposure experiments of tributyltin (TBT) and triphenyltin (TPhT) were conducted with abalone, Haliotis gigantea. Nominal concentrations of 100 ng TBT/l and 100 ng TPhT/l caused significant spermatogenesis in ovaries of exposed females. There were also significantly more contracted primary oocytes observed in females exposed to either TBT or TPhT than controls. The incidence of two types of unknown cells was also significant in females exposed to TPhT. No significant histological changes were observed in testis of exposed males. This ovarian spermatogenesis caused by TBT and/or TPhT resembles gastropod imposex. Remarkably high concentrations of TBT and TPhT were observed in the head (including central nervous system ganglia), compared to muscles concentrations. Accumulation of TBT and TPhT in the head may disturb reproductive hormonal regulators through neuropeptides released from ganglia. This, as well as possible aromatase inhibition, may be one of the inducers for spermatogenesis in the abalone ovaries.

  15. A comparison of biomarker responses in juvenile diploid and ...

    EPA Pesticide Factsheets

    Influence of waterborne butachlor (BUC), a commonly used pesticide, on morphometric, biochemical, and molecular biomarkers was evaluated in juvenile, full sibling, diploid and triploid African catfish (Clarias gariepinus). Fish were exposed for 21 days to one of three concentrations of BUC [mean measured µg/L: 22, 44 or 60]. Unexposed (control) triploids were heavier and longer and had higher visceral-somatic index (VSI) than diploids. Also, they had lighter liver weight (HSI) and showed lower transcript levels of brain gonadotropin-releasing hormone (GnRH), aromatase (cyp191b) and fushi tarazu-factor (ftz-f1), and plasma testosterone levels than diploids. Butachlor treatments had no effects, in either diploid or triploid fish, on VSI, HSI, weight or length changes, condition factor (CF), levels of plasma testosterone, 17-β estradiol (E2), cortisol, cholesterol, or mRNA levels of brain tryptophan hydroxylase (tph2), forkhead box L2 (foxl2), and 11 β-hydroxysteroid dehydrogenase type 2 (11β-hsd2). Expressions of cyp191b and ftz-f1 in triploids were upregulated by the two highest concentrations of BUC. In diploid fish, however, exposures to all BUC concentrations decreased GnRH transcription and the medium BUC concentration decreased ftz-f1 transcription. Substantial differences between ploidies in basal biomarker responses are consistent with the reported impaired reproductive axis in triploid C. gariepinus. Furthermore, the present study showed the low impac

  16. Activation of the Serotonin Pathway is Associated with Poor Outcome in COPD Exacerbation: Results of a Long-Term Cohort Study.

    PubMed

    Meier, Marc A; Ottiger, Manuel; Vögeli, Alaadin; Steuer, Christian; Bernasconi, Luca; Thomann, Robert; Christ-Crain, Mirjam; Henzen, Christoph; Hoess, Claus; Zimmerli, Werner; Huber, Andreas; Mueller, Beat; Schuetz, Philipp

    2017-06-01

    Indoleamine 2,3-dioxygenase (IDO) metabolizes tryptophan to kynurenine. An increase of its activity is associated with severity in patients with pneumonia. In chronic obstructive pulmonary disease (COPD) patients, an elevation of serotonin has been reported. Experimental models showed that cigarette smoke inhibits monoamine oxidase (MAO) leading to higher levels of serotonin. We investigated the prognostic ability of tryptophan, serotonin, kynurenine, IDO, and tryptophan hydroxylase (TPH) to predict short- and long-term outcomes in patients with a COPD exacerbation. We measured tryptophan, serotonin, and kynurenine on admission plasma samples in patients with a COPD exacerbation from a previous trial by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). IDO and TPH were calculated as ratios of kynurenine over tryptophan, and serotonin over tryptophan, respectively. We studied their association with parameters measured in clinical routine at emergency department admission representing inflammation (C-reactive protein [CRP]), infection (procalcitonin [PCT]), oxygenation (SpO 2 ), as well as patients' clinical outcome, confirmed by structured phone interviews. Mortality in the 149 included patients was 53.7% within six years of follow-up. While IDO activity showed strong positive correlations, tryptophan was negatively correlated with CRP and PCT. For 30-day adverse outcome defined as death and/or intensive care unit (ICU) admission, a multivariate regression analysis adjusted for age and comorbidities found strong associations for IDO activity (adjusted odds ratios of 31.4 (95%CI 1.1-857), p = 0.041) and TPH (adjusted odds ratios 27.0 (95%CI 2.2-327), p = 0.010). TPH also showed a significant association with mortality at 18 months, (hazard ratio 2.61 (95%CI 1.2-5.8), p = 0.020). In hospitalized patients with a COPD exacerbation, higher IDO and TPH activities independently predicted adverse short-term outcomes and TPH levels were also predictive of 18-month mortality. Whether therapeutic modulation of the serotonin pathway has positive effects on outcome needs further investigation.

  17. Association between tryptophan hydroxylase-2 genotype and the antidepressant effect of citalopram and paroxetine on immobility time in the forced swim test in mice.

    PubMed

    Kulikov, Alexander V; Tikhonova, Maria A; Osipova, Daria V; Kulikov, Victor A; Popova, Nina K

    2011-10-01

    Tryptophan hydroxylase-2 (TPH2) is the rate limiting enzyme of serotonin synthesis in the brain. The 1473G allele of the C1473G polymorphism in mTPH2 gene is associated with reduced enzyme activity and serotonin synthesis rate in the mouse brain. Here, the influence of the 1473G allele on the antidepressant effect of selective serotonin reuptake inhibitors (SSRIs), citalopram (2.5 or 5.0mg/kg) and paroxetine (5.0 or 10.0mg/kg), in the forced swim test was studied using B6-1473G and B6-1473C congenic mouse lines with the 1473G (decreased TPH2 activity) or 1473C (normal TPH2 activity) alleles, respectively, transferred to the genome of C57BL/6 mouse strain. Paroxetine (5.0 or 10.0mg/kg) and citalopram (2.5 or 5.0mg/kg) decreased immobility time in B6-1473C mice, while both doses of paroxetine and 2.5mg/kg of citaloprame did not alter immobility time in B6-1473G mice. However, 5.0mg/kg of citalopram reduced immobility in B6-1473G mice. The results provided genetic evidence of moderate association between 1473G allele and reduced sensitivity to SSRIs in mice. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Effects of combined treatment with mephedrone and methamphetamine or 3,4-methylenedioxymethamphetamine on serotonin nerve endings of the hippocampus

    PubMed Central

    Angoa-Pérez, Mariana; Kane, Michael J.; Herrera-Mundo, Nieves; Francescutti, Dina M.; Kuhn, Donald M.

    2013-01-01

    Aims Mephedrone is a stimulant drug of abuse with close structural and mechanistic similarities to methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA). Although mephedrone does not damage dopamine nerve endings it increases the neurotoxicity of amphetamine, methamphetamine and MDMA. The effects of mephedrone on serotonin (5HT) nerve endings are not fully understood, with some investigators reporting damage while others conclude it does not. Presently, we investigate if mephedrone given alone or with methamphetamine or MDMA damages 5HT nerve endings of the hippocampus. Main methods The status of 5HT nerve endings in hippocampus of female C57BL mice was assessed through measures of 5HT by HPLC and by immunoblot analysis of serotonin transporter (SERT) and tryptophan hydroxylase 2 (TPH2), selective markers of 5HT nerve endings. Astrocytosis was assessed through measures of glial fibrillary acidic protein (GFAP) (immunoblotting) and microglial activation was determined by histochemical staining with Isolectin B4. Key findings Mephedrone alone did not cause persistent reductions in the levels of 5HT, SERT or TPH2. Methamphetamine and MDMA alone caused mild reductions in 5HT but did not change SERT and TPH2 levels. Combined treatment with mephedrone and methamphetamine or MDMA did not change the status of 5HT nerve endings to an extent that was different from either drug alone. Significance Mephedrone does not cause toxicity to 5HT nerve endings of the hippocampus. When co-administered with methamphetamine or MDMA, drugs that are often co-abused with mephedrone by humans, toxicity is not increased as is the case for dopamine nerve endings when these drugs are taken together. PMID:23892197

  19. Genetics of Aggression in Alzheimer’s Disease (AD)

    PubMed Central

    Lukiw, Walter J.; Rogaev, Evgeny I.

    2017-01-01

    Alzheimer’s disease (AD) is a terminal, age-related neurological syndrome exhibiting progressive cognitive and memory decline, however AD patients in addition exhibit ancillary neuropsychiatric symptoms (NPSs) and these include aggression. In this communication we provide recent evidence for the mis-regulation of a small family of genes expressed in the human hippocampus that appear to be significantly involved in expression patterns common to both AD and aggression. DNA array- and mRNA transcriptome-based gene expression analysis and candidate gene association and/or genome-wide association studies (CGAS, GWAS) of aggressive attributes in humans have revealed a surprisingly small subset of six brain genes that are also strongly associated with altered gene expression patterns in AD. These genes encoded on five different chromosomes (chr) include the androgen receptor (AR; chrXq12), brain-derived neurotrophic factor (BDNF; chr11p14.1), catechol-O-methyl transferase (COMT; chr22q11.21), neuronal specific nitric oxide synthase (NOS1; chr12q24.22), dopamine beta-hydroxylase (DBH chr9q34.2) and tryptophan hydroxylase (TPH1, chr11p15.1 and TPH2, chr12q21.1). Interestingly, (i) the expression of three of these six genes (COMT, DBH, NOS1) are highly variable; (ii) three of these six genes (COMT, DBH, TPH1) are involved in DA or serotonin metabolism, biosynthesis and/or neurotransmission; and (iii) five of these six genes (AR, BDNF, COMT, DBH, NOS1) have been implicated in the development, onset and/or propagation of schizophrenia. The magnitude of the expression of genes implicated in aggressive behavior appears to be more pronounced in the later stages of AD when compared to MCI. These recent genetic data further indicate that the extent of cognitive impairment may have some bearing on the degree of aggression which accompanies the AD phenotype. PMID:28443016

  20. Rapid Estimation of TPH Reduction in Oil-Contaminated Soils Using the MED Method

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Edenborn, H.M.; Zenone, V.A.

    2007-09-01

    Oil-contaminated soil and sludge generated during federal well plugging activities in northwestern Pennsylvania are currently remediated on small landfarm sites in lieu of more expensive landfill disposal. Bioremediation success at these sites in the past has been gauged by the decrease in total petroleum hydrocarbon (TPH) concentrations to less than 10,000 mg/kg measured using EPA Method 418.1. We tested the “molarity of ethanol droplet” (MED) water repellency test as a rapid indicator of TPH concentration in soil at one landfarm near Bradford, PA. MED was estimated by determining the minimum ethanol concentration (0 – 6 M) required to penetrate air-driedmore » and sieved soil samples within 10 sec. TPH in soil was analyzed by rapid fluorometric analysis of methanol soil extracts, which correlated well with EPA Method 1664. Uncontaminated landfarm site soil amended with increasing concentrations of waste oil sludge showed a high correlation between MED and TPH. MED values exceeded the upper limit of 6 M as TPH estimates exceed ca. 25,000 mg/kg. MED and TPH at the land farm were sampled monthly during summer months over two years in a grid pattern that allowed spatial comparisons of site remediation effectiveness. MED and TPH decreased at a constant rate over time and remained highly correlated. Inexpensive alternatives to reagent-grade ethanol gave comparable results. The simple MED approach served as an inexpensive alternative to the routine laboratory analysis of TPH during the monitoring of oily waste bioremediation at this landfarm site.« less

  1. A novel role for antizyme inhibitor 2 as a regulator of serotonin and histamine biosynthesis and content in mouse mast cells.

    PubMed

    Acosta-Andrade, Carlos; Lambertos, Ana; Urdiales, José L; Sánchez-Jiménez, Francisca; Peñafiel, Rafael; Fajardo, Ignacio

    2016-10-01

    Antizymes and antizyme inhibitors are key regulatory proteins of polyamine levels by affecting ornithine decarboxylase and polyamine uptake. Our previous studies indicated a metabolic interplay among polyamines, histamine and serotonin in mast cells, and demonstrated that polyamines are present in mast cell secretory granules, being important for histamine storage and serotonin levels. Recently, the novel antizyme inhibitor-2 (AZIN2) was proposed as a local regulator of polyamine biosynthesis in association with mast cell serotonin-containing granules. To gain insight into the role of AZIN2 in the biosynthesis and storage of serotonin and histamine, we have generated bone marrow derived mast cells (BMMCs) from both wild-type and transgenic Azin2 hypomorphic mice, and have analyzed polyamines, serotonin and histamine contents, and some elements of their metabolisms. Azin2 hypomorphic BMMCs did not show major mast cell phenotypic alterations as judged by morphology and specific mast cell proteases. However, compared to wild-type controls, these cells showed reduced spermidine and spermine levels, and diminished growth rate. Serotonin levels were also reduced, whereas histamine levels tended to increase. Accordingly, tryptophan hydroxylase-1 (TPH1; the key enzyme for serotonin biosynthesis) mRNA expression and protein levels were reduced, whereas histidine decarboxylase (the enzyme responsible for histamine biosynthesis) enzymatic activity was increased. Furthermore, microphtalmia-associated transcription factor, an element involved in the regulation of Tph1 expression, was reduced. Taken together, our results show, for the first time, an element of polyamine metabolism -AZIN2-, so far described as exclusively devoted to the control of polyamine concentrations, involved in regulating the biosynthesis and content of other amines like serotonin and histamine.

  2. Bioremediation of hydrocarbon degradation in a petroleum-contaminated soil and microbial population and activity determination.

    PubMed

    Wu, Manli; Li, Wei; Dick, Warren A; Ye, Xiqiong; Chen, Kaili; Kost, David; Chen, Liming

    2017-02-01

    Bioremediation of hydrocarbon degradation in petroleum-polluted soil is carried out by various microorganisms. However, little information is available for the relationships between hydrocarbon degradation rates in petroleum-contaminated soil and microbial population and activity in laboratory assay. In a microcosm study, degradation rate and efficiency of total petroleum hydrocarbons (TPH), alkanes, and polycyclic aromatic hydrocarbons (PAH) in a petroleum-contaminated soil were determined using an infrared photometer oil content analyzer and a gas chromatography mass spectrometry (GC-MS). Also, the populations of TPH, alkane, and PAH degraders were enumerated by a modified most probable number (MPN) procedure, and the hydrocarbon degrading activities of these degraders were determined by the Biolog (MT2) MicroPlates assay. Results showed linear correlations between the TPH and alkane degradation rates and the population and activity increases of TPH and alkane degraders, but no correlation was observed between the PAH degradation rates and the PAH population and activity increases. Petroleum hydrocarbon degrading microbial population measured by MPN was significantly correlated with metabolic activity in the Biolog assay. The results suggest that the MPN procedure and the Biolog assay are efficient methods for assessing the rates of TPH and alkane, but not PAH, bioremediation in oil-contaminated soil in laboratory. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Lack of association between TPH2 gene polymorphisms with major depressive disorder in multiethnic Malaysian population.

    PubMed

    Nazree, Nur Elia; Loke, Ai Chin; Zainal, Nor Zuraida; Mohamed, Zahurin

    2015-03-01

    Numerous association studies of candidate genes studies with major depressive disorder (MDD) have been conducted for many years; however, the evidence of association between genes and the risk of developing MDD still remains inconclusive. In this study, we aimed to investigate the association between the tryptophan hydroxylase 2 (TPH2) gene and MDD in three ethnic groups (Malay, Chinese and Indian) within the Malaysian population. Two hundred and sixty five MDD patients who fulfilled the Diagnostic and Statistical Manual of Mental Disorders-IV criteria for MDD and 332 healthy controls were recruited for the study. All cases and controls were then genotyped for TPH2 polymorphisms rs1386494, rs1386495 and rs7305115. Single locus analysis in pooled and ethnically stratified subjects revealed no association between each of the three variants of the TPH2 gene with susceptibility to MDD. Strong linkage disequilibrium was detected between rs1386495 and rs1386494 in pooled subjects; however, no significant association was found in the haplotype analysis. In this study, we suggest that in both the Chinese and Indian populations, gender distribution differ significantly between cases and controls, showing that women are more at risk of developing MDD compared with men. Therefore, we suggest that the occurrence of MDD in both Chinese and Indians in the Malaysian population may be influenced by gender. Copyright © 2013 Wiley Publishing Asia Pty Ltd.

  4. TPH2 -703G/T SNP may have important effect on susceptibility to suicidal behavior in major depression.

    PubMed

    Yoon, Ho-Kyoung; Kim, Yong-Ku

    2009-04-30

    Serotonergic system-related genes can be good candidate genes for both major depressive disorder (MDD) and suicidal behavior. In this study, we aimed to investigate the association of serotonin 2A receptor gene -1438A/G SNP (HTR2A -1438A/G), tryptophan hydroxylase 2 gene -703G/T SNP (TPH2 -703G/T) and serotonin 1A receptor C-1019G (HTR1A C-1019G) with suicidal behavior. One hundred and eighty one suicidal depressed patients and 143 non-suicidal depressed patients who met DSM-IV criteria for major depressive disorder were recruited from patients who were admitted to Korea University Ansan Hospital. One hundred seventy six normal controls were healthy volunteers who were recruited by local advertisement. Patients and normal controls were genotyped for HTR2A -1438A/G, TPH2 -703G/T and 5-HT1A C-1019G. The suicidal depressed patients were evaluated by the lethality of individual suicide attempts using Weisman and Worden's risk-rescue rating (RRR) and the Lethality Suicide Attempt Rating Scale-updated (LSARS-II). In order to assess the severity of depressive symptoms of patients, Hamilton's Depression Rating Scale (HDRS) was administered. Genotype and allele frequencies were compared between groups by chi(2) statistics. Association of genotype of the candidate genes with the lethality of suicidal behavior was examined with ANOVA by comparing the mean scores of LSARS and RRR according to the genotype. There were statistically significant differences in the genotype distributions and allele frequencies of TPH2 -703G/T between the suicidal depressive group and the normal control group. The homozygous allele G (G/G genotype) frequency was significantly higher in suicidal depressed patients than in controls. However, no differences in either genotype distribution or in allele frequencies of HTR2A -1438A/G and HTR1A C-1019G were observed between the suicidal depressed patients, the non-suicidal depressed patients, and the normal controls. There were no differences in the lethality of suicidal behavior in suicidal depressed patients according to the genotypes of three polymorphisms. Our results suggest that TPH2 -703G/T SNP may have an important effect on susceptibility to suicidal behavior. Furthermore, an increased frequency of G allele of TPH2 SNP may be associated with elevated suicidal behavior itself rather than with the diagnosis of major depression and may increase risk of suicidality, independent of diagnosis.

  5. Treadmill exercise ameliorates social isolation-induced depression through neuronal generation in rat pups.

    PubMed

    Cho, Jung-Wan; Jung, Sun-Young; Lee, Sang-Won; Lee, Sam-Jun; Seo, Tae-Beom; Kim, Young-Pyo; Kim, Dae-Young

    2017-12-01

    Social isolation is known to induce emotional and behavioral changes in animals and humans. The effect of treadmill exercise on depression was investigated using social isolated rat pups. The rat pups in the social isolation groups were housed individually. The rat pups in the exercise groups were forced to run on treadmill for 30 min once a day from postnatal day 21 to postnatal day 34. In order to evaluate depression state of rat pups, forced swimming test was performed. Newly generated cells in the hippocampal dentate gyrus were determined by 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry. We examined the expression of 5-hydroxytryptamine (5-HT) and tryptophan hydroxylase (TPH) in the dorsal raphe using immunofluorescence. The expression of brain-derived neurotrophic factor (BDNF) and tyrosine kinase B (TrkB) was detected by Western blot analysis. The present results demonstrated that social isolation increased resting time and decreased mobility time. Expression of 5-HT and TPH in the dorsal raphe and expression of BDNF and TrkB in the hippocampus were decreased by social isolation. The number of BrdU-positive cells in the hippocampal dentate gyrus was suppressed by social isolation. Treadmill exercise decreased resting time and increased mobility in the social isolated rat pups. Expression of 5-HT, TPH, BDNF, and TrkB was increased by treadmill exercise. The present results suggested that treadmill exercise may ameliorates social isolation-induced depression through increasing neuronal generation.

  6. No effect of C1473G polymorphism in the tryptophan hydroxylase 2 gene on the response of the brain serotonin system to chronic fluoxetine treatment in mice.

    PubMed

    Bazhenova, Ekaterina Y; Sinyakova, Nadezhda A; Kulikova, Elizabeth A; Kazarinova, Irina A; Bazovkina, Daria V; Gainetdinov, Raul R; Kulikov, Alexander V

    2017-07-13

    Selective serotonin reuptake inhibitors (SSRIs) are antidepressants that block serotonin transporter (SERT) and increase serotonin (5-HT) level in the synaptic cleft. The interaction between SERT and the key enzyme of 5-HT synthesis in the brain, tryptophan hydroxylase 2 (TPH2), is essential to maintain the brain 5-HT level. The G allele of C1473G polymorphism in Tph2 gene decreases enzyme activity by half in mouse brain. Here we studied effect of C1473G polymorphism on the reaction of brain 5-HT system to chronic fluoxetine treatment (120mg/l in drinking water, for 3 weeks) in adult males of the congenic B6-1473C and B6-1473G mouse lines with high and low enzyme activity, respectively. The polymorphism did not affect the levels of 5-HT, its metabolite, 5-hydroxyindoleacetic acid (5-HIAA) and Tph2 gene mRNA in the brain. Fluoxetine significantly attenuated 5-HT levels in the cortex and striatum, 5-HIAA concentrations in the cortex, hippocampus, striatum and midbrain, and Tph2 gene expression in the midbrain. However, we did not observed any effect of the genotype x treatment interaction on these neurochemical characteristics. Therefore, C1473G polymorphism does not seem to play an essential role in the reaction of the brain 5-HT system to chronic fluoxetine treatment. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Tryptophan Hydroxylase 2 haplotype association with borderline personality disorder and aggression in a sample of patients with personality disorders and healthy controls

    PubMed Central

    Perez-Rodriguez, M. Mercedes; Weinstein, Shauna; New, Antonia S.; Bevilacqua, Laura; Yuan, Qiaoping; Zhou, Zhifeng; Hodgkinson, Colin; Goodman, Marianne; Koenigsberg, Harold W.; Goldman, David; Siever, Larry J.

    2010-01-01

    Background There is decreased serotonergic function in impulsive aggression and borderline personality disorder (BPD), and genetic association studies suggest a role of serotonergic genes in impulsive aggression and BPD. Only one study has analyzed the association between the tryptophan-hydroxylase 2 (TPH2) gene and BPD. A TPH2 “risk” haplotype has been described that is associated with anxiety, depression and suicidal behavior. Methods We assessed the relationship between the previously identified “risk” haplotype at the TPH2 locus and BPD diagnosis, impulsive aggression, affective lability, and suicidal/parasuicidal behaviors, in a well-characterized clinical sample of 103 healthy controls (HCs) and 251 patients with personality disorders (109 with BPD). A logistic regression including measures of depression, affective lability and aggression scores in predicting “risk” haplotype was conducted. Results The prevalence of the “risk” haplotype was significantly higher in patients with BPD compared to HCs. Those with the “risk” haplotype have higher aggression and affect lability scores and more suicidal/parasuicidal behaviors than those without it. In the logistic regression model, affect lability was the only significant predictor and it correctly classified 83.1% of the subjects as “risk” or “non-risk” haplotype carriers. Conclusions We found an association between the previously described TPH2 “risk” haplotype and BPD diagnosis, affective lability, suicidal/parasuicidal behavior, and aggression scores. PMID:20451217

  8. Effect of gas velocity and influent concentration on biofiltration of gasoline off-gas from soil vapor extraction.

    PubMed

    Namkoong, Wan; Park, Joon-Seok; VanderGheynst, Jean S

    2004-11-01

    This study was conducted to evaluate the effects of gas inlet concentration and velocity on the biofiltration of gasoline vapor. Gasoline vapor was treated using a compost biofilter operated in an upflow mode for about 3 months. The inlet concentration of gasoline total petroleum hydrocarbon (TPH) ranged from about 300 to 7000 mgm(-3) and gas was injected at velocities of 6 and 15 mh(-1) (empty bed residence time (EBRT)=10 and 4 min, respectively). The maximum elimination capacities of TPH at 6 and 15 mh(-1) found in this research were over 24 and 19 gm(-3) of filling material h(-1), respectively. TPH removal data was fit using a first-order kinetic relationship. In the low concentration range of 300-3000 mg m(-3), the first-order kinetic constants varied between about 0.10 and 0.29 min(-1) regardless of gas velocities. At TPH concentrations greater than 3000 mgm(-3), the first-order kinetic constants were about 0.09 and 0.07 min(-1) at gas velocities of 6 mh(-1) and 15 mh(-1), respectively. To evaluate microbial dynamics, dehydrogenase activity, CO2 generation and microbial species diversity were analyzed. Dehydrogenase activity could be used as an indicator of microbial activity. TPH removal corresponded well with CO2 evolution. The average CO2 recovery efficiency for the entire biofilter ranged between 60% and 70%. When the gas velocity was 6 mh(-1), most of the microbial activity and TPH removal occurred in the first quarter of the biofilter. However, when the gas velocity was 15 mh(-1), the entire column contributed to removal. Spatial and temporal variations in the biofilter microbial population were also observed. Nearly 60% of the colonies isolated from the compost media prior to biofiltration were Bacillus. After 90 days of biofiltration, the predominant species in the lower portion (0-50 cm) of the filter were Rhodococcus, while Pseudomonas and Acinetobacter dominated the upper portion (75-100 cm). copyright 2004 Elsevier Ltd.

  9. Petroleum hydrocarbon contaminations in the intertidal seawater after the Hebei Spirit oil spill--effect of tidal cycle on the TPH concentrations and the chromatographic characterization of seawater extracts.

    PubMed

    Kim, Moonkoo; Hong, Sang Hee; Won, Jongho; Yim, Un Hyuk; Jung, Jee-Hyun; Ha, Sung Yong; An, Joon Geon; Joo, Changkyu; Kim, Eunsic; Han, Gi Myung; Baek, Seongho; Choi, Hyun-Woo; Shim, Won Joon

    2013-02-01

    In December 2007, the oil tanker Hebei Spirit released approximately 12,547,000 L of crude oil off the west coast of Korea, impacting more than 375 km of coastline. The seawater TPH concentrations immediately after the spill ranged from 1.5 to 7310 μg L⁻¹, with an average of 732 μg L⁻¹. The concentrations appeared to decrease drastically to 2.0-224 μg L⁻¹ in one month after the spill. The TPH concentrations in seawater fluctuated with time thereafter because of the remobilization of oil by continuing shoreline cleanup activities and subsequent wave/tidal actions. Seawater TPH concentrations were much higher during high tide than during low tide due to the resuspension of stranded oil. The variation of TPH levels in seawater also matched the spring-neap tidal cycle in the study areas for the first three weeks of the study. Comparisons of the gas chromatograms of the seawater with the water accommodated fraction and the cargo oil indicated that seawater samples were contaminated mainly by the dispersed droplets of spilled oil. One year of monitoring revealed that the oil content in seawater had clearly decreased at most sites, although some regional fluctuations of oil contamination were noted until June 2008. Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. Effect of tryptophan hydroxylase gene polymorphism on aggression in major depressive disorder and undifferentiated somatoform disorder.

    PubMed

    Koh, Kyung Bong; Kim, Chan Hyung; Choi, Eun Hee; Lee, Young-joon; Seo, Won Youl

    2012-05-01

    Aggression and anger have been linked with depression, and anger suppression has been linked with somatic symptoms of somatoform disorders. However, the relationship between aggression or anger and genes in patients with depression and somatoform disorders has not been clearly elucidated. The objective of this study was to examine the effect of serotonin-related gene polymorphism on aggression in depressive disorders and somatoform disorders. A serotonin-related polymorphic marker was assessed by using single nucleotide polymorphism (SNP) genotyping. 106 outpatients with major depressive disorder (MDD), 102 outpatients with undifferentiated somatoform disorder, and 133 healthy subjects were enrolled between October 2005 and May 2008. Diagnoses were made according to the Korean version of the Structured Clinical Interview Schedule for DSM-IV. The allele and genotype frequencies of tryptophan hydroxylase-1 (TPH1) A218C were compared between groups. The Hamilton Depression Rating Scale and the Aggression Questionnaire were used for psychological assessment. Each of the 2 disorder groups scored significantly higher on all the Aggression Questionnaire subscales and on the total Aggression Questionnaire score than the healthy subjects (P < .001). Patients with MDD had significantly higher frequencies of TPH1 C allele (P = .0002) and CC homozygote (P = .0003) than healthy subjects, regardless of sex and age. However, no significant differences were found in TPH1 C allele and CC homozygote frequencies between the undifferentiated somatoform disorder patients and the healthy subjects. TPH1 CC homozygote in the MDD group scored significantly higher in terms of verbal aggression (P = .03) and total Aggression Questionnaire score (P = .04) than A-carrier genotypes, regardless of sex and age. However, no significant differences were found in the scores of all the Aggression Questionnaire subscales and the total Aggression Questionnaire score between TPH1 CC homozygote and A-carrier genotypes in the undifferentiated somatoform disorder group and the control group, respectively. Aggression in MDD patients is more susceptible to an excess of TPH1 CC homozygote than in undifferentiated somatoform disorder patients, though the 2 disorders are high risk groups for aggression. In addition, TPH1 gene is most likely to have a shared effect on aggression and MDD. © Copyright 2012 Physicians Postgraduate Press, Inc.

  11. [Effects and Biological Response on Bioremediation of Petroleum Contaminated Soil].

    PubMed

    Yang, Qian; Wu, Man-li; Nie, Mai-qian; Wang, Ting-ting; Zhang, Ming-hui

    2015-05-01

    Bioaugmentation and biostimulation were used to remediate petroleum-contaminated soil which were collected from Zichang city in North of Shaanxi. The optimal bioremediation method was obtained by determining the total petroleum hydrocarbon(TPH) using the infrared spectroscopy. During the bioremediation, number of degrading strains, TPH catabolic genes, and soil microbial community diversity were determined by Most Probable Number (MPN), polymerase chain reaction (PCR) combined agarose electrophoresis, and PCR-denaturing gradient electrophoresis (DGGE). The results in different treatments showed different biodegradation effects towards total petroleum hydrocarbon (TPH). Biostimulation by adding N and P to soils achieved the best degradation effects towards TPH, and the bioaugmentation was achieved by inoculating strain SZ-1 to soils. Further analysis indicated the positive correlation between catabolic genes and TPH removal efficiency. During the bioremediation, the number of TPH and alkanes degrading strains was higher than the number of aromatic degrading strains. The results of PCR-DGGE showed microbial inoculums could enhance microbial community functional diversity. These results contribute to understand the ecologically microbial effects during the bioremediation of petroleum-polluted soil.

  12. Anthocyanins, phenolics, and antioxidant capacity in diverse small fruits: vaccinium, rubus, and ribes.

    PubMed

    Moyer, Richard A; Hummer, Kim E; Finn, Chad E; Frei, Balz; Wrolstad, Ronald E

    2002-01-30

    Fruits from 107 genotypes of Vaccinium L., Rubus L., and Ribes L., were analyzed for total anthocyanins (ACY), total phenolics (TPH), and antioxidant capacities as determined by oxygen radical absorbing capacity (ORAC) and ferric reducing antioxidant power (FRAP). Fruit size was highly correlated (r = 0.84) with ACY within Vaccinium corymbosum L., but was not correlated to ACY across eight other Vaccinium species, or within 27 blackberry hybrids. Certain Vaccinium and Ribes fruits with pigmented flesh were lower in ACY, TPH, ORAC, and FRAP compared to those values in berries with nonpigmented flesh. ORAC values ranged from 19 to 131 micromol Trolox equivalents/g in Vaccinium, from 13 to 146 in Rubus, and from 17 to 116 in Ribes. Though ACY may indicate TPH, the range observed in ACY/TPH ratios precludes prediction of ACY from TPH and vice versa for a single genotype. In general, TPH was more highly correlated to antioxidant capacity than ACY was. This study demonstrates the wide diversity of phytochemical levels and antioxidant capacities within and across three genera of small fruit.

  13. Case-control and family-based association studies of candidate genes in autistic disorder and its endophenotypes: TPH2 and GLO1.

    PubMed

    Sacco, Roberto; Papaleo, Veruska; Hager, Jorg; Rousseau, Francis; Moessner, Rainald; Militerni, Roberto; Bravaccio, Carmela; Trillo, Simona; Schneider, Cindy; Melmed, Raun; Elia, Maurizio; Curatolo, Paolo; Manzi, Barbara; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Reichelt, Karl-Ludvig; Persico, Antonio M

    2007-03-08

    The TPH2 gene encodes the enzyme responsible for serotonin (5-HT) synthesis in the Central Nervous System (CNS). Stereotypic and repetitive behaviors are influenced by 5-HT, and initial studies report an association of TPH2 alleles with childhood-onset obsessive-compulsive disorder (OCD) and with autism. GLO1 encodes glyoxalase I, the enzyme which detoxifies alpha-oxoaldehydes such as methylglyoxal in all living cells. The A111E GLO1 protein variant, encoded by SNP C419A, was identified in autopsied autistic brains and proposed to act as an autism susceptibility factor. Hyperserotoninemia, macrocephaly, and peptiduria represent some of the best-characterized endophenotypes in autism research. Family-based and case-control association studies were performed on clinical samples drawn from 312 simplex and 29 multiplex families including 371 non-syndromic autistic patients and 156 unaffected siblings, as well as on 171 controls. TPH2 SNPs rs4570625 and rs4565946 were genotyped using the TaqMan assay; GLO1 SNP C419A was genotyped by PCR and allele-specific restriction digest. Family-based association analyses were performed by TDT and FBAT, case-control by chi2, endophenotypic analyses for 5-HT blood levels, cranial circumference and urinary peptide excretion rates by ANOVA and FBAT. TPH2 alleles and haplotypes are not significantly associated in our sample with autism (rs4570625: TDT P = 0.27, and FBAT P = 0.35; rs4565946: TDT P = 0.45, and FBAT P = 0.55; haplotype P = 0.84), with any endophenotype, or with the presence/absence of prominent repetitive and stereotyped behaviors (motor stereotypies: P = 0.81 and 0.84, verbal stereotypies: P = 0.38 and 0.73 for rs4570625 and rs4565946, respectively). Also GLO1 alleles display no association with autism (191 patients vs 171 controls, P = 0.36; TDT P = 0.79, and FBAT P = 0.37), but unaffected siblings seemingly carry a protective gene variant marked by the A419 allele (TDT P < 0.05; patients vs unaffected siblings TDT and FBAT P < 0.00001). TPH2 gene variants are unlikely to contribute to autism or to the presence/absence of prominent repetitive behaviors in our sample, although an influence on the intensity of these behaviors in autism cannot be excluded. GLO1 gene variants do not confer autism vulnerability in this sample, but allele A419 apparently carries a protective effect, spurring interest into functional correlates of the C419A SNP.

  14. Case-control and family-based association studies of candidate genes in autistic disorder and its endophenotypes: TPH2 and GLO1

    PubMed Central

    Sacco, Roberto; Papaleo, Veruska; Hager, Jorg; Rousseau, Francis; Moessner, Rainald; Militerni, Roberto; Bravaccio, Carmela; Trillo, Simona; Schneider, Cindy; Melmed, Raun; Elia, Maurizio; Curatolo, Paolo; Manzi, Barbara; Pascucci, Tiziana; Puglisi-Allegra, Stefano; Reichelt, Karl-Ludvig; Persico, Antonio M

    2007-01-01

    Background The TPH2 gene encodes the enzyme responsible for serotonin (5-HT) synthesis in the Central Nervous System (CNS). Stereotypic and repetitive behaviors are influenced by 5-HT, and initial studies report an association of TPH2 alleles with childhood-onset obsessive-compulsive disorder (OCD) and with autism. GLO1 encodes glyoxalase I, the enzyme which detoxifies α-oxoaldehydes such as methylglyoxal in all living cells. The A111E GLO1 protein variant, encoded by SNP C419A, was identifed in autopsied autistic brains and proposed to act as an autism susceptibility factor. Hyperserotoninemia, macrocephaly, and peptiduria represent some of the best-characterized endophenotypes in autism research. Methods Family-based and case-control association studies were performed on clinical samples drawn from 312 simplex and 29 multiplex families including 371 non-syndromic autistic patients and 156 unaffected siblings, as well as on 171 controls. TPH2 SNPs rs4570625 and rs4565946 were genotyped using the TaqMan assay; GLO1 SNP C419A was genotyped by PCR and allele-specific restriction digest. Family-based association analyses were performed by TDT and FBAT, case-control by χ2, endophenotypic analyses for 5-HT blood levels, cranial circumference and urinary peptide excretion rates by ANOVA and FBAT. Results TPH2 alleles and haplotypes are not significantly associated in our sample with autism (rs4570625: TDT P = 0.27, and FBAT P = 0.35; rs4565946: TDT P = 0.45, and FBAT P = 0.55; haplotype P = 0.84), with any endophenotype, or with the presence/absence of prominent repetitive and stereotyped behaviors (motor stereotypies: P = 0.81 and 0.84, verbal stereotypies: P = 0.38 and 0.73 for rs4570625 and rs4565946, respectively). Also GLO1 alleles display no association with autism (191 patients vs 171 controls, P = 0.36; TDT P = 0.79, and FBAT P = 0.37), but unaffected siblings seemingly carry a protective gene variant marked by the A419 allele (TDT P < 0.05; patients vs unaffected siblings TDT and FBAT P < 0.00001). Conclusion TPH2 gene variants are unlikely to contribute to autism or to the presence/absence of prominent repetitive behaviors in our sample, although an influence on the intensity of these behaviors in autism cannot be excluded. GLO1 gene variants do not confer autism vulnerability in this sample, but allele A419 apparently carries a protective effect, spurring interest into functional correlates of the C419A SNP. PMID:17346350

  15. Platelet serotonin promotes the recruitment of neutrophils to sites of acute inflammation in mice

    PubMed Central

    Suidan, Georgette L.; Demers, Melanie; Herr, Nadine; Carbo, Carla; Brill, Alexander; Cifuni, Stephen M.; Mauler, Maximilian; Cicko, Sanja; Bader, Michael; Idzko, Marco; Bode, Christoph

    2013-01-01

    The majority of peripheral serotonin is stored in platelets, which secrete it on activation. Serotonin releases Weibel-Palade bodies (WPBs) and we asked whether absence of platelet serotonin affects neutrophil recruitment in inflammatory responses. Tryptophan hydroxylase (Tph)1–deficient mice, lacking non-neuronal serotonin, showed mild leukocytosis compared with wild-type (WT), primarily driven by an elevated neutrophil count. Despite this, 50% fewer leukocytes rolled on unstimulated mesenteric venous endothelium of Tph1−/− mice. The velocity of rolling leukocytes was higher in Tph1−/− mice, indicating fewer selectin-mediated interactions with endothelium. Stimulation of endothelium with histamine, a secretagogue of WPBs, or injection of serotonin normalized the rolling in Tph1−/− mice. Diminished rolling in Tph1−/− mice resulted in reduced firm adhesion of leukocytes after lipopolysaccharide treatment. Blocking platelet serotonin uptake with fluoxetine in WT mice reduced serum serotonin by > 80% and similarly reduced leukocyte rolling and adhesion. Four hours after inflammatory stimulation, neutrophil extravasation into lung, peritoneum, and skin wounds was reduced in Tph1−/− mice, whereas in vitro neutrophil chemotaxis was independent of serotonin. Survival of lipopolysaccharide-induced endotoxic shock was improved in Tph1−/− mice. In conclusion, platelet serotonin promotes the recruitment of neutrophils in acute inflammation, supporting an important role for platelet serotonin in innate immunity. PMID:23243271

  16. Do your extractable TPH concentrations represent dissolved petroleum? An update on applied research

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zemo, D.A.

    1997-12-31

    Elevated concentrations of {open_quotes}dissolved-phase{close_quotes} extractable total petroleum hydrocarbons (TPH) in groundwater samples can be a significant impediment to site closure in states that regulate groundwater using TPH criteria. These analytical results are inconsistent with petroleum chemistry because of the relatively low water solubility of petroleum products. This paper presents an update of our research into the source of medium- to high-boiling TPH detections in groundwater samples and application of the results to multiple projects. This work follows from a 1995 publication in which positive interferences to the Method 8015M (GC-FID) TPH measurement by soluble, non-petroleum hydrocarbons resulting from intrinsic bioremediationmore » or non-dissolved petroleum adhered to particulates was described. The 1995 paper was largely theoretical and focused on one case study. Since 1995, we have evaluated the source of TPH detections in groundwater at numerous petroleum sites and have demonstrated the significance of interferences to the Method 8015M measurement to the California regulatory community. Our work has shown conclusively that elevated concentrations of extractable TPH are not representative of dissolved petroleum constituents. We have shown that a sample cleanup prior to analysis using silica gel cleanup (to remove polar non-petroleum hydrocarbons) and/or laboratory filtration (to reduce petroleum-affected particulates) is required to overcome the false positives caused by interferences to the Method 8015M measurement.« less

  17. Selective removal of caffeine from tea extracts using macroporous crosslinked polyvinyl alcohol adsorbents.

    PubMed

    Ma, Ning; Wang, Peng; Kong, Xia; Shi, Rongfu; Yuan, Zhi; Wang, Chunhong

    2012-01-01

    The hydrolysis reaction of ester groups in vinyl acetate (VAc) was used to introduce hydroxyl groups into the matrix of a macroporous adsorbent, which was itself prepared by free radical suspension copolymerization of triallyl isocyanurate (TAIC) and VAc. Therefore, the copolymerization incompatibility between the hydrophilic and the hydrophobic monomer was overcome successfully and the hydrophobic matrix of the polymeric adsorbent containing a polyvinyl alcohol (PVA) segment was obtained. Introduction of the PVA segment decreased the hydrophobic adsorption affinity of the adsorbent while producing the hydrogen-bonding interaction. When isolating the two active components, polyphenols (TPh) and caffeine (CAF), from green tea extracts, this polymeric adsorbent, namely poly(TAIC-co-VA), exhibited good adsorption selectivity towards TPh over CAF. The adsorption mechanism leading to this selectivity involved a hydrophobic interaction mechanism for CAF and multiple weak hydrophobic and hydrogen-bonding interactions for TPh. The adsorption thermodynamics for TPh on poly(TAIC-co-VA) were studied. The effects of adsorbent structure and gradient desorption conditions on isolation were investigated. The result showed that adsorbent, with 20% TAIC content, was able to efficiently remove CAF from different tea extracts with different ratios of TPh and CAF. Finally, almost no CAF was detected in the TPh fraction and the recovery of TPh was greater than 95%. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. [Influence of chronic alcohol treatment on the expression of the Bdnf, Bax, Bcl-xL, and CASP3 genes in the mouse brain: Role of the C1473G polymorphism in the gene encoding tryptophan hydroxylase 2].

    PubMed

    Bazovkina, D V; Tsybko, A S; Filimonova, E A; Ilchibaeva, T V; Naumenko, V S

    2016-01-01

    Tryptophan hydroxylase 2 (Tph-2) is the key enzyme in serotonin biosynthesis. Serotonin is one of the main neurotransmitters involved in the regulation of various physiological functions and behavior patterns. The influence of chronic ethanol consumption on the expression of the Bdnf, Bax, Bcl-xL, and CASP3 genes was studied in the brain structures of B6-1473C (C/C) and B6-1473G (G/G) mice that had been obtained on the base of the C57BL/6 strain. The strains differed in the genotype for the C1473G single nucleotide polymorphism in the Tph-2 gene and in Tph-2 enzyme activity. It was found that chronic alcohol treatment led to a significant increase in the expression of the Bdnf gene in the midbrain of B6-1473G mice, but not in B6-1473С. Chronic alcohol treatment considerably decreased the expression of the ultimate brain apoptosis effector, caspase 3, in the frontal cortex, but increased it in the hippocampus of B6-1473G mice. At the same time, chronic ethanol administration reduced the level of the antiapoptotic Bcl-xL mRNA in the midbrain of B6-1473C mice. Thus, the C1473G polymorphism in the Tph-2 gene considerably influenced the changes in the expression patterns of genes involved in the regulation of neurogenesis and neural apoptosis induced by chronic ethanol treatment.

  19. Phytoremediation potential and ecological and phenological changes of native pioneer plants from weathered oil spill-impacted sites at tropical wetlands.

    PubMed

    Palma-Cruz, Felipe de J; Pérez-Vargas, Josefina; Rivera Casado, Noemí Araceli; Gómez Guzmán, Octavio; Calva-Calva, Graciano

    2016-08-01

    Pioneer native plant species from weathered oil spill-affected sites were selected to study their potential for phytoremediation on the basis of their ecological and phenological changes during the phytoremediation process. Experiments were conducted in field and in greenhouse. In field, native plants from aged oil spill-impacted sites with up 400 g of weathered petroleum hydrocarbons per kilogram soil were selected. In the impacted sites, the principal dominant plant species with potential for hydrocarbons removal were Cyperus laxus, Cyperus esculentus, and Ludwigia peploides. In greenhouse, the phenology of the selected plant species was drastically affected by the hydrocarbons level above 325 g total petroleum hydrocarbons (TPH) per kilogram soil after 2 years of phytoremediation of soils from the aged oil spill-impacted sites. From the phytoremediation treatments, a mix-culture of C. laxus, C. esculentus, and L. peploides in soil containing 325 g TPH/kg soil, from which 20.3 % were polyaromatic hydrocarbons (PAH) and 34.2 % were asphaltenes (ASF), was able to remove up 93 % of the TPH, while in unvegetated soil the TPH removal was 12.6 %. Furthermore, evaluation of the biodiversity and life forms of plant species in the impacted sites showed that phytoremediation with C. esculentus, alone or in a mix-culture with C. laxus and L. peploides, reduces the TPH to such extent that the native plant community was progressively reestablished by replacing the cultivated species resulting in the ecological recovery of the affected soil. These results demonstrate that native Cyperus species from weathered oil spill-affected sites, specifically C. esculentus and C. laxus, alone or in a mix-culture, have particular potential for phytoremediation of soils from tropical wetlands contaminated with weathered oil hydrocarbons.

  20. All genes encoding enzymes participating in melatonin biosynthesis in the chicken pineal gland are transcribed rhythmically.

    PubMed

    Adamska, I; Marhelava, K; Walkiewicz, D; Kedzierska, U; Markowska, M; Majewski, P M

    2016-08-01

    Our recent research on the pineal gland of young chickens confirmed that three genes encoding enzymes involved in pineal melatonin biosynthesis, tryptophan hydroxylase 1 (Tph1), arylalkylamine-N-acetyltransferase (Aanat) and acetylserotonin O-methyltransferase (Asmt), are transcribed rhythmically under light:dark (L:D) 12:12 conditions in vivo. Additionally, in the pineal gland of maturing chickens, the dopa decarboxylase (Ddc) gene is transcribed rhythmically at a specific stage of the developmental process. Therefore, the aim of the present study was to verify whether all of these genes are transcribed rhythmically in vivo under constant darkness (D:D) and in pinealocyte cultures under both L:D and D:D. Experiments were performed on chickens maintained under L:D 12:12 conditions. Chickens at 15 days of age were divided into two groups; chickens from the first group remained under the same conditions, whereas those from the second group were kept in darkness. Subsequently, 16-day-old animals were sacrificed every 2 hours over a 24-h period. For the in vitro experiments, 16-day-old chickens were sacrificed at ZT 6, and their pineal glands were isolated. Pineal cultures were maintained for up to two days in L:D conditions. Then, the pinealocyte cultures were divided into two groups: the first remained under L:D conditions, whereas the second was transferred to D:D conditions. Pinealocytes were subsequently collected every 2 hours over a 24-h period. Transcription was evaluated using the RT-qPCR method, and the rhythm percentage was calculated through Cosinor analysis. The mRNA levels of all genes examined were rhythmic under all conditions. Moreover, in silico analysis of the promoters of all of the genes examined revealed the presence of enhancer box sequences in all of the promoters as well as DBP/E4BP4 binding elements in the promoters of Tph1 and Asmt. This suggests that these genes may all be regulated transcriptionally by the molecular clock mechanism and may be considered clock as controlled genes.

  1. The effects of brain serotonin deficiency on behavioural disinhibition and anxiety-like behaviour following mild early life stress.

    PubMed

    Sachs, Benjamin D; Rodriguiz, Ramona M; Siesser, William B; Kenan, Alexander; Royer, Elizabeth L; Jacobsen, Jacob P R; Wetsel, William C; Caron, Marc G

    2013-10-01

    Aberrant serotonin (5-HT) signalling and exposure to early life stress have both been suggested to play a role in anxiety- and impulsivity-related behaviours. However, whether congenital 5-HT deficiency × early life stress interactions influence the development of anxiety- or impulsivity-like behaviour has not been established. Here, we examined the effects of early life maternal separation (MS) stress on anxiety-like behaviour and behavioural disinhibition, a type of impulsivity-like behaviour, in wild-type (WT) and tryptophan hydroxylase 2 (Tph2) knock-in (Tph2KI) mice, which exhibit ~60-80% reductions in the levels of brain 5-HT due to a R439H mutation in Tph2. We also investigated the effects of 5-HT deficiency and early life stress on adult hippocampal neurogenesis, plasma corticosterone levels and several signal transduction pathways in the amygdala. We demonstrate that MS slightly increases anxiety-like behaviour in WT mice and induces behavioural disinhibition in Tph2KI animals. We also demonstrate that MS leads to a slight decrease in cell proliferation within the hippocampus and potentiates corticosterone responses to acute stress, but these effects are not affected by brain 5-HT deficiency. However, we show that 5-HT deficiency leads to significant alterations in SGK-1 and GSK3β signalling and NMDA receptor expression in the amygdala in response to MS. Together, these findings support a potential role for 5-HT-dependent signalling in the amygdala in regulating the long-term effects of early life stress on anxiety-like behaviour and behavioural disinhibition.

  2. Genetic, Psychological, and Personal Network Factors Associated With Changes in Binge Drinking Over 2 Years Among Mexican Heritage Adolescents in the USA.

    PubMed

    Song, Sunmi; Marcum, Christopher Steven; Wilkinson, Anna V; Shete, Sanjay; Koehly, Laura M

    2018-04-24

    Despite prevalent binge drinking and alcohol-dependent symptoms among Hispanics, few studies have examined how multidimensional factors influence Hispanic adolescents' binge drinking. Purpose This study examines the effects of genetic, psychological, and social network factors on binge drinking over time among Mexican heritage adolescents in the USA and whether there are correlations among genetic variants that are associated with binge drinking and psychological and network characteristics. Mexican heritage adolescents (n = 731) participated in a longitudinal study, which included genetic testing at baseline, alcohol use assessments at first and second follow-ups, and questionnaires on sensation seeking, impulsivity, and peer and family network characteristics at second follow-up. Logistic regression and Spearman correlation analyses were performed. After adjusting for demographic characteristics, underlying genetic clustering, and binge drinking at first follow-up, two genetic variants on tryptophan hydroxylase 2 (TPH2; rs17110451, rs7963717), sensation seeking and impulsivity, and having a greater fraction of peers who drink or encourage drinking alcohol were associated with greater risk whereas another genetic variant on TPH2 (rs11178999) and having a greater fraction of close family relationships were associated with reduced risk for binge drinking at second follow-up. Genetic variants in TPH1 (rs591556) were associated with sensation seeking and impulsivity, while genetic variants in TPH2 (rs17110451) were associated with the fraction of drinkers in family. Results reveal that genetic variants in the serotonin pathway, behavioral disinhibition traits, and social networks exert joint influences on binge drinking in Mexican heritage adolescents in the USA.

  3. Enhancement of Gastric Ulcer Healing and Angiogenesis by Hepatocyte Growth Factor Gene Mediated by Attenuated Salmonella in Rats.

    PubMed

    Ha, Xiaoqin; Peng, Junhua; Zhao, Hongbin; Deng, Zhiyun; Dong, Juzi; Fan, Hongyan; Zhao, Yong; Li, Bing; Feng, Qiangsheng; Yang, Zhihua

    2017-02-01

    The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 10⁹ cfu), vehicle (TP, 1 × 10⁹ cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm², which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm², respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P < 0.05). The microvessel density in the ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm², which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site.

  4. Neuronal serotonin regulates growth of the intestinal mucosa in mice.

    PubMed

    Gross, Erica R; Gershon, Michael D; Margolis, Kara G; Gertsberg, Zoya V; Li, Zhishan; Cowles, Robert A

    2012-08-01

    The enteric abundance of serotonin (5-HT), its ability to promote proliferation of neural precursors, and reports that 5-HT antagonists affect crypt epithelial proliferation led us to investigate whether 5-HT affects growth and maintenance of the intestinal mucosa in mice. cMice that lack the serotonin re-uptake transporter (SERTKO mice) and wild-type mice were given injections of selective serotonin re-uptake inhibitors (gain-of-function models). We also analyzed mice that lack tryptophan hydroxylase-1 (TPH1KO mice, which lack mucosal but not neuronal 5-HT) and mice deficient in tryptophan hydroxylase-2 (TPH2KO mice, which lack neuronal but not mucosal 5-HT) (loss-of-function models). Wild-type and SERTKO mice were given ketanserin (an antagonist of the 5-HT receptor, 5-HT(2A)) or scopolamine (an antagonist of the muscarinic receptor). 5-HT(2A) receptors and choline acetyltransferase were localized by immunocytochemical analysis. Growth of the mucosa and proliferation of mucosal cells were significantly greater in SERTKO mice and in mice given selective serotonin re-uptake inhibitors than in wild-type mice, but were diminished in TPH2KO (but not in TPH1KO) mice. Ketanserin and scopolamine each prevented the ability of SERT knockout or inhibition to increase mucosal growth and proliferation. Cholinergic submucosal neurons reacted with antibodies against 5-HT(2A). 5-HT promotes growth and turnover of the intestinal mucosal epithelium. Surprisingly, these processes appear to be mediated by neuronal, rather than mucosal, 5-HT. The 5-HT(2A) receptor activates cholinergic neurons, which provide a muscarinic innervation to epithelial effectors. Copyright © 2012 AGA Institute. Published by Elsevier Inc. All rights reserved.

  5. Bioremediation of petroleum hydrocarbon contaminated soil by Rhodobacter sphaeroides biofertilizer and plants.

    PubMed

    Jiao, Haihua; Luo, Jinxue; Zhang, Yiming; Xu, Shengjun; Bai, Zhihui; Huang, Zhanbin

    2015-09-01

    Bio-augmentation is a promising technique for remediation of polluted soils. This study aimed to evaluate the bio-augmentation effect of Rhodobacter sphaeroides biofertilizer (RBF) on the bioremediation of total petroleum hydrocarbons (TPH) contaminated soil. A greenhouse pot experiment was conducted over a period of 120 days, three methods for enhancing bio-augmentation were tested on TPH contaminated soils, including single addition RBF, planting, and combining of RBF and three crop species, such as wheat (W), cabbage (C) and spinach (S), respectively. The results demonstrated that the best removal of TPH from contaminated soil in the RBF bio-augmentation rhizosphere soils was found to be 46.2%, 65.4%, 67.5% for W+RBF, C+RBF, S+RBF rhizosphere soils respectively. RBF supply impacted on the microbial community diversity (phospholipid fatty acids, PLFA) and the activity of soil enzymes, such as dehydrogenase (DH), alkaline phosphatase (AP) and urease (UR). There were significant difference among the soil only containing crude oil (CK), W, C and S rhizosphere soils and RBF bio-augmentation soils. Moreover, the changes were significantly distinct depended on crops species. It was concluded that the RBF is a valuable material for improving effect of remediation of TPH polluted soils.

  6. Treatment of coal gasification wastewater by membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system.

    PubMed

    Jia, Shengyong; Han, Hongjun; Hou, Baolin; Zhuang, Haifeng; Fang, Fang; Zhao, Qian

    2014-12-01

    A laboratory-scale membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system was developed to treat coal gasification wastewater to enhance the COD, total phenols (TPh), NH4+ removals and migrate the membrane fouling. Since the MBR–PAC system operated with PAC dosage of 4 g L−1, the maximum removal efficiencies of COD, TPh and NH4+ reached 93%, 99% and 63%, respectively with the corresponding influent concentrations of 2.27 g L−1, 497 mg L−1 and 164 mg N L−1; the PAC extraction efficiencies of COD, TPh and NH4+ were 6%, 3% and 13%, respectively; the transmembrane pressure decreased 34% with PAC after 50 d operation. The results demonstrate that PAC played a key role in the enhancement of biodegradability and mitigation of membrane fouling.

  7. Solar-chemical treatment of groundwater contaminated with petroleum at gas station sites: ex situ remediation using solar/TiO(2) photocatalysis and Solar Photo-Fenton.

    PubMed

    Cho, Ii-Hyoung; Kim, Young-Gyu; Yang, Jae-Kyu; Lee, Nae-Hyun; Lee, Seung-Mok

    2006-01-01

    Groundwater samples contaminated by BTEX (benzene, toluene, ethylbenzene, xylene isomers and TPHs (total petroleum hydrocarbons) were treated with advanced oxidation processes (AOPs), such as TiO(2) photocatalysis and Fe(2+)/H(2)O(2) exposed to solar light (37 degrees N and 128 degrees E) with an average intensity of 1.7 mW/cm(2) at 365 nm. These AOP processes showed feasibility in the treatment of groundwater contaminated with BTEX, TPH and TOC (Total Organic Carbon). Outdoor field tests showed that the degradation efficiency of each contaminant was higher in the Fe(2+)/H(2)O(2) system without solar light compared to the TiO(2)/solar light and H(2)O(2)/solar light systems. However, the TiO(2)/solar light and the Fe(2+)/H(2)O(2)/solar light systems showed significantly enhanced efficiencies in the degradation of BTEX, TPH and TOC with the additional use of H(2)O(2). Near complete degradation of BTEX and TPH was observed within 2 and 4 hrs, respectively, however, that of TOC was slower. Without pretreatment of the groundwater, fouling of the TiO(2), due to the ionic species present, was observed within 1 hr of operation, which resulted in the inhibition of further BTEX, TPH and TOC destruction. The degradation rate of n-alkanes with carbon numbers ranging from C10 to C15 was relatively greater than that of n-alknaes with carbon numbers ranging from C16 to C20. From this work, the AOP process (Fe(2+)/H(2)O(2)/solar light and TiO(2)/H(2)O(2)/solar light) illuminated with solar light was identified as an effective ex situ technique in the remediation of groundwater contaminated with petroleum.

  8. Effect of phloretin on the permeability of thin lipid membranes

    PubMed Central

    1976-01-01

    Phloretin dramatically increases cation conductances and decreases anion conductances of membranes treated with ion carriers (nonactin, valinomycin, carbonyl-cyanide-m-chlorophenylhydrazone [CCCP], and Hg(C6F5)2) or lipophilic ions (tetraphenylarsonium [tphAs+] and tetraphenylborate [TPhB-]). For example, on phosphatidylethanolamine membranes, 10(-4) M phloretin increases K+ -nonactin and TPhAs+ conductances and decreases CCCP- and TPhB- conductances 10(3)-fold; on lecithin: cholesterol membranes, it increases K+-nonactin conductance 10(5)-fold and decreases CCCP- conductance 10(3)-fold. Similar effects are obtained with p- and m-nitrophenol at 10(-2) M. These effects are produced by the un-ionized form of phloretin and the nitrophenols. We believe that phloretin, which possesses a large dipole moment, adsorbs and orients at the membrane surface to introduce a dipole potential of opposite polarity to the preexisting positive one, thus increasing the partition coefficient of cations into the membrane interior and decreasing the partition coefficient of anions. (Phloretin may also increase the fluidity of cholesterol-containing membranes; this is manifested by its two- to three-fold increase in nonelectrolyte permeability and its asymmetrical effect on cation and anion conductances in cholesterol-containing membranes.) It is possible that pholoretin's inhibition of chloride, urea, and glucose transport in biological membranes results from the effects of these intense intrafacial dipole fields on the translocator(s) of these molecules. PMID:946975

  9. Polymerization Stress Development in Dental Composites: Effect of Cavity Design Factor

    PubMed Central

    Antonucci, Joseph M.; Giuseppetti, Anthony A.; O’Donnell, Justin N.R.; Schumacher, Gary E.; Skrtic, Drago

    2009-01-01

    The objective of the study was to assess the effect of the cavity design factor (C-factor) on polymerization stress development (PSD) in resin composites. An experimental resin (BT resin) was prepared, which contained 2,2-bis[p-(2’-hydroxy-3’-methacryloxypropoxy)phenylene]propane (B) and triethylene glycol dimethacrylate (T) in 1:1 mass ratio, and an activator for visible light polymerization. An experimental composite with demonstrated remineralizing potential was also formulated by inclusion into the BT resin of zirconia-hybridized amorphous calcium phosphate (ACP) filler at a mass fraction of 40 % (BT/ACP composite). A commercial glass-filled composite (TPH) was used as a control. To assess the effect of the test geometry on PSD, C-factor was systematically varied between 0.8 and 6.0 by varying the height of the cylindrical composite specimens. The measured PSD values obtained by cantilever beam tensometry for specimens with variable C-factors were normalized for mass to specimens with a C-factor of 1.33 (h=2.25 mm) as controls to give calculated PSD values. Degrees of vinyl conversions (DC) attained in the TPH control and in the experimental BT/ACP composites were measured by near-infrared spectroscopy. In both the TPH and BT/ACP composite series, PSDcalc increased with the increasing C-factor, confirming the hypothesis that the C-factor value influences PSD values. The higher PSDmeas and PSDcalc values for the experimental BT/ACP composite compared to the commercial TPH composite probably reflect differences in the type and mass of the resin and filler phases in the two types of composite. These differences also account for the observed variation (21 %) in DC attained in a BT/ACP composite 2 h after cure (69.5 %) and in the DC of the TPH composite (57.5 %) having the same C-factor. The cavity design factor seems to play a key role in influencing the PSD of bonded composites, but other factors such as composite mass and composition also must be considered for their effects on PSD. PMID:26413236

  10. Serotonin synthesis rate and the tryptophan hydroxylase-2: G-703T polymorphism in social anxiety disorder.

    PubMed

    Furmark, Tomas; Marteinsdottir, Ina; Frick, Andreas; Heurling, Kerstin; Tillfors, Maria; Appel, Lieuwe; Antoni, Gunnar; Hartvig, Per; Fischer, Håkan; Långström, Bengt; Eriksson, Elias; Fredrikson, Mats

    2016-10-01

    It is disputed whether anxiety disorders, like social anxiety disorder, are characterized by serotonin over- or underactivity. Here, we evaluated whether our recent finding of elevated neural serotonin synthesis rate in patients with social anxiety disorder could be reproduced in a separate cohort, and whether allelic variation in the tryptophan hydroxylase-2 (TPH2) G-703T polymorphism relates to differences in serotonin synthesis assessed with positron emission tomography. Eighteen social anxiety disorder patients and six healthy controls were scanned during 60 minutes in a resting state using positron emission tomography and 5-hydroxy-L-[β -(11)C]tryptophan, [(11)C]5-HTP, a substrate of the second enzymatic step in serotonin synthesis. Parametric images were generated, using the reference Patlak method, and analysed using Statistical Parametric Mapping (SPM8). Blood samples for genotyping of the TPH2 G-703T polymorphism were obtained from 16 social anxiety disorder patients (T carriers: n=5, GG carriers: n=11). A significantly elevated [(11)C]5-HTP accumulation rate, indicative of enhanced decarboxylase activity and thereby serotonin synthesis capacity, was detected in social anxiety disorder patients compared with controls in the hippocampus and basal ganglia nuclei and, at a more lenient (uncorrected) statistical threshold, in the amygdala and anterior cingulate cortex. In patients, the serotonin synthesis rate in the amygdala and anterior cingulate cortex was significantly elevated in TPH2 T carriers in comparison with GG homozygotes. Our results support that social anxiety disorder entails an overactive presynaptic serotonergic system that, in turn, seems functionally influenced by the TPH2 G-703T polymorphism in emotionally relevant brain regions. © The Author(s) 2016.

  11. Mast Cells and Serotonin Synthesis Modulate Chagas Disease in the Colon: Clinical and Experimental Evidence.

    PubMed

    Kannen, Vinicius; Sakita, Juliana Y; Carneiro, Zumira A; Bader, Michael; Alenina, Natalia; Teixeira, Regina R; de Oliveira, Enio C; Brunaldi, Mariângela O; Gasparotto, Bianca; Sartori, Daniela C; Fernandes, Cleverson R; Silva, João S; Andrade, Marcus V; Silva, Wilson A; Uyemura, Sergio A; Garcia, Sérgio B

    2018-06-01

    Trypanosoma cruzi (T. cruzi) infects millions of Latin Americans each year and can induce chagasic megacolon. Little is known about how serotonin (5-HT) modulates this condition. Aim We investigated whether 5-HT synthesis alters T. cruzi infection in the colon. Forty-eight paraffin-embedded samples from normal colon and chagasic megacolon were histopathologically analyzed (173/2009). Tryptophan hydroxylase 1 (Tph1) knockout (KO) mice and c-Kit W-sh mice underwent T. cruzi infection together with their wild-type counterparts. Also, mice underwent different drug treatments (16.1.1064.60.3). In both humans and experimental mouse models, the serotonergic system was activated by T. cruzi infection (p < 0.05). While treating Tph1KO mice with 5-HT did not significantly increase parasitemia in the colon (p > 0.05), rescuing its synthesis promoted trypanosomiasis (p < 0.01). T. cruzi-related 5-HT release (p < 0.05) seemed not only to increase inflammatory signaling, but also to enlarge the pericryptal macrophage and mast cell populations (p < 0.01). Knocking out mast cells reduced trypanosomiasis (p < 0.01), although it did not further alter the neuroendocrine cell number and Tph1 expression (p > 0.05). Further experimentation revealed that pharmacologically inhibiting mast cell activity reduced colonic infection (p < 0.01). A similar finding was achieved when 5-HT synthesis was blocked in c-Kit W-sh mice (p > 0.05). However, inhibiting mast cell activity in Tph1KO mice increased colonic trypanosomiasis (p < 0.01). We show that mast cells may modulate the T. cruzi-related increase of 5-HT synthesis in the intestinal colon.

  12. A gene-environment investigation on personality traits in two independent clinical sets of adult patients with personality disorder and attention deficit/hyperactive disorder.

    PubMed

    Jacob, Christian P; Nguyen, Thuy Trang; Dempfle, Astrid; Heine, Monika; Windemuth-Kieselbach, Christine; Baumann, Katarina; Jacob, Florian; Prechtl, Julian; Wittlich, Maike; Herrmann, Martin J; Gross-Lesch, Silke; Lesch, Klaus-Peter; Reif, Andreas

    2010-06-01

    While an interactive effect of genes with adverse life events is increasingly appreciated in current concepts of depression etiology, no data are presently available on interactions between genetic and environmental (G x E) factors with respect to personality and related disorders. The present study therefore aimed to detect main effects as well as interactions of serotonergic candidate genes (coding for the serotonin transporter, 5-HTT; the serotonin autoreceptor, HTR1A; and the enzyme which synthesizes serotonin in the brain, TPH2) with the burden of life events (#LE) in two independent samples consisting of 183 patients suffering from personality disorders and 123 patients suffering from adult attention deficit/hyperactivity disorder (aADHD). Simple analyses ignoring possible G x E interactions revealed no evidence for associations of either #LE or of the considered polymorphisms in 5-HTT and TPH2. Only the G allele of HTR1A rs6295 seemed to increase the risk of emotional-dramatic cluster B personality disorders (p = 0.019, in the personality disorder sample) and to decrease the risk of anxious-fearful cluster C personality disorders (p = 0.016, in the aADHD sample). We extended the initial simple model by taking a G x E interaction term into account, since this approach may better fit the data indicating that the effect of a gene is modified by stressful life events or, vice versa, that stressful life events only have an effect in the presence of a susceptibility genotype. By doing so, we observed nominal evidence for G x E effects as well as main effects of 5-HTT-LPR and the TPH2 SNP rs4570625 on the occurrence of personality disorders. Further replication studies, however, are necessary to validate the apparent complexity of G x E interactions in disorders of human personality.

  13. Lack of Tryptophan Hydroxylase-1 in Mice Results in Gait Abnormalities

    PubMed Central

    Suidan, Georgette L.; Vanderhorst, Veronique; Hampton, Thomas G.; Wong, Siu Ling; Voorhees, Jaymie R.; Wagner, Denisa D.

    2013-01-01

    The role of peripheral serotonin in nervous system development is poorly understood. Tryptophan hydroxylase-1 (TPH1) is expressed by non-neuronal cells including enterochromaffin cells of the gut, mast cells and the pineal gland and is the rate-limiting enzyme involved in the biosynthesis of peripheral serotonin. Serotonin released into circulation is taken up by platelets via the serotonin transporter and stored in dense granules. It has been previously reported that mouse embryos removed from Tph1-deficient mothers present abnormal nervous system morphology. The goal of this study was to assess whether Tph1-deficiency results in behavioral abnormalities. We did not find any differences between Tph1-deficient and wild-type mice in general motor behavior as tested by rotarod, grip-strength test, open field and beam walk. However, here we report that Tph1 (−/−) mice display altered gait dynamics and deficits in rearing behavior compared to wild-type (WT) suggesting that tryptophan hydroxylase-1 expression has an impact on the nervous system. PMID:23516593

  14. Lack of tryptophan hydroxylase-1 in mice results in gait abnormalities.

    PubMed

    Suidan, Georgette L; Duerschmied, Daniel; Dillon, Gregory M; Vanderhorst, Veronique; Hampton, Thomas G; Wong, Siu Ling; Voorhees, Jaymie R; Wagner, Denisa D

    2013-01-01

    The role of peripheral serotonin in nervous system development is poorly understood. Tryptophan hydroxylase-1 (TPH1) is expressed by non-neuronal cells including enterochromaffin cells of the gut, mast cells and the pineal gland and is the rate-limiting enzyme involved in the biosynthesis of peripheral serotonin. Serotonin released into circulation is taken up by platelets via the serotonin transporter and stored in dense granules. It has been previously reported that mouse embryos removed from Tph1-deficient mothers present abnormal nervous system morphology. The goal of this study was to assess whether Tph1-deficiency results in behavioral abnormalities. We did not find any differences between Tph1-deficient and wild-type mice in general motor behavior as tested by rotarod, grip-strength test, open field and beam walk. However, here we report that Tph1 (-/-) mice display altered gait dynamics and deficits in rearing behavior compared to wild-type (WT) suggesting that tryptophan hydroxylase-1 expression has an impact on the nervous system.

  15. [Use of Leersia hexandra (Poaceae) for soil phytoremediation in soils contaminated with fresh and weathered oil].

    PubMed

    Arias-Trinidad, Alfredo; Rivera-Cruz, María del Carmen; Roldán-Garrigós, Antonio; Aceves-Navarro, Lorenzo Armando; Quintero-Lizaola, Roberto; Hernández-Guzmán, Javier

    2017-03-01

    The oil industry has generated chronic oil spills and their accumulation in wetlands of the state of Tabasco, in Southeastern Mexico. Waterlogging is a factor that limits the use of remediation technologies because of its high cost and low levels of oil degradation. However, Leersia hexandra is a grass that grows in these contaminated areas with weathered oil. The aim of the study was to evaluate the bacteria density, plant biomass production and phytoremediation of L. hexandra in contaminated soil. For this, two experiments in plastic tunnel were performed with fresh (E1) and weathered petroleum (E2) under waterlogging experimental conditions. The E1 was based on eight doses: 6 000, 10 000, 30 000, 60 000, 90 000, 120 000, 150 000 and 180 000 mg.kg-1 dry basis (d. b.) of total petroleum hydrocarbons fresh (TPH-F), and the E2, that evaluated five doses: 14 173, 28 400, 50 598, 75 492 and 112 142 mg. kg-1 d. b. of total petroleum hydrocarbons weathered (TPH-W); a control treatment with 2 607 mg.kg-1 d. b. was used. Each experiment, with eight replicates per treatment, evaluated after three and six months: a) microbial density of total free-living nitrogen-fixing bacteria (NFB) of Azospirillum (AZP) and Azotobacter group (AZT), for viable count in serial plate; b) dry matter production (DMP), quantified gravimetrically as dry weight of L. hexandra; and c) the decontamination percentage of hydrocarbons (PDH) by Soxhlet extraction. In soil with TPH-F, the NFB, AZP y AZT populations were stimulated five times more than the control both at the three and six months; however, concentrations of 150 000 and 180 000 mg.kg-1 d. b. inhibited the bacterial density between 70 and 89 %. Likewise, in soil with TPH-W, the FNB, AZP and AZT inhibitions were 90 %, with the exception of the 14 173 mg.kg-1 d. b. treatment, which stimulated the NFB and AZT in 2 and 0.10 times more than the control, respectively. The DMP was continued at the six months in the experiments, with values of 63 and 89 g in fresh and weathered petroleum, respectively; had no significant differences with the control (p≤0.05). The PDH reached values of 66 to 87 % both TPH-F and TPH-W at six months, respectively. These results demonstrated the ability the L. hexandra rhizosphere to stimulate the high NFB density, vegetal biomass production and phytoremediation of contaminated soils (with fresh and weathered petroleum), in a tropical waterlogging environment.

  16. Petroleum-degrading microbial numbers in rhizosphere and non-rhizosphere crude oil-contaminated soil.

    PubMed

    Kirkpatrick, W D; White, P M; Wolf, D C; Thoma, G J; Reynolds, C M

    2008-01-01

    Phytoremediation can be a cost-effective and environmentally acceptable method to clean up crude oil-contaminated soils in situ. Our research objective was to determine the effects of nitrogen (N) additions and plant growth on the number of total hydrocarbon (TH)-, alkane-, and polycyclic aromatic hydrocarbon (PAH)-degrading microorganisms in weathered crude oil-contaminated soil. A warm-season grass, sudangrass (Sorghum sudanense (Piper) Stapf), was grown for 7 wk in soil with a total petroleum hydrocarbon (TPH) level of 16.6 g TPH/kg soil. Nitrogen was added based upon TPH-C:added total N (TPH-C:TN) ratios ranging from 44:1 to 11:1. Unvegetated and unamended controls were also evaluated. The TH-, alkane-, and PAH-degrading microbial numbers per gram of dry soil were enumerated from rhizosphere and non-rhizosphere soil for vegetated pots and non-rhizosphere soil populations were enumerated from non-vegetated pots. Total petroleum-degrading microbial numbers were also calculated for each pot. The TH-, alkane-, and PAH-degrading microbial numbers per gram of dry soil in the sudangrass rhizosphere were 3.4, 2.6, and 4.8 times larger, respectively, than those in non-rhizosphere soil across all N rates. The presence of sudangrass resulted in significantly more TH-degrading microorganisms per pot when grown in soil with a TPH-C:TN ratio of 11:1 as compared to the control. Increased plant root growth in a crude oil-contaminated soil and a concomitant increase in petroleum-degrading microbial numbers in the rhizosphere have the potential to enhance phytoremediation.

  17. Three year field test of a plant growth promoting rhizobacteria enhanced phytoremediation system at a land farm for treatment of hydrocarbon waste.

    PubMed

    Gurska, Jolanta; Wang, Wenxi; Gerhardt, Karen E; Khalid, Aaron M; Isherwood, David M; Huang, Xiao-Dong; Glick, Bernard R; Greenberg, Bruce M

    2009-06-15

    Phytoremediation of total petroleum hydrocarbons (TPH) has the potential to be a sustainable waste management technology if it can be proven to be effective in the field. Over the past decade, our laboratory has developed a system which utilizes plant growth promoting rhizobacteria (PGPR) enhanced phytoremediation (PEP) that, following extensive greenhouse testing, was shown to be effective at remediating TPH from soils. This system consists of physical soil manipulation and plant growth following seed inoculation with PGPR. PGPR elicit biomass increases, particularly in roots, by minimizing plant stress in highly contaminated soils. Extensive development of the root system enhances degradation of contaminants by the plants and supports an active rhizosphere that effectively promotes TPH degradation by a broad microbial consortium. Following promising greenhouse trials, field tests of PEP were performed over a period of three years at a Southern Ontario site (approximately 130 g kg(-1) TPH) used for land farming of refinery hydrocarbon waste for many years. The low molecular weight fractions (the Canadian Council of Ministers of the Environment (CCME) fractions 1 and 2) were removed through land farming and bioremediation; the high molecular weight, recalcitrant fractions (CCME fractions 3 and 4) remained at high levels in the soil. Using PEP, we substantially remediated fractions 3 and 4, and lowered TPH from 130 g kg(-1) to approximately 50 g kg(-1) over a three year period. The amount of plant growth and extent of oil remediation were consistently enhanced by PGPR.

  18. Assessment of soil pollution based on total petroleum hydrocarbons and individual oil substances.

    PubMed

    Pinedo, J; Ibáñez, R; Lijzen, J P A; Irabien, Á

    2013-11-30

    Different oil products like gasoline, diesel or heavy oils can cause soil contamination. The assessment of soils exposed to oil products can be conducted through the comparison between a measured concentration and an intervention value (IV). Several national policies include the IV based on the so called total petroleum hydrocarbons (TPH) measure. However, the TPH assessment does not indicate the individual substances that may produce contamination. The soil quality assessment can be improved by including common hazardous compounds as polycyclic aromatic hydrocarbons (PAHs) and aromatic volatile hydrocarbons like benzene, toluene, ethylbenzene and xylenes (BTEX). This study, focused on 62 samples collected from different sites throughout The Netherlands, evaluates TPH, PAH and BTEX concentrations in soils. Several indices of pollution are defined for the assessment of individual variables (TPH, PAH, B, T, E, and X) and multivariables (MV, BTEX), allowing us to group the pollutants and simplify the methodology. TPH and PAH concentrations above the IV are mainly found in medium and heavy oil products such as diesel and heavy oil. On the other hand, unacceptable BTEX concentrations are reached in soils contaminated with gasoline and kerosene. The TPH assessment suggests the need for further action to include lighter products. The application of multivariable indices allows us to include these products in the soil quality assessment without changing the IV for TPH. This work provides useful information about the soil quality assessment methodology of oil products in soils, focussing the analysis into the substances that mainly cause the risk. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Valorisation of tuna processing waste biomass for recovery of functional and antioxidant peptides using enzymatic hydrolysis and membrane fractionation process.

    PubMed

    Saidi, Sami; Ben Amar, Raja

    2016-10-01

    The enzymatic hydrolysis using Prolyve BS coupled to membrane process (Ultrafiltration (UF) and nanofiltration (NF)) is a means of biotransformation of tuna protein waste to Tuna protein hydrolysate (TPH) with higher added values. This method could be an effective solution for the production of bioactive compounds used in various biotechnological applications and minimizing the pollution problems generated by the seafood processing industries. The amino acid composition, functional and antioxidant properties of produced TPH were evaluated. The results show that the glutamic acid, aspartic acid, glycine, alaline, valine and leucine were the major amino acids detected in the TPH profile. After membrane fractionation process, those major amino acids were concentrated in the NF retentate (NFR). The NFR and NF permeate (NFP) have a higher protein solubility (>95 %) when compared to TPH (80 %). Higher oil and water binding capacity were observed in TPH and higher emulsifying and foam stability was found in UF retentate. The NFP showed the highest DPPH radical scavenging activity (65 %). The NFR contained antioxidant amino acid (30.3 %) showed the highest superoxide radical and reducing power activities. The TPH showed the highest iron chelating activity (75 %) compared to other peptide fractions. The effect of the membrane fractionation on the molecular weight distribution of the peptide and their bioactivities was underlined. We concluded that the TPH is a valuable source of bioactive peptides and their peptide fractions may serve as useful ingredients for application in food industry and formulation of nutritional products.

  20. Decoy receptor 3 regulates the expression of tryptophan hydroxylase 1 in rheumatoid synovial fibroblasts.

    PubMed

    Maeda, Toshihisa; Miura, Yasushi; Fukuda, Koji; Hayashi, Shinya; Kurosaka, Masahiro

    2015-10-01

    Decoy receptor 3 (DcR3) is expressed in rheumatoid arthritis fibroblast‑like synoviocytes (RA‑FLS) and downregulates the expression of tryptophan hydroxylase 1 (TPH1), which is the rate‑limiting enzyme in serotonin synthesis. The aim of the present study was to determine the specificity of the effects of DcR3 on TPH1 in RA‑FLS, and therefore determine whether DcR3 had the potential to modulate the pathogenesis of RA. The present study also aimed to compare the effects of DcR3 and inflammatory cytokines on the expression of TPH1 in RA‑FLS and osteoarthritis (OA)‑FLS. Primary cultured RA‑ or OA‑FLS were incubated with 1.0 µg/ml DcR3‑Fc protein or 1.0 µg/ml control immunoglobulin G (IgG)1 for 12 h, or with 1.0 ng/ml tumor necrosis factor (TNF)α, 1.0 ng/ml interleukin (IL)‑1β or serum‑free Opti‑MEM only, for 24 h. The relative mRNA expression levels of TPH1 were subsequently quantified using reverse transcription‑polymerase chain reaction. The expression of serotonin in RA or OA synovial tissue was detected using immunohistochemistry. The mRNA expression of TPH1 was observed in both RA‑ and OA‑FLS and was significantly decreased following treatment with DcR3 in the RA‑FLS, however, not in the OA‑FLS. The mRNA expression of TPH1 was significantly decreased following treatment with TNFα or IL‑1β in both the RA‑ and OA‑FLS. The expression of serotonin in the multi‑layered lining synovial cells of RA and the outer layer lining synovial cells of OA was detected using immunohistochemistry. The present study is the first, to the best of our knowledge, to demonstrate that the expression of TPH1 in FLS is downregulated by inflammatory cytokines, and that DcR3 suppressed the expression of TPH1 in RA‑FLS in a disease‑specific manner. These results suggested that synovial serotonin may be involved in the pathogenesis of RA, and that TPH1 and DcR3 may be potential therapeutic targets for the treatment of RA.

  1. Technical note: nitrogen fertilization effects on the degradation of aged diesel oil in composted drilling wastes.

    PubMed

    Choi, Woo-Jung; Chang, Scott X

    2009-07-01

    Hydrocarbon-contaminated wastes generated from oil and gas drilling activities may be used as a soil amendment once composted and further decomposition of residual hydrocarbons can be accomplished after the composts are applied to soils. To test if N fertilization may enhance hydrocarbon decomposition, we investigated the effects of N application on hydrocarbon degradation in different-aged composts (1-, 2-, 3-, and 4-year-old composts, coded as 1Y, 2Y, 3Y, and 4Y composts, respectively) through a pot experiment planted with white spruce (Picea glauca [Moench] Voss) seedlings. The percentage degradation of total petroleum hydrocarbon (TPH, C11 to C40) in the composts without N fertilization was correlated to initial NH4+ concentrations (R = 0.99, P < 0.001). The percentage degradation of TPH was highest in the 3Y compost (41.1%) that had an initial level of 325.3 mg NH4+ -N kg(-1) and the lowest in the IY compost (9.3%) that had an initial level of 8.3 mg NH4+ -N kg(-1). The degradation of TPH was enhanced by Nfertilization in the 1Y (from 9.3 to 15.3%) and 4Y composts (from 14.3 to 22.6%) that had low initial NH4+ concentrations. Our results show that application of NH4+ -based fertilizers may enhance the degradation of TPH when initial NH4+ concentrations in the compost are low.

  2. The effect of higher ATP cost of contraction on the metabolic response to graded exercise in patients with chronic obstructive pulmonary disease.

    PubMed

    Layec, Gwenael; Haseler, Luke J; Richardson, Russell S

    2012-03-01

    To better understand the metabolic implications of a higher ATP cost of contraction in chronic obstructive pulmonary disease (COPD), we used (31)P-magnetic resonance spectroscopy ((31)P-MRS) to examine muscle energetics and pH in response to graded exercise. Specifically, in six patients and six well-matched healthy controls, we determined the intracellular threshold for pH (T(pH)) and inorganic phosphate-to-phosphocreatine ratio (T(Pi/PCr)) during progressive dynamic plantar flexion exercise with work rate expressed as both absolute and relative intensity. Patients with COPD displayed a lower peak power output (WRmax) compared with controls (controls 25 ± 4 W, COPD 15 ± 5 W, P = 0.01) while end-exercise pH (controls 6.79 ± 0.15, COPD 6.76 ± 0.21, P = 0.87) and PCr consumption (controls 82 ± 10%, COPD 70 ± 18%, P = 0.26) were similar between groups. Both T(pH) and T(Pi/PCr) occurred at a significantly lower absolute work rate in patients with COPD compared with controls (controls: 14.7 ± 2.4 W for T(pH) and 15.3 ± 2.4 W for T(Pi/PCr); COPD: 9.7 ± 4.5 W for T(pH) and 10.0 ± 4.6 W for T(Pi/PCr), P < 0.05), but these thresholds occurred at the same percentage of WRmax (controls: 63 ± 11% WRmax for T(pH) and 67 ± 18% WRmax for T(Pi/PCr); COPD: 59 ± 9% WRmax for T(pH) and 61 ± 12% WRmax for T(Pi/PCr), P > 0.05). Indexes of mitochondrial function, the PCr recovery time constant (controls 42 ± 7 s, COPD 45 ± 11 s, P = 0.66) and the PCr resynthesis rate (controls 105 ± 21%/min, COPD 91 ± 31%/min, P = 0.43) were similar between groups. In combination, these results reveal that when energy demand is normalized to WRmax, as a consequence of higher ATP cost of contraction, patients with COPD display the same metabolic pattern as healthy subjects, suggesting that skeletal muscle energy production is well preserved in these patients.

  3. Application of large volume injection GC-MS to analysis of organic compounds in the extracts and leachates of municipal solid waste incineration fly ash

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Korenkova, Eva; Matisova, Eva; Slobodnik, Jaroslav

    2006-07-01

    Organic solvent and water extracts of fly ash from a Milan (Italy) municipal solid waste incinerator (MSWI) were analyzed by large volume injection-gas chromatography-mass spectrometry (LVI-GC-MS) with programmable temperature vaporizer (PTV). Using injection volumes of 10-100 {mu}l, typically over a hundred compounds were detected in organic solvent extracts and ca. 35% of them could be tentatively identified from their electron impact ionization mass spectra. A protocol for the determination of the maximum amount of a potential environmental pollutant available for leaching (availability test) was developed for four selected target compounds: pentachlorobenzene (PeCB), hexachlorobenzene (HxCB), o-terphenyl (o-TPH) and m-terphenyl (m-TPH). Keymore » parameters, extraction time and liquid-to-solid ratio (L/S), were studied in more detail. Recoveries of PeCB, HxCB and o-TPH spiked into the fly ash samples at two concentration levels ranged from 38% to 53% for freshly spiked and from 14% to 40% for 40-day aged fly ash. Recoveries of m-TPH were 8% to 11% from freshly spiked and less than 3% from aged spiked fly ash. The native amounts in Milan MSWI fly ash, determined in an interlaboratory exercise using the developed protocol, were 31 ng/g PeCB, 34 ng/g HxCB, 72 ng/g o-TPH and 4.4 ng/g m-TPH. A separate methodology was developed for the determination of compounds extracted from fly ash by water (leaching test). Following 8-h sonication at L/S 20, the leached amounts of PeCB, HxCB and o-TPH were 1.1, 3.1 and 6.0 ng/g fly ash, respectively.« less

  4. Ex situ bioremediation of oil-contaminated soil.

    PubMed

    Lin, Ta-Chen; Pan, Po-Tsen; Cheng, Sheng-Shung

    2010-04-15

    An innovative bioprocess method, Systematic Environmental Molecular Bioremediation Technology (SEMBT) that combines bioaugmentation and biostimulation with a molecular monitoring microarray biochip, was developed as an integrated bioremediation technology to treat S- and T-series biopiles by using the landfarming operation and reseeding process to enhance the bioremediation efficiency. After 28 days of the bioremediation process, diesel oil (TPH(C10-C28)) and fuel oil (TPH(C10-C40)) were degraded up to approximately 70% and 63% respectively in the S-series biopiles. When the bioaugmentation and biostimulation were applied in the beginning of bioremediation, the microbial concentration increased from approximately 10(5) to 10(6) CFU/g dry soil along with the TPH biodegradation. Analysis of microbial diversity in the contaminated soils by microarray biochips revealed that Acinetobacter sp. and Pseudomonas aeruginosa were the predominant groups in indigenous consortia, while the augmented consortia were Gordonia alkanivorans and Rhodococcus erythropolis in both series of biopiles during bioremediation. Microbial respiration as influenced by the microbial activity reflected directly the active microbial population and indirectly the biodegradation of TPH. Field experimental results showed that the residual TPH concentration in the complex biopile was reduced to less than 500 mg TPH/kg dry soil. The above results demonstrated that the SEMBT technology is a feasible alternative to bioremediate the oil-contaminated soil. Crown Copyright 2009. Published by Elsevier B.V. All rights reserved.

  5. TPH detection in groundwater: Identification and elimination of positive interferences

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zemo, D.A.; Synowiec, K.A.

    1996-01-01

    Groundwater assessment programs frequently require total petroleum hydrocarbon (TPH) analyses (Methods 8015M and 418.1). TPH analyses are often unreliable indicators of water quality because these methods are not constituent-specific and are vulnerable to significant sources of positive interferences. These positive interferences include: (a) non-dissolved petroleum constituents; (b) soluble, non-petroleum hydrocarbons (e.g., biodegradation products); and (c) turbidity, commonly introduced into water samples during sample collection. In this paper, we show that the portion of a TPH concentration not directly the result of water-soluble petroleum constituents can be attributed solely to these positive interferences. To demonstrate the impact of these interferences, wemore » conducted a field experiment at a site affected by degraded crude oil. Although TPH was consistently detected in groundwater samples, BTEX was not detected. PNAs were not detected, except for very low concentrations of fluorene (<5 ug/1). Filtering and silica gel cleanup steps were added to sampling and analyses to remove particulates and biogenic by-products. Results showed that filtering lowered the Method 8015M concentrations and reduced the Method 418.1 concentrations to non-detectable. Silica gel cleanup reduced the Method 8015M concentrations to non-detectable. We conclude from this study that the TPH results from groundwater samples are artifacts of positive interferences caused by both particulates and biogenic materials and do not represent dissolved-phase petroleum constituents.« less

  6. TPH detection in groundwater: Identification and elimination of positive interferences

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zemo, D.A.; Synowiec, K.A.

    1996-12-31

    Groundwater assessment programs frequently require total petroleum hydrocarbon (TPH) analyses (Methods 8015M and 418.1). TPH analyses are often unreliable indicators of water quality because these methods are not constituent-specific and are vulnerable to significant sources of positive interferences. These positive interferences include: (a) non-dissolved petroleum constituents; (b) soluble, non-petroleum hydrocarbons (e.g., biodegradation products); and (c) turbidity, commonly introduced into water samples during sample collection. In this paper, we show that the portion of a TPH concentration not directly the result of water-soluble petroleum constituents can be attributed solely to these positive interferences. To demonstrate the impact of these interferences, wemore » conducted a field experiment at a site affected by degraded crude oil. Although TPH was consistently detected in groundwater samples, BTEX was not detected. PNAs were not detected, except for very low concentrations of fluorene (<5 ug/1). Filtering and silica gel cleanup steps were added to sampling and analyses to remove particulates and biogenic by-products. Results showed that filtering lowered the Method 8015M concentrations and reduced the Method 418.1 concentrations to non-detectable. Silica gel cleanup reduced the Method 8015M concentrations to non-detectable. We conclude from this study that the TPH results from groundwater samples are artifacts of positive interferences caused by both particulates and biogenic materials and do not represent dissolved-phase petroleum constituents.« less

  7. Aqueous solubility calculation for petroleum mixtures in soil using comprehensive two-dimensional gas chromatography analysis data.

    PubMed

    Mao, Debin; Lookman, Richard; Van De Weghe, Hendrik; Vanermen, Guido; De Brucker, Nicole; Diels, Ludo

    2009-04-03

    An assessment of aqueous solubility (leaching potential) of soil contaminations with petroleum hydrocarbons (TPH) is important in the context of the evaluation of (migration) risks and soil/groundwater remediation. Field measurements using monitoring wells often overestimate real TPH concentrations in case of presence of pure oil in the screened interval of the well. This paper presents a method to calculate TPH equilibrium concentrations in groundwater using soil analysis by high-performance liquid chromatography followed by comprehensive two-dimensional gas chromatography (HPLC-GCXGC). The oil in the soil sample is divided into 79 defined hydrocarbon fractions on two GCXGC color plots. To each of these fractions a representative water solubility is assigned. Overall equilibrium water solubility of the non-aqueous phase liquid (NAPL) present in the sample and the water phase's chemical composition (in terms of the 79 fractions defined) are then calculated using Raoult's law. The calculation method was validated using soil spiked with 13 different TPH mixtures and 1 field-contaminated soil. Measured water solubilities using a column recirculation equilibration experiment agreed well to calculated equilibrium concentrations and water phase TPH composition.

  8. Leucanthemum vulgare Lam. crude oil phytoremediation.

    PubMed

    Noori, Azam; Maivan, Hassan Zare; Alaie, Ebrahim; Newman, Lee A

    2015-06-29

    Sites with crude oil pollution have been successfully treated using phytoremediation, but expanding the range of plants that can be used and understanding how exposure impacts the plants are two areas of study that are important to continue. Leucanthemum vulgare has been shown to grow well under a variety of stressful conditions. To examine L. vulgare's ability to both survive crude oil exposure and to reduce crude oil concentrations in soil, plants were placed in soil containing 0, 2.5%, 5%, 7.5% or 10% w/w crude oil. Total Petroleum Hydrocarbons (TPH) concentration, peroxidase and catalase activity, proline and phenol content in roots and leaves were determined at the start of planting and every 2 months for six months. L. vulgare roots were successfully colonized with mycorrhizae under all conditions. Results showed positive correlation between antioxidant compound concentration and crude oil contamination. Also, a significant reduction occurred in TPH content of soil over time in planted pots as compared to controls. The lowest TPH content was recorded after 6 months under all treatments. Results showed L. vulgare could survive crude oil exposure and enhance reducing of crude oil from soil.

  9. C1473G polymorphism in mouse tph2 gene is linked to tryptophan hydroxylase-2 activity in the brain, intermale aggression, and depressive-like behavior in the forced swim test.

    PubMed

    Osipova, Daria V; Kulikov, Alexander V; Popova, Nina K

    2009-04-01

    Tryptophan hydroxylase-2 (TPH2) is the rate-limiting enzyme of brain serotonin synthesis. The C1473G polymorphism in the mouse tryptophan hydroxylase-2 gene affects the enzyme's activity. In the present study, we investigated the linkage between the C1473G polymorphism, enzyme activity in the brain, and behavior in the forced swim, intermale aggression, and open field tests using mice of the C57BL/6 (C/C) and CC57BR/Mv (G/G) strains and the B6-1473C (C/C) and B6-1473G (G/G) lines created by three successive backcrossings on C57BL/6. Mice of the CC57BR/Mv strain had decreased brain enzyme activity, aggression intensity, and immobility in the forced swim test, but increased locomotor activity and time spent in the central part of the open field arena compared with animals of the C57BL/6 strain. Mice of the B6-1473G line homozygous for the 1473G allele had lower TPH2 activity in the brain, aggression intensity, and immobility time in the forced swim test compared with animals of the B6-1473C line homozygous for the 1473C allele. No differences were found between the B6-1473G and B6-1473C mice in locomotor activity and time spent in the central part of the arena in the open field test. Thus, the C1473G polymorphism is involved in the determination of TPH2 activity and is linked to aggression intensity and forced-swim immobility in mice. At the same time, the polymorphism does not affect locomotion and anxiety-related behavior in the open field test. The B6-1473C and B6-1473G mice represent a valuable experimental model for investigating molecular mechanisms of serotonin-related behavior.

  10. Chronic low dose ovine corticotropin releasing factor or urocortin II into the rostral dorsal raphe alters exploratory behavior and serotonergic gene expression in specific subregions of the dorsal raphe

    PubMed Central

    Clark, Michael S.; McDevitt, Ross A.; Hoplight, Blair J.; Neumaier, John F.

    2007-01-01

    Corticotropin releasing factor (CRF) family peptides play key roles in integrating neural responses to stress. Both major CRF receptors have been pharmacologically identified in the dorsal raphe nucleus (DRN), a stress sensitive and internally heterogeneous nucleus supplying many forebrain regions with serotonergic input. Despite the involvement of chronic stress and serotonergic dysfunction in human mood and anxiety disorders, little is known about the effects of chronic CRF receptor activation on the DRN. We infused ovine CRF (1ng/hr), urocortin II (UCNII, 1ng/hr), or vehicle alone into rat DRN over 6 days. During infusion, animals were allowed to freely explore an open field for 15 minutes on each of two days, with the addition of a novel object on the second day. Following behavioral testing, 5-HT1A, 5-HT1B, serotonin transporter (SERT), and tryptophan hydroxylase-2 (Tph2) expression were examined through the DRN by in situ hybridization. Ovine CRF infusion resulted in significantly decreased novel object touches, climbs, as well as increased latency to first novel object contact. UCNII had a similar but less dramatic effect, decreasing only climbing behavior. Both ovine CRF and UCNII blunted the decrease in corner time expected on re-exposure to the open field. Both peptides also produced regionally specific changes in gene expression: 5-HT1A expression was increased 30% in the mid-rostral ventromedial DRN, while SERT was decreased by 30% in the mid-caudal shell dorsomedial DRN. There also appeared to be a shift in the relative level of Tph2 expression between the ventromedial and core dorsomedial DRN at the mid-rostral level. Changes in 5-HT1A, SERT, and relative Tph2 mRNA abundance were correlated with novel object exploration. These findings suggest chronic intra-DRN administration of CRF agonists decreases exploratory behavior, while producing subregionally limited changes in serotonergic gene expression. These studies may be relevant to mechanisms underlying behavioral changes after chronic stress. PMID:17467184

  11. Assessment of organic pollutants in the offshore sediments of Dubai, United Arab Emirates

    NASA Astrophysics Data System (ADS)

    Darwish, H. A. Al; El-Gawad, E. A. Abd; Mohammed, F. H.; Lotfy, M. M.

    2005-08-01

    Fifteen stations (st) were selected along Dubai coastal region to delineate the distribution and the source of total petroleum hydrocarbon (TPH), total organic carbon (TOC), total Kjeldhal nitrogen (TKN), polycyclic aromatic hydrocarbon (PAHs) and polychlorinated biphenyls. The concentrations of TPH fluctuated between 2 μg g -1 and 48018 μg g -1 and the values of TOC were in the range of 0.16-5.9 wt%, while TPAHs ranged from 0.09 μg g -1 to 161.72 μg g -1. On the other hand, TPCBs showed values between 0.8 μg kg-1 and 93.3 μg kg-1 and TKN values varied from 218 μg g-1 to 2457 μg g -1. Distribution of oil and organic compounds in Dubai sediments are safe compared with previous studies except for limited areas at the northeastern offshore. These readings are probably due to: (1) presence of commercial or industrial ports, dry docks and fishing harbours and (2) population centers mainly concentrated at the northern part of the study area. Results indicate that TOC can be used as indicator of oil pollution only in heavily oiled sediments. The highest values of TOC, TPH, TPAHs and TPCBs corresponded to the stations covered with fine sand, due to adsorption properties and larger surface area. The evaporation of low boiling point compounds from surface layers led to enrichment of sediments with the thick residual. Al-Hamriya St 3 exhibited the highest values of TPH, TOC, TPAHs and TPCBs and the second highest value of TKN.

  12. INNOVATIVE TECHNOLOGY VERIFICATION REPORT " ...

    EPA Pesticide Factsheets

    The EnSys Petro Test System developed by Strategic Diagnostics Inc. (SDI), was demonstrated under the U.S. Environmental Protection Agency Superfund Innovative Technology Evaluation Program in June 2000 at the Navy Base Ventura County site in Port Hueneme, California. The purpose of the demonstration was to collect reliable performance and cost data for the EnSys Petro Test System and six other field measurement devices for total petroleum hydrocarbons (TPH) in soil. In addition to assessing ease of device operation, the key objectives of the demonstration included determining the (1) method detection limit, (2) accuracy and precision, (3) effects of interferents and soil moisture content on TPH measurement, (4) sample throughput, and (5) TPH measurement costs for each device. The demonstration involved analysis of both performance evaluation samples and environmental samples collected in four areas contaminated with gasoline, diesel, or other petroleum products. The performance and cost results for a given field measurement device were compared to those for an off-site laboratory reference method,

  13. INNOVATIVE TECHNOLOGY VERIFICATION REPORT " ...

    EPA Pesticide Factsheets

    The Synchronous Scanning Luminoscope (Luminoscope) developed by the Oak Ridge National Laboratory in collaboration with Environmental Systems Corporation (ESC) was demonstrated under the U.S. Environmental Protection Agency Superfund Innovative Technology Evaluation Program in June 2000 at the Navy Base Ventura County site in Port Hueneme, California. The purpose of the demonstration was to collect reliable performance and cost data for the Luminoscope and six other field measurement devices for total petroleum hydrocarbons (TPH) in soil. In addition to assessing ease of device operation, the key objectives of the demonstration included determining the (1) method detection limit, (2) accuracy and precision, (3) effects of interferents and soil moisture content on TPH measurement, (4) sample throughput, and (5) TPH measurement costs for each device. The demonstration involved analysis of both performance evaluation samples and environmental samples collected in five areas contaminated with gasoline, diesel, lubricating oil, or other petroleum products. The performance and cost results for a given field measurement device were compared to those for an off-site laboratory reference method,

  14. Assessment of soil-gas contamination at three former fuel-dispensing sites, Fort Gordon, Georgia, 2010—2011

    USGS Publications Warehouse

    Caldwell, Andral W.; Falls, W. Fred; Guimaraes, Wladmir B.; Ratliff, W. Hagan; Wellborn, John B.; Landmeyer, James E.

    2012-01-01

    Soil gas was assessed for contaminants at three former fuel-dispensing sites at Fort Gordon, Georgia, from October 2010 to September 2011. The assessment included delineation of organic contaminants using soil-gas samplers collected from the former fuel-dispensing sites at 8th Street, Chamberlain Avenue, and 12th Street. This assessment was conducted to provide environmental contamination data to Fort Gordon personnel pursuant to requirements for the Resource Conservation and Recovery Act Part B Hazardous Waste Permit process. Soil-gas samplers installed and retrieved during June and August 2011 at the 8th Street site had detections above the method detection level (MDL) for the mass of total petroleum hydrocarbons (TPH), benzene, toluene, ortho-xylene, undecane, tridecane, pentadecane, and chloroform. Total petroleum hydrocarbons soil-gas mass exceeded the MDL of 0.02 microgram in 54 of the 55 soil-gas samplers. The highest detection of TPH soil-gas mass was 146.10 micrograms, located in the central part of the site. Benzene mass exceeded the MDL of 0.01 microgram in 23 soil-gas samplers, whereas toluene was detected in only 10 soil-gas samplers. Ortho-xylene was detected above the MDL in only one soil-gas sampler. The highest soil-gas mass detected for undecane, tridecane, and pentadecane was located in the northeastern corner of the 8th Street site. Chloroform mass greater than the MDL of 0.01 microgram was detected in less than one-third of the soil-gas samplers. Soil-gas masses above the MDL were identified for TPH, gasoline-related compounds, diesel-range alkanes, trimethylbenzenes, naphthalene, 2-methyl-napthalene, octane, and tetrachloroethylene for the July 2011 soil-gas survey at the Chamberlain Avenue site. All 30 of the soil-gas samplers contained TPH mass above the MDL. The highest detection of TPH mass, 426.36 micrograms, was for a soil-gas sampler located near the northern boundary of the site. Gasoline-related compounds and diesel-range alkanes were detected in multiple soil-gas samplers, and the highest detections of these compounds were located near the central part of the site near existing, nonoperational, fuel-dispensing pumps. Trimethylbenzenes were detected in less than half of the soil-gas samplers. Naphthalene soil-gas mass was detected above the MDL in 10 soil-gas samplers, whereas 2-methyl-napthalene was detected above the MDL in half of the soil-gas samplers. Octane mass was detected above the MDL in one soil-gas sampler located near the central part of the site. Tetrachloroethylene soil-gas mass was detected above the MDL in more than half of the soil-gas samplers, and the highest tetrachloroethylene soil-gas mass of 0.90 microgram was located in the northeastern part of the site. Soil-gas samplers collected at the 12th Street site during July 2011 contained soil-gas mass above the MDL for TPH, toluene, undecane, tridecane, and pentadecane (diesel-range alkanes), trichloroethylene, 1,4-dichlorobenzene, chloroform, and 1,2,4-trimethylbenzene. The highest detected TPH mass was 24.37 micrograms in a soil-gas sampler located in the northern part of the site. The highest detection of toluene soil-gas mass was from a soil-gas sampler located near the southern boundary of the site. The diesel-range alkanes were detected above the MDL in five soil-gas samplers; the highest detection of soil-gas diesel mass, 0.65 microgram, was located in the southern part of the site. Trichloroethylene and 1,4-dichlorobenzene were detected above the MDL in the northern part of the site in one soil-gas sampler that also had one of the highest detections of TPH. Chloroform was detected above the MDL in three soil-gas samplers, whereas 1,2,4-trimethylbenzene soil-gas mass was detected above the MDL in two soil-gas samplers.

  15. Microbial activity and community composition during bioremediation of diesel-oil-contaminated soil: effects of hydrocarbon concentration, fertilizers, and incubation time.

    PubMed

    Margesin, Rosa; Hämmerle, Marion; Tscherko, Dagmar

    2007-02-01

    We investigated the influence of three factors-diesel oil concentration [2500, 5000, 10,000, 20,000 mg total petroleum hydrocarbons (TPH) kg(-1) soil], biostimulation (unfertilized, inorganic fertilization with NPK nutrients, or oleophilic fertilization with Inipol EAP22), and incubation time-on hydrocarbon removal, enzyme activity (lipase), and microbial community structure [phospholipid fatty acids (PLFA)] in a laboratory soil bioremediation treatment. Fertilization enhanced TPH removal and lipase activity significantly (P < or = 0.001). The higher the initial contamination, the more marked was the effect of fertilization. Differences between the two fertilizers were not significant (P > 0.05). Microbial communities, as assessed by PLFA patterns, were primarily influenced by the TPH content, followed by fertilization, and the interaction of these two factors, whereas incubation time was of minor importance. This was demonstrated by three-factorial analysis of variance and multidimensional scaling analysis. Low TPH content had no significant effect on soil microbial community, independent of the treatment. High TPH content generally resulted in increased PLFA concentrations, whereby a significant increase in microbial biomass with time was only observed with inorganic fertilization, whereas oleophilic fertilization (Inipol EAP22) tended to inhibit microbial activity and to reduce PLFA contents with time. Among bacteria, PLFA indicative of the Gram-negative population were significantly (P < or = 0.05) increased in soil samples containing high amounts of diesel oil and fertilized with NPK after 21-38 days of incubation at 20 degrees C. The Gram-positive population was not significantly influenced by TPH content or biostimulation treatment.

  16. Phase transition for the system of finite volume in the ϕ4 theory in the Tsallis nonextensive statistics

    NASA Astrophysics Data System (ADS)

    Ishihara, Masamichi

    2018-04-01

    We studied the effects of nonextensivity on the phase transition for the system of finite volume V in the ϕ4 theory in the Tsallis nonextensive statistics of entropic parameter q and temperature T, when the deviation from the Boltzmann-Gibbs (BG) statistics, |q ‑ 1|, is small. We calculated the condensate and the effective mass to the order q ‑ 1 with the normalized q-expectation value under the free particle approximation with zero bare mass. The following facts were found. The condensate Φ divided by v, Φ/v, at q (v is the value of the condensate at T = 0) is smaller than that at q‧ for q > q‧ as a function of Tph/v which is the physical temperature Tph divided by v. The physical temperature Tph is related to the variation of the Tsallis entropy and the variation of the internal energies, and Tph at q = 1 coincides with T. The effective mass decreases, reaches minimum, and increases after that, as Tph increases. The effective mass at q > 1 is lighter than the effective mass at q = 1 at low physical temperature and heavier than the effective mass at q = 1 at high physical temperature. The effects of the nonextensivity on the physical quantity as a function of Tph become strong as |q ‑ 1| increases. The results indicate the significance of the definition of the expectation value, the definition of the physical temperature, and the constraints for the density operator, when the terms including the volume of the system are not negligible.

  17. Stress-induced activation of the brainstem Bcl-xL gene expression in rats treated with fluoxetine: correlations with serotonin metabolism and depressive-like behavior.

    PubMed

    Shishkina, Galina T; Kalinina, Tatyana S; Berezova, Inna V; Dygalo, Nikolay N

    2012-01-01

    Mechanisms underlying stress-induced depression and antidepressant drug action were shown to involve alterations in serotonergic (5-HT) neurotransmission and expression of genes coding for proteins associated with neurotrophic signaling pathways and cell-survival in the hippocampus and cortex. Expression of these genes in the brainstem containing 5-HT neurons may also be related to vulnerability or resilience to stress-related psychopathology. Here we investigated 5-HT markers and expression of genes for Brain-Derived Neurotrophic Factor (BDNF) and apoptotic proteins in the brainstem in relation to swim stress-induced behavioral despair. We found that anti-apoptotic Bcl-xL gene is sensitive to stress during the course of fluoxetine administration. Responsiveness of this gene to stress appeared concomitantly with an antidepressant-like effect of fluoxetine in the forced swim test. Bcl-xL transcript levels showed negative correlations with duration of immobility in the test and 5-HT turnover in the brainstem. In contrast, BDNF and pro-apoptotic protein Bax mRNA levels were unchanged by either fluoxetine or stress, suggesting specificity of Bcl-xL gene responses to these treatments. We also found that the levels of mRNAs for tryptophan hydroxylase-2 (TPH2) and 5-HT transporter (5-HTT) were significantly down-regulated following prolonged treatment with fluoxetine, but were not affected by stress. Unlike TPH2 and 5-HTT, 5-HT1A receptor mRNA levels were not altered by fluoxetine but significantly increased in response to swim stress. These data show that long-term fluoxetine treatment leads to changes in 5-HT and Bcl-xL responses to stress associated with antidepressant-like effects of the drug. This article is part of a Special Issue entitled 'Anxiety and Depression'. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. The C1473G polymorphism in the Tryptophan hydroxylase-2 gene: involvement in ethanol-related behavior in mice.

    PubMed

    Bazovkina, Darya V; Lichman, Daria V; Kulikov, Alexander V

    2015-03-04

    Tryptophan hydroxylase-2 (Tph2) is the rate limiting enzyme of serotonin synthesis in the brain. The functional (C1473G) polymorphism in the mouse Tph2 gene affecting the enzymatic activity was suspected to be involved in behavioral actions of ethanol (EtOH). Congenic B6-1473C (C/C) and B6-1473G (G/G) lines bred from C57BL/6 mice were not different in EtOH-induced sleep time and hypothermia. B6-1473C mice displayed increased EtOH preference on the second and third days compared to that of the first day, but no differences in this parameter was found across genotypes. Both lines demonstrated the same responsiveness to hypothermic and hypnotic effect of acute EtOH treatment after repeated alcohol exposure. However, acute EtOH administration led to reduction of locomotor activity in B6-1473C, but not in B6-1473G animals and to increase of time spent in the center of open-field arena in B6-1473G, but not in B6-1473C mice. Thus, the present study indicates the involvement of C1473G polymorphism in mTph2 gene in the regulation of EtOH-induced effects on locomotor activity and anxiety-like behavior in mice. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  19. The Concept and Economics of RDF-3 (Refuse Derived Fuel) Utilization in a Navy Size Pulverized Coal Boiler.

    DTIC Science & Technology

    1983-05-01

    ELE ENTM. PRO ECT. TASK AREA 4 WORK LIN IT kuldElS WASTE ENERGY TECHNOLOGY CORPORATION Y0817-006-01-211 Bedford, MA 01730 I P CONTROLLING OFFICE NAME...Louis Miller-Hoft. 150 Augers 1 50 Missouri Live bottom, Tons TPH rectangular Ames Atlas. 500 Sweep bucket 4 14 Iowa Tons and drag con - TPH veyor Each...Monroe County a. Trailers 17 Hyd. Rams NewYork Tons Each b. Atlas 450 Sweep bucket 8 6 Tons and drag con - TPH veyor Each . Milwaukee Atlas 900 Sweep

  20. Risk-Based Evaluation of Total Petroleum Hydrocarbons in Vapor Intrusion Studies

    PubMed Central

    Brewer, Roger; Nagashima, Josh; Kelley, Michael; Heskett, Marvin; Rigby, Mark

    2013-01-01

    This paper presents a quantitative method for the risk-based evaluation of Total Petroleum Hydrocarbons (TPH) in vapor intrusion investigations. Vapors from petroleum fuels are characterized by a complex mixture of aliphatic and, to a lesser extent, aromatic compounds. These compounds can be measured and described in terms of TPH carbon ranges. Toxicity factors published by USEPA and other parties allow development of risk-based, air and soil vapor screening levels for each carbon range in the same manner as done for individual compounds such as benzene. The relative, carbon range makeup of petroleum vapors can be used to develop weighted, site-specific or generic screening levels for TPH. At some critical ratio of TPH to a targeted, individual compound, the overwhelming proportion of TPH will drive vapor intrusion risk over the individual compound. This is particularly true for vapors associated with diesel and other middle distillate fuels, but can also be the case for low-benzene gasolines or even for high-benzene gasolines if an adequately conservative, target risk is not applied to individually targeted chemicals. This necessitates a re-evaluation of the reliance on benzene and other individual compounds as a stand-alone tool to evaluate vapor intrusion risk associated with petroleum. PMID:23765191

  1. Chronic central serotonin depletion attenuates ventilation and body temperature in young but not adult Tph2 knockout rats

    PubMed Central

    Kaplan, Kara; Echert, Ashley E.; Massat, Ben; Puissant, Madeleine M.; Palygin, Oleg; Geurts, Aron M.

    2016-01-01

    Genetic deletion of brain serotonin (5-HT) neurons in mice leads to ventilatory deficits and increased neonatal mortality during development. However, it is unclear if the loss of the 5-HT neurons or the loss of the neurochemical 5-HT led to the observed physiologic deficits. Herein, we generated a mutant rat model with constitutive central nervous system (CNS) 5-HT depletion by mutation of the tryptophan hydroxylase 2 (Tph2) gene in dark agouti (DATph2−/−) rats. DATph2−/− rats lacked TPH immunoreactivity and brain 5-HT but retain dopa decarboxylase-expressing raphe neurons. Mutant rats were also smaller, had relatively high mortality (∼50%), and compared with controls had reduced room air ventilation and body temperatures at specific postnatal ages. In adult rats, breathing at rest and hypoxic and hypercapnic chemoreflexes were unaltered in adult male and female DATph2−/− rats. Body temperature was also maintained in adult DATph2−/− rats exposed to 4°C, indicating unaltered ventilatory and/or thermoregulatory control mechanisms. Finally, DATph2−/− rats treated with the 5-HT precursor 5-hydroxytryptophan (5-HTP) partially restored CNS 5-HT and showed increased ventilation (P < 0.05) at a developmental age when it was otherwise attenuated in the mutants. We conclude that constitutive CNS production of 5-HT is critically important to fundamental homeostatic control systems for breathing and temperature during postnatal development in the rat. PMID:26869713

  2. Assessment of Hyporheic Zone, Flood-Plain, Soil-Gas, Soil, and Surface-Water Contamination at the McCoys Creek Chemical Training Area, Fort Gordon, Georgia, 2009-2010

    USGS Publications Warehouse

    Guimaraes, Wladmir B.; Falls, W. Fred; Caldwell, Andral W.; Ratliff, W. Hagan; Wellborn, John B.; Landmeyer, James E.

    2011-01-01

    The U.S. Geological Survey, in cooperation with the U.S. Department of the Army Environmental and Natural Resources Management Office of the U.S. Army Signal Center and Fort Gordon, Georgia, assessed the hyporheic zone, flood plain, soil gas, soil, and surface water for contaminants at the McCoys Creek Chemical Training Area (MCTA) at Fort Gordon, from October 2009 to September 2010. The assessment included the detection of organic contaminants in the hyporheic zone, flood plain, soil gas, and surface water. In addition, the organic contaminant assessment included the analysis of organic compounds classified as explosives and chemical agents in selected areas. Inorganic contaminants were assessed in soil and surface-water samples. The assessment was conducted to provide environmental contamination data to the U.S. Army at Fort Gordon pursuant to requirements of the Resource Conservation and Recovery Act Part B Hazardous Waste Permit process. Ten passive samplers were deployed in the hyporheic zone and flood plain, and total petroleum hydrocarbons (TPH) and octane were detected above the method detection level in every sampler. Other organic compounds detected above the method detection level in the hyporheic zone and flood-plain samplers were trichloroethylene, and cis- and trans- 1, 2-dichloroethylene. One trip blank detected TPH below the method detection level but above the nondetection level. The concentrations of TPH in the samplers were many times greater than the concentrations detected in the blank; therefore, all other TPH concentrations detected are considered to represent environmental conditions. Seventy-one soil-gas samplers were deployed in a grid pattern across the MCTA. Three trip blanks and three method blanks were used and not deployed, and TPH was detected above the method detection level in two trip blanks and one method blank. Detection of TPH was observed at all 71 samplers, but because TPH was detected in the trip and method blanks, TPH was censored and, therefore, only 7 of the 71 samplers were reported as detecting TPH. In addition, benzene, toluene, ethylbenzene, and total xylene were detected above the method detection level in 22 samplers. Other compounds detected above the method detection level included naphthalene, octane, undecane, tridecane, 1,2,4-trimethylbenzene, trichloroethylene, perchloroethylene, chloroform, and 1,4-dichlorobenzene. Subsequent to the soil-gas survey, five locations with elevated contaminant mass were selected and a passive sampler was deployed at those locations to detect the presence of organic compounds classified as explosives or chemical agents. No explosives or chemical agents were detected above the method detection level, but some compounds were detected below the method detection level but above the nondetection level. Dimethyl disulfide, benzothiazole, chloroacetophenones, and para-chlorophenyl methyl sulfide were all detected below the method detection level but above the nondetection level. The compounds 2,4-dinitrotoluene, and para-chlorophenyl methyl sulfone were detected in samplers but also were detected in trip blanks and are not considered as present in the MCTA. The same five locations that were selected for sampling of explosives and chemical agents were selected for soil sampling. Metal concentrations in composite soil samples collected at five locations from land surface to a depth of 6 inches did not exceed the U.S. Environmental Protection Agency Regional Screening Levels for Industrial Soil. Concentrations in some compounds were higher than the South Carolina Department of Health and Environmental Control background levels for nearby South Carolina, including aluminum, arsenic, barium, beryllium, chromium, copper, iron, lead, manganese, nickel, and potassium. A surface-water sample was collected from McCoys Creek and analyzed for volatile organic compounds, semivolatile organic compounds, and inorganic compounds (metals). No volatile organic compounds and (or) semivolatile organic compounds were detected at levels above the maximum contaminant level of the U.S. Environmental Protection Agency (USEPA) National Primary Drinking Water Standard, and no inorganic compounds exceeded the maximum contaminant level of the USEPA National Primary Drinking Water Standard or the Georgia In-Stream Water-Quality Standard. Iron was the only inorganic compound detected in the surface-water sample (578 micrograms per liter) that exceeded the USEPA National Secondary Drinking Water Standard of 300 micrograms per liter.

  3. Field Application of Modified In Situ Soil Flushing in Combination with Air Sparging at a Military Site Polluted by Diesel and Gasoline in Korea

    PubMed Central

    Lee, Hwan; Lee, Yoonjin; Kim, Jaeyoung; Kim, Choltae

    2014-01-01

    In this study the full-scale operation of soil flushing with air sparging to improve the removal efficiency of petroleum at depths of less than 7 m at a military site in Korea was evaluated. The target area was polluted by multiple gasoline and diesel fuel sources. The soil was composed of heterogeneous layers of granules, sand, silt and clay. The operation factors were systemically assessed using a column test and a pilot study before running the full-scale process at the site. The discharged TPH and BTEX (benzene, toluene, ethylbenzene, and xylenes) concentrations in the water were highest at 20 min and at a rate of 350 L/min, which was selected as the volume of air for the full-scale operation in the pilot air sparging test. The surfactant-aid condition was 1.4 times more efficient than the non-surfactant condition in the serial operations of modified soil flushing followed by air sparging. The hydraulic conductivity (3.13 × 10−3 cm/s) increased 4.7 times after the serial operation of both processes relative to the existing condition (6.61 × 10−4 cm/s). The removal efficiencies of TPH were 52.8%, 57.4%, and 61.8% for the soil layers at 6 to 7, 7 to 8 and 8 to 9 m, respectively. Therefore, the TPH removal was improved at depth of less than 7 m by using this modified remediation system. The removal efficiencies for the areas with TPH and BTEX concentrations of more than 500 and 80 mg/kg, were 55.5% and 92.9%, respectively, at a pore volume of 2.9. The total TPH and BTEX mass removed during the full-scale operation was 5109 and 752 kg, respectively. PMID:25166919

  4. The role of the serotonergic system in suicidal behavior

    PubMed Central

    Sadkowski, Marta; Dennis, Brittany; Clayden, Robert C; ElSheikh, Wala; Rangarajan, Sumathy; DeJesus, Jane; Samaan, Zainab

    2013-01-01

    Serotonin is a widely investigated neurotransmitter in several psychopathologies, including suicidal behavior (SB); however, its role extends to several physiological functions involving the nervous system, as well as the gastrointestinal and cardiovascular systems. This review summarizes recent research into ten serotonergic genes related to SB. These genes – TPH1, TPH2, SLC6A4, SLC18A2, HTR1A, HTR1B, HTR2A, DDC, MAOA, and MAOB – encode proteins that are vital to serotonergic function: tryptophan hydroxylase; the serotonin transporter 5-HTT; the vesicular transporter VMAT2; the HTR1A, HTR1B, and HTR2A receptors; the L-amino acid decarboxylase; and the monoamine oxidases. This review employed a systematic search strategy and a narrative research methodology to disseminate the current literature investigating the link between SB and serotonin. PMID:24235834

  5. Use of two-surfactants mixtures to attain specific HLB values for assisted TPH-diesel biodegradation.

    PubMed

    Torres, Luis G; Rojas, Neftalí; Iturbe, Rosario

    2004-01-01

    In a surfactant assisted biodegradation process, the choice of surfactant(s) is of crucial importance. The question is: does the type of surfactant (i.e. chemical family) affect the biodegradation process at fixed hidrophillic-lypofillic balance (HLB) values? Microcosm assessments were developed using contaminated soil, with around of 5000 mg/kg of hydrocarbons as TPH-diesel. Mixtures of three nonionic surfactants were employed to get a wide range of specific HLB values. Tween20 and Span20 were mixed in the appropriate proportions to get HLB values between 8.6 and 16.7. Tween/Span60 mixtures reached HLB values between 4.7 and 14.9. Finally, Tween/Span80 combinations yielded HLB values between 4.3 and 15. TPH-diesel biodegradation was measured at the beginning, and after 8 weeks, as well as the FCU/gr(soil), as a measure of microorganisms' development during the biodegradation period. A second aim of this work was to assess the use of guar gum as a biodegradation enhancer instead of synthetic products. The conclusions of this work are that surfactant chemical family, and not only the HLB value clearly affects the assisted biodegradation rate. Surfactant's synergism was clearly observed. Regarding the use of guar gum, no biodegradation enhancement was observed for the three assessed concentrations, i.e., 2, 20, and 200 mg/kg, respectively. On the contrary, TPH-diesel removal was lower as the gum concentration increased. It is quite possible that guar gum was used as a microbial substrate.

  6. Serotonin Control of Thermotaxis Memory Behavior in Nematode Caenorhabditis elegans

    PubMed Central

    Guo, Yuling; Wang, Daoyong; Li, Chaojun; Wang, Dayong

    2013-01-01

    Caenorhabditis elegans is as an ideal model system for the study of mechanisms underlying learning and memory. In the present study, we employed C. elegans assay system of thermotaxis memory to investigate the possible role of serotonin neurotransmitter in memory control. Our data showed that both mutations of tph-1, bas-1, and cat-4 genes, required for serotonin synthesis, and mutations of mod-5 gene, encoding a serotonin reuptake transporter, resulted in deficits in thermotaxis memory behavior. Exogenous treatment with serotonin effectively recovered the deficits in thermotaxis memory of tph-1 and bas-1 mutants to the level of wild-type N2. Neuron-specific activity assay of TPH-1 suggests that serotonin might regulate the thermotaxis memory behavior by release from the ADF sensory neurons. Ablation of ADF sensory neurons by expressing a cell-death activator gene egl-1 decreased the thermotaxis memory, whereas activation of ADF neurons by expression of a constitutively active protein kinase C homologue (pkc-1(gf)) increased the thermotaxis memory and rescued the deficits in thermotaxis memory in tph-1 mutants. Moreover, serotonin released from the ADF sensory neurons might act through the G-protein-coupled serotonin receptors of SER-4 and SER-7 to regulate the thermotaxis memory behavior. Genetic analysis implies that serotonin might further target the insulin signaling pathway to regulate the thermotaxis memory behavior. Thus, our results suggest the possible crucial role of serotonin and ADF sensory neurons in thermotaxis memory control in C. elegans. PMID:24223727

  7. 76 FR 48931 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-09

    ... or TPH organization is appropriate and reasonable to ensure it is in congruence with that level of... Broker fee to each Trading Permit Holder or TPH organizations is reasonable to ensure it is in congruence...

  8. Photoenhanced toxicity of weathered oil to Mysidopsis bahia

    USGS Publications Warehouse

    Cleveland, L.; Little, E.E.; Calfee, R.D.; Barron, M.G.

    2000-01-01

    The toxicity of a water-accommodated fraction (WAF) prepared from weathered oil was assessed in a 7-day static renewal test with Mysidopsis bahia. Weathered oil was collected from the 5 x monitoring well at the Guadalupe oil field. Solar ultraviolet and visible light intensities were measured in various habitats in the vicinity of the weathered oil sample collection site, and the resultant measurements were used to produce laboratory light treatments that were representative of the on-site quality and intensity of natural solar radiation. Each of five WAF dilutions and a control without WAF was tested under three different simulated solar radiation intensities. During the test, survival and growth of the mysids, irradiance, and total petroleum hydrocarbon (TPH) concentrations in the test treatments were measured. Significant increases (P ??? 0.05) in mortality occurred among mysids exposed to 0.57 and 1.30 mg TPH/l and the effects were potentiated as irradiance increased. Seven-day LC50 (0.92-0.42 mg TPH/l) and LC20 (0.58-0.15 mg TPH/l) values decreased as the simulated solar irradiance increased. Calculated EC20 and EC50 values for mysid growth indicate that surviving mysids exposed to 0.1-1.0 mg TPH/l would incur significant reductions (P ??? 0.05) in productivity (biomass). Results of the present study indicate that effects elicited through the interaction of WAF of weathered oil and solar radiation will substantially increase the toxicity of weathered oil. Further, the photomediated effects of petroleum compounds measured as TPH on mysid survival and growth demonstrate a need to consider the interactions of ultraviolet light and contaminant to avoid under estimating toxicity that might occur in the environment. (C) 2000 Elsevier Science B.V.

  9. Differential effect of the 5-HTT gene-linked polymorphic region on emotional eating during stress exposure following tryptophan challenge.

    PubMed

    Markus, C Rob; Verschoor, Ellen; Smeets, Tom

    2012-04-01

    Stress and negative moods, which are thought to be partly mediated by reduced brain serotonin function, often increase emotional eating in dieting women (restrainers). Because the short (S) allele polymorphism in the serotonin transporter gene (5-HTTLPR) is associated with serotonin dysfunction, S allele compared to long (L) allele 5-HTTLPR genotypes may be more susceptible to stress-induced emotional eating. Consequently, serotonin challenge via tryptophan (TRP)-rich protein hydrolysate (TPH) may alleviate stress-induced emotional eating particularly in S/S allele carriers. We tested whether acute stress affects emotional eating in women with high or low dietary restraints depending on their 5-HTTLPR genotype and TPH intake. Nineteen female subjects who were homozygous for the short-allele 5-HTTLPR genotype (S'/S'=S/L(G), L(G)/L(G): restrainers vs. nonrestrainers) and 23 female subjects who were homozygous for the long-allele 5-HTTLPR genotype (L'/L'=L(A)/L(A): restrainers vs. nonrestrainers) were tested in a double-blind, placebo-controlled crossover study of stress-induced emotional eating following intake of TPH or a placebo. TPH intake significantly increased the plasma TRP/large neutral amino acid ratio (P<.0001) in the L'/L' group (70%) compared to the S'/S' group (30%). TPH reduced food intake in both groups, but in the L'/L' group, it also reduced stress-induced negative mood (P=.037) and the desire for sweet, high-fat foods (P=.011) regardless of dietary restraint. Since TPH caused a greater increase in the plasma TRP/large neutral amino acid ratio in the L'/L' group compared to S'/S' group, the exclusive beneficial effects of L'/L' genotype may be due to enhanced brain 5-HT function. Copyright © 2012 Elsevier Inc. All rights reserved.

  10. Monitoring biodegradation of diesel fuel in bioventing processes using in situ respiration rate.

    PubMed

    Lee, T H; Byun, I G; Kim, Y O; Hwang, I S; Park, T J

    2006-01-01

    An in situ measuring system of respiration rate was applied for monitoring biodegradation of diesel fuel in a bioventing process for bioremediation of diesel contaminated soil. Two laboratory-scale soil columns were packed with 5 kg of soil that was artificially contaminated by diesel fuel as final TPH (total petroleum hydrocarbon) concentration of 8,000 mg/kg soil. Nutrient was added to make a relative concentration of C:N:P = 100:10:1. One soil column was operated with continuous venting mode, and the other one with intermittent (6 h venting/6 h rest) venting mode. On-line O2 and CO2 gas measuring system was applied to measure O2 utilisation and CO2 production during biodegradation of diesel for 5 months. Biodegradation rate of TPH was calculated from respiration rate measured by the on-line gas measuring system. There were no apparent differences between calculated biodegradation rates from two columns with different venting modes. The variation of biodegradation rates corresponded well with trend of the remaining TPH concentrations comparing other biodegradation indicators, such as C17/pristane and C18/phytane ratio, dehydrogenase activity, and the ratio of hydrocarbon utilising bacteria to total heterotrophic bacteria. These results suggested that the on-line measuring system of respiration rate would be applied to monitoring biodegradation rate and to determine the potential applicability of bioventing process for bioremediation of oil contaminated soil.

  11. Molecular Regulation of Sexual Preference Revealed by Genetic Studies of 5-HT in the Brain of Male Mice

    PubMed Central

    Liu, Yan; Jiang, Yun’ai; Si, Yunxia; Kim, Ji-Young; Chen, Zhou-Feng; Rao, Yi

    2014-01-01

    To whom should a male directs his mating? While it is a critical social interaction, little is known about molecular and cellular mechanisms controlling mammalian sexual preference. Here we report that the neurotransmitter 5-HT is required for male sexual preference. Male mice lacking central serotonergic neurons lost sexual preference but were not generally defective in olfaction. A role for 5-hydroxytryptamine (5-HT) was demonstrated by the phenotype of mice unable to synthesize 5-HT in the brain when lacking tryptophan hydroxylase 2 (Tph2). 5-hydroxytryptophan (5-HTP) injection rescued the phenotype of adult Tph2 knockout mice within 35 minutes. These results indicate that 5-HT and serotonergic neurons in the adult brain regulate mammalian sexual preference. PMID:21441904

  12. Bioremediation of Crude Oil Contaminated Desert Soil: Effect of Biostimulation, Bioaugmentation and Bioavailability in Biopile Treatment Systems

    PubMed Central

    Benyahia, Farid; Embaby, Ahmed Shams

    2016-01-01

    This work was aimed at evaluating the relative merits of bioaugmentation, biostimulation and surfactant-enhanced bioavailability of a desert soil contaminated by crude oil through biopile treatment. The results show that the desert soil required bioaugmentation and biostimulation for bioremediation of crude oil. The bioaugmented biopile system led to a total petroleum hydrocarbon (TPH) reduction of 77% over 156 days while the system with polyoxyethylene (20) sorbitan monooleate (Tween 80) gave a 56% decrease in TPH. The biostimulated system with indigenous micro-organisms gave 23% reduction in TPH. The control system gave 4% TPH reduction. The addition of Tween 80 led to a respiration rate that peaked in 48 days compared to 88 days for the bioaugmented system and respiration declined rapidly due to nitrogen depletion. The residual hydrocarbon in the biopile systems studied contained polyaromatics (PAH) in quantities that may be considered as hazardous. Nitrogen was found to be a limiting nutrient in desert soil bioremediation. PMID:26891314

  13. Bioremediation of Crude Oil Contaminated Desert Soil: Effect of Biostimulation, Bioaugmentation and Bioavailability in Biopile Treatment Systems.

    PubMed

    Benyahia, Farid; Embaby, Ahmed Shams

    2016-02-15

    This work was aimed at evaluating the relative merits of bioaugmentation, biostimulation and surfactant-enhanced bioavailability of a desert soil contaminated by crude oil through biopile treatment. The results show that the desert soil required bioaugmentation and biostimulation for bioremediation of crude oil. The bioaugmented biopile system led to a total petroleum hydrocarbon (TPH) reduction of 77% over 156 days while the system with polyoxyethylene (20) sorbitan monooleate (Tween 80) gave a 56% decrease in TPH. The biostimulated system with indigenous micro-organisms gave 23% reduction in TPH. The control system gave 4% TPH reduction. The addition of Tween 80 led to a respiration rate that peaked in 48 days compared to 88 days for the bioaugmented system and respiration declined rapidly due to nitrogen depletion. The residual hydrocarbon in the biopile systems studied contained polyaromatics (PAH) in quantities that may be considered as hazardous. Nitrogen was found to be a limiting nutrient in desert soil bioremediation.

  14. Serotonergic system antagonists target breast tumor initiating cells and synergize with chemotherapy to shrink human breast tumor xenografts

    PubMed Central

    Gwynne, William D; Hallett, Robin M; Girgis-Gabardo, Adele; Bojovic, Bojana; Dvorkin-Gheva, Anna; Aarts, Craig; Dias, Kay; Bane, Anita; Hassell, John A

    2017-01-01

    Breast tumors comprise an infrequent tumor cell population, termed breast tumor initiating cells (BTIC), which sustain tumor growth, seed metastases and resist cytotoxic therapies. Hence therapies are needed to target BTIC to provide more durable breast cancer remissions than are currently achieved. We previously reported that serotonergic system antagonists abrogated the activity of mouse BTIC resident in the mammary tumors of a HER2-overexpressing model of breast cancer. Here we report that antagonists of serotonin (5-hydroxytryptamine; 5-HT) biosynthesis and activity, including US Federal Food and Drug Administration (FDA)-approved antidepressants, targeted BTIC resident in numerous breast tumor cell lines regardless of their clinical or molecular subtype. Notably, inhibitors of tryptophan hydroxylase 1 (TPH1), required for 5-HT biosynthesis in select non-neuronal cells, the serotonin reuptake transporter (SERT) and several 5-HT receptors compromised BTIC activity as assessed by functional sphere-forming assays. Consistent with these findings, human breast tumor cells express TPH1, 5-HT and SERT independent of their molecular or clinical subtype. Exposure of breast tumor cells ex vivo to sertraline (Zoloft), a selective serotonin reuptake inhibitor (SSRI), reduced BTIC frequency as determined by transplanting drug-treated tumor cells into immune-compromised mice. Moreover, another SSRI (vilazodone; Viibryd) synergized with chemotherapy to shrink breast tumor xenografts in immune-compromised mice by inhibiting tumor cell proliferation and inducing their apoptosis. Collectively our data suggest that antidepressants in combination with cytotoxic anticancer therapies may be an appropriate treatment regimen for testing in clinical trials. PMID:28404880

  15. Serotonergic system antagonists target breast tumor initiating cells and synergize with chemotherapy to shrink human breast tumor xenografts.

    PubMed

    Gwynne, William D; Hallett, Robin M; Girgis-Gabardo, Adele; Bojovic, Bojana; Dvorkin-Gheva, Anna; Aarts, Craig; Dias, Kay; Bane, Anita; Hassell, John A

    2017-05-09

    Breast tumors comprise an infrequent tumor cell population, termed breast tumor initiating cells (BTIC), which sustain tumor growth, seed metastases and resist cytotoxic therapies. Hence therapies are needed to target BTIC to provide more durable breast cancer remissions than are currently achieved. We previously reported that serotonergic system antagonists abrogated the activity of mouse BTIC resident in the mammary tumors of a HER2-overexpressing model of breast cancer. Here we report that antagonists of serotonin (5-hydroxytryptamine; 5-HT) biosynthesis and activity, including US Federal Food and Drug Administration (FDA)-approved antidepressants, targeted BTIC resident in numerous breast tumor cell lines regardless of their clinical or molecular subtype. Notably, inhibitors of tryptophan hydroxylase 1 (TPH1), required for 5-HT biosynthesis in select non-neuronal cells, the serotonin reuptake transporter (SERT) and several 5-HT receptors compromised BTIC activity as assessed by functional sphere-forming assays. Consistent with these findings, human breast tumor cells express TPH1, 5-HT and SERT independent of their molecular or clinical subtype. Exposure of breast tumor cells ex vivo to sertraline (Zoloft), a selective serotonin reuptake inhibitor (SSRI), reduced BTIC frequency as determined by transplanting drug-treated tumor cells into immune-compromised mice. Moreover, another SSRI (vilazodone; Viibryd) synergized with chemotherapy to shrink breast tumor xenografts in immune-compromised mice by inhibiting tumor cell proliferation and inducing their apoptosis. Collectively our data suggest that antidepressants in combination with cytotoxic anticancer therapies may be an appropriate treatment regimen for testing in clinical trials.

  16. Bioremediation of diesel-polluted soil using biostimulation as post-treatment after oxidation with Fenton-like reagents: assays in a pilot plant.

    PubMed

    Silva-Castro, Gloria Andrea; Rodelas, Belén; Perucha, Carlos; Laguna, Jaime; González-López, Jesús; Calvo, Concepción

    2013-02-15

    The present study focuses on the remediation of diesel-polluted soil using modified Fenton treatment coupled with inorganic NPK fertilizer ("Fenton+NPK"). Studies were carried out in a pilot plant containing 1 m(3) of sandy soil contaminated with 20,000 mg kg(-1) of diesel, placed outdoors at a temperature ranging between 5 and 10 °C. Results showed that NPK-fertilizer as post-treatment stimulated culturable degrading bacteria and enhanced dehydrogenase activity. Fenton+NPK treatment increased total petroleum hydrocarbon (TPH) removal efficacy. Natural attenuation removed 49% of TPH in the surface layer, 23% of TPH in the non-saturated layer and 4% of the TPH in the saturated layer, while the percentage removed of TPH after Fenton+NPK treatment was 58%, 57% and 32% respectively. The results from our study showed that, immediately after soil contamination, occurred a specialization and differentiation of the bacterial community, but after this initial modification, no significant changes of bacterial diversity was observed under natural attenuation conditions. In contrast, when the Fenton's reagent was applied a reduction of the bacterial biodiversity was observed. However, the post-biostimulation did enhance the degrading microbiota and stimulated their degrading biological activity. In conclusion, biostimulation, as a post-treatment step in chemical oxidation, is an effective solution to remediate hydrocarbon-polluted sites. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Toxicity testing of crude oil and related compounds using early life stages of the crimson-spotted rainbowfish (Melanotaenia fluviatilis).

    PubMed

    Pollino, Carmel A; Holdway, Douglas A

    2002-07-01

    The toxicity of petroleum hydrocarbons to marine aquatic organisms has been widely investigated; however, the effects on freshwater environments have largely been ignored. In the Australian freshwater environment, the potential impacts of petroleum hydrocarbons are virtually unknown. The toxicity of crude oil and related compounds were measured in the sensitive early life stages of the crimson-spotted rainbowfish (Melanotaenia fluviatilis). Waterborne petroleum hydrocarbons crossed the chorion of embryonic rainbowfish, reducing survival and hatchability. Acute exposures resulted in developmental abnormalities at and above 0.5 mg/L total petroleum hydrocarbons (TPH). Deformities included pericardial edema, disturbed axis formation, and abnormal jaw development. When assessing the acute toxicities of the water-accommodated fraction (WAF) of crude oil, dispersants, dispersant-oil mixtures, and naphthalene to larval rainbowfish, the lowest to highest 96-h median lethal concentrations for day of hatch larvae were naphthalene (0.51 mg/L), dispersed crude oil WAF (DCWAF)-9527 (0.74 mg/L TPH), WAF (1.28 mg/L TPH), DCWAF-9500 (1.37 mg/L TPH), Corexit 9500 (14.5 mg/L TPH), and Corexit 9527 (20.1 mg/L). Using naphthalene as a reference toxicant, no differences were found between the sensitivities of larval rainbowfish collected from adults exposed to petroleum hydrocarbons during embryonic development and those collected from unexposed adults.

  18. Biodegradation of aged diesel in diverse soil matrixes: impact of environmental conditions and bioavailability on microbial remediation capacity.

    PubMed

    Sutton, Nora B; van Gaans, Pauline; Langenhoff, Alette A M; Maphosa, Farai; Smidt, Hauke; Grotenhuis, Tim; Rijnaarts, Huub H M

    2013-07-01

    While bioremediation of total petroleum hydrocarbons (TPH) is in general a robust technique, heterogeneity in terms of contaminant and environmental characteristics can impact the extent of biodegradation. The current study investigates the implications of different soil matrix types (anthropogenic fill layer, peat, clay, and sand) and bioavailability on bioremediation of an aged diesel contamination from a heterogeneous site. In addition to an uncontaminated sample for each soil type, samples representing two levels of contamination (high and low) were also used; initial TPH concentrations varied between 1.6 and 26.6 g TPH/kg and bioavailability between 36 and 100 %. While significant biodegradation occurred during 100 days of incubation under biostimulating conditions (64.4-100 % remediation efficiency), low bioavailability restricted full biodegradation, yielding a residual TPH concentration. Respiration levels, as well as the abundance of alkB, encoding mono-oxygenases pivotal for hydrocarbon metabolism, were positively correlated with TPH degradation, demonstrating their usefulness as a proxy for hydrocarbon biodegradation. However, absolute respiration and alkB presence were dependent on soil matrix type, indicating the sensitivity of results to initial environmental conditions. Through investigating biodegradation potential across a heterogeneous site, this research illuminates the interplay between soil matrix type, bioavailability, and bioremediation and the implications of these parameters for the effectiveness of an in situ treatment.

  19. Is there a link between Depressive Disorders and Tryptophan Hydroxylase 1 (TPH1) Gene Polymorphism? - Study from a Distressed Area, Kashmir (India)

    PubMed Central

    Tarfarosh, Shah Faisal Ahmad; Dar, Mohammad Maqbool; Hussain, Arshad; Shoib, Sheikh; Shah, Tabindah; Shah, Sahil; Manzoor, Mushbiq

    2016-01-01

    Background The progress that man has made in all domains of life, during all these years of reign over the earth, is utterly remarkable. However, it always came at a price. Each epoch of progress has seen human beings inflicted with trauma and cynical consequences. During the last two decades, Kashmiri (Indian) people have experienced continuous violence, a reign of terror, and political turmoil. Each of these disastrous events has contributed to the increase in psychiatric disorders in this part of the world, especially major depressive disorders. We can observe that besides the environmental influences, gene polymorphism also plays a crucial role in the development of depressive disorders. The role of Tryptophan Hydroxylase 1 (TPH1) gene is implicated in various psychiatric disorders, including depression. However, no study has investigated TPH1 A779C gene polymorphism in depressive disorders in a distressed society like Kashmir (India). Aims To study TPH1 A779C single nucleotide polymorphism in depressive disorders in Kashmiri (Indian) population. Materials and Methods Two hundred and forty patients diagnosed with depressive disorder, and 160 unrelated healthy volunteers (control), were studied in a case-control study design. Polymorphism was determined using polymerase chain reaction (PCR) and agarose gel electrophoresis, after digestion with HAP II enzyme. Genotypes and allele frequencies were compared using Chi-square tests, Fisher’s exact test, odds ratio, 95% confidence interval (C.I.) and a p-value of <0.05 was considered to be statistically significant. Results The mean age ± standard deviation (SD) of depression and control group was 32.02±10.99 and 31.75±9.93, respectively (p= 0.512). It was found that the patients from depression group had AA genotype (51.7%) in comparison to control group (17.5%) and these results were statistically significant (p≤0.0001). Calculation of allelic frequency revealed a stronger association of A allele with depression group (70.83%) than with the control group (41.25%), and it was also found to be statistically significant (p≤0.0001) with C.I. of 3.459 (1.909-6.266). Conclusion TPH1 A779C A gene was found to be associated with a major depressive disorder (MDD) in Kashmiri (Indian) population. There were high HAM-A as well as HAM-D scores in depressive patients of Kashmir (India). PMID:27672527

  20. Glucocorticoids Inhibit Basal and Hormone-Induced Serotonin Synthesis in Pancreatic Beta Cells

    PubMed Central

    Hasni Ebou, Moina; Singh-Estivalet, Amrit; Launay, Jean-Marie; Callebert, Jacques; Tronche, François; Ferré, Pascal; Gautier, Jean-François; Guillemain, Ghislaine; Bréant, Bernadette

    2016-01-01

    Diabetes is a major complication of chronic Glucocorticoids (GCs) treatment. GCs induce insulin resistance and also inhibit insulin secretion from pancreatic beta cells. Yet, a full understanding of this negative regulation remains to be deciphered. In the present study, we investigated whether GCs could inhibit serotonin synthesis in beta cell since this neurotransmitter has been shown to be involved in the regulation of insulin secretion. To this aim, serotonin synthesis was evaluated in vitro after treatment with GCs of either islets from CD1 mice or MIN6 cells, a beta-cell line. We also explored the effect of GCs on the stimulation of serotonin synthesis by several hormones such as prolactin and GLP 1. We finally studied this regulation in islet in two in vivo models: mice treated with GCs and with liraglutide, a GLP1 analog, and mice deleted for the glucocorticoid receptor in the pancreas. We showed in isolated islets and MIN6 cells that GCs decreased expression and activity of the two key enzymes of serotonin synthesis, Tryptophan Hydroxylase 1 (Tph1) and 2 (Tph2), leading to reduced serotonin contents. GCs also blocked the induction of serotonin synthesis by prolactin or by a previously unknown serotonin activator, the GLP-1 analog exendin-4. In vivo, activation of the Glucagon-like-Peptide-1 receptor with liraglutide during 4 weeks increased islet serotonin contents and GCs treatment prevented this increase. Finally, islets from mice deleted for the GR in the pancreas displayed an increased expression of Tph1 and Tph2 and a strong increased serotonin content per islet. In conclusion, our results demonstrate an original inhibition of serotonin synthesis by GCs, both in basal condition and after stimulation by prolactin or activators of the GLP-1 receptor. This regulation may contribute to the deleterious effects of GCs on beta cells. PMID:26901633

  1. Application of an in-situ soil sampler for assessing subsurface biogeochemical dynamics in a diesel-contaminated coastal site during soil flushing operations.

    PubMed

    Kwon, Man Jae; O'Loughlin, Edward J; Ham, Baknoon; Hwang, Yunho; Shim, Moojoon; Lee, Soonjae

    2018-01-15

    Subsurface biogeochemistry and contaminant dynamics during the remediation of diesel-contamination by in-situ soil flushing were investigated at a site located in a coastal region. An in-situ sampler containing diesel-contaminated soils separated into two size fractions (<0.063- and <2-mm) was utilized in two monitoring wells: DH1 (located close to the injection and extraction wells for in-situ soil flushing) and DH2 (located beyond sheet piles placed to block the transport of leaked diesel). Total petroleum hydrocarbon (TPH) concentrations and biogeochemical properties were monitored both in soil and groundwater for six months. A shift occurred in the groundwater type from Ca-HCO 3 to Na-Cl due to seawater intrusion during intense pumping, while the concentrations of Ni, Cu, Co, V, Cr, and Se increased substantially following surfactant (TWEEN 80) injection. The in-situ sampler with fine particles was more sensitive to variations in conditions during the remedial soil flushing process. In both wells, soil TPH concentrations in the <0.063-mm fraction were much higher than those in the <2-mm fraction. Increases in soil TPH in DH1 were consistent with the expected outcomes following well pumping and surfactant injection used to enhance TPH extraction. However, the number of diesel-degrading microorganisms decreased after surfactant injection. 16S-rRNA gene-based analysis also showed that the community composition and diversity depended on both particle size and diesel contamination. The multidisciplinary approach to the contaminated site assessments showed that soil flushing with surfactant enhanced diesel extraction, but negatively impacted in-situ diesel biodegradation as well as groundwater quality. The results also suggest that the in-situ sampler can be an effective monitoring tool for subsurface biogeochemistry as well as contaminant dynamics. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Assessment of soil-gas contamination at the 17th Street landfill, Fort Gordon, Georgia, 2011

    USGS Publications Warehouse

    Falls, W. Fred; Caldwell, Andral W.; Guimaraes, Wladmir G.; Ratliff, W. Hagan; Wellborn, John B.; Landmeyer, James E.

    2012-01-01

    Assessments of contaminants in soil gas were conducted in two study areas at Fort Gordon, Georgia, in July and August of 2011 to supplement environmental contaminant data for previous studies at the 17th Street landfill. The two study areas include northern and eastern parts of the 17th Street landfill and the adjacent wooded areas to the north and east of the landfill. These study areas were chosen because of their close proximity to the surface water in Wilkerson Lake and McCoys Creek. A total of 48 soil-gas samplers were deployed for the July 28 to August 3, 2011, assessment in the eastern study area. The assessment mostly identified detections of total petroleum hydrocarbons (TPH), and gasoline- and diesel-range compounds, but also identified the presence of chlorinated solvents in six samplers, chloroform in three samplers, 2-methyl naphthalene in one sampler, and trimethylbenzene in one sampler. The TPH masses exceeded 0.02 microgram (μg) in all 48 samplers and exceeded 0.9 μg in 24 samplers. Undecane, one of the three diesel-range compounds used to calculate the combined mass for diesel-range compounds, was detected in 17 samplers and is the second most commonly detected compound in the eastern study area, exceeded only by the number of TPH detections. Six samplers had detections of toluene, but other gasoline compounds were detected with toluene in three of the samplers, including detections of ethylbenzene, meta- and para-xylene, and octane. All detections of chlorinated organic compounds had soil-gas masses equal to or less than 0.08 μg, including three detections of trichloroethene, three detections of perchloroethene, three chloroform detections, one 1,4-dichlorobenzene detection, and one 1,1,2-trichloroethane detection. Three methylated compounds were detected in the eastern study area, but were detected at or below method detection levels. A total of 32 soil-gas samplers were deployed for the August 11–24, 2011, assessment in the northern study area. All samplers in the survey had detections of TPH, but only eight of the samplers had detections of TPH greater than 0.9 mg. Four samplers had TPH detections greater than 9 mg; the only other fuel-related compounds detected in these four samplers included toluene in three of the samplers and undecane in the fourth sampler. Three samplers deployed along the western margin of the northern landfill had detections of both diesel-and gasoline-related compounds; however, the diesel-related compounds were detected at or below method detection levels. Seven samplers in the northern study area had detections of chlorinated compounds, including three perchloroethene detections, three chloroform detections, and one 1,4-dichloro-benzene detection. One sampler on the western margin of the landfill had detections of 1,2,4-trimethylbenzene and 1,3,5-tr-methylbenene below method detection levels.

  3. 76 FR 5415 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Order Approving...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-31

    ... Rule Change Regarding Rule 4.20--Anti-Money Laundering Compliance Program January 25, 2011. I... Rule 4.20, Anti-Money Laundering Compliance Program, to require all Trading Permit Holders or TPH... or TPH organization's existence to ensure anti-money laundering compliance is in place and...

  4. SETTING A GOAL FOR TPH THAT ELIMINATES FREE PRODUCT IN WELLS

    EPA Science Inventory

    The U.S. EPA expects that remedial action at UST sites will reduce free product in monitoring wells to the maximum extent practicable. How much active treatment is necessary to ensure that free product will not accumulate in a monitoring well? The concentration of TPH that will...

  5. Chemical composition, antioxidant capacity and content of phenolic compounds in meals collected in hospitals in Bolivia and Sweden.

    PubMed

    Tejeda, L; Dębiec, M; Nilsson, L; Peñarrieta, J M; Alvarado, J A

    2012-01-01

    The objective of this study was to evaluate the proximal composition, as well as Total Antioxidant Capacity (TAC) and Total Phenols (TPH) in meals that represent a complex food matrix, from different hospitals in Bolivia and Sweden. Protein, fat, ash, dietary fiber and carbohydrate contents were measured in 29 samples: 20 from two Bolivian hospitals and 9 from the university hospital in Lund, Sweden. The antioxidant capacity was measured by three spectrophotometric methods: the ferric reducing antioxidant power (FRAP) method, the 2, 2'- azinobis-3-ethylbenzotiazoline-6-sulfonic acid (ABTS) method and Total Phenolic Compounds (TPH) using the Folin-Ciocalteu reagent. The results show that fat, protein, carbohydrate and dietary fiber in Bolivian and Swedish hospital meals are following internationally established recommendations. Regarding the main courses, TPH contents in both countries were in the same range. However, TAC and dietary fiber content were higher in Swedish meals than in Bolivian meals and the TAC was far lower, in both cases, in comparison with the value obtained from individual food items reported from literature. The results show that antioxidant levels can be easily overestimated by considering only individual uncooked ingredients. An interesting consideration is, the fiber content in the meals, which can be an important source of antioxidants and non-extractable phenolic compounds.

  6. [Determination of organotin compounds in textile auxiliaries by gas chromatography-mass spectrometry].

    PubMed

    Li, Yanming; Hu, Yongjie; Liu, Jinhua; Guo, Yuliang; Wang, Guiqin

    2011-04-01

    A gas chromatography-mass spectrometry (GC-MS) method has been developed for the determination of dibutyltin (DBT), tributyltin (TBT) and triphenyltin (TPhT) in textile auxiliaries. The sample was first extracted with n-hexane in acetate buffer solution (pH 4.0) under ultrasonication (for hydrophobic sample) or oscillation extraction (for hydrophilic sample) and then derivatized with sodium tetraethylborate in tetrahydrofuran. The derivative was determined by GC-MS in selected ion monitoring (SIM) mode. The separation and quantification were achieved using a Rxi-5 ms silica capillary column (30 m x 0.25 mm x 0.25 microm). The linear ranges were 0.1-8.0 mg/L for both DBT and TBT, and 0.1-4.0 mg/L for TPhT. There were good linear relationships between the peak area and concentration in the linear ranges and the correlation coefficients (r2) were 0.9994-0.9998. The detection limits (LOD) were from 0.003 mg/L to 0.005 mg/L. The average recoveries of these organotin compounds at the three spiked levels of 4.0, 10.0 and 40.0 mg/kg were 92.6%-108.0% with the relative standard deviations (RSDs) of 2.5%-10.2%. The method is simple and accurate for simultaneous analysis of the DBT, TBT and TPhT in textile auxiliaries.

  7. Estradiol or fluoxetine alters depressive behavior and tryptophan hydroxylase in rat raphe.

    PubMed

    Yang, Fu-Zhong; Wu, Yan; Zhang, Wei-Guo; Cai, Yi-Yun; Shi, Shen-Xun

    2010-03-10

    The effects of 17beta-estradiol and fluoxetine on behavior of ovariectomized rats subjected to the forced swimming test and the expression of tryptophan hydroxylase (TPH) in dorsal and median raphe were investigated, respectively through time sampling technique of behavior scoring and immunohistochemistry. Both estradiol and fluoxetine increased swimming and decreased immobility in the forced swimming test. The forced swimming stress decreased integrated optical density of TPH-positive regions in dorsal and median raphe. Both estradiol and fluoxetine administration prevented integrated optical density of TPH-positive regions from being decreased by forced swimming stress. These observations suggest that both estradiol and fluoxetine have protective bearing on ovariectomized rats enduring forced swimming stress.

  8. Altered serotonin physiology in human breast cancers favors paradoxical growth and cell survival.

    PubMed

    Pai, Vaibhav P; Marshall, Aaron M; Hernandez, Laura L; Buckley, Arthur R; Horseman, Nelson D

    2009-01-01

    The breast microenvironment can either retard or accelerate the events associated with progression of latent cancers. However, the actions of local physiological mediators in the context of breast cancers are poorly understood. Serotonin (5-HT) is a critical local regulator of epithelial homeostasis in the breast and other organs. Herein, we report complex alterations in the intrinsic mammary gland serotonin system of human breast cancers. Serotonin biosynthetic capacity was analyzed in human breast tumor tissue microarrays using immunohistochemistry for tryptophan hydroxylase 1 (TPH1). Serotonin receptors (5-HT1-7) were analyzed in human breast tumors using the Oncomine database. Serotonin receptor expression, signal transduction, and 5-HT effects on breast cancer cell phenotype were compared in non-transformed and transformed human breast cells. In the context of the normal mammary gland, 5-HT acts as a physiological regulator of lactation and involution, in part by favoring growth arrest and cell death. This tightly regulated 5-HT system is subverted in multiple ways in human breast cancers. Specifically, TPH1 expression undergoes a non-linear change during progression, with increased expression during malignant progression. Correspondingly, the tightly regulated pattern of 5-HT receptors becomes dysregulated in human breast cancer cells, resulting in both ectopic expression of some isoforms and suppression of others. The receptor expression change is accompanied by altered downstream signaling of 5-HT receptors in human breast cancer cells, resulting in resistance to 5-HT-induced apoptosis, and stimulated proliferation. Our data constitutes the first report of direct involvement of 5-HT in human breast cancer. Increased 5-HT biosynthetic capacity accompanied by multiple changes in 5-HT receptor expression and signaling favor malignant progression of human breast cancer cells (for example, stimulated proliferation, inappropriate cell survival). This occurs through uncoupling of serotonin from the homeostatic regulatory mechanisms of the normal mammary epithelium. The findings open a new avenue for identification of diagnostic and prognostic markers, and valuable new therapeutic targets for managing breast cancer.

  9. 77 FR 34422 - Self-Regulatory Organizations; Chicago Stock Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-11

    ... Clearing and Outsourcing Solutions, Inc. (``Apex Clearing''), as a CHX Participant and TPH. The text of... as it pertains to the application approval process of a CHX Participant \\5\\ and TPH \\6\\ in order to...'').\\9\\ \\8\\ Prior to the Transaction, Apex Clearing's name was Ridge Clearing & Outsourcing Solutions...

  10. INNOVATIVE TECHNOLOGY VERIFICATION REPORT "FIELD MEASUREMENT TECHNOLOGIES FOR TOTAL PETROLEUM HYDROCARBONS IN SOIL" WILKS ENTERPRISE, INC. INFRACAL TOG/TPH ANALYZER

    EPA Science Inventory


    The hifracal' TOG/TPH Analyzer developed by Wilks Enterprise, Inc. (Wilks), was demonstrated under the U.S. Environmental Protection Agency Superfund Innovative Technology Evaluation Program in June 2000 at the Navy Base Ventura County site in Port Hueneme, California. The pu...

  11. Hydrocarbon Degradation and Lead Solubility in a Soil Polluted with Lead and Used Motor Oil Treated by Composting and Phytoremediation.

    PubMed

    Escobar-Alvarado, L F; Vaca-Mier, M; López, R; Rojas-Valencia, M N

    2018-02-01

    Used lubricant oils and metals can be common soil pollutants in abandoned sites. When soil is contaminated with various hazardous wastes, the efficiency of biological treatments could be affected. The purpose of this work was to investigate the effect of combining phytoremediation and composting on the efficiency of hydrocarbon degradation and lead solubility in a soil contaminated with 31,823 mg/kg of total petroleum hydrocarbon (TPH) from used motor oil and 8260 mg/kg of lead. Mexican cactus (Opuntia ficus indica) and yard trimmings were added in the composting process, and lucerne (Medicago sativa) was used in the phytoremediation process. After a 9 week composting process, only 13% of the initial TPH concentration was removed. The following 20 week phytoremediation process removed 48% of TPH. The highest TPH degradation percentage (66%), was observed in the experiment with phytoremediation only. This work demonstrates sustainable technologies, such as biological treatments, represent low-cost options for remediation; however, they are not frequently used because they require long periods of time for success.

  12. Assessment of soil-gas, soil, and water contamination at the former 19th Street landfill, Fort Gordon, Georgia, 2009-2010

    USGS Publications Warehouse

    Falls, W. Fred; Caldwell, Andral W.; Guimaraes, Wladmir B.; Ratliff, W. Hagan; Wellborn, John B.; Landmeyer, James E.

    2011-01-01

    Soil gas, soil, and water were assessed for organic and inorganic constituents at the former 19th Street landfill at Fort Gordon, Georgia, from February to September 2010. Passive soil-gas samplers were analyzed to evaluate organic constituents in the hyporheic zone and flood plain of a creek and soil gas within the estimated boundaries of the former landfill. Soil and water samples were analyzed to evaluate inorganic constituents in soil samples, and organic and inorganic constituents in the surface water of a creek adjacent to the landfill, respectively. This assessment was conducted to provide environmental constituent data to Fort Gordon pursuant to requirements of the Resource Conservation and Recovery Act Part B Hazardous Waste Permit process. The passive soil-gas samplers deployed in the water-saturated hyporheic zone and flood plain of the creek adjacent to the former landfill indicated the presence of total petroleum hydrocarbon (TPH) and octane above method detection levels in groundwater beneath the creek bed and flood plain at all 12 soil-gas sampler locations. The TPH concentrations ranged from 51.4 to 81.4 micrograms per liter. Octane concentrations ranged from 1.78 to 2.63 micrograms per liter. These detections do not clearly identify specific source areas in the former landfill; moreover, detections of TPH and octane in a soil-gas sampler installed at a seep on the western bank of the creek indicated the potential for these constituents to be derived from source areas outside the estimated boundaries of the former landfill. A passive soil-gas sampler survey was conducted in the former landfill from June 30 to July 5, 2010, and involved 56 soil-gas samplers that were analyzed for petroleum and halogenated compounds not classified as chemical agents or explosives. The TPH soil-gas mass exceeded 2.0 micrograms in 21 samplers. Most noticeable are the two sites with TPH detections which are located in and near the hyporheic zone and are likely to affect the creek. However, most TPH detections were located in and immediately adjacent to a debris field located within the former landfill and in areas where debris was not visible, including the northwestern and southeastern parts of the study area. Two of the four soil-gas samplers installed within a former military training area adjacent to the landfill also had TPH detections above the method detection level. Benzene, toluene, ethylbenzene, and xylene (as combined BTEX mass) were detected at 0.02 microgram or greater in three soil-gas samplers installed at the northwestern boundary and in five samplers installed in the southeastern part of the study area. There was no BTEX mass detected above the method detection level in samplers installed in the debris field. Toluene was the most frequently detected BTEX compound. Compounds indicative of diesel-range organics were detected above 0.04 microgram in 12 soil-gas samplers and had a distribution similar to that of TPH, including being detected in the debris field. Undecane was the most frequently detected diesel compound. Chloroform and naphthalene were detected in eight and two soil-gas samplers, respectively. Five soil-gas samplers deployed during September 2010 were analyzed for organic compounds classified as chemical agents and explosives, but none exceeded the method detection levels. Five composite soil samples collected from within the estimated boundaries of the former landfill were analyzed for 35 inorganic constituents, but none of the constituents detected exceeded regional screening levels for industrial soils. The sample collected in the debris field exceeded background levels for aluminum, barium, calcium, chromium, lead, nickel, potassium, sodium, and zinc. Three surface-water samples were collected in September 2010 from a stormwater outfall culvert that drains to the creek and from the open channel of the creek at upstream and downstream locations relative to the outfall. Toluene was detected at 0.661 mi

  13. Sorption and desorption of organophosphate esters with different hydrophobicity by soils.

    PubMed

    Cristale, Joyce; Álvarez-Martín, Alba; Rodríguez-Cruz, Sonia; Sánchez-Martín, María J; Lacorte, Silvia

    2017-12-01

    Organophosphate esters (OPEs) are ubiquitous contaminants with potentially hazardous effects on both the environment and human health. Knowledge about the soil sorption-desorption process of organic chemicals is important in order to understand their fate, mobility, and bioavailability, enabling an estimation to be made of possible risks to the environment and biota. The aim of this study was to use the batch equilibrium technique to evaluate the sorption-desorption behavior of seven OPEs (TCEP, TCPP, TBEP, TDCP, TBP, TPhP, and EHDP) in soils with distinctive characteristics (two unamended soils and a soil amended with sewage sludge). The equilibrium concentrations of the OPEs were determined by high performance liquid chromatography coupled to a triple quadrupole mass spectrometer (HPLC-MS/MS). All the compounds were sorbed by the soils, and soil organic carbon (OC) played an important role in this process. The sorption of the most soluble OPEs (TCEP, TCPP, and TBEP) depended on soil OC content, although desorption was ≥ 58.1%. The less water-soluble OPEs (TDCP, TBP, TPhP, and EHDP) recorded total sorption (100% for TPhP and EHDP) or very high sorption (≥ 34.9%) by all the soils and were not desorbed, which could be explained by their highly hydrophobic nature, as indicated by the logarithmic octanol/water partition coefficient (K ow ) values higher than 3.8, resulting in a high affinity for soil OC. The results of the sorption-desorption of the OPEs by soils with different characteristics highlighted the influence of these compounds' physicochemical properties and the content and nature of soil OC in this process.

  14. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Yingjie, E-mail: yzx@ansto.gov.au; Karatchevtseva, Inna; Bhadbhade, Mohan

    With the coordination of dimethylformamide (DMF), two new uranium(VI) complexes with either 4-hydroxybenzoic acid (H{sub 2}phb) or terephthalic acid (H{sub 2}tph) have been synthesized under solvothermal conditions and structurally characterized. [(UO{sub 2}){sub 2}(Hphb){sub 2}(phb)(DMF)(H{sub 2}O){sub 3}]·4H{sub 2}O (1) has a dinuclear structure constructed with both pentagonal and hexagonal bipyramidal uranium polyhedra linked through a µ{sub 2}-bridging ligand via both chelating carboxylate arm and alcohol oxygen bonding, first observation of such a coordination mode of 4-hydroxybenzoate for 5 f ions. [(UO{sub 2})(tph)(DMF)] (2) has a three-dimensional (3D) framework built with pentagonal bipyramidal uranium polyhedra linked with µ{sub 4}-terephthalate ligands. The 3Dmore » channeled structure is facilitated by the unique carboxylate bonding with nearly linear C–O–U angles and the coordination of DMF molecules. The presence of phb ligands in different coordination modes, uranyl ions in diverse environments and DMF in complex 1, and tph ligand, DMF and uranyl ion in complex 2 has been confirmed by Raman spectroscopy. In addition, their thermal stability and photoluminescence properties have been investigated. - Graphical abstract: With the coordination of dimethylformamide, two new uranyl complexes with either 4-hydroxybenzoate or terephthalate have been synthesized under solvothermal conditions and structurally characterized. - Highlights: • Solvent facilitates the synthesis of two new uranium(VI) complexes. • A dinuclear complex with both penta- and hexagonal bipyramidal uranium polyhedral. • A unique µ{sub 2}-bridging mode of 4-hydroxybenzoate via alcohol oxygen for 5 f ions. • A 3D framework with uranium polyhedra and µ{sub 4}-terephthalate ligands. • Vibration modes and photoluminescence properties are reported.« less

  15. Rapid prediction of total petroleum hydrocarbons concentration in contaminated soil using vis-NIR spectroscopy and regression techniques.

    PubMed

    Douglas, R K; Nawar, S; Alamar, M C; Mouazen, A M; Coulon, F

    2018-03-01

    Visible and near infrared spectrometry (vis-NIRS) coupled with data mining techniques can offer fast and cost-effective quantitative measurement of total petroleum hydrocarbons (TPH) in contaminated soils. Literature showed however significant differences in the performance on the vis-NIRS between linear and non-linear calibration methods. This study compared the performance of linear partial least squares regression (PLSR) with a nonlinear random forest (RF) regression for the calibration of vis-NIRS when analysing TPH in soils. 88 soil samples (3 uncontaminated and 85 contaminated) collected from three sites located in the Niger Delta were scanned using an analytical spectral device (ASD) spectrophotometer (350-2500nm) in diffuse reflectance mode. Sequential ultrasonic solvent extraction-gas chromatography (SUSE-GC) was used as reference quantification method for TPH which equal to the sum of aliphatic and aromatic fractions ranging between C 10 and C 35 . Prior to model development, spectra were subjected to pre-processing including noise cut, maximum normalization, first derivative and smoothing. Then 65 samples were selected as calibration set and the remaining 20 samples as validation set. Both vis-NIR spectrometry and gas chromatography profiles of the 85 soil samples were subjected to RF and PLSR with leave-one-out cross-validation (LOOCV) for the calibration models. Results showed that RF calibration model with a coefficient of determination (R 2 ) of 0.85, a root means square error of prediction (RMSEP) 68.43mgkg -1 , and a residual prediction deviation (RPD) of 2.61 outperformed PLSR (R 2 =0.63, RMSEP=107.54mgkg -1 and RDP=2.55) in cross-validation. These results indicate that RF modelling approach is accounting for the nonlinearity of the soil spectral responses hence, providing significantly higher prediction accuracy compared to the linear PLSR. It is recommended to adopt the vis-NIRS coupled with RF modelling approach as a portable and cost effective method for the rapid quantification of TPH in soils. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Development, optimization, validation and application of faster gas chromatography - flame ionization detector method for the analysis of total petroleum hydrocarbons in contaminated soils.

    PubMed

    Zubair, Abdulrazaq; Pappoe, Michael; James, Lesley A; Hawboldt, Kelly

    2015-12-18

    This paper presents an important new approach to improving the timeliness of Total Petroleum Hydrocarbon (TPH) analysis in the soil by Gas Chromatography - Flame Ionization Detector (GC-FID) using the CCME Canada-Wide Standard reference method. The Canada-Wide Standard (CWS) method is used for the analysis of petroleum hydrocarbon compounds across Canada. However, inter-laboratory application of this method for the analysis of TPH in the soil has often shown considerable variability in the results. This could be due, in part, to the different gas chromatography (GC) conditions, other steps involved in the method, as well as the soil properties. In addition, there are differences in the interpretation of the GC results, which impacts the determination of the effectiveness of remediation at hydrocarbon-contaminated sites. In this work, multivariate experimental design approach was used to develop and validate the analytical method for a faster quantitative analysis of TPH in (contaminated) soil. A fractional factorial design (fFD) was used to screen six factors to identify the most significant factors impacting the analysis. These factors included: injection volume (μL), injection temperature (°C), oven program (°C/min), detector temperature (°C), carrier gas flow rate (mL/min) and solvent ratio (v/v hexane/dichloromethane). The most important factors (carrier gas flow rate and oven program) were then optimized using a central composite response surface design. Robustness testing and validation of model compares favourably with the experimental results with percentage difference of 2.78% for the analysis time. This research successfully reduced the method's standard analytical time from 20 to 8min with all the carbon fractions eluting. The method was successfully applied for fast TPH analysis of Bunker C oil contaminated soil. A reduced analytical time would offer many benefits including an improved laboratory reporting times, and overall improved clean up efficiency. The method was successfully applied for the analysis of TPH of Bunker C oil in contaminated soil. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  17. Effect of glial cell line-derived neurotrophic factor on behavior and key members of the brain serotonin system in mouse strains genetically predisposed to behavioral disorders.

    PubMed

    Naumenko, Vladimir S; Bazovkina, Daria V; Semenova, Alina A; Tsybko, Anton S; Il'chibaeva, Tatyana V; Kondaurova, Elena M; Popova, Nina K

    2013-12-01

    The effect of glial cell line-derived neurotrophic factor (GDNF) on behavior and on the serotonin (5-HT) system of a mouse strain predisposed to depressive-like behavior, ASC/Icg (Antidepressant Sensitive Cataleptics), in comparison with the parental "nondepressive" CBA/Lac mice was studied. Within 7 days after acute administration, GDNF (800 ng, i.c.v.) decreased cataleptic immobility but increased depressive-like behavioral traits in both investigated mouse strains and produced anxiolytic effects in ASC mice. The expression of the gene encoding the key enzyme for 5-HT biosynthesis in the brain, tryptophan hydroxylase-2 (Tph-2), and 5-HT1A receptor gene in the midbrain as well as 5-HT2A receptor gene in the frontal cortex were increased in GDNF-treated ASC mice. At the same time, GDNF decreased 5-HT1A and 5-HT2A receptor gene expression in the hippocampus of ASC mice. GDNF failed to change Tph2, 5-HT1A , or 5-HT2A receptor mRNA levels in CBA mice as well as 5-HT transporter gene expression and 5-HT1A and 5-HT2A receptor functional activity in both investigated mouse strains. The results show 1) a GDNF-induced increase in the expression of key genes of the brain 5-HT system, Tph2, 5-HT1A , and 5-HT2A receptors, and 2) significant genotype-dependent differences in the 5-HT system response to GDNF treatment. The data suggest that genetically defined cross-talk between neurotrophic factors and the brain 5-HT system underlies the variability in behavioral response to GDNF. Copyright © 2013 Wiley Periodicals, Inc.

  18. Influence of subinhibitory antibiotic concentration on Streptococcus pyogenes adherence and biofilm production.

    PubMed

    Šmitran, Aleksandra; Vuković, Dragana; Opavski, Nataša; Gajić, Ina; Marinković, Jelena; Božić, Ljiljana; Živanović, Irena; Kekić, Dušan; Popović, Sunčica; Ranin, Lazar

    2018-06-01

    In this study, the focus was on the effects of sub-MICs of the antibiotics on adherence, hydrophobicity, and biofilm formation by two groups of Streptococcus pyogenes strains, which were responsible for different clinical cases. The aim of this study was to explore the effects of sub-MICs of penicillin, ceftriaxone, erythromycin, and clindamycin on adherence, surface hydrophobicity, and biofilm biomass in two selected collections of group A streptococcus (GAS): strains isolated from carriers (CA) and strains isolated from patients with tonsillopharyngitis (TPh). Isolates were tested for hydrophobicity to xylene, adherence, and biofilm production in uncoated microtiter plates before and after treatment with 1/2 and 1/4 MICs of antibiotics. Penicillin reduced adherence and biofilm production in TPh strains, whereas ceftriaxone diminished adherence and biofilm formation in CA group. On the contrary, clindamycin enhanced adherence and biofilm production in both groups of strains. Erythromycin did not significantly alter adherence, but triggered biofilm production in both groups of isolates. Hydrophobicity of both groups of strains was significantly reduced after exposure to all antibiotics. Beta-lactams displayed anti-biofilm activity; penicillin diminished both adherence and biofilm production in TPh strains, whereas ceftriaxone reduced it in strains isolated from CA.

  19. Total petroleum hydrocarbons and heavy metals in the surface sediments of Bohai Bay, China: long-term variations in pollution status and adverse biological risk.

    PubMed

    Zhou, Ran; Qin, Xuebo; Peng, Shitao; Deng, Shihuai

    2014-06-15

    Surface sediments collected from 2001 to 2011 were analyzed for total petroleum hydrocarbons (TPH) and five heavy metals. The sediment concentration ranges of TPH, Zn, Cu, Pb, Cd and Hg were 6.3-535 μg/g, 58-332 μg/g, 7.2-63 μg/g, 4.3-138 μg/g, 0-0.98μg/g, and 0.10-0.68 μg/g, respectively. These results met the highest marine sediment quality standards in China, indicating that the sediment was fairly clean. However, based on the effects range-median (ERM) quotient method, the calculated values for all of the sampling sites were higher than 0.10, suggesting that there was a potential adverse biological risk in Bohai Bay. According to the calculated results, the biological risk decreased from 2001 to 2007 and increased afterwards. High-risk sites were mainly distributed along the coast. This study suggests that anthropogenic influences might be responsible for the potential risk of adverse biological effects from TPH and heavy metals in Bohai Bay. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Peripheral Serotonin Regulates Maternal Calcium Trafficking in Mammary Epithelial Cells during Lactation in Mice

    PubMed Central

    Laporta, Jimena; Keil, Kimberly P.; Vezina, Chad M.; Hernandez, Laura L.

    2014-01-01

    Lactation is characterized by massive transcellular flux of calcium, from the basolateral side of the mammary alveolar epithelium (blood) into the ductal lumen (milk). Regulation of calcium transport during lactation is critical for maternal and neonatal health. The monoamine serotonin (5-HT) is synthesized by the mammary gland and functions as a homeostatic regulation of lactation. Genetic ablation of tryptophan hydroxylase 1 (Tph1), which encodes the rate-limiting enzyme in non-neuronal serotonin synthesis, causes a deficiency in circulating serotonin. As a consequence maternal calcium concentrations decrease, mammary epithelial cell morphology is altered, and cell proliferation is decreased during lactation. Here we demonstrate that serotonin deficiency decreases the expression and disrupts the normal localization of calcium transporters located in the apical (PMCA2) and basolateral (CaSR, ORAI-1) membranes of the lactating mammary gland. In addition, serotonin deficiency decreases the mRNA expression of calcium transporters located in intracellular compartments (SERCA2, SPCA1 and 2). Mammary expression of serotonin receptor isoform 2b and its downstream pathways (PLCβ3, PKC and MAP-ERK1/2) are also decreased by serotonin deficiency, which might explain the numerous phenotypic alterations described above. In most cases, addition of exogenous 5-hydroxy-L-tryptophan to the Tph1 deficient mice rescued the phenotype. Our data supports the hypothesis that serotonin is necessary for proper mammary gland structure and function, to regulate blood and mammary epithelial cell transport of calcium during lactation. These findings can be applicable to the treatment of lactation-induced hypocalcemia in dairy cows and can have profound implications in humans, given the wide-spread use of selective serotonin reuptake inhibitors as antidepressants during pregnancy and lactation. PMID:25299122

  1. Bond strength to dentin with artificial carious lesions: influence of caries detecting dye.

    PubMed

    Palma, R G; Turbino, M L; Matson, E; Powers, J M

    1998-06-01

    To evaluate the influence of dyes for caries detection on tensile bond strength of adhesive materials to artificial carious dentin. Buccal and lingual enamel of human molars were removed leaving intact dentin surfaces. The entire surface of each specimen was covered with nail varnish, keeping a window area of 4 x 4 mm. Artificial carious lesions were induced with acidified gel. Three dyes (0.5% basic fuchsin; Caries Finder and Cari-D-Tect) were used according to manufacturers' recommendations. Specimens were etched with 35% phosphoric acid for 20 s, washed and dried, leaving a wet dentin surface. The adhesive system (Prime & Bond 2.0) was applied in two layers and light-cured. Restorative materials (TPH Spectrum, Dyract, Advance) were bonded using a 3-mm diameter inverted-cone mold. Control groups were made without dye. Eight samples were tested for each group. After 24 hrs of storage in distilled water, the samples were debonded using a testing machine at 0.5 mm/min crosshead speed. ANOVA and Tukey-Kramer test showed that TPH Spectrum (0.73 MPa) and Dyract (0.74 MPa) had similar bond strengths, and both were higher than Advance (0.0 MPa), which was statistically different (P < 0.01). The use of the dyes did not cause any changes in tensile bond strength for any tested materials.

  2. 77 FR 20675 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Order Approving...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-05

    ... overlap. In addition, unrelated FLEX Orders may not be submitted to the electronic book for the duration... available an electronic book). Public customers and non-TPH broker-dealers RFR responses and FLEX Orders... AIM Auction price locks a public customer or non-TPH broker-dealer order in the electronic book on the...

  3. The relation of serotonin-related gene and COMT gene polymorphisms with criminal behavior in schizophrenic disorder.

    PubMed

    Koh, Kyung Bong; Choi, Eun Hee; Lee, Young-joon; Han, Mooyoung; Choi, Sang-Sup; Kim, So Won; Lee, Min Goo

    2012-02-01

    It has been suggested that patients with schizophrenia might be involved in criminal behavior, such as homicidal and violent behavior. However, the relationship between criminal behavior and genes in patients with schizophrenia has not been clearly elucidated. The objective of this study was to examine the relation between criminal behavior and serotonin-related gene or catechol-O-methyltransferase (COMT) gene polymorphisms in patients with schizophrenia. Serotonin-related and COMT polymorphic markers were assessed by using single nucleotide polymorphism (SNP) genotyping. Ninety-nine crime-related inpatients with schizophrenia (57 homicidal and 42 nonhomicidal violent) and 133 healthy subjects were enrolled between October 2005 and May 2008. Diagnoses were made according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria. The genotype frequencies of tryptophan hydroxylase-1 (TPH1) A218C and COMT V158M were compared between groups. The TPH1 CC genotype had 2.7-fold higher odds of crime-related schizophrenia compared with A-carrier genotype after the analysis was controlled for sex and age (OR, 2.69; 95% CI, 1.22 - 5.91; P = .01). In addition, the TPH1 CC genotype had 3.4-fold higher odds of homicidal schizophrenia compared with A-carrier genotype after the analysis was controlled for sex and age (OR, 3.38; 95% CI, 1.40 - 8.18; P = .007). However, no significant differences were found in the frequencies of genotype of COMT polymorphism between criminal schizophrenics and healthy subjects, nor were any significant differences found between nonhomicidal schizophrenics and healthy subjects. These results indicate that the TPH1 CC recessive genotype is likely to be a genetic risk factor for criminal behavior, especially homicidal behavior in patients with schizophrenia. However, COMT gene polymorphisms were not associated with criminal behavior in schizophrenic patients. © Copyright 2012 Physicians Postgraduate Press, Inc.

  4. Synthetic olive mill wastewater treatment by Fenton's process in batch and continuous reactors operation.

    PubMed

    Esteves, Bruno M; Rodrigues, Carmen S D; Madeira, Luís M

    2017-11-04

    Degradation of total phenol (TPh) and organic matter, (expressed as total organic carbon TOC), of a simulated olive mill wastewater was evaluated by the Fenton oxidation process under batch and continuous mode conditions. A mixture of six phenolic acids usually found in these agro-industrial wastewaters was used for this purpose. The study focused on the optimization of key operational parameters of the Fenton process in a batch reactor, namely Fe 2+ dosage, hydrogen peroxide concentration, pH, and reaction temperature. On the assessment of the process efficiency, > 99% of TPh and > 56% of TOC removal were attained when [Fe 2+ ] = 100 ppm, [H 2 O 2 ] = 2.0 g/L, T = 30 °C, and initial pH = 5.0, after 300 min of reaction. Under those operational conditions, experiments on a continuous stirred-tank reactor (CSTR) were performed for different space-time values (τ). TOC and TPh removals of 47.5 and 96.9%, respectively, were reached at steady-state (for τ = 120 min). High removal of COD (> 75%) and BOD 5 (> 70%) was achieved for both batch and CSTR optimum conditions; analysis of the BOD 5 /COD ratio also revealed an increase in the effluent's biodegradability. Despite the high removal of lumped parameters, the treated effluent did not met the Portuguese legal limits for direct discharge of wastewaters into water bodies, which indicates that coupled chemical-biological process may be the best solution for real olive mill wastewater treatment.

  5. Impact of environmental adaptation on tear film assessments.

    PubMed

    Fagehi, R

    2018-03-01

    The purpose of this study was to investigate the effect of ocular environmental adaptation on clinical tear film assessment. Thirty subjects (male, mean age 23±2.5) participated in this study. A number of clinical tear film tests were applied, including: fluorescein tear break-up time (FTBUT), Schirmer test and tear prism height test (TPH). The tear physiology of each subject was evaluated twice, once immediately when they arrived from the external environment, and then after 30minutes adaptation in the exam room environment. The mean values were: Schirmer test A (22.1±2.99), Schirmer test B (24.2±2.63), FTBUT A (8.00±1.94), FTBUT B (9.13±2.04), TPH A (0.179±0.026) and TPH B* (0.187±0.023). Statistical testing using Wilcoxon-signed rank test showed a significant difference between the Schirmer test results measured at the different times (P=0.008). Also, the FTBUT and tear prism height test results showed significant differences between the two evaluation times, (P=0.001, 0.011, respectively) (A: tear assessed when the subject comes from the outside environment, B: tear film assessed after 30min adaptation in the clinical environment). This study showed a significant difference between the tear film test results evaluated when the subjects were assessed immediately from the outside environment and after an adaptation time in the clinic environment. Practitioners must consider the effect of differences between external and clinical environment adaptation on clinical tear film physiology. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  6. Discovery of novel transcripts of the human tissue kallikrein (KLK1) and kallikrein-related peptidase 2 (KLK2) in human cancer cells, exploiting Next-Generation Sequencing technology.

    PubMed

    Adamopoulos, Panagiotis G; Kontos, Christos K; Scorilas, Andreas

    2018-03-31

    Tissue kallikrein, kallikrein-related peptidases (KLKs), and plasma kallikrein form the largest group of serine proteases in the human genome, sharing many structural and functional properties. Several KLK transcripts have been found aberrantly expressed in numerous human malignancies, confirming their prognostic or/and diagnostic values. However, the process of alternative splicing can now be studied in-depth due to the development of Next-Generation Sequencing (NGS). In the present study, we used NGS to discover novel transcripts of the KLK1 and KLK2 genes, after nested touchdown PCR. Bioinformatics analysis and PCR experiments revealed a total of eleven novel KLK transcripts (two KLK1 and nine KLK2 transcripts). In addition, the expression profiles of each novel transcript were investigated with nested PCR experiments using variant-specific primers. Since KLKs are implicated in human malignancies, qualifying as potential biomarkers, the quantification of the presented novel transcripts in human samples may have clinical applications in different types of cancer. Copyright © 2018. Published by Elsevier Inc.

  7. Stress-Induced Depression Is Alleviated by Aerobic Exercise Through Up-Regulation of 5-Hydroxytryptamine 1A Receptors in Rats.

    PubMed

    Kim, Tae Woon; Lim, Baek Vin; Baek, Dongjin; Ryu, Dong-Soo; Seo, Jin Hee

    2015-03-01

    Stress is associated with depression, which induces many psychiatric disorders. Serotonin, also known as 5-hydroxy-tryptamine (5-HT), acts as a biochemical messenger and regulator in the brain. It also mediates several important physiological functions. Depression is closely associated with an overactive bladder. In the present study, we investigated the effect of treadmill exercise on stress-induced depression while focusing on the expression of 5-HT 1A (5-H1A) receptors in the dorsal raphe. Stress was induced by applying a 0.2-mA electric foot shock to rats. Each set of electric foot shocks comprised a 6-second shock duration that was repeated 10 times with a 30-second interval. Three sets of electric foot shocks were applied each day for 7 days. For the confirmation of depressive state, a forced swimming test was performed. To visualize the expression of 5-HT and tryptophan hydroxylase (TPH), immunohistochemistry for 5-HT and TPH in the dorsal raphe was performed. Expression of 5-H1A receptors was determined by western blot analysis. A depressive state was induced by stress, and treadmill exercise alleviated the depression symptoms in the stress-induced rats. Expressions of 5-HT, TPH, and HT 1A in the dorsal raphe were reduced by the induction of stress. Treadmill exercise increased 5-HT, TPH, and HT 1A expressions in the stress-induced rats. Treadmill exercise enhanced 5-HT synthesis through the up-regulation of 5-HT1A receptors, and improved the stress-induced depression. In the present study, treadmill exercise improved depression symptoms by enhancing 5-HT1A receptor expression. The present results suggest that treadmill exercise might be helpful for the alleviation of overactive bladder and improve sexual function.

  8. Impact of tributyltin and triphenyltin on ivory shell (Babylonia japonica) populations.

    PubMed

    Horiguchi, Toshihiro; Kojima, Mitsuhiro; Hamada, Fumihiko; Kajikawa, Akira; Shiraishi, Hiroaki; Morita, Masatoshi; Shimizu, Makoto

    2006-04-01

    We histopathologically examined gonads and chemically determined organotin compounds in tissues of the ivory shell, Babylonia japonica. Imposex (a superimposition of male-type genital organs on females) occurred in approximately 80-90% of B. japonica specimens that we examined, with the penis and vas deferens both well developed. No oviduct blockage by vas deferens formation was observed. Ovarian spermatogenesis and suppressed ovarian maturation were observed in the females that exhibited imposex, although no histopathological abnormalities were found in males. Tissue distributions of organotin compounds [tributyltin (TBT), triphenyltin (TPhT), and their metabolites] were different for butyltins and phenyltins; a remarkably high accumulation of TBT was observed in the ctenidium, osphradium, and heart, whereas high concentrations of TPhT were detected in the ovary and digestive gland. More than one-third of TBT accumulated in the digestive glands of both males and females, followed by the testis, ctenidium, muscle, and heart tissues in males and in the muscle, ovary, ctenidium, and head tissues (including the central nervous system ganglia) in females. In both males and females, more than half of total TPhT accumulated in the digestive glands, followed by the gonads. The next highest values were in the muscle, ctenidium, and heart tissues in males and in the muscle, oviduct, and head tissues in females. Both TBT and TPhT concentrations in the gonads were positively correlated with penis length in females. Our findings strongly suggest that reproductive failure in adult females accompanied by imposex, possibly induced by TBT and TPhT from antifouling paints, may have caused the marked decline of B. japonica populations in Japan.

  9. 78 FR 43953 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-22

    ... CBOE and CBOE Stock Exchange, LLC (``CBSX''). In addition, the PULSe workstation provides a user with... the national best bid or offer (``NBBO''), regardless of size or time, but allows any user to manually... available to Non-TPH User A. To the extent that orders originating from Non-TPH User A's PULSe workstation...

  10. 76 FR 1203 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-07

    ... elect to have CBOE perform certain marketing services and/or billing services on behalf of the... Participating CBOE TPH, if a Participating Non-CBOE TPH elects to have CBOE perform marketing services on its... default destination for an order to be the U.S. options exchange with the best bid or offer, except CBOE...

  11. Bioremediation and phytoremediation of total petroleum hydrocarbons (TPH) under various conditions.

    PubMed

    McIntosh, Patrick; Schulthess, Cristian P; Kuzovkina, Yulia A; Guillard, Karl

    2017-08-03

    Remediation of contaminated soils is often studied using fine-textured soils rather than low-fertility sandy soils, and few studies focus on recontamination events. This study compared aerobic and anaerobic treatments for remediation of freshly introduced used motor oil on a sandy soil previously phytoremediated and bioacclimated (microorganisms already adapted in the soil environment) with some residual total petroleum hydrocarbon (TPH) contamination. Vegetated and unvegetated conditions to remediate anthropogenic fill containing residual TPH that was spiked with nonaqueous phase liquids (NAPLs) were evaluated in a 90-day greenhouse pot study. Vegetated treatments used switchgrass (Panicum virgatum). The concentration of aerobic bacteria were orders of magnitude higher in vegetated treatments compared to unvegetated. Nevertheless, final TPH concentrations were low in all saturated soil treatments, and high in the presence of switchgrass. Concentrations were also low in unvegetated pots with fertilizer. Acclimated indigenous microbial communities were shown to be more effective in breaking down hydrocarbons than introducing microbes from the addition of plant treatments in sandy soils. Remediation of fresh introduced NAPLs on pre-phytoremediated and bioacclimated soil was most efficient in saturated, anaerobic environments, probably due to the already pre-established microbial associations, easily bioavailable contaminants, and optimized soil conditions for microbial establishment and survival.

  12. Toxicity assessment for petroleum-contaminated soil using terrestrial invertebrates and plant bioassays.

    PubMed

    Hentati, Olfa; Lachhab, Radhia; Ayadi, Mariem; Ksibi, Mohamed

    2013-04-01

    The assessment of soil quality after a chemical or oil spill and/or remediation effort may be measured by evaluating the toxicity of soil organisms. To enhance our understanding of the soil quality resulting from laboratory and oil field spill remediation, we assessed toxicity levels by using earthworms and springtails testing and plant growth experiments. Total petroleum hydrocarbons (TPH)-contaminated soil samples were collected from an oilfield in Sfax, Tunisia. Two types of bioassays were performed. The first assessed the toxicity of spiked crude oil (API gravity 32) in Organization for Economic Co-operation and Development artificial soil. The second evaluated the habitat function through the avoidance responses of earthworms and springtails and the ability of Avena sativa to grow in TPH-contaminated soils diluted with farmland soil. The EC50 of petroleum-contaminated soil for earthworms was 644 mg of TPH/kg of soil at 14 days, with 67 % of the earthworms dying after 14 days when the TPH content reached 1,000 mg/kg. The average germination rate, calculated 8 days after sowing, varied between 64 and 74 % in low contaminated soils and less than 50 % in highly contaminated soils.

  13. Life cycle of petroleum biodegradation metabolite plumes, and implications for risk management at fuel release sites.

    PubMed

    Zemo, Dawn A; O'Reilly, Kirk T; Mohler, Rachel E; Magaw, Renae I; Espino Devine, Catalina; Ahn, Sungwoo; Tiwary, Asheesh K

    2017-07-01

    This paper summarizes the results of a 5-y research study of the nature and toxicity of petroleum biodegradation metabolites in groundwater at fuel release sites that are quantified as diesel-range "Total Petroleum Hydrocarbons" (TPH; also known as TPHd, diesel-range organics (DRO), etc.), unless a silica gel cleanup (SGC) step is used on the sample extract prior to the TPH analysis. This issue is important for site risk management in regulatory jurisdictions that use TPH as a metric; the presence of these metabolites may preclude site closure even if all other factors can be considered "low-risk." Previous work has shown that up to 100% of the extractable organics in groundwater at petroleum release sites can be biodegradation metabolites. The metabolites can be separated from the hydrocarbons by incorporating an SGC step; however, regulatory agency acceptance of SGC has been inconsistent because of questions about the nature and toxicity of the metabolites. The present study was conducted to answer these specific questions. Groundwater samples collected from source and downgradient wells at fuel release sites were extracted and subjected to targeted gas chromatography-mass spectrometry (GC-MS) and nontargeted two-dimensional gas chromatography with time-of-flight mass spectrometry (GC×GC-MS) analyses, and the metabolites identified in each sample were classified according to molecular structural classes and assigned an oral reference dose (RfD)-based toxicity ranking. Our work demonstrates that the metabolites identified in groundwater at biodegrading fuel release sites are in classes ranked as low toxicity to humans and are not expected to pose significant risk to human health. The identified metabolites naturally attenuate in a predictable manner, with an overall trend to an increasingly higher proportion of organic acids and esters, and a lower human toxicity profile, and a life cycle that is consistent with the low-risk natural attenuation paradigm adopted by many regulatory agencies for petroleum release sites. Integr Environ Assess Manag 2017;13:714-727. © 2016 The Authors. Integrated Environmental Assessment and Management Published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC). © 2016 The Authors. Integrated Environmental Assessment and Management Published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

  14. Improvement in pinch function after surgical treatment for thumb in the plane of the hand.

    PubMed

    Iba, K; Wada, T; Aoki, M; Yamashita, T

    2012-02-01

    Thumb in the plane of the hand (TPH) is a congenital deformity in which the nail plane of the radial-most digit is parallel to that of other digits, but structurally the digit retains the characteristics of a thumb. Four hands from four patients were retrospectively diagnosed as having TPH, with the underlying congenital conditions being symbrachydactyly, cleft hand and constriction band syndrome. Thumb web-plasty was carried out in all hands; one required additional rotation osteotomy of the metacarpal. Postoperative hand function was markedly improved and pinch function was possible in all cases. In most cases of TPH, the thumb has the potential to act in opposition, indicating that a thumb web-plasty is worth considering as an initial procedure.

  15. Development of an Efficient Bacterial Consortium for the Potential Remediation of Hydrocarbons from Contaminated Sites

    PubMed Central

    Patowary, Kaustuvmani; Patowary, Rupshikha; Kalita, Mohan C.; Deka, Suresh

    2016-01-01

    The intrinsic biodegradability of hydrocarbons and the distribution of proficient degrading microorganisms in the environment are very crucial for the implementation of bioremediation practices. Among others, one of the most favorable methods that can enhance the effectiveness of bioremediation of hydrocarbon-contaminated environment is the application of biosurfactant producing microbes. In the present study, the biodegradation capacities of native bacterial consortia toward total petroleum hydrocarbons (TPH) with special emphasis to poly aromatic hydrocarbons were determined. The purpose of the study was to isolate TPH degrading bacterial strains from various petroleum contaminated soil of Assam, India and develop a robust bacterial consortium for bioremediation of crude oil of this native land. From a total of 23 bacterial isolates obtained from three different hydrocarbons contaminated samples five isolates, namely KS2, PG1, PG5, R1, and R2 were selected as efficient crude oil degraders with respect to their growth on crude oil enriched samples. Isolates KS2, PG1, and R2 are biosurfactant producers and PG5, R1 are non-producers. Fourteen different consortia were designed involving both biosurfactant producing and non-producing isolates. Consortium 10, which comprises two Bacillus strains namely, Bacillus pumilus KS2 and B. cereus R2 (identified by 16s rRNA sequencing) has shown the best result in the desired degradation of crude oil. The consortium showed degradation up to 84.15% of TPH after 5 weeks of incubation, as revealed from gravimetric analysis. FTIR (Fourier transform infrared) and GCMS (Gas chromatography-mass spectrometer) analyses were correlated with gravimetric data which reveals that the consortium has removed a wide range of petroleum hydrocarbons in comparison with abiotic control including different aliphatic and aromatic hydrocarbons. PMID:27471499

  16. BiVO4/WO3/SnO2 Double-Heterojunction Photoanode with Enhanced Charge Separation and Visible-Transparency for Bias-Free Solar Water-Splitting with a Perovskite Solar Cell.

    PubMed

    Baek, Ji Hyun; Kim, Byeong Jo; Han, Gill Sang; Hwang, Sung Won; Kim, Dong Rip; Cho, In Sun; Jung, Hyun Suk

    2017-01-18

    Coupling dissimilar oxides in heterostructures allows the engineering of interfacial, optical, charge separation/transport and transfer properties of photoanodes for photoelectrochemical (PEC) water splitting. Here, we demonstrate a double-heterojunction concept based on a BiVO 4 /WO 3 /SnO 2 triple-layer planar heterojunction (TPH) photoanode, which shows simultaneous improvements in the charge transport (∼93% at 1.23 V vs RHE) and transmittance at longer wavelengths (>500 nm). The TPH photoanode was prepared by a facile solution method: a porous SnO 2 film was first deposited on a fluorine-doped tin oxide (FTO)/glass substrate followed by WO 3 deposition, leading to the formation of a double layer of dense WO 3 and a WO 3 /SnO 2 mixture at the bottom. Subsequently, a BiVO 4 nanoparticle film was deposited by spin coating. Importantly, the WO 3 /(WO 3 +SnO 2 ) composite bottom layer forms a disordered heterojunction, enabling intimate contact, lower interfacial resistance, and efficient charge transport/transfer. In addition, the top BiVO 4 /WO 3 heterojunction layer improves light absorption and charge separation. The resultant TPH photoanode shows greatly improved internal quantum efficiency (∼80%) and PEC water oxidation performance (∼3.1 mA/cm 2 at 1.23 V vs RHE) compared to the previously reported BiVO 4 /WO 3 photoanodes. The PEC performance was further improved by a reactive-ion etching treatment and CoO x electrocatalyst deposition. Finally, we demonstrated a bias-free and stable solar water-splitting by constructing a tandem PEC device with a perovskite solar cell (STH ∼3.5%).

  17. [Serotonergic genes in the development of anxiety/depression-like state and pathology of aggressive behavior in male mice: RNA-seq data].

    PubMed

    Kudryavtseva, N N; Smagin, D A; Kovalenko, I L; Galyamina, A G; Vishnivetskaya, G B; Babenko, V N; Orlov, Yu L

    2017-01-01

    In course of daily agonistic interactions, mice tend to stratify into those with chronic social defeats and those that repeatedly display aggression, which lead to the development of mixed anxiety/depression-like state and the pathology of aggressive behavior, respectively. Using the data of whole transcriptome analysis (RNA-seq), the changes in the expression of serotonergic genes involved in the synthesis, inactivation, and reception of serotonin, as well as of the Creb1 (transcription factor) gene and the Bdnf (brain-derived neurotrophic factor) gene were detected in the striatum (STR), ventral tegmental area (VTA), midbrain raphe nuclei (MRN), hypothalamus (HYP), and hippocampus (HIP) of defeated and aggressive male mice. In mice of both groups, the Tph2, Ddc, Slc6a4, Htr2a, Htr3a, Htr5b, Slc18a2, and Bdnf genes were downregulated in the MRN and the Tph2, Ddc, and Slc6a4 genes were upregulated in the VTA. These changes were more significant in defeated mice. The Htr5b gene has first been shown to be involved in mechanisms of depression and pathology of aggressive behavior. In the defeated mice, the expression levels of the Htr4 and Aldh1b1 genes were increased in the MRN, and expression levels of the Maob, Htr4, Htr1a, and Slc18a2 genes were increased in the VTA, while the expression level of the Htr3a gene was decreased. In the HYP of aggressive mice the Maoa, Htr2a, Htr2c, and Creb1 genes were downregulated and the Htr6 gene was upregulated. In the defeated mice, the Maoa and Creb1 genes were downregulated and the Htr6 and Aldh1b1 genes were upregulated in the HYP. In the STR, the Htr1a gene was downregulated and the Htr7 and Bdnf genes were upregulated. The Htr1b gene was upregulated in the HIP. The coexpression of dopaminergic and serotonergic genes in the MRN and VTA in the control of pathological behaviors is discussed. Thus, the complex pattern of differential expression of serotonergic genes in brain regions developing under repeated agonistic interactions in mice in dependence on behavioral pathology have been observed.

  18. C1473G polymorphism in mouse tryptophan hydroxylase-2 gene in the regulation of the reaction to emotional stress.

    PubMed

    Bazhenova, Ekaterina Y; Bazovkina, Daria V; Kulikova, Elizabeth A; Fursenko, Dariya V; Khotskin, Nikita V; Lichman, Daria V; Kulikov, Alexander V

    2017-02-15

    Neurotransmitter serotonin (5-HT) is involved in the regulation of stress response. Tryptophan hydroxylase-2 (TPH2) is the key enzyme of serotonin (5-HT) synthesis in the brain. C1473G polymorphism in Tph2 gene is the main factor defining the enzyme activity in the brain of laboratory mice. The effect of interaction between C1473G polymorphism and 30min restriction stress on the behavior in the open field test, c-Fos gene expression and 5-HT metabolism in the brain in adult male of B6-1473C and B6-1473G congenic mouse lines with high and low TPH2 activity was investigated. A significant effect of genotype x stress interaction on c-Fos mRNA in the hypothalamus (F 1,21 =10.66, p<0.001) and midbrain (F 1,21 =9.18, p<0.01) was observed. The stress-induced rise of c-Fos mRNA in these structures is more intensive in B6-1473G than in B6-1473C mice. A marked effect of genotype x stress interaction on 5-HT level in the cortex (F 1,18 =9.38, p<0.01) and 5-HIAA/5-HT turnover rate in the hypothalamus (F 1,18 =9.01, p<0.01) was revealed. The restriction significantly decreased 5-HT level in the cortex (p<0.01) and increased 5-HIAA/5-HT rate (p<0.001) in the hypothalamus in B6-1473C mice, but not in B6-1473G mice. The present result is the first experimental evidence that C1473G polymorphism is involved in the regulation of the reaction to emotional stress in mice. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Serotonin passes through myoendothelial gap junctions to promote pulmonary arterial smooth muscle cell differentiation.

    PubMed

    Gairhe, Salina; Bauer, Natalie N; Gebb, Sarah A; McMurtry, Ivan F

    2012-11-01

    Myoendothelial gap junctional signaling mediates pulmonary arterial endothelial cell (PAEC)-induced activation of latent TGF-β and differentiation of cocultured pulmonary arterial smooth muscle cells (PASMCs), but the nature of the signal passing from PAECs to PASMCs through the gap junctions is unknown. Because PAECs but not PASMCs synthesize serotonin, and serotonin can pass through gap junctions, we hypothesized that the monoamine is the intercellular signal. We aimed to determine whether PAEC-derived serotonin mediates PAEC-induced myoendothelial gap junction-dependent activation of TGF-β signaling and differentiation of PASMCs. Rat PAECs and PASMCs were monocultured or cocultured with (touch) or without (no-touch) direct cell-cell contact. In all cases, tryptophan hydroxylase 1 (Tph1) transcripts were expressed predominantly in PAECs. Serotonin was detected by immunostaining in both PAECs and PASMCs in PAEC/PASMC touch coculture but was not found in PASMCs in either PAEC/PASMC no-touch coculture or in PASMC/PASMC touch coculture. Furthermore, inhibition of gap junctions but not of the serotonin transporter in PAEC/PASMC touch coculture prevented serotonin transfer from PAECs to PASMCs. Inhibition of serotonin synthesis pharmacologically or by small interfering RNAs to Tph1 in PAECs inhibited the PAEC-induced activation of TGF-β signaling and differentiation of PASMCs. We concluded that serotonin synthesized by PAECs is transferred through myoendothelial gap junctions into PASMCs, where it activates TGF-β signaling and induces a more differentiated phenotype. This finding suggests a novel role of gap junction-mediated intercellular serotonin signaling in regulation of PASMC phenotype.

  20. Synergistic effects of bioremediation and electrokinetics in the remediation of petroleum-contaminated soil.

    PubMed

    Guo, Shuhai; Fan, Ruijuan; Li, Tingting; Hartog, Niels; Li, Fengmei; Yang, Xuelian

    2014-08-01

    The present study evaluated the coupling interactions between bioremediation (BIO) and electrokinetics (EK) in the remediation of total petroleum hydrocarbons (TPH) by using bio-electrokinetics (BIO-EK) with a rotatory 2-D electric field. The results demonstrated an obvious positive correlation between the degradation extents of TPH and electric intensity both in the EK and BIO-EK tests. The use of BIO-EK showed a significant improvement in degradation of TPH as compared to BIO or EK alone. The actual degradation curve in BIO-EK tests fitted well with the simulated curve obtained by combining the degradation curves in BIO- and EK-only tests during the first 60 d, indicating a superimposed effect of biological degradation and electrochemical stimulation. The synergistic effect was particularly expressed during the later phase of the experiment, concurrent with changes in the microbial community structure. The community composition changed mainly according to the duration of the electric field, leading to a reduction in diversity. No significant spatial shifts in microbial community composition and bacterial numbers were detected among different sampling positions. Soil pH was uniform during the experimental process, soil temperature showed no variations between the soil chambers with and without an electric field. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Total petroleum hydrocarbon degradation by hybrid electrobiochemical reactor in oilfield produced water.

    PubMed

    Mousa, Ibrahim E

    2016-08-15

    The crude oil drilling and extraction operations are aimed to maximize the production may be counterbalanced by the huge production of contaminated produced water (PW). PW is conventionally treated through different physical, chemical, and biological technologies. The efficiency of suggested hybrid electrobiochemical (EBC) methods for the simultaneous removal of total petroleum hydrocarbon (TPH) and sulfate from PW generated by petroleum industry is studied. Also, the factors that affect the stability of PW quality are investigated. The results indicated that the effect of biological treatment is very important to keep control of the electrochemical by-products and more TPH removal in the EBC system. The maximum TPH and sulfate removal efficiency was achieved 75% and 25.3%, respectively when the detention time was about 5.1min and the energy consumption was 32.6mA/cm(2). However, a slight increasing in total bacterial count was observed when the EBC compact unit worked at a flow rate of average 20L/h. Pseudo steady state was achieved after 30min of current application in the solution. Also, the results of the study indicate that when the current intensity was increased above optimum level, no significant results occurred due to the release of gases. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Impact of Tributyltin and Triphenyltin on Ivory Shell (Babylonia japonica) Populations

    PubMed Central

    Horiguchi, Toshihiro; Kojima, Mitsuhiro; Hamada, Fumihiko; Kajikawa, Akira; Shiraishi, Hiroaki; Morita, Masatoshi; Shimizu, Makoto

    2006-01-01

    We histopathologically examined gonads and chemically determined organotin compounds in tissues of the ivory shell, Babylonia japonica. Imposex (a superimposition of male-type genital organs on females) occurred in approximately 80–90% of B. japonica specimens that we examined, with the penis and vas deferens both well developed. No oviduct blockage by vas deferens formation was observed. Ovarian spermatogenesis and suppressed ovarian maturation were observed in the females that exhibited imposex, although no histopathological abnormalities were found in males. Tissue distributions of organotin compounds [tributyltin (TBT), triphenyltin (TPhT), and their metabolites] were different for butyltins and phenyltins; a remarkably high accumulation of TBT was observed in the ctenidium, osphradium, and heart, whereas high concentrations of TPhT were detected in the ovary and digestive gland. More than one-third of TBT accumulated in the digestive glands of both males and females, followed by the testis, ctenidium, muscle, and heart tissues in males and in the muscle, ovary, ctenidium, and head tissues (including the central nervous system ganglia) in females. In both males and females, more than half of total TPhT accumulated in the digestive glands, followed by the gonads. The next highest values were in the muscle, ctenidium, and heart tissues in males and in the muscle, oviduct, and head tissues in females. Both TBT and TPhT concentrations in the gonads were positively correlated with penis length in females. Our findings strongly suggest that reproductive failure in adult females accompanied by imposex, possibly induced by TBT and TPhT from antifouling paints, may have caused the marked decline of B. japonica populations in Japan. PMID:16818241

  3. The effects of child maltreatment on early signs of antisocial behavior: Genetic moderation by Tryptophan Hydroxylase, Serotonin Transporter, and Monoamine Oxidase-A-Genes

    PubMed Central

    Cicchetti, Dante; Rogosch, Fred A.; Thibodeau, Eric

    2013-01-01

    Gene-environment interaction effects in predicting antisocial behavior in late childhood were investigated among maltreated and nonmaltreated low-income children (N = 627, M age = 11.27). Variants in three genes, TPH1, 5-HTTLPR, and MAOA uVNTR, were examined. In addition to child maltreatment status, we also considered the impact of maltreatment subtypes, developmental timing of maltreatment, and chronicity. Indicators of antisocial behavior were obtained from self-, peer-, and adult counselor-reports. In a series of ANCOVAs, child maltreatment and its parameters demonstrated strong main effects on early antisocial behavior as assessed by all forms of report. Genetic effects operated primarily in the context of gene-environment interactions, moderating the impact of child maltreatment on outcomes. Across the three genes, among nonmaltreated children no differences in antisocial behavior were found based on genetic variation. In contrast, among maltreated children specific polymorphisms of TPH1, 5-HTTLPR, and MAOA were each related to heightened self-report of antisocial behavior; the interaction of 5-HTTLPR and developmental timing of maltreatment also indicated more severe antisocial outcomes for children with early onset and recurrent maltreatment based on genotype. TPH1 and 5-HTTLPR interacted with maltreatment subtype to predict peer-report of antisocial behavior; genetic variation contributed to larger differences in antisocial behavior among abused children. TPH1 and 5-HTTLPR polymorphisms also moderated the effects of maltreatment subtype on adult report of antisocial behavior; again genetic effects were strongest for children who were abused. Additionally, TPH1 moderated the effect of developmental timing of maltreatment and chronicity on adult report of antisocial behavior. The findings elucidate how genetic variation contributes to identifying which maltreated children are most vulnerable to antisocial development. PMID:22781862

  4. 5' diversity of human hepatic PXR (NR1I2) transcripts and identification of the major transcription initiation site.

    PubMed

    Kurose, Kouichi; Koyano, Satoru; Ikeda, Shinobu; Tohkin, Masahiro; Hasegawa, Ryuichi; Sawada, Jun-Ichi

    2005-05-01

    The human pregnane X receptor (PXR) is a crucial regulator of the genes encoding several major cytochrome P450 enzymes and transporters, such as CYP3A4 and MDR1, but its own transcriptional regulation remains unclear. To elucidate the transcriptional mechanisms of human PXR gene, we first endeavored to identify the transcription initiation site of human PXR using 5'-RACE. Five types of 5'-variable transcripts (a, b, c, d, and e) with common exon 2 sequence were found, and comparison of these sequences with the genomic sequence suggested that their 5' diversity is derived from initiation by alternative promoters and alternative splicing. None of the exons found in our study contain any new in-frame coding regions. Newly identified introns IVS-a and IVS-b were found to have CT-AC splice sites that do not follow the GT-AG rule of conventional donor and acceptor splice sites. Of the five types of 5' variable transcripts identified, RT-PCR showed that type-a was the major transcript type. Four transcription initiation sites (A-D) for type-a transcript were identified by 5'-RACE using GeneRacer RACE Ready cDNA (human liver) constructed by the oligo-capping method. Putative TATA boxes were located approximately 30 bp upstream from the transcriptional start sites of the major transcript (C) and the longest minor transcript (A) expressed in the human liver. These results indicate that the initiation of transcription of human PXR is more complex than previously reported.

  5. Molecular genetics of the platelet serotonin system in first-degree relatives of patients with autism

    PubMed Central

    Cross, Sarah; Kim, Soo-Jeong; Weiss, Lauren A.; Delahanty, Ryan J.; Sutcliffe, James S.; Leventhal, Bennett L.; Cook, Edwin H.; Veenstra-VanderWeele, Jeremy

    2009-01-01

    Elevated platelet serotonin (5-HT) is found in a subset of children with autism and in some of their first-degree relatives. Indices of the platelet serotonin system, including whole blood serotonin (5-HT), 5-HT binding affinity for the serotonin transporter (Km), 5-HT uptake (Vmax), and lysergic acid diethylamide (LSD) receptor binding, were previously studied in twenty-four first-degree relatives of probands with autism, half of whom were selected for elevated whole blood 5-HT levels. All subjects were then genotyped for selected polymorphisms at the SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 loci. Previous studies allowed an a priori prediction of SLC6A4 haplotypes that separated the subjects into three groups that showed significantly different 5-HT binding affinity (Km, p = 0.005) and 5-HT uptake rate (Vmax, p = 0.046). Genotypes at four individual polymorphisms in SLC6A4 were not associated with platelet 5-HT indices. Haplotypes at SLC6A4 and individual genotypes of polymorphisms at SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 showed no significant association with whole blood 5-HT. Haplotype analysis of two polymorphisms in TPH1 revealed a nominally significant association with whole blood 5-HT (p = 0.046). These initial studies of indices of the 5-HT system with several SNPs at loci in this system generate hypotheses for testing in other samples. PMID:17406648

  6. Insights into microbial communities mediating the bioremediation of hydrocarbon-contaminated soil from an Alpine former military site.

    PubMed

    Siles, José A; Margesin, Rosa

    2018-05-01

    The study of microbial communities involved in soil bioremediation is important to identify the specific microbial characteristics that determine improved decontamination rates. Here, we characterized bacterial, archaeal, and fungal communities in terms of (i) abundance (using quantitative PCR) and (ii) taxonomic diversity and structure (using Illumina amplicon sequencing) during the bioremediation of long-term hydrocarbon-contaminated soil from an Alpine former military site during 15 weeks comparing biostimulation (inorganic NPK fertilization) vs. natural attenuation and considering the effect of temperature (10 vs. 20 °C). Although a considerable amount of total petroleum hydrocarbon (TPH) loss could be attributed to natural attenuation, significantly higher TPH removal rates were obtained with NPK fertilization and at increased temperature, which were related to the stimulation of the activities of indigenous soil microorganisms. Changing structures of bacterial and fungal communities significantly explained shifts in TPH contents in both natural attenuation and biostimulation treatments at 10 and 20 °C. However, archaeal communities, in general, and changing abundances and diversities in bacterial and fungal communities did not play a decisive role on the effectiveness of soil bioremediation. Gammaproteobacteria and Bacteroidia classes, within bacterial community, and undescribed/novel groups, within fungal community, proved to be actively involved in TPH removal in natural attenuation and biostimulation at both temperatures.

  7. Live fate-mapping of joint-associated fibroblasts visualizes expansion of cell contributions during zebrafish fin regeneration.

    PubMed

    Tornini, Valerie A; Thompson, John D; Allen, Raymond L; Poss, Kenneth D

    2017-08-15

    The blastema is a mass of progenitor cells responsible for regeneration of amputated salamander limbs and fish fins. Previous studies have indicated that resident cell sources producing the blastema contribute lineage-restricted progeny to regenerating tissue. However, these studies have labeled general cell types rather than granular cell subpopulations, and they do not explain the developmental transitions that must occur for distal structures to arise from cells with proximal identities in the appendage stump. Here, we find that regulatory sequences of tph1b , which encodes an enzyme that synthesizes serotonin, mark a subpopulation of fibroblast-like cells restricted to the joints of uninjured adult zebrafish fins. Amputation stimulates serotonin production in regenerating fin fibroblasts, yet targeted tph1b mutations abrogating this response do not disrupt fin regeneration. In uninjured animals, tph1b -expressing cells contribute fibroblast progeny that remain restricted to joints throughout life. By contrast, upon amputation, tph1b + joint cells give rise to fibroblasts that distribute across the entire lengths of regenerating fin rays. Our experiments visualize and quantify how incorporation into an appendage blastema broadens the progeny contributions of a cellular subpopulation that normally has proximodistal restrictions. © 2017. Published by The Company of Biologists Ltd.

  8. Altered serotonin physiology in human breast cancers favors paradoxical growth and cell survival

    PubMed Central

    2009-01-01

    Introduction The breast microenvironment can either retard or accelerate the events associated with progression of latent cancers. However, the actions of local physiological mediators in the context of breast cancers are poorly understood. Serotonin (5-HT) is a critical local regulator of epithelial homeostasis in the breast and other organs. Herein, we report complex alterations in the intrinsic mammary gland serotonin system of human breast cancers. Methods Serotonin biosynthetic capacity was analyzed in human breast tumor tissue microarrays using immunohistochemistry for tryptophan hydroxylase 1 (TPH1). Serotonin receptors (5-HT1-7) were analyzed in human breast tumors using the Oncomine database. Serotonin receptor expression, signal transduction, and 5-HT effects on breast cancer cell phenotype were compared in non-transformed and transformed human breast cells. Results In the context of the normal mammary gland, 5-HT acts as a physiological regulator of lactation and involution, in part by favoring growth arrest and cell death. This tightly regulated 5-HT system is subverted in multiple ways in human breast cancers. Specifically, TPH1 expression undergoes a non-linear change during progression, with increased expression during malignant progression. Correspondingly, the tightly regulated pattern of 5-HT receptors becomes dysregulated in human breast cancer cells, resulting in both ectopic expression of some isoforms and suppression of others. The receptor expression change is accompanied by altered downstream signaling of 5-HT receptors in human breast cancer cells, resulting in resistance to 5-HT-induced apoptosis, and stimulated proliferation. Conclusions Our data constitutes the first report of direct involvement of 5-HT in human breast cancer. Increased 5-HT biosynthetic capacity accompanied by multiple changes in 5-HT receptor expression and signaling favor malignant progression of human breast cancer cells (for example, stimulated proliferation, inappropriate cell survival). This occurs through uncoupling of serotonin from the homeostatic regulatory mechanisms of the normal mammary epithelium. The findings open a new avenue for identification of diagnostic and prognostic markers, and valuable new therapeutic targets for managing breast cancer. PMID:19903352

  9. Response of soil microorganisms to radioactive oil waste: results from a leaching experiment

    NASA Astrophysics Data System (ADS)

    Galitskaya, P.; Biktasheva, L.; Saveliev, A.; Ratering, S.; Schnell, S.; Selivanovskaya, S.

    2015-01-01

    Oil wastes produced in large amounts in the processes of oil extraction, refining, and transportation are of great environmental concern because of their mutagenicity, toxicity, high fire hazardousness, and other properties. About 40% of these wastes contain radionuclides; however, the effects of oil products and radionuclides on soil microorganisms are frequently studied separately. The toxicity and effects on various microbial parameters of raw waste (H) containing 575 g of total petroleum hydrocarbons (TPH) kg-1 waste, 4.4 kBq kg-1 of 226Ra, 2.8 kBq kg-1 of 232Th, and 1.3 kBq kg-1 of 40K and its treated variant (R) (1.6 g kg-1 of TPH, 7.9 kBq kg-1 of 226Ra, 3.9 kBq kg-1 of 232Th, and 183 kBq kg-1 of 40K) were estimated in a leaching column experiment to separate the effects of hydrocarbons from those of radioactive elements. The disposal of H waste samples on the soil surface led to an increase of the TPH content in soil: it became 3.5, 2.8, and 2.2 times higher in the upper (0-20 cm), middle (20-40 cm), and lower (40-60 cm) layers respectively. Activity concentrations of 226Ra and 232Th increased in soil sampled from both H- and R-columns in comparison to their concentrations in control soil. The activity concentrations of these two elements in samples taken from the upper and middle layers were much higher for the R-column compared to the H-column, despite the fact that the amount of waste added to the columns was equalized with respect to the activity concentrations of radionuclides. The H waste containing both TPH and radionuclides affected the functioning of the soil microbial community, and the effect was more pronounced in the upper layer of the column. Metabolic quotient and cellulase activity were the most sensitive microbial parameters as their levels were changed 5-1.4 times in comparison to control ones. Changes of soil functional characteristics caused by the treated waste containing mainly radionuclides were not observed. PCR-SSCP (polymerase chain reaction - single strand conformation polymorphism) analysis followed by MDS (metric multidimensional scaling) and clustering analysis revealed that the shifts in microbial community structure were affected by both hydrocarbons and radioactivity.

  10. Biodegradation of effluent contaminated with diesel fuel and gasoline.

    PubMed

    Vieira, P A; Vieira, R B; de França, F P; Cardoso, V L

    2007-02-09

    We studied the effects of fuel concentration (diesel and gasoline), nitrogen concentration and culture type on the biodegradation of synthetic effluent similar to what was found at inland fuel distribution terminals. An experimental design with two levels and three variables (2(3)) was used. The mixed cultures used in this study were obtained from lake with a history of petroleum contamination and were named culture C(1) (collected from surface sediment) and C(2) (collected from a depth of approximately 30cm). Of the parameters studied, the ones that had the greatest influence on the removal of total petroleum hydrocarbons (TPH) were a nitrogen concentration of 550mg/L and a fuel concentration of 4% (v/v) in the presence of culture C(1). The biodegradability study showed a TPH removal of 90+/-2% over a process period of 49 days. Analysis using gas chromatography identified 16 hydrocarbons. The aromatic compounds did not degrade as readily as the other hydrocarbons that were identified.

  11. Mice genetically depleted of brain serotonin display social impairments, communication deficits and repetitive behaviors: possible relevance to autism.

    PubMed

    Kane, Michael J; Angoa-Peréz, Mariana; Briggs, Denise I; Sykes, Catherine E; Francescutti, Dina M; Rosenberg, David R; Kuhn, Donald M

    2012-01-01

    Autism is a complex neurodevelopmental disorder characterized by impaired reciprocal social interaction, communication deficits and repetitive behaviors. A very large number of genes have been linked to autism, many of which encode proteins involved in the development and function of synaptic circuitry. However, the manner in which these mutated genes might participate, either individually or together, to cause autism is not understood. One factor known to exert extremely broad influence on brain development and network formation, and which has been linked to autism, is the neurotransmitter serotonin. Unfortunately, very little is known about how alterations in serotonin neuronal function might contribute to autism. To test the hypothesis that serotonin dysfunction can contribute to the core symptoms of autism, we analyzed mice lacking brain serotonin (via a null mutation in the gene for tryptophan hydroxylase 2 (TPH2)) for behaviors that are relevant to this disorder. Mice lacking brain serotonin (TPH2-/-) showed substantial deficits in numerous validated tests of social interaction and communication. These mice also display highly repetitive and compulsive behaviors. Newborn TPH2-/- mutant mice show delays in the expression of key developmental milestones and their diminished preference for maternal scents over the scent of an unrelated female is a forerunner of more severe socialization deficits that emerge in weanlings and persist into adulthood. Taken together, these results indicate that a hypo-serotonin condition can lead to behavioral traits that are highly characteristic of autism. Our findings should stimulate new studies that focus on determining how brain hyposerotonemia during critical neurodevelopmental periods can alter the maturation of synaptic circuits known to be mis-wired in autism and how prevention of such deficits might prevent this disorder.

  12. Water-soluble organic carbon, dicarboxylic acids, ketoacids, and α-dicarbonyls in the tropical Indian aerosols

    NASA Astrophysics Data System (ADS)

    Pavuluri, Chandra Mouli; Kawamura, Kimitaka; Swaminathan, T.

    2010-06-01

    Tropical aerosol (PM10) samples (n = 49) collected from southeast coast of India were studied for water-soluble dicarboxylic acids (C2-C12), ketocarboxylic acids (C2-C9), and α-dicarbonyls (glyoxal and methylglyoxal), together with analyses of total carbon (TC) and water-soluble organic carbon (WSOC). Their distributions were characterized by a predominance of oxalic acid followed by terephthalic (t-Ph), malonic, and succinic acids. Total concentrations of diacids (227-1030 ng m-3), ketoacids (16-105 ng m-3), and dicarbonyls (4-23 ng m-3) are comparative to those from other Asian megacities such as Tokyo and Hong Kong. t-Ph acid was found as the second most abundant diacid in the Chennai aerosols. This feature has not been reported previously in atmospheric aerosols. t-Ph acid is most likely derived from the field burning of plastics. Water-soluble diacids were found to contribute 0.4%-3% of TC and 4%-11% of WSOC. Based on molecular distributions and backward air mass trajectories, we found that diacids and related compounds in coastal South Indian aerosols are influenced by South Asian and Indian Ocean monsoons. Organic aerosols are also suggested to be significantly transported long distances from North India and the Middle East in early winter and from Southeast Asia in late winter, but some originate from photochemical reactions over the Bay of Bengal. In contrast, the Arabian Sea, Indian Ocean, and South Indian continent are suggested as major source regions in summer. We also found daytime maxima of most diacids, except for C9 and t-Ph acids, which showed nighttime maxima in summer. Emissions from marine and terrestrial plants, combined with land/sea breezes and in situ photochemical oxidation, are suggested especially in summer as an important factor that controls the composition of water-soluble organic aerosols over the southeast coast of India. Regional emissions from anthropogenic sources are also important in megacity Chennai, but their influence is weakened due to the dispersion caused by dynamic land/sea breeze on the coast.

  13. San Francisco Deep Ocean Dredged Material Disposal Site (SF-DODS) Monitoring Program. Physical, Chemical, and Benthic Community Monitoring

    DTIC Science & Technology

    2003-08-29

    analyzed for total volatile solids, total organic carbon, oil and grease/total petroleum hydrocarbons , grain size distribution, metals, polycyclic...TBT Tri-Butyltin TOC Total Organic Carbon TPCB Total Polychlorinated Biphenyls TPH Total Petroleum Hydrocarbons USACE U.S. Army Corps of Engineers U.S...Health PQL Practical Quantitation Limit RCRA Resource Conservation and Recovery Act SIM Selected Ion Monitoring TPH Total Petroleum Hydrocarbons tr Trace

  14. Effects of chemical additives on hydrocarbon disappearance and biodegradation in freshwater marsh microcosms.

    PubMed

    Nyman, J A; Klerks, P L; Bhattacharyya, S

    2007-09-01

    We determined how a cleaner and a dispersant affected hydrocarbon biodegradation in wetland soils dominated by the plant Panicum hemitomon, which occurs throughout North and South America. Microcosms received no hydrocarbons, South Louisiana crude, or diesel; and no additive, a dispersant, or a cleaner. We determined the concentration of four total petroleum hydrocarbon (TPH) measures and 43 target hydrocarbons in water and sediment fractions 1, 7, 31, and 186 days later. Disappearance was distinguished from biodegradation via hopane-normalization. After 186 days, TPH disappearance ranged from 24% to 97%. There was poor correlation among the four TPH measures, which indicated that each quantified a different suite of hydrocarbons. Hydrocarbon disappearance and biodegradation were unaltered by these additives under worse-case scenarios. Any use of these additives must generate benefits that outweigh the lack of effect on biodegradation demonstrated in this report, and the increase in toxicity that we reported earlier.

  15. Degradation of Total Petroleum Hydrocarbon (TPH) in Contaminated Soil Using Bacillus pumilus MVSV3.

    PubMed

    Varma, Surendra Sheeba; Lakshmi, Mahalingam Brinda; Rajagopal, Perumalsam; Velan, Manickam

    2017-01-01

     A study on bioremediation of soil contaminated with petroleum sludge was performed using Bacillus pumilus/MVSV3 (Accession number JN089707). In this study, 5 kg of agricultural soil was mixed well with 5% oil sludge and fertilizers containing nitrogen, phosphorus and potassium (N:P:K). The treatment resulted in 97% removal of total petroleum hydrocarbon (TPH) in 122 d in bacteria mixed contaminated soil when compared to 12% removal of TPH in uninoculated contaminated soil. The population of the microorganism remained stable after introduced into the oil environment. The physical and chemical parameters of the soil mixed with sludge showed variation indicating improvement and the pH level decreased during the experiment period. Elemental analysis and Gas Chromatography-Mass Spectroscopy (GC-MS) analysis revealed the bacterial ability to degrade oil sludge components. Growth experiments with Trigonellafoenumgraecum (Fenugreek) showed the applicability of bioremediated soil for the production.

  16. Temporal and spatial trends of total petroleum hydrocarbons and heavy metals in the surface sediment of Caofeidian Sea Area, China from 2011 to 2016

    NASA Astrophysics Data System (ADS)

    Huang, Wei

    2018-05-01

    The temporal and spatial distribution of total petroleum hydrocarbons (TPH) and four heavy metals in the surface sediments of Caofeidian Sea Area during 2011–2016 was investigated. The sediment concentration of TPH, Cu, Zn, Pb and Cd were 10.07-186.4 mg/L, 16.5-84.9 mg/L, 11.1-135 mg/L, 6.8-24.6 mg/L, and 0.07-0.199 mg/L, respectively. The pollution level in Caofeidian sea area is lower than those in other area in China. These results reached the highest marine sediment quality standards in China, indicating that the sediment was fairly clean. In addition, TPH at all stations decreased during 2011-2016. The highest values obtained were at stations near the port areas and estuary region.

  17. Stress-Induced Depression Is Alleviated by Aerobic Exercise Through Up-Regulation of 5-Hydroxytryptamine 1A Receptors in Rats

    PubMed Central

    Kim, Tae Woon; Lim, Baek Vin; Baek, Dongjin; Ryu, Dong-Soo; Seo, Jin Hee

    2015-01-01

    Purpose: Stress is associated with depression, which induces many psychiatric disorders. Serotonin, also known as 5-hydroxy-tryptamine (5-HT), acts as a biochemical messenger and regulator in the brain. It also mediates several important physiological functions. Depression is closely associated with an overactive bladder. In the present study, we investigated the effect of treadmill exercise on stress-induced depression while focusing on the expression of 5-HT 1A (5-H1A) receptors in the dorsal raphe. Methods: Stress was induced by applying a 0.2-mA electric foot shock to rats. Each set of electric foot shocks comprised a 6-second shock duration that was repeated 10 times with a 30-second interval. Three sets of electric foot shocks were applied each day for 7 days. For the confirmation of depressive state, a forced swimming test was performed. To visualize the expression of 5-HT and tryptophan hydroxylase (TPH), immunohistochemistry for 5-HT and TPH in the dorsal raphe was performed. Expression of 5-H1A receptors was determined by western blot analysis. Results: A depressive state was induced by stress, and treadmill exercise alleviated the depression symptoms in the stress-induced rats. Expressions of 5-HT, TPH, and HT 1A in the dorsal raphe were reduced by the induction of stress. Treadmill exercise increased 5-HT, TPH, and HT 1A expressions in the stress-induced rats. Conclusions: Treadmill exercise enhanced 5-HT synthesis through the up-regulation of 5-HT1A receptors, and improved the stress-induced depression. In the present study, treadmill exercise improved depression symptoms by enhancing 5-HT1A receptor expression. The present results suggest that treadmill exercise might be helpful for the alleviation of overactive bladder and improve sexual function. PMID:25833478

  18. Organotins in North Sea brown shrimp (Crangon crangon L.) after implementation of the TBT ban.

    PubMed

    Verhaegen, Y; Monteyne, E; Neudecker, T; Tulp, I; Smagghe, G; Cooreman, K; Roose, P; Parmentier, K

    2012-03-01

    The organotin (OT) compounds tributyltin (TBT) and triphenyltin (TPhT) are potent biocides that have been used ubiquitously in antifouling paints and pesticides since the mid-1970s. These biocides are extremely toxic to marine life, particularly marine gastropod populations. The European Union therefore took measures to reduce the use of TBT-based antifouling paints on ships and ultimately banned these paints in 2003. Despite sufficient data on OT concentrations in marine gastropods, data are scarce for other species such as the North Sea brown shrimp (Crangon crangon), a dominant crustacean species in North Sea inshore benthic communities. The present study provides the first spatial overview of OT concentrations in North Sea brown shrimp. We have compared these data with historical concentrations in shrimp as well as with sediment concentrations. We have also addressed the effect on the shrimp stock and any human health risks associated with the OT concentrations found. TBT and TPhT in shrimp tail muscle ranged from 4 to 124 and from 1 to 24 μg kg(-1) DW, respectively. High levels are accumulated in estuarine areas and are clearly related with sediment concentrations (biota-sediment accumulation factor ~10). Levels have decreased approximately 10-fold since the ban took effect, coinciding with a recovery of the shrimp stock after 30 years of gradual regression. Furthermore, the OT levels found in brown shrimp no longer present a human health risk. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. Simple surface foam application enhances bioremediation of oil-contaminated soil in cold conditions.

    PubMed

    Jeong, Seung-Woo; Jeong, Jongshin; Kim, Jaisoo

    2015-04-09

    Landfarming of oil-contaminated soil is ineffective at low temperatures, because the number and activity of micro-organisms declines. This study presents a simple and versatile technique for bioremediation of diesel-contaminated soil, which involves spraying foam on the soil surface without additional works such as tilling, or supply of water and air. Surfactant foam containing psychrophilic oil-degrading microbes and nutrients was sprayed twice daily over diesel-contaminated soil at 6 °C. Removal efficiencies in total petroleum hydrocarbon (TPH) at 30 days were 46.3% for landfarming and 73.7% for foam-spraying. The first-order kinetic biodegradation rates for landfarming and foam-spraying were calculated as 0.019 d(-1) and 0.044 d(-1), respectively. Foam acted as an insulating medium, keeping the soil 2 °C warmer than ambient air. Sprayed foam was slowly converted to aqueous solution within 10-12h and infiltrated the soil, providing microbes, nutrients, water, and air for bioaugmentation. Furthermore, surfactant present in the aqueous solution accelerated the dissolution of oil from the soil, resulting in readily biodegradable aqueous form. Significant reductions in hydrocarbon concentration were simultaneously observed in both semi-volatile and non-volatile fractions. As the initial soil TPH concentration increased, the TPH removal rate of the foam-spraying method also increased. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Comparative bioremediation of soils contaminated with diesel oil by natural attenuation, biostimulation and bioaugmentation.

    PubMed

    Bento, Fatima M; Camargo, Flávio A O; Okeke, Benedict C; Frankenberger, William T

    2005-06-01

    Bioremediation of diesel oil in soil can occur by natural attenuation, or treated by biostimulation or bioaugmentation. In this study we evaluated all three technologies on the degradation of total petroleum hydrocarbons (TPH) in soil. In addition, the number of diesel-degrading microorganisms present and microbial activity as indexed by the dehydrogenase assay were monitored. Soils contaminated with diesel oil in the field were collected from Long Beach, California, USA and Hong Kong, China. After 12 weeks of incubation, all three treatments showed differing effects on the degradation of light (C12-C23) and heavy (C23-C40) fractions of TPH in the soil samples. Bioaugmentation of the Long Beach soil showed the greatest degradation in the light (72.7%) and heavy (75.2%) fractions of TPH. Natural attenuation was more effective than biostimulation (addition of nutrients), most notably in the Hong Kong soil. The greatest microbial activity (dehydrogenase activity) was observed with bioaugmentation of the Long Beach soil (3.3-fold) and upon natural attenuation of the Hong Kong sample (4.0-fold). The number of diesel-degrading microorganisms and heterotrophic population was not influenced by the bioremediation treatments. Soil properties and the indigenous soil microbial population affect the degree of biodegradation; hence detailed site specific characterization studies are needed prior to deciding on the proper bioremediation method.

  1. Air National Guard Installation Restoration Protram. Site Investigation Report: Georgia Air National Guard, Savannah, Georgia

    DTIC Science & Technology

    1992-01-01

    except TPH, which was detected at 0.06 mg/l in Monitor Well 01-MW-02. Some metals (arsenic, cadmium , chromium, lead, silver, and zinc ) were detected at...extraction. Trace quantities of some priority pollutant metals were detected in the surface water samples. Arsenic, cadmium , and zinc were detected at...storage tank. TPH was detected in all five groundwater samples. Arsenic, beryllium, cadmium , chromium, copper, lead, nickel, silver, and zinc were also

  2. Feasibility of oxidation-biodegradation serial foam spraying for total petroleum hydrocarbon removal without soil disturbance.

    PubMed

    Bajagain, Rishikesh; Park, Yoonsu; Jeong, Seung-Woo

    2018-06-01

    This study evaluated surface foam spraying technology, which avoids disturbing the soil, to deliver chemical oxidant and oil-degrading microbes to unsaturated soil for 30 days. Hydrogen peroxide foam was sprayed once onto diesel contaminated soil for oxidation of soil total petroleum hydrocarbon (TPH). Periodic bioaugmentation foam was sprayed every three days for biodegradation of soil TPH. Foam spraying employing oxidation-bioaugmentation serial application significantly reduced soil TPH concentrations to 550 mg·kg -1 from an initial 7470 mg·kg -1 . This study selected an optimal hydrogen peroxide concentration of 5%, which is capable of treating diesel oil contaminated soil following biodegradation without supplementary iron. Application of hydrogen peroxide by foam spraying increased the infiltration of hydrogen peroxide into the unsaturated soil. Surface foam spraying provided the aqueous phase of remediation agents evenly to the unsaturated soil and resulted in relatively similar soil water content throughout the soil. The easy and even infiltration of remediation reagents increased their contact with contaminants, resulting in enhanced oxidation and biodegradation. Fractional analysis of TPH showed C18-C22 present in diesel as biodegradation recalcitrant hydrocarbons. Recalcitrant hydrocarbons were reduced by 92% using oxidation-biodegradation serial foam, while biodegradation alone only reduced the recalcitrant fraction by 25%. Copyright © 2018 Elsevier B.V. All rights reserved.

  3. Serotonin transporter protects the placental cells against apoptosis in caspase 3-independent pathway.

    PubMed

    Hadden, Coedy; Fahmi, Tariq; Cooper, Anthonya; Savenka, Alena V; Lupashin, Vladimir V; Roberts, Drucilla J; Maroteaux, Luc; Hauguel-de Mouzon, Sylvie; Kilic, Fusun

    2017-12-01

    Serotonin (5-HT) and its specific transporter, SERT play important roles in pregnancy. Using placentas dissected from 18d gestational SERT-knock out (KO), peripheral 5-HT (TPH1)-KO, and wild-type (WT) mice, we explored the role of 5-HT and SERT in placental functions in detail. An abnormal thick band of fibrosis and necrosis under the giant cell layer in SERT-KO placentas appeared only moderately in TPH1-KO and minimally present in WT placentas. The majority of the changes were located at the junctional zone of the placentas in SERT. The etiology of these findings was tested with TUNEL assays. The placentas from SERT-KO and TPH1-KO showed 49- and 8-fold increase in TUNEL-positive cells without a concurrent change in the DNA repair or cell proliferation compared to WT placentas. While the proliferation rate in the embryos of TPH1-KO mice was 16-fold lower than the rate in gestational age matched embryos of WT or SERT-KO mice. These findings highlight an important role of continuous 5-HT signaling on trophoblast cell viability. SERT may contribute to protecting trophoblast cells against cell death via terminating the 5-HT signaling which changes cell death ratio in trophoblast as well as proliferation rate in embryos. However, the cell death in SERT-KO placentas is in caspase 3-independent pathway. © 2017 Wiley Periodicals, Inc.

  4. Bioremediation of petroleum-contaminated soil using aged refuse from landfills.

    PubMed

    Liu, Qingmei; Li, Qibin; Wang, Ning; Liu, Dan; Zan, Li; Chang, Le; Gou, Xuemei; Wang, Peijin

    2018-05-10

    This study explored the effects and mechanisms of petroleum-contaminated soil bioremediation using aged refuse (AR) from landfills. Three treatments of petroleum-contaminated soil (47.28 mg·g -1 ) amended with AR, sterilized aged refuse (SAR) and petroleum-contaminated soil only (as a control) were tested. During 98 days of incubation, changes in soil physicochemical properties, residual total petroleum hydrocarbon (TPH), biodegradation kinetics, enzyme activities and the microbial community were investigated. The results demonstrated that AR was an effective soil conditioner and biostimulation agent that could comprehensively improve the quality of petroleum-contaminated soil and promote microbial growth, with an 74.64% TPH removal rate, 22.36 day half-life for SAR treatment, compared with the control (half-life: 138.63 days; TPH removal rate: 22.40%). In addition, the petroleum-degrading bacteria isolation results demonstrated that AR was also a petroleum-degrading microbial agent containing abundant microorganisms. AR addition significantly improved both the biotic and abiotic conditions of petroleum-contaminated soil without other additives. The cooperation of conditioner addition, biostimulation and bioaugmentation in AR treatment led to better bioremediation effects (half-life: 13.86 days; TPH removal rate: 89.83%). In conclusion, AR amendment is a cost-effective, easy-to-use method facilitating in situ large-scale application while simultaneously recycling huge amounts of AR from landfills. Copyright © 2018 Elsevier Ltd. All rights reserved.

  5. A field trial for an ex-situ bioremediation of a drilling mud-polluted site.

    PubMed

    Rojas-Avelizapa, N G; Roldán-Carrillo, T; Zegarra-Martínez, H; Muñoz-Colunga, A M; Fernández-Linares, L C

    2007-01-01

    The remediation of drilling mud-polluted sites in the Southeast of Mexico is a top priority for Mexican oil industry. The objective of this work was to find a technology to remediate these sites. A field trial was performed by composting in biopiles, where four 1ton soil-biopiles were established, one treatment in triplicate and one unamended biopile. Amended biopiles were added with nutrients to get a C/N/P ratio of 100/3/0.5 plus a bulking agent (straw) at a soil/straw ratio of 97/3. Moisture content was maintained around 30-35%. Results showed that, after 180 d, total petroleum hydrocarbon (TPH) concentrations decreased from 99300+/-23000mgTPHkg(-1) soil to 5500+/-770mgTPHkg(-1) for amended biopiles and to 22900+/-7800mgTPHkg(-1) for unamended biopile. An undisturbed soil control showed no change in TPH concentrations. Gas chromatographic analysis showed residual alkyl dibenzothiophene type compounds. Highest bacterial counts were observed during the first 30 d which correlated with highest TPH removal, whereas fungal count increased at the end of the experimentation period. Results suggested an important role of the straw, nutrient addition and water content in stimulating aerobic microbial activity and thus hydrocarbon removal. This finding opens an opportunity to remediate old polluted sites with recalcitrant and high TPH concentration.

  6. Rehabilitation of Seven (7) Hydrocarbon Contaminated Sites in a Brackish Water/Lagoon Environment in South Trinidad

    NASA Astrophysics Data System (ADS)

    Mohammed, Avryl; Ramnath, Kelvin; Dyal, Shyam; Lalla, Francesca; Roopchand, Jaipersad

    2007-12-01

    The Petroleum Company of Trinidad and Tobago Limited operates in a wide diversity of tropical habitats in South Trinidad one of which is a brackish water environment known as the Godineau Swamp. Historically this field was operated by predecessor multinational companies, who at that time employed operational practices based on the absence of legal requirements, that were not environmentally considerate. Following a detailed environmental audit of the field (also known as the Oropouche Field), seven (7) contaminated sites were found, that presented a risk to the lagoon and its associated mangrove swamp ecology. Remediation of the seven (7) sites was done in two (2) phases; phase 1 being sampling and characterization of the waste inclusive of migration and phase 2 the actual on-site soil remediation. Phase 1 conducted during the period December 2004 to February 2005, indicated a total of 19,484 m3 of contaminated material with TPH being the main contaminant. The average concentration of TPH was 3.25%. Phase 2 remediation was initiated in October 2005 and involved the following three (3) aspects to achieve a TPH concentration of less than 1%: ▪ Preparation of waste remediation sites adjacent to contaminated sites and excavation and spreading onto cells ▪ Bioremediation onsite using naturally occurring bacteria and rototilling ▪ Rehabilitation and closure of the site following accepted lab results. The benefits of conducting this project in the petroleum industry are to ensure compliance to the national Sensitive Areas Rules and Draft Waste Management Rules, conformance to ISO 14001 Certification requirements and conservation of biodiversity in the mangrove swamp.

  7. Expression of cerebral serotonin related to anxiety-like behaviors in C57BL/6 offspring induced by repeated subcutaneous prenatal exposure to low-dose lipopolysaccharide

    PubMed Central

    Hsueh, Pei-Tan; Wang, Hsuan-Han; Liu, Chiu-Lin; Ni, Wei-Fen

    2017-01-01

    Prenatal exposure to lipopolysaccharide (LPS), which likely occurs due to infection or contact with environmental allergens during pregnancy, is a proposed risk factor that induces anxiety- and autism spectrum disorder-like behaviors in offspring. However, the molecular and behavioral changes in offspring after maternal immune activation have not been completely identified. We hypothesized that a subcutaneous injection of LPS in a pregnant mouse would induce changes in cerebral serotonin (5-HT) in parallel to the appearance of anxiety-like behaviors in the dam’s offspring. After LPS injections (total, 100 μg/Kg), the time spent in the central region during the open field test and the number of times that the mice moved between the light and dark boxes and between the open and closed arms on the elevated plus maze test revealed anxiety-like behaviors in offspring at 5, 6 and 9 weeks of age. The mRNA expression levels of tph2 (5-HT synthesizing enzyme) and slc6a4 (5-HT transporter) were down-regulated in both adolescent (5 weeks of age) and adult (8 weeks of age) brains. Immunohistochemistry revealed that the numbers and sizes of tph2-expressing cells were notably decreased in the raphe nuclei of the midbrain of adults. Moreover, compared with controls (phosphate-buffered saline-treated offspring), the cerebral 5-HT concentration at adolescence and adulthood in LPS-induced offspring was significantly decreased. We concluded that maternal immune activation induced by exposure to a low dose of LPS decreased cerebral 5-HT levels in parallel to the down-regulation of the tph2 and slc6a4 genes and in conjunction with anxiety-like behaviors in offspring. PMID:28650979

  8. Genetic and Non-genetic Factors Associated With Constipation in Cancer Patients Receiving Opioids.

    PubMed

    Laugsand, Eivor A; Skorpen, Frank; Kaasa, Stein; Sabatowski, Rainer; Strasser, Florian; Fayers, Peter; Klepstad, Pål

    2015-06-18

    To examine whether the inter-individual variation in constipation among patients receiving opioids for cancer pain is associated with genetic or non-genetic factors. Cancer patients receiving opioids were included from 17 centers in 11 European countries. Intensity of constipation was reported by 1,568 patients on a four-point categorical scale. Non-genetic factors were included as covariates in stratified regression analyses on the association between constipation and 75 single-nucleotide polymorphisms (SNPs) within 15 candidate genes related to opioid- or constipation-signaling pathways (HTR3E, HTR4, HTR2A, TPH1, ADRA2A, CHRM3, TACR1, CCKAR, KIT, ARRB2, GHRL, ABCB1, COMT, OPRM1, and OPRD1). The non-genetic factors significantly associated with constipation were type of laxative, mobility and place of care among patients receiving laxatives (N=806), in addition to Karnofsky performance status and presence of metastases among patients not receiving laxatives (N=762) (P<0.01). Age, gender, body mass index, cancer diagnosis, time on opioids, opioid dose, and type of opioid did not contribute to the inter-individual differences in constipation. Five SNPs, rs1800532 in TPH1, rs1799971 in OPRM1, rs4437575 in ABCB1, rs10802789 in CHRM3, and rs2020917 in COMT were associated with constipation (P<0.01). Only rs2020917 in COMT passed the Benjamini-Hochberg criterion for a 10% false discovery rate. Type of laxative, mobility, hospitalization, Karnofsky performance status, presence of metastases, and five SNPs within TPH1, OPRM1, ABCB1, CHRM3, and COMT may contribute to the variability in constipation among cancer patients treated with opioids. Knowledge of these factors may help to develop new therapies and to identify patients needing a more individualized approach to treatment.

  9. Molecular genetics of the platelet serotonin system in first-degree relatives of patients with autism.

    PubMed

    Cross, Sarah; Kim, Soo-Jeong; Weiss, Lauren A; Delahanty, Ryan J; Sutcliffe, James S; Leventhal, Bennett L; Cook, Edwin H; Veenstra-Vanderweele, Jeremy

    2008-01-01

    Elevated platelet serotonin (5-hydroxytryptamine, 5-HT) is found in a subset of children with autism and in some of their first-degree relatives. Indices of the platelet serotonin system, including whole blood 5-HT, 5-HT binding affinity for the serotonin transporter (K(m)), 5-HT uptake (V(max)), and lysergic acid diethylamide (LSD) receptor binding, were previously studied in 24 first-degree relatives of probands with autism, half of whom were selected for elevated whole blood 5-HT levels. All subjects were then genotyped for selected polymorphisms at the SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 loci. Previous studies allowed an a priori prediction of SLC6A4 haplotypes that separated the subjects into three groups that showed significantly different 5-HT binding affinity (K(m), p=0.005) and 5-HT uptake rate (V(max), p=0.046). Genotypes at four individual polymorphisms in SLC6A4 were not associated with platelet 5-HT indices. Haplotypes at SLC6A4 and individual genotypes of polymorphisms at SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 showed no significant association with whole blood 5-HT. Haplotype analysis of two polymorphisms in TPH1 revealed a nominally significant association with whole blood 5-HT (p=0.046). These initial studies of indices of the 5-HT system with several single-nucleotide polymorphisms at loci in this system generate hypotheses for testing in other samples.

  10. Effect of sex steroid hormones on the number of serotonergic neurons in rat dorsal raphe nucleus.

    PubMed

    Kunimura, Yuyu; Iwata, Kinuyo; Iijima, Norio; Kobayashi, Makito; Ozawa, Hitoshi

    2015-05-06

    Disorders caused by the malfunction of the serotonergic system in the central nervous system show sex-specific prevalence. Many studies have reported a relationship between sex steroid hormones and the brain serotonergic system; however, the interaction between sex steroid hormones and the number of brain neurons expressing serotonin has not yet been elucidated. In the present study, we determined whether sex steroid hormones altered the number of serotonergic neurons in the dorsal raphe nucleus (DR) of adult rat brains. Animals were divided into five groups: ovariectomized (OVX), OVX+low estradiol (E2), OVX+high E2, castrated males, and intact males. Antibodies against 5-hydroxytryptamine (5-HT, serotonin) and tryptophan hydroxylase (Tph), an enzyme for 5-HT synthesis, were used as markers of 5-HT neurons, and the number of 5-HT-immunoreactive (ir) or Tph-ir cells was counted. We detected no significant differences in the number of 5-HT-ir or Tph-ir cells in the DR among the five groups. By contrast, the intensity of 5-HT-ir showed significant sex differences in specific subregions of the DR independent of sex steroid levels, suggesting that the manipulation of sex steroid hormones after maturation does not affect the number and intensive immunostaining of serotonergic neurons in rat brain. Our results suggest that, the sexual dimorphism observed in the serotonergic system is due to factors such as 5-HT synthesis, transportation, and degradation but not to the number of serotonergic neurons. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  11. Characterization of herpes simplex virus type 2 latency-associated transcription in human sacral ganglia and in cell culture.

    PubMed

    Croen, K D; Ostrove, J M; Dragovic, L; Straus, S E

    1991-01-01

    The ability of herpes simplex virus type 2 (HSV-2) to establish latency in and reactivate from sacral dorsal root sensory ganglia is the basis for recurrent genital herpes. The expression of HSV-2 genes in latently infected human sacral ganglia was investigated by in situ hybridization. Hybridizations with a probe from the long repeat region of HSV-2 revealed strong nuclear signals overlying neurons in sacral ganglia from five of nine individuals. The RNA detected overlaps with the transcript for infected cell protein O but in the opposite, or "anti-sense," orientation. These observations mimic those made previously with HSV-1 in human trigeminal ganglia and confirm the recent findings during latency in HSV-2-infected mice and guinea pigs. Northern hybridization of RNA from infected Vero cells showed that an HSV-2 latency-associated transcript was similar in size to the larger (1.85 kb) latency transcript of HSV-1. Thus, HSV-1 and HSV-2 latency in human sensory ganglia are similar, if not identical, in terms of their cellular localization and pattern of transcription.

  12. NAD+ protects against EAE by regulating CD4+ T-cell differentiation

    PubMed Central

    Tullius, Stefan G.; Biefer, Hector Rodriguez Cetina; Li, Suyan; Trachtenberg, Alexander J.; Edtinger, Karoline; Quante, Markus; Krenzien, Felix; Uehara, Hirofumi; Yang, Xiaoyong; Kissick, Haydn T.; Kuo, Winston P.; Ghiran, Ionita; de la Fuente, Miguel A.; Arredouani, Mohamed S.; Camacho, Virginia; Tigges, John C.; Toxavidis, Vasilis; El Fatimy, Rachid; Smith, Brian D.; Vasudevan, Anju; ElKhal, Abdallah

    2014-01-01

    CD4+ T cells are involved in the development of autoimmunity, including multiple sclerosis (MS). Here we show that nicotinamide adenine dinucleotide (NAD+) blocks experimental autoimmune encephalomyelitis (EAE), a mouse model of MS, by inducing immune homeostasis through CD4+IFNγ+IL-10+ T cells and reverses disease progression by restoring tissue integrity via remyelination and neuroregeneration. We show that NAD+ regulates CD4+ T-cell differentiation through tryptophan hydroxylase-1 (Tph1), independently of well-established transcription factors. In the presence of NAD+, the frequency of T-bet−/− CD4+IFNγ+ T cells was twofold higher than wild-type CD4+ T cells cultured in conventional T helper 1 polarizing conditions. Our findings unravel a new pathway orchestrating CD4+ T-cell differentiation and demonstrate that NAD+ may serve as a powerful therapeutic agent for the treatment of autoimmune and other diseases. PMID:25290058

  13. Characterization of a spliced variant of human IRF-3 promoter and its regulation by the transcription factor Sp1.

    PubMed

    Ren, Wei; Zhu, Liang-Hua; Xu, Hua-Guo; Jin, Rui; Zhou, Guo-Ping

    2012-06-01

    Interferon regulatory factor 3 (IRF-3), an essential transcriptional regulator of the interferon genes, plays an important role in host defense against viral and microbial infection as well as in cell growth regulation. Promoter plays a crucial role in gene transcription. We have reported the characterization of the wide type of human IRF-3 promoter, but the characterization of the spliced variant of human IRF-3 Int2V1 promoter has not been systematically analyzed. To observe the spliced variant of human IRF-3 promoter, we have cloned the human IRF-3 gene promoter region containing 300 nucleotides upstream the transcription start site (TSS). Transient transfection of 5' deleted promoter-reporter constructs and luciferase assay illustrated the region -159/-100 relative to the TSS is sufficient for full promoter activity. This region contains GATA1 and specific protein-1 (Sp1) transcription factor binding sites. Interestingly, mutation of this Sp1 site reduced the promoter activity by 50%. However, overexpression of Sp1 increased the transcription activity by 2.4-fold. These results indicated that the spliced variant of human IRF-3 gene core promoter was located within the region -159/-100 relative to the TSS. Sp1 transcription factor upregulates the spliced variant of human IRF-3 gene promoter.

  14. Eicosapentaenoic and docosahexaenoic acids have different effects on peripheral phospholipase A2 gene expressions in acute depressed patients.

    PubMed

    Su, Kuan-Pin; Yang, Hui-Ting; Chang, Jane Pei-Chen; Shih, Yin-Hua; Guu, Ta-Wei; Kumaran, Satyanarayanan Senthil; Gałecki, Piotr; Walczewska, Anna; Pariante, Carmine M

    2018-01-03

    Omega-3 polyunsaturated fatty acids (PUFAs) have been proven critical in the development and management of major depressive disorder (MDD) by a number of epidemiological, clinical and preclinical studies, but the molecular mechanisms underlying this therapeutic action are yet to be understood. Although eicosapentaenoic acid (EPA) seems to be the active component of omega-3 PUFAs' antidepressant effects, the biological research about the difference of specific genetic regulations between EPA and docosahexaenoic acid (DHA), the two main components of omega-3 PUFAs, is still lacking in human subjects. We conducted a 12-week randomized-controlled trial comparing the effects of EPA and DHA on gene expressions of phospholipase A2 (cPLA2) and cyclooxygenase-2 (COX2), serotonin transporter (5HTT), and Tryptophan hydroxylase 2 (TPH-2) in 27 MDD patients. In addition, the erythrocyte PUFA compositions and the candidate gene expressions were also compared between these 27 MDD patients and 22 healthy controls. EPA was associated with a significant decrease in HAM-D scores (CI: -13 to -21, p<0.001) and significant increases in erythrocyte levels of EPA (CI: +1.0% to +2.9%, p=0.001) and DHA (CI: +2.9% to +5.6%, p=0.007). DHA treatment was associated with a significant decrease in HAM-D scores (CI: -6 to -14, p<0.001) and a significant increase in DHA levels (CI: +0.2% to +2.3%, p=0.047), but not of EPA levels. The cPLA2 gene expression levels were significantly increased in patients received EPA (1.9 folds, p=0.038), but not DHA (1.08 folds, p=0.92). There was a tendency for both EPA and DHA groups to decrease COX-2 gene expressions. The gene expressions of COX-2, cPLA2, TPH-2 and 5-HTT did not differ between MDD cases and healthy controls. EPA differentiates from DHA in clinical antidepressant efficacy and in upregulating cPLA2 gene regulations, which supports the clinical observation showing the superiority of EPA's antidepressant effects. ClinicalTrials.gov identifier: NCT02615405. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Slurry-phase biodegradation of weathered oily sludge waste.

    PubMed

    Machín-Ramírez, C; Okoh, A I; Morales, D; Mayolo-Deloisa, K; Quintero, R; Trejo-Hernández, M R

    2008-01-01

    We assessed the biodegradation of a typical oily sludge waste (PB401) in Mexico using several regimes of indigenous microbial consortium and relevant bioremediation strategies in slurry-phase system. Abiotic loss of total petroleum hydrocarbons (TPH) in the PB401 was insignificant, and degradation rates under the various treatment conditions ranged between 666.9 and 2168.7 mg kg(-1) day(-1) over a 15 days reaction period, while viable cell count peaked at between log(10)5.7 and log(10)7.4 cfu g(-1). Biostimulation with a commercial fertilizer resulted in 24% biodegradation of the TPH in the oily waste and a corresponding peak cell density of log(10)7.4 cfu g(-1). Addition of non-indigenous adapted consortium did not appear to enhance the removal of TPH from the oily waste. It would appear that the complexities of the components of the alkylaromatic fraction of the waste limited biodegradation rate even in a slurry system.

  16. Association study of the tryptophan hydroxylase gene and bipolar affective disorder using family-based internal controls.

    PubMed

    Rietschel, M; Schorr, A; Albus, M; Franzek, E; Kreiner, R; Held, T; Knapp, M; Müller, D J; Schulze, T G; Propping, P; Maier, W; Nöthen, M M

    2000-06-12

    The tryptophan hydroxylase (TPH) gene encodes for the rate-limiting enzyme of the serotonin metabolism and, therefore, has to be considered a major candidate for association studies in affective disorders. Recently, an association between this gene and bipolar affective disorder has been reported in a French population. We sought to replicate this finding in a German sample. Allele frequencies of a biallelic polymorphism (A218C) of the TPH gene were determined in 95 bipolar I patients and their parents. Preferential transmission of alleles from heterozygous parents to bipolar offspring was tested with the "transmission disequilibrium test" (TDT), which eliminates the contribution of population stratification to an association finding. Our sample yielded a power >90% to detect the originally reported effect. Neither allele 218A nor allele 218C were preferentially transmitted from heterozygous parents to bipolar offspring. Our results, therefore, do not support the hypothesis that the TPH gene is involved in the etiology of bipolar disorder.

  17. Solving a supply chain scheduling problem with non-identical job sizes and release times by applying a novel effective heuristic algorithm

    NASA Astrophysics Data System (ADS)

    Pei, Jun; Liu, Xinbao; Pardalos, Panos M.; Fan, Wenjuan; Wang, Ling; Yang, Shanlin

    2016-03-01

    Motivated by applications in manufacturing industry, we consider a supply chain scheduling problem, where each job is characterised by non-identical sizes, different release times and unequal processing times. The objective is to minimise the makespan by making batching and sequencing decisions. The problem is formalised as a mixed integer programming model and proved to be strongly NP-hard. Some structural properties are presented for both the general case and a special case. Based on these properties, a lower bound is derived, and a novel two-phase heuristic (TP-H) is developed to solve the problem, which guarantees to obtain a worst case performance ratio of ?. Computational experiments with a set of different sizes of random instances are conducted to evaluate the proposed approach TP-H, which is superior to another two heuristics proposed in the literature. Furthermore, the experimental results indicate that TP-H can effectively and efficiently solve large-size problems in a reasonable time.

  18. Purinergic receptor immunoreactivity in the rostral ventromedial medulla.

    PubMed

    Close, L N; Cetas, J S; Heinricher, M M; Selden, N R

    2009-01-23

    The rostral ventromedial medulla (RVM) has long been recognized to play a pivotal role in nociceptive modulation. Pro-nociception within the RVM is associated with a distinct functional class of neurons, ON-cells that begin to discharge immediately before nocifensive reflexes. Anti-nociceptive function within the RVM, including the analgesic response to opiates, is associated with another distinct class, OFF-cells, which pause immediately prior to nocifensive reflexes. A third class of RVM neurons, NEUTRAL-cells, does not alter firing in association with nocifensive reflexes. ON-, OFF- and NEUTRAL-cells show differential responsiveness to various behaviorally relevant neuromodulators, including purinergic ligands. Iontophoresis of semi-selective P2X ligands, which are associated with nociceptive transmission in the spinal cord and dorsal root ganglia, preferentially activate ON-cells. By contrast, P2Y ligands activate OFF-cells and P1 ligands suppress the firing of NEUTRAL cells. The current study investigates the distribution of P2X, P2Y and P1 receptor immunoreactivity in RVM neurons of Sprague-Dawley rats. Co-localization with tryptophan hydroxylase (TPH), a well-established marker for serotonergic neurons was also studied. Immunoreactivity for the four purinergic receptor subtypes examined was abundant in all anatomical subdivisions of the RVM. By contrast, TPH-immunoreactivity was restricted to a relatively small subset of RVM neurons concentrated in the nucleus raphe magnus and pallidus, as expected. There was a significant degree of co-localization of each purinergic receptor subtype with TPH-immunoreactivity. This co-localization was most pronounced for P2Y1 receptor immunoreactivity, although this was the least abundant among the different purinergic receptor subtypes examined. Immunoreactivity for multiple purinergic receptor subtypes was often co-localized in single neurons. These results confirm the physiological finding that purinergic receptors are widely expressed in the RVM. Purinergic neurotransmission in this region may play an important role in nociception and/or nociceptive modulation, as at other levels of the neuraxis.

  19. Boosting serotonin in the brain: is it time to revamp the treatment of depression?

    PubMed

    Torrente, Mariana P; Gelenberg, Alan J; Vrana, Kent E

    2012-05-01

    Abnormalities in serotonin systems are presumably linked to various psychiatric disorders including schizophrenia and depression. Medications intended for these disorders aim to either block the reuptake or the degradation of this neurotransmitter. In an alternative approach, efforts have been made to enhance serotonin levels through dietary manipulation of precursor levels with modest clinical success. In the last 30 years, there has been little improvement in the pharmaceutical management of depression, and now is the time to revisit therapeutic strategies for the treatment of this disease. Tryptophan hydroxylase (TPH) catalyzes the first and rate-limiting step in the biosynthesis of serotonin. A recently discovered isoform, TPH2, is responsible for serotonin biosynthesis in the brain. Learning how to activate this enzyme (and its polymorphic versions) may lead to a new, more selective generation of antidepressants, able to regulate the levels of serotonin in the brain with fewer side effects.

  20. Optimal conditions for bioremediation of oily seawater.

    PubMed

    Zahed, Mohammad Ali; Aziz, Hamidi Abdul; Isa, Mohamed Hasnain; Mohajeri, Leila; Mohajeri, Soraya

    2010-12-01

    To determine the influence of nutrients on the rate of biodegradation, a five-level, three-factor central composite design (CCD) was employed for bioremediation of seawater artificially contaminated with crude oil. Removal of total petroleum hydrocarbons (TPH) was the dependent variable. Samples were extracted and analyzed according to US-EPA protocols. A significant (R(2)=0.9645, P<0.0001) quadratic polynomial mathematical model was generated. Removal from samples not subjected to optimization and removal by natural attenuation were 53.3% and 22.6%, respectively. Numerical optimization was carried out based on desirability functions for maximum TPH removal. For an initial crude oil concentration of 1g/L supplemented with 190.21 mg/L nitrogen and 12.71 mg/L phosphorus, the Design-Expert software predicted 60.9% hydrocarbon removal; 58.6% removal was observed in a 28-day experiment. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  1. Integrated cistromic and expression analysis of amplified NKX2-1 in lung adenocarcinoma identifies LMO3 as a functional transcriptional target

    PubMed Central

    Watanabe, Hideo; Francis, Joshua M.; Woo, Michele S.; Etemad, Banafsheh; Lin, Wenchu; Fries, Daniel F.; Peng, Shouyong; Snyder, Eric L.; Tata, Purushothama Rao; Izzo, Francesca; Schinzel, Anna C.; Cho, Jeonghee; Hammerman, Peter S.; Verhaak, Roel G.; Hahn, William C.; Rajagopal, Jayaraj; Jacks, Tyler; Meyerson, Matthew

    2013-01-01

    The NKX2-1 transcription factor, a regulator of normal lung development, is the most significantly amplified gene in human lung adenocarcinoma. To study the transcriptional impact of NKX2-1 amplification, we generated an expression signature associated with NKX2-1 amplification in human lung adenocarcinoma and analyzed DNA-binding sites of NKX2-1 by genome-wide chromatin immunoprecipitation. Integration of these expression and cistromic analyses identified LMO3, itself encoding a transcription regulator, as a candidate direct transcriptional target of NKX2-1. Further cistromic and overexpression analyses indicated that NKX2-1 can cooperate with the forkhead box transcription factor FOXA1 to regulate LMO3 gene expression. RNAi analysis of NKX2-1-amplified cells compared with nonamplified cells demonstrated that LMO3 mediates cell survival downstream from NKX2-1. Our findings provide new insight into the transcriptional regulatory network of NKX2-1 and suggest that LMO3 is a transcriptional signal transducer in NKX2-1-amplified lung adenocarcinomas. PMID:23322301

  2. A new paradigm for transcription factor TFIIB functionality

    PubMed Central

    Gelev, Vladimir; Zabolotny, Janice M.; Lange, Martin; Hiromura, Makoto; Yoo, Sang Wook; Orlando, Joseph S.; Kushnir, Anna; Horikoshi, Nobuo; Paquet, Eric; Bachvarov, Dimcho; Schaffer, Priscilla A.; Usheva, Anny

    2014-01-01

    Experimental and bioinformatic studies of transcription initiation by RNA polymerase II (RNAP2) have revealed a mechanism of RNAP2 transcription initiation less uniform across gene promoters than initially thought. However, the general transcription factor TFIIB is presumed to be universally required for RNAP2 transcription initiation. Based on bioinformatic analysis of data and effects of TFIIB knockdown in primary and transformed cell lines on cellular functionality and global gene expression, we report that TFIIB is dispensable for transcription of many human promoters, but is essential for herpes simplex virus-1 (HSV-1) gene transcription and replication. We report a novel cell cycle TFIIB regulation and localization of the acetylated TFIIB variant on the transcriptionally silent mitotic chromatids. Taken together, these results establish a new paradigm for TFIIB functionality in human gene expression, which when downregulated has potent anti-viral effects. PMID:24441171

  3. Petroleum Hydrocarbon Fingerprints of Water and Sediment Samples of Buffalo River Estuary in the Eastern Cape Province, South Africa

    PubMed Central

    Okoh, A. I.

    2017-01-01

    Petroleum hydrocarbon status of the Buffalo River Estuary in East London, South Africa, was evaluated from January to May, 2016. Surface water and sediment samples were collected from five points in the estuary and extracted using standard methods. The extracts were subsequently analyzed by gas chromatography-flame ionization detection. Results showed that total petroleum hydrocarbon (TPH) varied from 7.65 to 477 μg/L in the water and 12.59 to 1,100 mg/kg in the sediments, with mean values of 146.50 ± 27.96 μg/L and 209.81 ± 63.82 mg/kg, respectively. Concentrations of TPH in the sediments correlated significantly with organic carbon (OC) in both seasons. TPH and OC levels were slightly lower in summer than in autumn in the two environmental matrices, and the average amount of TPH in the water samples collected from all the sampling stations was generally lower than the EU standard limit of 300 μg/L. However, the levels in the sediments exceeded the EGASPIN target value (50 mg/kg) for mineral oil but were below the intervention value (5,000 mg/kg), indicating a serious impact of industrial growth and urbanization on the area, although the n-alkane ratios and indexes used for source tracking revealed excessive flow from both natural and anthropogenic sources. PMID:28638675

  4. Relationships between nuclear magnetic resonance parameters used to characterize weathering spilled oil and soil toxicity in central Patagonia.

    PubMed

    Ríos, Stella Maris; Barquin, Mercedes; Katusich, Ofelia; Nudelman, Norma

    2014-01-01

    Oil spill in the Central Patagonian zone was studied to evaluate if any relationship exists between the parameters used to characterize weathering spilled oil and soil toxicity for two plant species and to evaluate if the phytotoxicity to local species would be a good index for the soil contamination. Nuclear magnetic resonance (NMR) structural indexes and column chromatography compositional indexes were determined to characterize the oil spill in the soil samples. Bioassays were also carried out using Lactuca sativa L (reference) and Atriplex lampa (native species) as test organisms. Measurements of the total petroleum hydrocarbon (TPH) and the electrical conductivity (EC) of the soil were carried out to evaluate the effect on the bioassays. The principal components analysis of the parameters determined by NMR, compositional indexes, EC, TPH, and toxicology data shows that the first three principal components accounted for the 78% of the total variance (40%, 25%, and 13% for the first, second, and third PC, respectively). A good agreement was found between information obtained by compositional indexes and NMR structural indexes. Soil toxicity increases with the increase of EC and TPH. Other factors, such as, the presence of branched and aromatic hydrocarbons is also significant. The statistical evaluation showed that the Euclidean distances (3D) between the background and each one of the samples might be a better indicator of the soil contamination, compared with chemical criterion of TPH.

  5. Homeoprotein Six2 promotes breast cancer metastasis via transcriptional and epigenetic control of E-cadherin expression

    PubMed Central

    Wang, Chu-An; Drasin, David; Pham, Catherine; Jedlicka, Paul; Zaberezhnyy, Vadym; Guney, Michelle; Li, Howard; Nemenoff, Raphael; Costello, James C.; Tan, Aik-Choon; Ford, Heide L.

    2014-01-01

    Misexpression of developmental transcription factors occurs often in human cancers, where embryonic programs may be reinstated in a context that promotes or sustains malignant development. In this study, we report the involvement of the kidney development transcription factor Six2 in the metastatic progression of human breast cancer. We found that Six2 promoted breast cancer metastasis by a novel mechanism involving both transcriptional and epigenetic regulation of E-cadherin. Downregulation of E-cadherin by Six2 was necessary for its ability to increase soft agar growth and in vivo metastasis in an immune competent mouse model of breast cancer. Mechanistic investigations showed that Six2 represses E-cadherin expression by upregulating Zeb2, in part through a microRNA-mediated mechanism, and by stimulating promoter methylation of the E-cadherin gene (Cdh1). Clinically, SIX2 expression correlated inversely with CDH1 expression in human breast cancer specimens, corroborating the disease relevance of their interaction. Our findings establish Six2 as a regulator of metastasis in human breast cancers and demonstrate an epigenetic function for SIX family transcription factors in metastatic progression through the regulation of E-cadherin. PMID:25348955

  6. Differential Transcription Factor Use by the KIR2DL4 Promoter Under Constitutive and IL-2/15-Treated Conditions

    PubMed Central

    Presnell, Steven R.; Zhang, Lei; Chlebowy, Corrin N.; Al-Attar, Ahmad; Lutz, Charles T.

    2012-01-01

    KIR2DL4 is unique among human KIR genes in expression, cellular localization, structure, and function, yet the transcription factors required for its expression have not been identified. Using mutagenesis, electrophoretic mobility shift assay, and co-transfection assays, we identified two redundant Runx binding sites in the 2DL4 promoter as essential for constitutive 2DL4 transcription, with contributions by a CRE site and initiator elements. IL-2-and IL-15-stimulated human NK cell lines increased 2DL4 promoter activity, which required functional Runx, CRE, and Ets sites. Chromatin immunoprecipitation experiments show that Runx3 and Ets1 bind the 2DL4 promoter in situ. 2DL4 promoter activity had similar transcription factor requirements in T cells. Runx, CRE, and Ets binding motifs are present in 2DL4 promoters from across primate species, but other postulated transcription factor binding sites are not preserved. Differences between 2DL4 and clonally-restricted KIR promoters suggest a model that explains the unique 2DL4 expression pattern in human NK cells. PMID:22467658

  7. TPH2 G/T Polymorphism Is Associated with Hyperphagia, IQ, and Internalizing Problems in Prader-Willi Syndrome

    ERIC Educational Resources Information Center

    Dykens, Elisabeth M.; Roof, Elizabeth; Bittel, Douglas; Butler, Merlin G.

    2011-01-01

    Background: Prader-Willi syndrome (PWS) is a genetic, neurodevelopmental disorder characterized by intellectual disabilities, growth hormone dysregulation, hyperphagia, increased risks of morbid obesity, compulsive behaviors, and irritability. As aberrant serotonergic functioning is strongly implicated in PWS, we examined associations between the…

  8. Metabolic Pathway Signatures Associated with Urinary Metabolite Biomarkers Differentiate Bladder Cancer Patients from Healthy Controls.

    PubMed

    Kim, Won Tae; Yun, Seok Joong; Yan, Chunri; Jeong, Pildu; Kim, Ye Hwan; Lee, Il Seok; Kang, Ho Won; Park, Sunghyouk; Moon, Sung Kwon; Choi, Yung Hyun; Choi, Young Deuk; Kim, Isaac Yi; Kim, Jayoung; Kim, Wun Jae

    2016-07-01

    Our previous high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry study identified bladder cancer (BCA)-specific urine metabolites, including carnitine, acylcarnitines, and melatonin. The objective of the current study was to determine which metabolic pathways are perturbed in BCA, based on our previously identified urinary metabolome. A total of 135 primary BCA samples and 26 control tissue samples from healthy volunteers were analyzed. The association between specific urinary metabolites and their related encoding genes was analyzed. Significant alterations in the carnitine-acylcarnitine and tryptophan metabolic pathways were detected in urine specimens from BCA patients compared to those of healthy controls. The expression of eight genes involved in the carnitine-acylcarnitine metabolic pathway (CPT1A, CPT1B, CPT1C, CPT2, SLC25A20, and CRAT) or tryptophan metabolism (TPH1 and IDO1) was assessed by RT-PCR in our BCA cohort (n=135). CPT1B, CPT1C, SLC25A20, CRAT, TPH1, and IOD1 were significantly downregulated in tumor tissues compared to normal bladder tissues (p<0.05 all) of patients with non-muscle invasive BCA, whereas CPT1B, CPT1C, CRAT, and TPH1 were downregulated in those with muscle invasive BCA (p<0.05), with no changes in IDO1 expression. Alterations in the expression of genes associated with the carnitine-acylcarnitine and tryptophan metabolic pathways, which were the most perturbed pathways in BCA, were determined.

  9. Diametral tensile strength of two dental composites when immersed in ethanol, distilled water and artificial saliva.

    PubMed

    Rehman, Abdur; Amin, Faiza; Abbas, Muhammad

    2014-11-01

    To examine the effect of distilled water, artificial saliva and ethanol on the tensile strength of direct tooth-coloured restorative material. The study was conducted at Dr. Ishrat ul Ebad Khan Institute of Oral Health Sciences, Dow University of Health Sciences (DUHS), Karachi, from April 2011 to September 2012. The testing was performed at the Pakistan Council of Scientific and Industrial Research (PCSIR) laboratories. Two composite resins Filtek Z250 and Spectrum TPH were tested. Specimens (13 mm x 3 mm x 2 mm) of each material were prepared in the stainless steel mould according to the manufacturers' instructions and distributed into 3 equal groups: one immersed in distilled water, the other in artificial saliva, and the last one in ethanol for 24 hours. Tensile strength was determined after 24 hours in universal Instron Testing Machine. There were 72 specimens in all; 36 (50%) each for Filtek Z250 and Spectrum TPH. The three sub-groups in each case had 12 (33.3%) specimens. For the Filtek Z250, there was no statistically significant difference between immersion in distilled water and artificial saliva, but the ethanol group presented lower tensile strength (p<0.05). For the Spectrum TPH, samples immersed in ethanol and artificial saliva presented lower tensile strength compared to distilled water (p<0.05). The tested composite resins were affected by the immersion media and adversely affected the mechanical properties of composite resins.

  10. SURFACE DEGRADATION OF COMPOSITE RESINS BY ACIDIC MEDICINES AND pH-CYCLING

    PubMed Central

    Valinoti, Ana Carolina; Neves, Beatriz Gonçalves; da Silva, Eduardo Moreira; Maia, Lucianne Cople

    2008-01-01

    This study evaluated the effects of acidic medicines (Dimetapp® and Claritin®), under pH-cycling conditions, on the surface degradation of four composite resins (microhybrid: TPH, Concept, Opallis and Nanofilled: Supreme). Thirty disc-shaped specimens (Ø = 5.0 mm / thickness = 2.0 mm) of each composite were randomly assigned to 3 groups (n = 10): a control and two experimental groups, according to the acidic medicines evaluated. The specimens were finished and polished with aluminum oxide discs, and the surface roughness was measured by using a profilometer. After the specimens were submitted to a pH-cycling regimen and immersion in acidic medicines for 12 days, the surface roughness was measured again. Two specimens for each material and group were analyzed by scanning electron microscopy (SEM) before and after pH-cycling. Data were analyzed by the Student's-t test, ANOVA, Duncan's multiple range test and paired t-test (α=0.05). Significant increase in roughness was found only for TPH in the control group and TPH and Supreme immersed in Claritin® (p<0.05). SEM analyses showed that the 4 composite resins underwent erosion and surface degradation after being subjected to the experimental conditions. In conclusion, although the roughness was slightly affected, the pH-cycling and acidic medicines caused surface degradation of the composite resins evaluated. Titratable acidity seemed to play a more crucial role on surface degradation of composite resins than pH. PMID:19089257

  11. Decabromodiphenyl Ether (DecaBDE) in Electrical and Electronic Equipment in Japan: Stock, Emission, and Substitution Evaluation.

    PubMed

    Xue, Mianqiang; Zhou, Liang; Kojima, Naoya; Machimura, Takashi; Tokai, Akihiro

    2017-11-21

    DecaBDE has been widely used as flame retardant in electrical and electronic equipment (EEE). It has recently been listed in Annex A of the Stockholm Convention. The time series flow, stock, and emission of DecaBDE in EEE in Japan were quantified. On this basis, a risk/risk trade-off analysis of substituting DecaBDE with triphenyl phosphate (TPhP) that is one possible phosphorus-based alternative was conducted. The stock of DecaBDE reached a maximum of ∼42 000 t in 1995. Even though the demand flow was negligible in 2030, the stock was modeled to be still ∼470 t. The outflow of DecaBDE, from the use phase to the disposal phase, peaked at ∼4500 t/yr. in 2001. The DecaBDE emission to atmosphere was mainly derived from the production phase before 1990. The use phase became the largest contributor to the total emission from 1995 to 2000. Whereas the disposal phase dominated the total emission from 2000 onward. In the substitution analysis, a trade-off between human and ecological health effect was revealed in case of replacing DecaBDE with TPhP. This study attempted to give an overall picture of DecaBDE application at national level providing insights into relevant environmental policy making.

  12. Radiopacity of Methacrylate and Silorane Composite Resins Using a Digital Radiographic System.

    PubMed

    Firoozmand, Leily Macedo; Cordeiro, Mariana Gonçalves; Da Silva, Marcos André Dos Santos; De Jesus Tavarez, Rudys Rodolfo; Matos Maia Filho, Etevaldo

    The aim of this study was to evaluate the radiopacity of silorane and methacrylate resin composites, comparing them to the enamel, dentin, and aluminum penetrometer using a digital image. From six resin composites (Filtek ™ P90, Filtek Z350, Filtek Z350 XT flow, Tetric Ceram, TPH Spectrum, and SureFil SDR flow) cylindrical disks (5 × 1 mm) were made and radiographed by a digital method, together with a 15-step aluminum step-wedge and a 1 mm slice of human tooth. The degree of radiopacity of each image was quantified using digital image processing. The mean values of the shades of gray of the tested materials were measured and the equivalent width of aluminum was calculated for each resin. The results of our work yielded the following radiopacity values, given here in descending order: Tetric Ceram > TPH > SDR > Z350 > Z350 flow > P90 > enamel > dentin. The radiopacity of the materials was different both for the enamel and for the dentin, except for resin P90, which was no different than enamel. In conclusion, silorane-based resin exhibited a radiopacity higher than dentin and closest to the enamel; a large portion of the methacrylate-based flow and conventional resins demonstrated greater radiopacity in comparison to dentin and enamel.

  13. Installation of a stoker-coal preparation plant in Krakow, Poland. Technical progress report 7, October--December 1995

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rozelle, P.

    1995-12-31

    This report describes the progress made during this reporting period of a two-year project to demonstrate that the air pollution from a traveling-grate stoker being used to heat water at one of MPEC`s central heating plants in Krakow, Poland can be reduced significantly by (1) substituting the unwashed, unsized coal currently being used with a mechanically cleaned, double-sized stoker fuel and by (2) optimizing the operating parameters of the stoker. It is anticipated that these improvements will prove to be cost-effective and hence will be adopted by the other central heating plants in Krakow and, ideally, throughout Eastern European citiesmore » where coal continues to be the primary source of fuel. EFH Coal Company has formed a partnership with two Polish institutions--MPEC, a central heating company in Krakow, and Naftokrak-Naftobudowa, preparation plant designers and fabricators--for the execution of this effort. A long- term contract for the procurement of 750,000 tons of 20 mm. {times} 0 raw coal for the new plant has been negotiated with the Katowice Coal Holding Company. This long-term lease includes a site near the defunct Kazimierz-Julius preparation plant that has all of the infrastructure needed to build and operate the proposed 300 tph stoker coal preparation plant. The search for markets for utilizing surplus production from the new plant continues. Bid prices for a prefabricated (modular) 300-tph turnkey preparation plant delivered to Poland for preparing a stoker coal ranged from $3.2 to $3.5 million dollars (US). A commitment has been negotiated with Bank PKO S.A. to provide $2 million in cost-share financing toward the capital cost of the project. This sum, when added to the $2.4 million in DOE- BPU funds will be adequate to meet the $3 to $3.5 million needed to finance the purchase, erection and start-up of the 300 tph processing plant.« less

  14. Feasibility Process for Remediation of the Crude Oil Contaminated Soil

    NASA Astrophysics Data System (ADS)

    Keum, H.; Choi, H.; Heo, H.; Lee, S.; Kang, G.

    2015-12-01

    More than 600 oil wells were destroyed in Kuwait by Iraqi in 1991. During the war, over 300 oil lakes with depth of up to 2m at more than 500 different locations which has been over 49km2. Therefore, approximately 22 million m3was crude oil contaminated. As exposure of more than 20 years under atmospheric conditions of Kuwait, the crude oil has volatile hydrocarbons and covered heavy oily sludge under the crude oil lake. One of crude oil contaminated soil which located Burgan Oilfield area was collected by Kuwait Oil Company and got by H-plus Company. This contaminated soil has about 42% crude oil and could not biodegraded itself due to the extremely high toxicity. This contaminated soil was separated by 2mm sieve for removal oil sludge ball. Total petroleum hydrocarbons (TPH) was analysis by GC FID and initial TPH concentration was average 48,783 mg/kg. Ten grams of the contaminated soil replaced in two micro reactors with 20mL of bio surfactant produce microorganism. Reactor 1 was added 0.1g powder hemoglobin and other reactor was not added hemoglobin at time 0 day. Those reactors shake 120 rpm on the shaker for 7 days and CO2 produced about 150mg/L per day. After 7 days under the slurry systems, the rest days operated by hemoglobin as primary carbon source for enhanced biodegradation. The crude oil contaminated soil was degraded from 48,783mg/kg to 20,234mg/kg by slurry process and final TPH concentration degraded 11,324mg/kg for 21days. Therefore, highly contaminated soil by crude oil will be combined bio slurry process and biodegradation process with hemoglobin as bio catalytic source. Keywords: crude-oil contaminated soil, bio slurry, biodegradation, hemoglobin ACKOWLEDGEMENTS This project was supported by the Korea Ministry of Environment (MOE) GAIA Program

  15. The Tryptophan Hydroxylase Inhibitor LX1031 Shows Clinical Benefit in Patients With Nonconstipating Irritable Bowel Syndrome

    PubMed Central

    Brown, Philip M.; Drossman, Douglas A.; Wood, Alastair J. J.; Cline, Gary A.; Frazier, Kenny S.; Jackson, Jessica I.; Bronner, Johanna; Freiman, Joel; Zambrowicz, Brian; Sands, Arthur; Gershon, Michael D.

    2016-01-01

    BACKGROUND & AIMS Serotonin (5-hydroxytryptamine [5-HT]) has an important role in gastrointestinal function. LX1031 is an oral, locally acting, small molecule inhibitor of tryptophan hydroxylase (TPH). Local inhibition of TPH in the gastrointestinal tract might reduce mucosal production of serotonin (5-HT) and be used to treat patients with nonconstipating irritable bowel syndrome (IBS). METHODS We evaluated 2 dose levels of LX1031 (250 mg or 1000 mg, given 4 times/day) in a 28-day, multicenter, randomized, double-blind, placebo-controlled study of 155 patients with nonconstipating IBS. 5-hydroxyindoleacetic acid (5-HIAA), a biomarker of pharmacodynamic activity, was measured in urine samples at baseline (24 hours after LX1031 administration), and at weeks 4 and 6 (n = 76). RESULTS Each dose of LX1031 was safe and well-tolerated. The primary efficacy end point, relief of IBS pain and discomfort, improved significantly in patients given 1000 mg LX1031 (25.5%), compared with those given placebo, at week 1 (P = .018); with nonsignificant improvements at weeks 2, 3, and 4 (17.9%, 16.3%, and 11.6%, respectively). Symptom improvement correlated with a dose-dependent reduction in 5-HIAA, a marker for TPH inhibition, from baseline until week 4. This suggests the efficacy of LX1031 is related to the extent of inhibition of 5-HT biosynthesis. Stool consistency significantly improved, compared with the group given placebo, at weeks 1 and 4 (P < .01) and at week 2 (P < .001). CONCLUSIONS In a phase 2 study, LX1031 was well tolerated, relieving symptoms and increasing stool consistency in patients with nonconstipating IBS. Symptom relief was associated with reduced levels of 5-HIAA in urine samples. This marker might be used to identify patients with nonconstipating IBS who respond to inhibitors of 5-HT synthesis. PMID:21684281

  16. Identification and applications of the Petunia class II Act1/dTph1 transposable element system.

    PubMed

    Gerats, Tom; Zethof, Jan; Vandenbussche, Michiel

    2013-01-01

    Transposable genetic elements are considered to be ubiquitous. Despite this, their mutagenic capacity has been exploited in only a few species. The main plant species are maize, Antirrhinum, and Petunia. Representatives of all three major groups of class II elements, viz., hAT-, CACTA- and Mutator-like elements, have been identified in Petunia. Here we focus on the research "history" of the Petunia two-element Act1-dTph1 system and the development of its application in forward- and reverse-genetics studies.

  17. Serotonin and Early Cognitive Development: Variation in the Tryptophan Hydroxylase 2 Gene Is Associated with Visual Attention in 7-Month-Old Infants

    ERIC Educational Resources Information Center

    Leppanen, Jukka M.; Peltola, Mikko J.; Puura, Kaija; Mantymaa, Mirjami; Mononen, Nina; Lehtimaki, Terho

    2011-01-01

    Background: Allelic variation in the promoter region of a gene that encodes tryptophan hydroxylase isoform 2 (TPH2), a rate-limiting enzyme of serotonin synthesis in the central nervous system, has been associated with variations in cognitive function and vulnerability to affective spectrum disorders. Little is known about the effects of this gene…

  18. The T-Cell Oncogene Tal2 Is a Target of PU.1 and Upregulated during Osteoclastogenesis

    PubMed Central

    Courtial, Nadine; Mücke, Christian; Herkt, Stefanie; Kolodziej, Stephan; Hussong, Helge; Lausen, Jörn

    2013-01-01

    Transcription factors play a crucial role in regulating differentiation processes during human life and are important in disease. The basic helix-loop-helix transcription factors Tal1 and Lyl1 play a major role in the regulation of gene expression in the hematopoietic system and are involved in human leukemia. Tal2, which belongs to the same family of transcription factors as Tal1 and Lyl1, is also involved in human leukaemia. However, little is known regarding the expression and regulation of Tal2 in hematopoietic cells. Here we show that Tal2 is expressed in hematopoietic cells of the myeloid lineage. Interestingly, we found that usage of the Tal2 promoter is different in human and mouse cells. Two promoters, hP1 and hP2 drive Tal2 expression in human erythroleukemia K562 cells, however in mouse RAW cells only the mP1 promoter is used. Furthermore, we found that Tal2 expression is upregulated during oesteoclastogenesis. We show that Tal2 is a direct target gene of the myeloid transcription factor PU.1, which is a key transcription factor for osteoclast gene expression. Strikingly, PU.1 binding to the P1 promoter is conserved between mouse and human, but PU.1 binding to P2 was only detected in human K562 cells. Additionally, we provide evidence that Tal2 influences the expression of the osteoclastic differentiation gene TRACP. These findings provide novel insight into the expression control of Tal2 in hematopoietic cells and reveal a function of Tal2 as a regulator of gene expression during osteoclast differentiation. PMID:24086757

  19. Identification of the Transcriptional Targets of FOXP2, a Gene Linked to Speech and Language, in Developing Human Brain

    PubMed Central

    Spiteri, Elizabeth ; Konopka, Genevieve ; Coppola, Giovanni ; Bomar, Jamee ; Oldham, Michael ; Ou, Jing ; Vernes, Sonja C. ; Fisher, Simon E. ; Ren, Bing ; Geschwind, Daniel H. 

    2007-01-01

    Mutations in FOXP2, a member of the forkhead family of transcription factor genes, are the only known cause of developmental speech and language disorders in humans. To date, there are no known targets of human FOXP2 in the nervous system. The identification of FOXP2 targets in the developing human brain, therefore, provides a unique tool with which to explore the development of human language and speech. Here, we define FOXP2 targets in human basal ganglia (BG) and inferior frontal cortex (IFC) by use of chromatin immunoprecipitation followed by microarray analysis (ChIP-chip) and validate the functional regulation of targets in vitro. ChIP-chip identified 285 FOXP2 targets in fetal human brain; statistically significant overlap of targets in BG and IFC indicates a core set of 34 transcriptional targets of FOXP2. We identified targets specific to IFC or BG that were not observed in lung, suggesting important regional and tissue differences in FOXP2 activity. Many target genes are known to play critical roles in specific aspects of central nervous system patterning or development, such as neurite outgrowth, as well as plasticity. Subsets of the FOXP2 transcriptional targets are either under positive selection in humans or differentially expressed between human and chimpanzee brain. This is the first ChIP-chip study to use human brain tissue, making the FOXP2-target genes identified in these studies important to understanding the pathways regulating speech and language in the developing human brain. These data provide the first insight into the functional network of genes directly regulated by FOXP2 in human brain and by evolutionary comparisons, highlighting genes likely to be involved in the development of human higher-order cognitive processes. PMID:17999357

  20. Properties and reactivity of aquatic organic matter from an Amazonian floodplain system

    NASA Astrophysics Data System (ADS)

    Perez, M. A. P.; Benedetti, M. F.; Moreira-Turcq, P.

    2009-04-01

    The aim of this study was to characterize the nature of the bulk dissolved organic matter (DOM) in different types of environments in the Amazon River-floodplain system and determine the importance of two different fractions of dissolved organic matter onto adsorption processes that occurs through the transport of organic matter in the Amazon Basin. Seven samples were collected in the Amazon River - "Lago Grande de Curuai" floodplain system, in rising water levels cruise (March 2006). The samples were taken in the Amazon main stem, in white and black floodplain waters, and in the middle of a phytoplaktonic bloom. The bulk, dissolved (i.e. < 0.22 micrometer), hydrophobic (HPO) and transphilic (TPH) fractions extracted by XAD-8 and XAD-4 columns chromatography respectively were isolated. Organic carbon (OC) and total nitrogen (TN) concentrations, Specific UV absorbance (SUVA), Size-Exclusion Chromatography (SEC), d13C and d15N isotopes, and reactivity (acid-base titration) were characterized for these fractions. Adsorption experiments onto mineral phase from de surface sediment of the Curuai floodplain lake (rich in smectite and kaolinite) were realized with HPO and TPH fractions. The OC concentrations in the natural organic matter (Bulk and < 0.22 micrometer fractions) varied between 3.7-5.7 mg/L. The OC and TN concentrations varied between 510 - 528 mg C/g in the HPO fraction, and 408 - 462 mg C/g in the TPH compounds and between 14.3 - 17.6 mg N/g (HPO), and 22.1 - 30.0 mg N/g (TPH). The molecular weight of both fractions (HPO and TPH) didn't present significant variation. Both fractions presented high aromaticity and they were rich in carboxylic groups, although smaller values are systematically reported for the HPO fractions. The OM of the main stem was the most adsorbed, followed by the white water lake, the phytoplanktonic bloom, and black water lake sample. These results helped us to strengthen the hypothesis that the organic matter carried from the river and sediment in the floodplain is closely associated with mineral phase.

  1. Telomere-surrounding regions are transcription-permissive 3D nuclear compartments in human cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Quina, Ana Sofia; Instituto Gulbenkian de Ciencia, 2781-901 Oeiras; Parreira, Leonor

    2005-07-01

    Positioning of genes relative to nuclear heterochromatic compartments is thought to help regulate their transcriptional activity. Given that human subtelomeric regions are rich in highly expressed genes, we asked whether human telomeres are related to transcription-permissive nuclear compartments. To address this question, we investigated in the nuclei of normal human lymphocytes the spatial relations of two constitutively expressed genes (ACTB and RARA) and three nuclear transcripts (ACTB, IL2RA and TCRB) to telomeres and centromeres, as a function of gene activity and transcription levels. We observed that genes and gene transcripts locate close to telomere clusters and away from chromocenters uponmore » activation of transcription. These findings, together with the observation that SC35 domains, which are enriched in pre-mRNA processing factors, are in close proximity to telomeres, indicate that telomere-neighboring regions are permissive to gene expression in human cells. Therefore, the associations of telomeres observed in the interphase nucleus might contribute, as opposed to chromocenters, for the establishment of transcription-permissive 3D nuclear compartments.« less

  2. Treatment of petroleum drill cuttings using bioaugmentation and biostimulation supplemented with phytoremediation.

    PubMed

    Kogbara, Reginald B; Ogar, Innocent; Okparanma, Reuben N; Ayotamuno, Josiah M

    2016-07-28

    This study sought to compare the effectiveness of bioaugmentation and biostimulation, as well as the combination of both techniques, supplemented with phytoremediation, in the decontamination of petroleum drill cuttings. Drill cuttings with relatively low concentration of total petroleum hydrocarbons (TPH) and metals were mixed with soil in the ratio 5:1 and treated with three different combinations of the bioremediation options. Option A entailed bioaugmentation supplemented with phytoremediation. Option B had the combination of biostimulation and bioaugmentation supplemented with phytoremediation. While biostimulation supplemented with phytoremediation was deployed in option C. Option O containing the drill cuttings-soil mixture without treatment served as untreated control. Fertilizer application, tillage and watering were used for biostimulation treatment, while spent mushroom substrate (Pleurotus ostreatus) and elephant grass (Pennisetum purpureum) were employed for bioaugmentation and phytoremediation treatment, respectively. The drill cuttings-soil mixtures were monitored for TPH, organic carbon, total nitrogen, pH, metal concentrations, and fungal counts, over time. After 56 days of treatment, there was a decline in the initial TPH concentration of 4,114 mg kg(-1) by 5.5%, 68.3%, 75.6% and 48% in options O, A, B and C, respectively. Generally, higher TPH loss resulted from the phytoremediation treatment stage. The treated options also showed slight reductions in metal concentrations ranging from 0% to 16% of the initial low concentrations. The results highlight the effectiveness of bioaugmentation supplemented with phytoremediation. The combination of bioaugmentation and biostimulation supplemented with phytoremediation, however, may prove better in decontaminating petroleum drill cuttings to environmentally benign levels.

  3. Thumb in the plane of the hand: characterization and results of surgical treatment.

    PubMed

    Langer, Jakub S; Manske, Paul R; Steffen, Jennifer A; Hu, Calvin; Goldfarb, Charles

    2009-12-01

    The purpose of this retrospective investigation is to characterize a congenital deformity, the thumb in the plane of the hand (TPH), and to evaluate the results of abduction-rotation osteotomy of the thumb metacarpal with thumb web space deepening (WSD). We performed a comprehensive analysis of the medical records, hand therapy notes, and radiographs to evaluate clinical features of the TPH deformity. We evaluated clinical and radiographic outcomes and incidence of deformity recurrence after abduction-rotation osteotomy and thumb WSD. Thirteen patients (7 girls and 6 boys) with 14 affected hands treated with an abduction-rotation osteotomy of the thumb metacarpal and formation of a deepened thumb-index web space met inclusion criteria. All TPH deformities were associated with other congenital conditions, including symbrachydactyly, syndactyly, central deficiency, and ulnar deficiency. During the course of treatment, patients had a mean of 4 surgeries per hand; 3 hands required osteotomy revision with or without revision WSD, and 6 additional hands required revision of thumb WSD alone. None of the affected hands were capable of thumb opposition to any finger before surgery, whereas after surgery, all 14 hands could actively perform key pinch, and 9 of the 14 hands could actively oppose the thumb to at least 1 finger. The TPH deformity occurs in association with other congenital abnormalities of the hand. An abduction-rotation osteotomy of the thumb metacarpal with thumb WSD can restore thumb opposition and improve function; nonetheless, multiple surgical procedures are often required, and thumb function may remain limited. Therapeutic IV.

  4. Bioremediation assessment of diesel-biodiesel-contaminated soil using an alternative bioaugmentation strategy.

    PubMed

    Colla, Tatiana Simonetto; Andreazza, Robson; Bücker, Francielle; de Souza, Marcela Moreira; Tramontini, Letícia; Prado, Gerônimo Rodrigues; Frazzon, Ana Paula Guedes; Camargo, Flávio Anastácio de Oliveira; Bento, Fátima Menezes

    2014-02-01

    This study investigated the effectiveness of successive bioaugmentation, conventional bioaugmentation, and biostimulation of biodegradation of B10 in soil. In addition, the structure of the soil microbial community was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis. The consortium was inoculated on the initial and the 11th day of incubation for successive bioaugmentation and only on the initial day for bioaugmentation and conventional bioaugmentation. The experiment was conducted for 32 days. The microbial consortium was identified based on sequencing of 16S rRNA gene and consisted as Pseudomonas aeruginosa, Achromobacter xylosoxidans, and Ochrobactrum intermedium. Nutrient introduction (biostimulation) promoted a positive effect on microbial populations. The results indicate that the edaphic community structure and dynamics were different according to the treatments employed. CO2 evolution demonstrated no significant difference in soil microbial activity between biostimulation and bioaugmentation treatments. The total petroleum hydrocarbon (TPH) analysis indicated a biodegradation level of 35.7 and 32.2 % for the biostimulation and successive bioaugmentation treatments, respectively. Successive bioaugmentation displayed positive effects on biodegradation, with a substantial reduction in TPH levels.

  5. Performance assessment of pilot horizontal sub-surface flow constructed wetlands for removal of diesel from wastewater by Scirpus grossus.

    PubMed

    Al-Baldawi, Israa Abdulwahab; Sheikh Abdullah, Siti Rozaimah; Anuar, Nurina; Suja, Fatihah; Idris, Mushrifah

    2013-01-01

    One of the appropriate development technology options for the treatment of wastewater contaminated with diesel is constructed wetlands (CWs). Throughout 72 days of exposure, sampling was carried out for monitoring of physical parameters, plant growth and the efficiency of total petroleum hydrocarbon (TPH) removal, as an indication for diesel contamination, to assess the pilot-scale performance. Four pilot CWs with a horizontal sub-surface flow system were applied using the bulrush of Scirpus grossus. The CWs were loaded with different diesel concentrations of 0, 0.1, 0.2 and 0.25% (Vdiesel/Vwater). The TPH removal efficiencies were 82, 71, and 67% at the end of 72 days for diesel concentrations of 0.1, 0.2, and 0.25% respectively. In addition, the high removal efficiency of total suspended solids and chemical oxygen demand (COD) were 100 and 75.4% respectively, for a diesel concentration of 0.1%. It was concluded that S. grossus is a potential plant that can be used in a well-operated CW for restoring 0.1% diesel-contaminated water.

  6. Bioremediation of storage tank bottom sludge by using a two-stage composting system: Effect of mixing ratio and nutrients addition.

    PubMed

    Koolivand, Ali; Rajaei, Mohammad Sadegh; Ghanadzadeh, Mohammad Javad; Saeedi, Reza; Abtahi, Hamid; Godini, Kazem

    2017-07-01

    The effect of mixing ratio and nutrients addition on the efficiency of a two-stage composting system in removal of total petroleum hydrocarbons (TPH) from storage tank bottom sludge (STBS) was investigated. The system consisted of ten windrow piles as primary composting (PC) followed by four in-vessel reactors as secondary composting (SC). Various initial C/N/P and mixing ratios of STBS to immature compost (IC) were examined in the PC and SC for 12 and 6weeks, respectively. The removal rates of TPH in the two-stage system (93.72-95.24%) were higher than those in the single-stage one. Depending on the experiments, TPH biodegradation fitted to the first- and second-order kinetics with the rate constants of 0.051-0.334d -1 and 0.002-0.165gkg -1 d -1 , respectively. The bacteria identified were Pseudomonas sp., Bacillus sp., Klebsiella sp., Staphylococcus sp., and Proteus sp. The study verified that a two-stage composting system is effective in treating the STBS. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. PAH, PCB, TPH and mercury in surface sediments of the Delaware River Estuary and Delmarva Peninsula, USA.

    PubMed

    Kim, A W; Vane, C H; Moss-Hayes, V; Engelhart, S E; Kemp, A C

    2018-04-01

    Surface sediment concentrations of polycyclic aromatic hydrocarbons (PAH) and polychlorinated biphenyls (PCB), total petroleum hydrocarbons (TPH) and mercury, were compared from two areas with contrasting land use history, the industrial Delaware Estuary and the rural Delmarva Peninsula (USA). TPH in the Delaware (38-616mg/kg) and saturate/aromatic fractions suggested petroleum/industrial sources compared to biogenic sources in the Delmarva coastal control (<34-159mg/kg). Within the Delaware the ∑PAH18 ranged from 3749 to 22,324μg/kg with isomeric ratios indicative of petroleum combustion source/s, conversely, those in the Delmarva (5-2139μg/kg) also yielded relatively higher perylene that were consistent with natural background levels derived from vegetation/coal combustion source/s. ∑PCB(tri-hepta) concentrations in the Delmarva (0.6-6.5μg/kg) were less than the threshold effect concentration (TEC), whereas the Delaware had received much higher PCB loading (18.1-136.8μg/kg) as evidenced by a significantly higher amounts in some samples (>TEC). Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Soil sampling strategies for site assessments in petroleum-contaminated areas.

    PubMed

    Kim, Geonha; Chowdhury, Saikat; Lin, Yen-Min; Lu, Chih-Jen

    2017-04-01

    Environmental site assessments are frequently executed for monitoring and remediation performance evaluation purposes, especially in total petroleum hydrocarbon (TPH)-contaminated areas, such as gas stations. As a key issue, reproducibility of the assessment results must be ensured, especially if attempts are made to compare results between different institutions. Although it is widely known that uncertainties associated with soil sampling are much higher than those with chemical analyses, field guides or protocols to deal with these uncertainties are not stipulated in detail in the relevant regulations, causing serious errors and distortion of the reliability of environmental site assessments. In this research, uncertainties associated with soil sampling and sample reduction for chemical analysis were quantified using laboratory-scale experiments and the theory of sampling. The research results showed that the TPH mass assessed by sampling tends to be overestimated and sampling errors are high, especially for the low range of TPH concentrations. Homogenization of soil was found to be an efficient method to suppress uncertainty, but high-resolution sampling could be an essential way to minimize this.

  9. Petroleum residues degradation in laboratory-scale by rhizosphere bacteria isolated from the mangrove ecosystem

    NASA Astrophysics Data System (ADS)

    Rinanti, A.; Nainggolan, I. J.

    2018-01-01

    This research is about petroleum bioremediation experiment to obtain bacterial isolate from mangrove ecosystem which potentially degrade petroleum. It was conducted in an Erlenmeyer batch system filled with growth medium of Stone Mineral Salt Solution (SMSS) plus petroleum residue, placed in an incubator shaker with a rotation speed of 120 rpm, temperature 3000C, for 14 research days. Indigenous bacteria that have been isolated and identified from the roots of mangrove plants are Ochrobactrum anthropi and Bacillus sp., Ralstonia pickettii and Bacillus circulans. Those bacteriain both monoculture and consortium form (mixed culture) are incorporated into erlenmeyer as remediator agents. All bacteria can utilize hydrocarbon compounds, but Ralstonia pickettii and Bacillus circulans reached exponential phase faster with more cell count than other bacteria. Compared to single cultures, petroleum degradation by a bacterial consortium provides a higher TPH reduction efficiency, i.e. at 5%, 10%, and 15% of initial TPH of 94.4%, 72%, and 80.3%, respectively. This study proved that all bacteria could optimize hydrocarbon compounds up to 15% TPH load.

  10. Growth of four tropical tree species in petroleum-contaminated soil and effects of crude oil contamination.

    PubMed

    Pérez-Hernández, I; Ochoa-Gaona, S; Adams, R H; Rivera-Cruz, M C; Pérez-Hernández, V; Jarquín-Sánchez, A; Geissen, V; Martínez-Zurimendi, P

    2017-01-01

    Under greenhouse conditions, we evaluated establishment of four tree species and their capacity to degrade crude oil recently incorporated into the soil; the species were as follows: Cedrela odorata (tropical cedar), Haematoxylum campechianum (tinto bush), Swietenia macrophylla (mahogany), and Tabebuia rosea (macuilis). Three-month-old plants were planted in soil with three treatments of heavy petroleum and a control (C0 0 mg kg -1 ; C1 18,000 mg kg -1 ; C2 31,700 mg kg -1 ; C3 47,100 mg kg -1 ) with four repetitions per treatment and species; the experiment was carried out for 245 days. Height and biomass of all species significantly diminished as petroleum concentration increased, although plant survival was not affected. The quantity of colony-forming units (CFU) of rhizospheric bacteria varied among tree species and treatments; petroleum stimulated bacterial CFU for S. macrophylla. The number of fungi CFU for S. macrophylla and T. rosea was significantly greater in C0 than in soil with petroleum, but among species and among different concentrations, no significant differences were found. The greatest percentage of total petroleum hydrocarbon (TPH) degradation was found in C1 for soil without plants (45 %). Differences from the remaining treatments (petroleum concentrations in soil and plant species) were not significant (P < 0.05). Among all trees, H. campechianum had the greatest TPH degradation (32.5 % in C2). T. rosea (C1) and H. campechianum (C2) resulted in petroleum degradation at levels ranging from 20.5 to 32.5 %. On the basis of this experiment, the tree species used did not improve TPH degradation. However, all of them showed high rates of survival and vigor. So, as tree species provide goods and services, experiments with inoculation of hydrocarbonclastic microorganisms, addition of fertilizers, and mixture of tree and grasses are recommended.

  11. Genetic and Non-genetic Factors Associated With Constipation in Cancer Patients Receiving Opioids

    PubMed Central

    Laugsand, Eivor A; Skorpen, Frank; Kaasa, Stein; Sabatowski, Rainer; Strasser, Florian; Fayers, Peter; Klepstad, Pål

    2015-01-01

    Objectives: To examine whether the inter-individual variation in constipation among patients receiving opioids for cancer pain is associated with genetic or non-genetic factors. Methods: Cancer patients receiving opioids were included from 17 centers in 11 European countries. Intensity of constipation was reported by 1,568 patients on a four-point categorical scale. Non-genetic factors were included as covariates in stratified regression analyses on the association between constipation and 75 single-nucleotide polymorphisms (SNPs) within 15 candidate genes related to opioid- or constipation-signaling pathways (HTR3E, HTR4, HTR2A, TPH1, ADRA2A, CHRM3, TACR1, CCKAR, KIT, ARRB2, GHRL, ABCB1, COMT, OPRM1, and OPRD1). Results: The non-genetic factors significantly associated with constipation were type of laxative, mobility and place of care among patients receiving laxatives (N=806), in addition to Karnofsky performance status and presence of metastases among patients not receiving laxatives (N=762) (P<0.01). Age, gender, body mass index, cancer diagnosis, time on opioids, opioid dose, and type of opioid did not contribute to the inter-individual differences in constipation. Five SNPs, rs1800532 in TPH1, rs1799971 in OPRM1, rs4437575 in ABCB1, rs10802789 in CHRM3, and rs2020917 in COMT were associated with constipation (P<0.01). Only rs2020917 in COMT passed the Benjamini–Hochberg criterion for a 10% false discovery rate. Conclusions: Type of laxative, mobility, hospitalization, Karnofsky performance status, presence of metastases, and five SNPs within TPH1, OPRM1, ABCB1, CHRM3, and COMT may contribute to the variability in constipation among cancer patients treated with opioids. Knowledge of these factors may help to develop new therapies and to identify patients needing a more individualized approach to treatment. PMID:26087058

  12. An in vitro bioassay for xenobiotics using the SXR-driven human CYP3A4/lacZ reporter gene.

    PubMed

    Lee, Mi R; Kim, Yeon J; Hwang, Dae Y; Kang, Tae S; Hwang, Jin H; Lim, Chae H; Kang, Hyung K; Goo, Jun S; Lim, Hwa J; Ahn, Kwang S; Cho, Jung S; Chae, Kap R; Kim, Yong K

    2003-01-01

    The dose and time effect of nine xenobiotics, including 17beta-estradiol, corticosterone, dexamethasone, progesterone, nifedipine, bisphenol A, rifampicin, methamphetamine, and nicotine were investigated, in vitro, using human steroid and xenobiotics receptor (SXR)-binding sites on the human CYP3A4 promoter, which can enhance the linked lacZ reporter gene transcription. To test this, liver-specific SAP (human serum amyloid P component)-SXR (SAP/SXR) and human CYP3A4 promoter-regulated lacZ (hCYP3A4/lacZ) constructs were transiently transfected into HepG2 and NIH3T3 cells to compare the xenobiotic responsiveness between human and nonhuman cell lines. In the HepG2 cells, rifampicin, followed by corticosterone, nicotine, methamphetamine, and dexamethasone, exhibited enhanced levels of the lacZ transcript, whereas those of bisphenol A and nifedipine were found to be reduced. No significant responses were observed with 17beta-estradiol or progesterone. In addition, 17beta-estradiol and progesterone did not change the levels of the lacZ transcripts in the HepG2 cells, but did induce significant increases in the transcripts of the NIH3T3 cells. Treatment with corticosterone and dexamethasone, which were highly expressed in the HepG2 cells, did not affect the levels of the lacZ transcript in NIH3T3 cells. These results show that lacZ transcripts can be measured, rapidly and reproducibly, using reverse transcriptase-polymerase chain reaction (RT-PCR) based on the expression of the hCYP3A4/lacZ reporter gene, and was mediated by the SXR. Thus, this in vitro reporter gene bioassay is useful for measuring xenobiotic activities, and is a means to a better relevant bioassay, using human cells, human genes and human promoters, in order to get a closer look at actual human exposure.

  13. Analysis of Several PLA2 mRNA in Human Meningiomas

    PubMed Central

    Denizot, Yves; De Armas, Rafael; Durand, Karine; Robert, Sandrine; Moreau, Jean-Jacques; Caire, François; Weinbreck, Nicolas; Labrousse, François

    2009-01-01

    In view of the important oncogenic action of phospholipase A2(PLA2) we investigated PLA2 transcripts in human meningiomas. Real-time PCR was used to investigate PLA2 transcripts in 26 human meningioma tumors. Results indicated that three Ca2+-dependent high molecular weight PLA2 (PLA2-IVA, PLA2-IVB, PLA2-IVC), one Ca2+-independent high molecular weight PLA2 (PLA2-VI) and five low molecular weight secreted forms of PLA2 (PLA2-IB, PLA2-IIA, PLA2-III, PLA2-V, and PLA2-XII) are expressed with PLA2-IVA, PLA2-IVB, PLA2-VI, and PLA2-XIIA as the major expressed forms. PLA2-IIE, PLA2-IIF, PLA2-IVD, and PLA2-XIIB are not detected. Plasma (PLA2-VIIA) and intracellular (PLA2-VIIB) platelet-activating factor acetylhydrolase transcripts are expressed in human meningiomas. However no difference was found for PLA2 transcript amounts in relation to the tumor grade, the subtype of meningiomas, the presence of inflammatory infiltrated cells, of an associated edema, mitosis, brain invasion, vascularisation or necrosis. In conclusion numerous genes encoding multiples forms of PLA2 are expressed in meningiomas where they might act on the phospholipid remodeling and on the local eicosanoid and/or cytokine networks. PMID:20339511

  14. Natural attenuation of aged tar-oil in soils: A case study from a former gas production site

    NASA Astrophysics Data System (ADS)

    Ivanov, Pavel; Eickhorst, Thilo; Wehrer, Markus; Georgiadis, Anna; Rennert, Thilo; Eusterhues, Karin; Totsche, Kai Uwe

    2017-04-01

    Contamination of soils with tar oil occurred on many industrial sites in Europe. The main source of such contamination has been former manufactured gas plants (MGP). As many of them were destroyed during the World War II or abandoned in the second half of the XXth century, the contamination is depleted in volatile and degradable hydrocarbons (HC) but enriched in the heavy oil fractions due to aging processes. We studied a small tar-oil spill in a former MGP reservoir basin. The tar-oil had a total petroleum hydrocarbon (TPH) content of 245 mg/g. At the margin of the spill, vegetation has started to overgrow and intensively root the tar-oil layer. This zone comprised the uppermost 5-7 cm of our profile and contained 28 mg/g of TPH (A-layer)- The layer below the root zone (7-15 cm) was the most contaminated, with 90 mg/g TPH (B-layer). The layer underneath (15-22 cm) had smaller concentrations of 16 mg/g TPH (C-layer). Further down in the profile (D-layer) we found only slightly higher TPH content than in the control samples (1,4 mg/g vs 0,6 mg/g). The polycyclic aromatic hydrocarbons analysis showed the same distribution throughout all layers with highest contents of the PAHs with 4-6 condensed aromatic rings. Direct cell count and extraction of microbial biomass showed that the highly contaminated soil layers A and B had 2-3 times more bacteria than the control soils. CARD-FISH analysis revealed that in samples from layers A and B Archaea were more abundant (12% opposing to 6-7% in control soil). Analysis of bacteria (tested for Alpha-, Beta-, Gamma- and Epsilonproteobacteria and Actinobacteria) showed the dominance of Alphaproteobacteria in the layer A and C both beneath and above the most contaminated layer B. The primers covered the whole microbial consortia in these two layers, leaving almost no unidentified cells. In the most contaminated layer B Alphaproteobacteria amounted only to 20% of the microbial consortium, and almost 40% of the cells remained unidentified, suggesting the presence of other microorganisms using high-molecular weight HC as carbon source. All contaminated layers were found to be enriched in total Fe and both dithionite-extractable and oxalate-extractable Fe. Besides, siderite crystals were identified using FTIR microscopy. The presence of secondary crystalline and poorly crystalline Fe(III)-oxides and secondary Fe(II)-carbonates in the same horizons suggests simultaneous occurrence of oxic and anoxic zones within the porous system of the contaminated layers. Although HC pollution is often considered to inhibit microbial activity in soil, in our study the layers with highest TPH-amounts were the most "alive". We assume that aging processes (the sum of volatilization, dissolution, microbial degradation, chemical oxidation, polymerization and migration) and eventually a long-term microbial adaption to the HC carbon source resulted in the development of a microbial consortium, capable of transforming high-molecular weight HC. Presumably, iron-compounds in the tar oil act as an electron acceptor and trigger HC degradation. However, to unravel natural attenuation processes and degradation pathways it seems mandatory to take into account the soil structure and spatial distribution of microbes.

  15. RNA Sequencing Analysis Reveals Interactions between Breast Cancer or Melanoma Cells and the Tissue Microenvironment during Brain Metastasis

    PubMed Central

    Hosonaga, Mari; Koya, Ikuko

    2017-01-01

    Metastasis is the main cause of treatment failure and death in cancer patients. Metastasis of tumor cells to the brain occurs frequently in individuals with breast cancer, non–small cell lung cancer, or melanoma. Despite recent advances in our understanding of the causes and in the treatment of primary tumors, the biological and molecular mechanisms underlying the metastasis of cancer cells to the brain have remained unclear. Metastasizing cancer cells interact with their microenvironment in the brain to establish metastases. We have now developed mouse models of brain metastasis based on intracardiac injection of human breast cancer or melanoma cell lines, and we have performed RNA sequencing analysis to identify genes in mouse brain tissue and the human cancer cells whose expression is associated specifically with metastasis. We found that the expressions of the mouse genes Tph2, Sspo, Ptprq, and Pole as well as those of the human genes CXCR4, PLLP, TNFSF4, VCAM1, SLC8A2, and SLC7A11 were upregulated in brain tissue harboring metastases. Further characterization of such genes that contribute to the establishment of brain metastases may provide a basis for the development of new therapeutic strategies and consequent improvement in the prognosis of cancer patients. PMID:28210624

  16. Petroleum Hydrocarbon Profiles of Water and Sediment of Algoa Bay, Eastern Cape, South Africa

    PubMed Central

    Adeniji, Abiodun O.; Okoh, Omobola O.

    2017-01-01

    Petroleum hydrocarbon profiles of water and sediment samples of Algoa Bay in the Eastern Cape Province of South Africa were assessed using standard analytical procedures. Water (from surface and bottom levels) and sediment samples were collected from five locations in the bay from February to June 2016. Extraction of the petroleum hydrocarbons from the water and sediment samples collected was achieved using liquid-liquid and Soxhlet extraction techniques, respectively, followed by column clean up. Target compounds were analytically determined with gas chromatography–flame ionization detector (GC-FID) and quantified by integrating the areas of both the resolved and unresolved components. Physicochemical properties of the water samples were also determined on site using a SeaBird 19plusV2 CTD SBE 55 device. Estimated limit of detection, limit of quantitation and relative standard deviation for the 35 n-alkane standards ranged from 0.06 to 0.13 μg/L, 0.30 to 0.69 μg/L and 3.61 to 8.32%, respectively. Results showed that total petroleum hydrocarbon (TPH) varied from 45.07 to 307 μg/L in the water and 0.72 to 27.03 mg/kg in the sediments. The mean concentrations of TPH in both the water and sediment samples from Algoa Bay revealed a slight level of pollution. The diagnostic indices used showed that the hydrocarbons in the area were from both biogenic and anthropogenic sources. Hence, there is need for adequate regulation and control of all activities contributing to the levels of petroleum hydrocarbon in the marine environment for the safety of human, aquatic and wild lives in the area. PMID:29053634

  17. Petroleum Hydrocarbon Profiles of Water and Sediment of Algoa Bay, Eastern Cape, South Africa.

    PubMed

    Adeniji, Abiodun O; Okoh, Omobola O; Okoh, Anthony I

    2017-10-20

    Petroleum hydrocarbon profiles of water and sediment samples of Algoa Bay in the Eastern Cape Province of South Africa were assessed using standard analytical procedures. Water (from surface and bottom levels) and sediment samples were collected from five locations in the bay from February to June 2016. Extraction of the petroleum hydrocarbons from the water and sediment samples collected was achieved using liquid-liquid and Soxhlet extraction techniques, respectively, followed by column clean up. Target compounds were analytically determined with gas chromatography-flame ionization detector (GC-FID) and quantified by integrating the areas of both the resolved and unresolved components. Physicochemical properties of the water samples were also determined on site using a SeaBird 19plusV2 CTD SBE 55 device. Estimated limit of detection, limit of quantitation and relative standard deviation for the 35 n -alkane standards ranged from 0.06 to 0.13 μg/L, 0.30 to 0.69 μg/L and 3.61 to 8.32%, respectively. Results showed that total petroleum hydrocarbon (TPH) varied from 45.07 to 307 μg/L in the water and 0.72 to 27.03 mg/kg in the sediments. The mean concentrations of TPH in both the water and sediment samples from Algoa Bay revealed a slight level of pollution. The diagnostic indices used showed that the hydrocarbons in the area were from both biogenic and anthropogenic sources. Hence, there is need for adequate regulation and control of all activities contributing to the levels of petroleum hydrocarbon in the marine environment for the safety of human, aquatic and wild lives in the area.

  18. A combination of solvent extraction and freeze thaw for oil recovery from petroleum refinery wastewater treatment pond sludge.

    PubMed

    Hu, Guangji; Li, Jianbing; Hou, Haobo

    2015-01-01

    A combination of solvent extraction and freeze thaw was examined for recovering oil from the high-moisture petroleum refinery wastewater treatment pond sludge. Five solvents including cyclohexane (CHX), dichloromethane (DCM), methyl ethyl ketone (MEK), ethyl acetate (EA), and 2-propanol (2-Pro) were examined. It was found that these solvents except 2-Pro showed a promising oil recovery rate of about 40%, but the recycling of DCM solvent after oil extraction was quite low. Three solvents (CHX, MEK and EA) were then selected for examining the effect of freeze/thaw treatment on improving the quality of recovered oil. This treatment increased the total petroleum hydrocarbon (TPH) content in recovered oil from about 40% to 60% for both MEK and EA extractions, but little effect was observed for CHX extraction. Although the solid residue after oil recovery had a significantly decreased TPH content, a high concentration of heavy metals was observed, indicating that this residue may require proper management. In general, the combination of solvent extraction with freeze/thaw is effective for high-moisture oily hazardous waste treatment. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. TPH2 polymorphisms and alcohol-related suicide.

    PubMed

    Zupanc, Tomaž; Pregelj, Peter; Tomori, Martina; Komel, Radovan; Paska, Alja Videtič

    2011-02-18

    Substantial evidence from family, twin, and adoption studies corroborates implication of genetic and environmental factors, as well as their interactions, on suicidal behavior and alcoholism risk. Serotonergic disfunction seems to be involved in the pathophysiology of substance abuse, and has also an important role in suicidal behavior. Recent studies of the tryptophan hydroxylase 2 showed mild or no association with suicide and alcohol-related suicide. We performed SNP and alcohol analysis on 388 suicide victims and 227 controls. The results showed association between suicide (Pχ²=0.043) and alcohol-related suicide (Pχ²=0.021) for SNP Rs1843809. A tendency for association was determined also for polymorphism Rs1386493 (Pχ²=0.055) and alcohol-related suicide. Data acquired from psychological autopsies in a subsample of suicide victims (n=79) determined more impulsive behavior (Pχ²=0.016) and verbal aggressive behavior (Pχ²=0.025) in the subgroup with alcohol misuse or dependency. In conclusion, our results suggest implication of polymorphisms in suicide and alcohol-related suicide, but further studies are needed to clarify the interplay among serotonergic system disfunction, suicide, alcohol dependence, impulsivity and the role of TPH2 enzyme. © 2010 Elsevier Ireland Ltd. All rights reserved.

  20. Response of soil microorganisms to radioactive oil waste: results from a leaching experiment

    NASA Astrophysics Data System (ADS)

    Galitskaya, P.; Biktasheva, L.; Saveliev, A.; Ratering, S.; Schnell, S.; Selivanovskaya, S.

    2015-06-01

    Oil wastes produced in large amounts in the processes of oil extraction, refining, and transportation are of great environmental concern because of their mutagenicity, toxicity, high fire hazardousness, and hydrophobicity. About 40% of these wastes contain radionuclides; however, the effects of oil products and radionuclides on soil microorganisms are frequently studied separately. The effects on various microbial parameters of raw waste containing 575 g of total petroleum hydrocarbons (TPH) kg-1 waste, 4.4 of 226Ra, 2.8 of 232Th, and 1.3 kBq kg-1 of 40K and its treated variant (1.6 g kg-1 of TPH, 7.9 of 226Ra, 3.9 of 232Th, and 183 kBq kg-1 of 40K) were examined in a leaching column experiment to separate the effects of hydrocarbons from those of radioactive elements. The raw waste sample (H) was collected from tanks during cleaning and maintenance, and a treated waste sample (R) was obtained from equipment for oil waste treatment. Thermal steam treatment is used in the production yard to reduce the oil content. The disposal of H waste samples on the soil surface led to an increase in the TPH content in soil: it became 3.5, 2.8, and 2.2 times higher in the upper (0-20 cm), middle (20-40 cm), and lower (40-60cm) layers, respectively. Activity concentrations of 226Ra and 232Th increased in soil sampled from both H- and R- columns in comparison to their concentrations in control soil. The activity concentrations of these two elements in samples taken from the upper and middle layers were much higher for the R-column compared to the H-column, despite the fact that the amount of waste added to the columns was equalized with respect to the activity concentrations of radionuclides. The H waste containing both TPH and radionuclides affected the functioning of the soil microbial community, and the effect was more pronounced in the upper layer of the column. Metabolic quotient and cellulase activity were the most sensitive microbial parameters as their levels were changed 5-1.4 times in comparison to control ones. Changes in soil functional characteristics caused by the treated waste containing mainly radionuclides were not observed. PCR-SSCP (polymerase chain reaction - single strand conformation polymorphism) analysis followed by MDS (metric multidimensional scaling) and clustering analysis revealed that the shifts in microbial community structure were affected by both hydrocarbons and radioactivity. Thus, molecular methods permitted to reveal the effects on soil microbial community not only from hydrocarbons, which significantly altered functional characteristics of soil microbiome, but also from radioactive elements.

  1. Corrective Action Investigation Plan for Corrective Action Unit 214: Bunkers and Storage Areas Nevada Test Site, Nevada: Revision 0, Including Record of Technical Change No. 1 and No. 2

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office

    This Corrective Action Investigation Plan contains the U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office's approach to collect the data necessary to evaluate corrective action alternatives appropriate for the closure of Corrective Action Unit (CAU) 214 under the Federal Facility Agreement and Consent Order. Located in Areas 5, 11, and 25 of the Nevada Test Site, CAU 214 consists of nine Corrective Action Sites (CASs): 05-99-01, Fallout Shelters; 11-22-03, Drum; 25-99-12, Fly Ash Storage; 25-23-01, Contaminated Materials; 25-23-19, Radioactive Material Storage; 25-99-18, Storage Area; 25-34-03, Motor Dr/Gr Assembly (Bunker); 25-34-04, Motor Dr/Gr Assembly (Bunker); and 25-34-05, Motormore » Dr/Gr Assembly (Bunker). These sites are being investigated because existing information on the nature and extent of potential contamination is insufficient to evaluate and recommend corrective action alternatives (CAAs). The suspected contaminants and critical analyte s for CAU 214 include oil (total petroleum hydrocarbons-diesel-range organics [TPH-DRO], polychlorinated biphenyls [PCBs]), pesticides (chlordane, heptachlor, 4,4-DDT), barium, cadmium, chronium, lubricants (TPH-DRO, TPH-gasoline-range organics [GRO]), and fly ash (arsenic). The land-use zones where CAU 214 CASs are located dictate that future land uses will be limited to nonresidential (i.e., industrial) activities. The results of this field investigation will support a defensible evaluation of viable corrective action alternatives that will be presented in the corrective action decision document.« less

  2. Expression Profiling Smackdown: Human Transcriptome Array HTA 2.0 vs. RNA-Seq

    PubMed Central

    Palermo, Meghann; Driscoll, Heather; Tighe, Scott; Dragon, Julie; Bond, Jeff; Shukla, Arti; Vangala, Mahesh; Vincent, James; Hunter, Tim

    2014-01-01

    The advent of both microarray and massively parallel sequencing have revolutionized high-throughput analysis of the human transcriptome. Due to limitations in microarray technology, detecting and quantifying coding transcript isoforms, in addition to non-coding transcripts, has been challenging. As a result, RNA-Seq has been the preferred method for characterizing the full human transcriptome, until now. A new high-resolution array from Affymetrix, GeneChip Human Transcriptome Array 2.0 (HTA 2.0), has been designed to interrogate all transcript isoforms in the human transcriptome with >6 million probes targeting coding transcripts, exon-exon splice junctions, and non-coding transcripts. Here we compare expression results from GeneChip HTA 2.0 and RNA-Seq data using identical RNA extractions from three samples each of healthy human mesothelial cells in culture, LP9-C1, and healthy mesothelial cells treated with asbestos, LP9-A1. For GeneChip HTA 2.0 sample preparation, we chose to compare two target preparation methods, NuGEN Ovation Pico WTA V2 with the Encore Biotin Module versus Affymetrix's GeneChip WT PLUS with the WT Terminal Labeling Kit, on identical RNA extractions from both untreated and treated samples. These same RNA extractions were used for the RNA-Seq library preparation. All analyses were performed in Partek Genomics Suite 6.6. Expression profiles for control and asbestos-treated mesothelial cells prepared with NuGEN versus Affymetrix target preparation methods (GeneChip HTA 2.0) are compared to each other as well as to RNA-Seq results.

  3. CCAAT-binding factor regulates expression of the beta1 subunit of soluble guanylyl cyclase gene in the BE2 human neuroblastoma cell line

    NASA Technical Reports Server (NTRS)

    Sharina, Iraida G.; Martin, Emil; Thomas, Anthony; Uray, Karen L.; Murad, Ferid

    2003-01-01

    Soluble guanylyl cyclase (sGC) is a cytosolic enzyme producing the intracellular messenger cyclic guanosine monophosphate (cGMP) on activation with nitric oxide (NO). sGC is an obligatory heterodimer composed of alpha and beta subunits. We investigated human beta1 sGC transcriptional regulation in BE2 human neuroblastoma cells. The 5' upstream region of the beta1 sGC gene was isolated and analyzed for promoter activity by using luciferase reporter constructs. The transcriptional start site of the beta1 sGC gene in BE2 cells was identified. The functional significance of consensus transcriptional factor binding sites proximal to the transcriptional start site was investigated by site deletions in the 800-bp promoter fragment. The elimination of CCAAT-binding factor (CBF) and growth factor independence 1 (GFI1) binding cores significantly diminished whereas deletion of the NF1 core elevated the transcription. Electrophoretic mobility-shift assay (EMSA) and Western analysis of proteins bound to biotinated EMSA probes confirmed the interaction of GFI1, CBF, and NF1 factors with the beta1 sGC promoter. Treatment of BE2 cells with genistein, known to inhibit the CBF binding to DNA, significantly reduced protein levels of beta1 sGC by inhibiting transcription. In summary, our study represents an analysis of the human beta1 sGC promoter regulation in human neuroblastoma BE2 cells and identifies CBF as a critically important factor in beta1 sGC expression.

  4. Integrated expression analysis identifies transcription networks in mouse and human gastric neoplasia.

    PubMed

    Chen, Zheng; Soutto, Mohammed; Rahman, Bushra; Fazili, Muhammad W; Peng, DunFa; Blanca Piazuelo, Maria; Chen, Heidi; Kay Washington, M; Shyr, Yu; El-Rifai, Wael

    2017-07-01

    Gastric cancer (GC) is a leading cause of cancer-related deaths worldwide. The Tff1 knockout (KO) mouse model develops gastric lesions that include low-grade dysplasia (LGD), high-grade dysplasia (HGD), and adenocarcinomas. In this study, we used Affymetrix microarrays gene expression platforms for analysis of molecular signatures in the mouse stomach [Tff1-KO (LGD) and Tff1 wild-type (normal)] and human gastric cancer tissues and their adjacent normal tissue samples. Combined integrated bioinformatics analysis of mouse and human datasets indicated that 172 genes were consistently deregulated in both human gastric cancer samples and Tff1-KO LGD lesions (P < .05). Using Ingenuity pathway analysis, these genes mapped to important transcription networks that include MYC, STAT3, β-catenin, RELA, NFATC2, HIF1A, and ETS1 in both human and mouse. Further analysis demonstrated activation of FOXM1 and inhibition of TP53 transcription networks in human gastric cancers but not in Tff1-KO LGD lesions. Using real-time RT-PCR, we validated the deregulated expression of several genes (VCAM1, BGN, CLDN2, COL1A1, COL1A2, COL3A1, EpCAM, IFITM1, MMP9, MMP12, MMP14, PDGFRB, PLAU, and TIMP1) that map to altered transcription networks in both mouse and human gastric neoplasia. Our study demonstrates significant similarities in deregulated transcription networks in human gastric cancer and gastric tumorigenesis in the Tff1-KO mouse model. The data also suggest that activation of MYC, STAT3, RELA, and β-catenin transcription networks could be an early molecular step in gastric carcinogenesis. © 2017 Wiley Periodicals, Inc.

  5. Conserved roles of mouse DUX and human DUX4 in activating cleavage-stage genes and MERVL/HERVL retrotransposons.

    PubMed

    Hendrickson, Peter G; Doráis, Jessie A; Grow, Edward J; Whiddon, Jennifer L; Lim, Jong-Won; Wike, Candice L; Weaver, Bradley D; Pflueger, Christian; Emery, Benjamin R; Wilcox, Aaron L; Nix, David A; Peterson, C Matthew; Tapscott, Stephen J; Carrell, Douglas T; Cairns, Bradley R

    2017-06-01

    To better understand transcriptional regulation during human oogenesis and preimplantation development, we defined stage-specific transcription, which highlighted the cleavage stage as being highly distinctive. Here, we present multiple lines of evidence that a eutherian-specific multicopy retrogene, DUX4, encodes a transcription factor that activates hundreds of endogenous genes (for example, ZSCAN4, KDM4E and PRAMEF-family genes) and retroviral elements (MERVL/HERVL family) that define the cleavage-specific transcriptional programs in humans and mice. Remarkably, mouse Dux expression is both necessary and sufficient to convert mouse embryonic stem cells (mESCs) into 2-cell-embryo-like ('2C-like') cells, measured here by the reactivation of '2C' genes and repeat elements, the loss of POU5F1 (also known as OCT4) protein and chromocenters, and the conversion of the chromatin landscape (as assessed by transposase-accessible chromatin using sequencing (ATAC-seq)) to a state strongly resembling that of mouse 2C embryos. Thus, we propose mouse DUX and human DUX4 as major drivers of the cleavage or 2C state.

  6. Underwater Sediment Sampling Research

    DTIC Science & Technology

    2017-01-01

    resolved through further experimentation . Underwater Sediment Sampling Research vi UNCLAS//Public | CG-926 RDC | A. Hanson, et al. Public...Chemical Oceanographer, and In situ Chemical Analysis Subject Matter Expert (SME). 2 LABORATORY TEST SET UP The experimental research and laboratory... methodology involved using a fluorescence oil sensor (Turner Designs Cyclops-7) to measure the TPH contained in the interstitial waters (i.e., pore

  7. Cost Effective, Ultra Sensitive Groundwater Monitoring for Site Remediation and Management

    DTIC Science & Technology

    2015-05-01

    feasibility studies. ................... 30  Table 5. Compounds screened in the laboratory for IS2 sampling...tank SVOC semivolatile organic compound TCE trichloroethene TPH total petroleum hydrocarbon USEPA U.S. Environmental Protection Agency UST...underground storage tank V volt VOA volatile organic analysis VOC volatile organic compound Technical material contained in this report has

  8. Effects of humic acid on phytodegradation of petroleum hydrocarbons in soil simultaneously contaminated with heavy metals.

    PubMed

    Park, Soyoung; Kim, Ki Seob; Kim, Jeong-Tae; Kang, Daeseok; Sung, Kijune

    2011-01-01

    The use of humic acid (HA) to enhance the efficiency of phytodegradation of petroleum hydrocarbons in soil contaminated with diesel fuel was evaluated in this study. A sample of soil was artificially contaminated with commercially available diesel fuel to an initial total petroleum hydrocarbons (TPH) concentration of 2300 mg/kg and four heavy metals with concentrations of 400 mg/kg for Pb, 200 mg/kg for Cu, 12 mg/kg for Cd, and 160 mg/kg for Ni. Three plant species, Brassica campestris, Festuca arundinacea, and Helianthus annuus, were selected for the phytodegradation experiment. Percentage degradation of TPH in the soil in a control pot supplemented with HA increased to 45% from 30% without HA. The addition of HA resulted in an increases in the removal of TPH from the soil in pots planted with B. campestris, E arundinacea, and H. annuus, enhancing percentage degradation to 86%, 64%, and 85% from 45%, 54%, and 66%, respectively. The effect of HA was also observed in the degradation of n-alkanes within 30 days. The rates of removal of n-alkanes in soil planted with B. campestris and H. annuus were high for n-alkanes in the range of C11-C28. A dynamic increase in dehydrogenase activity was observed during the last 15 days of a 30-day experimental period in all the pots amended with HA. The enhanced biodegradation performance for TPHs observed might be due to an increase in microbial activities and bioavailable TPH in soils caused by combined effects of plants and HA. The results suggested that HA could act as an enhancing agent for phytodegradation of petroleum hydrocarbons in soil contaminated with diesel fuel and heavy metals.

  9. Mathematical model simulation of a diesel spill in the Potomac River

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Feng, S.S.; Nicolette, J.P.; Markarian, R.K.

    1995-12-31

    A mathematical modeling technique was used to simulate the transport and fate of approximately 400,000 gallons of spilled diesel fuel and its impact on the aquatic biota in the Potomac River and Sugarland Run. Sugarland Run is a tributary about 21 miles upstream from Washington, DC. The mass balance model predicted the dynamic (spatial and temporal) distribution of spilled oil. The distributions were presented in terms of surface oil slick and sheen, dissolved and undissolved total petroleum hydrocarbons (TPH) in the water surface, water column, river sediments, shoreline and atmosphere. The processes simulated included advective movement, dispersion, dissolution, evaporation, volatilization,more » sedimentation, shoreline deposition, biodegradation, and removal of oil from cleanup operations. The model predicted that the spill resulted in a water column dissolved TPH concentration range of 0.05 to 18.6 ppm in Sugarland Run. The spilled oil traveled 10 miles along Sugarland Run before it reached the Potomac River. At the Potomac River, the water column TPH concentration was predicted to have decreased to the range of 0.0 to 0.43 ppm. These levels were consistent with field samples. To assess biological injury, the model used 4, 8, 24, 48, and 96-hr LC values in computing the fish injury caused by the fuel oil. The model used the maximum running average of dissolved TPH and exposure time to predict levels of fish mortality in the range of 38 to 40% in Sugarland Run. This prediction was consistent with field fisheries surveys. The model also computed the amount of spilled oil that adsorbed and settled into the river sediments.« less

  10. Mesocosm trials of bioremediation of contaminated soil of a petroleum refinery: comparison of natural attenuation, biostimulation and bioaugmentation.

    PubMed

    Couto, M Nazaré P F S; Monteiro, Emanuela; Vasconcelos, M Teresa S D

    2010-08-01

    Contamination with petroleum hydrocarbons (PHC) is a global problem with environmental implications. Physico-chemical treatments can be used for soil cleanup, but they are expensive, and can have implications for soil structure and environment. Otherwise, biological remediation treatments are cost-effective and restore soil structure. Several remediation experiments have been carried out in the lab and in the field; however, there is the challenge to achieve as good or better results in the field as in the laboratory. In the ambit of a project aiming at investigating suitable biological remediation approaches for recovering a refinery contaminated soil, we present here results obtained in bioremediation trials. The approaches biostimulation and bioaugmentation were tested, in parallel, and compared with natural attenuation. For this purpose, mesocosm experiments were carried out inside the refinery area, which constitutes a real asset of this work. Soil contaminated with crude oil was excavated, re-contaminated with turbine oil, homogenised and used to fill several 0.5 m(3) high-density polyethylene containers. The efficiency of procedures as follows: (1) natural attenuation; (2) manual aeration; (3) biostimulation by adding (3.1) only nutrients; and (3.2) nutrients and a non-ionic surfactant; and (4) bioaugmentation in the presence of added (4.1) nutrients or (4.2) nutrients and a non-ionic surfactant were evaluated after a 9-month period of experiment. For bioaugmentation, a commercial bacterial product was used. In addition to physico-chemical characterization, initial and final soil contents in total petroleum hydrocarbons (TPH) (by Fourier transform infrared spectrophotometry) and the total number of bacteria (by total cell counts) were carried out. For TPH degradation evaluation the soil was divided in four fractions corresponding to different depths: 0-5; 5-10; 10-15; and 15-20 cm. Mean values of percentages of PHC degradation varied between 20 and 50% at surface and between 10 and 35% below 5-cm depth. Natural attenuation was as efficient as most of the tested treatments (about 30% TPH degradation) being exceeded only by bioaugmentation combined with nutrient and surfactant amendments (about 50% TPH degradation). Higher TPH degradation at surface suggests that a combination of sufficient dioxygen, propitious for aerobically degradation, with sunlight required for production of strong photochemical oxidants like ozone, contributed for enhancing degradation. Indeed, the atmosphere of the refineries is relatively rich in volatile organic compounds and nitrogen dioxide (a side-product of the combustion of residual volatile PHC released by the chimneys), which are precursors of O(3) and other photochemical oxidants produced in sunny days, which are very common in Portugal. The fact that natural attenuation was as efficient as most of the soil treatments tested was very probably a result of the presence, in the initial soil, of physiologically adapted native microorganisms, which could be efficient in degrading PHC. A cost-effective way to reduce half-life for the degradation of PHC of contaminated soil of the refinery will be a periodic revolving of the soil, like tillage, in order to expose to the oxidative atmosphere the different layers of contaminated soil. A combination of soil revolving with bioaugmentation together with nutrients and surfactant amendments may result in an additional improvement of PHC degradation rate. However, this last procedure will raise markedly the price of the remediation treatment.

  11. Differences and similarities in the serotonergic diathesis for suicide attempts and mood disorders: a 22-year longitudinal gene-environment study.

    PubMed

    Brezo, J; Bureau, A; Mérette, C; Jomphe, V; Barker, E D; Vitaro, F; Hébert, M; Carbonneau, R; Tremblay, R E; Turecki, G

    2010-08-01

    To investigate similarities and differences in the serotonergic diathesis for mood disorders and suicide attempts, we conducted a study in a cohort followed longitudinally for 22 years. A total of 1255 members of this cohort, which is representative of the French-speaking population of Quebec, were investigated. Main outcome measures included (1) mood disorders (bipolar disorder and major depression) and suicide attempts by early adulthood; (2) odds ratios and probabilities associated with 143 single nucleotide polymorphisms in 11 serotonergic genes, acting directly or as moderators in gene-environment interactions with childhood sexual or childhood physical abuse (CPA), and in gene-gene interactions; (3) regression coefficients for putative endophenotypes for mood disorders (childhood anxiousness) and suicide attempts (childhood disruptiveness). Five genes showed significant adjusted effects (HTR2A, TPH1, HTR5A, SLC6A4 and HTR1A). Of these, HTR2A variation influenced both suicide attempts and mood disorders, although through different mechanisms. In suicide attempts, HTR2A variants (rs6561333, rs7997012 and rs1885884) were involved through interactions with histories of sexual and physical abuse whereas in mood disorders through one main effect (rs9316235). In terms of phenotype-specific contributions, TPH1 variation (rs10488683) was relevant only in the diathesis for suicide attempts. Three genes contributed exclusively to mood disorders, one through a main effect (HTR5A (rs1657268)) and two through gene-environment interactions with CPA (HTR1A (rs878567) and SLC6A4 (rs3794808)). Childhood anxiousness did not mediate the effects of HTR2A and HTR5A on mood disorders, nor did childhood disruptiveness mediate the effects of TPH1 on suicide attempts. Of the serotonergic genes implicated in mood disorders and suicidal behaviors, four exhibited phenotype-specific effects, suggesting that despite their high concordance and common genetic determinants, suicide attempts and mood disorders may also have partially independent etiological pathways. To identify where these pathways diverge, we need to understand the differential, phenotype-specific gene-environment interactions such as the ones observed in the present study, using suitably powered samples.

  12. Influence of enamel surface preparation on composite bond strength.

    PubMed

    Matos, Adriana Bona; Tate, William H; Powers, John M

    2003-09-01

    To evaluate the influence of air-particle abrasion and treatments on in vitro tensile bond strength of resin composite bonded to human enamel was evaluated using a single-bottle adhesive. Human teeth were divided into 12 groups of three treatments (none, 27-microm Al2O3 air-particle abrasion, 50-microm Al2O3 air-particle abrasion) and four conditioners [none, phosphoric acid (PA), NRC (no-rinse conditioner), and PA/NRC]. Bonding agent (Prime & Bond NT) and a resin composite (TPH Spectrum) were applied as inverted cones. Specimens were stored for 24 hours at 37 degrees C and debonded in tension using a testing machine at 0.5 mm/minute. Phosphoric acid treatment used with Prime & Bond NT produced the best bond strengths (24 MPa) to enamel for surfaces treated with 27-microm air-particle abrasion and for surfaces not treated with air-particle abrasion (control). With one exception, air-particle abraded surfaces resulted in bond strengths between 9 to 16 MPa. NRC with or without the use of phosphoric acid in general did not improve tensile bond strength to enamel when compared to surfaces not treated with NRC.

  13. PXR (NR1I2): splice variants in human tissues, including brain, and identification of neurosteroids and nicotine as PXR activators.

    PubMed

    Lamba, Vishal; Yasuda, Kazuto; Lamba, Jatinder K; Assem, Mahfoud; Davila, Julio; Strom, Stephen; Schuetz, Erin G

    2004-09-15

    To gain insight on the expression of pregnane X receptor (PXR), we analyzed PXR.1 and PXR alternatively spliced transcripts in a panel of 36 human tissues. PXR.1 was expressed in many more tissues than previously determined, including human bone marrow and select regions of the human brain. In each of these tissues, we observed alternative splicing of various exons of PXR that generated multiple distinct PXR isoforms. The most abundant PXR alternative mRNA transcripts lacked 111 nucleotides, deleting 37 amino acids from the PXR LBD (PXR.2), or lacked 123 nt, deleting 41 amino acids from the PXR LBD (PXR.3). CYP3A4, a gene transcriptionally regulated by PXR, showed incomplete overlap with PXR in its tissue distribution. Quantitation of PXR mRNAs in human liver demonstrated that PXR.2 and PXR.3 represented 6.7% and 0.32% of total PXR mRNA transcripts. Brain expression of PXR prompted analysis of whether some brain acting chemicals were PXR ligands. The neurosteroids allopregnanolone and pregnanolone activated PXR and induced transcription of a CYP3A4-luciferase reporter. Nicotine, the psychoactive and addictive chemical in cigarettes, and a known inducer of brain CYP2B6, was an efficacious activator of PXR and inducer of CYP3A4 transcription. Because nicotine activation of PXR will enhance metabolism of nicotine to the non-psychoactive cotinine, these results provide one molecular mechanism for the development of tolerance to nicotine. Moreover, the identification of PXR in many human tissues, such as brain, and activation by tissue specific ligands (such as neurosteroids) suggests additional biological roles for this receptor in these tissues.

  14. Genetic polymorphisms and their association with brain and behavioural measures in heterogeneous stock mice

    PubMed Central

    Janecka, Magdalena; Marzi, Sarah J.; Parsons, Michael J.; Liu, Lin; Paya-Cano, Jose L.; Smith, Rebecca G.; Fernandes, Cathy; Schalkwyk, Leonard C.

    2017-01-01

    Although the search for quantitative trait loci for behaviour remains a considerable challenge, the complicated genetic architecture of quantitative traits is beginning to be understood. The current project utilised heterogeneous stock (HS) male mice (n = 580) to investigate the genetic basis for brain weights, activity, anxiety and cognitive phenotypes. We identified 126 single nucleotide polymorphisms (SNPs) in genes involved in regulation of neurotransmitter systems, nerve growth/death and gene expression, and subsequently investigated their associations with changes in behaviour and/or brain weights in our sample. We found significant associations between four SNP-phenotype pairs, after controlling for multiple testing. Specificity protein 2 (Sp2, rs3708840), tryptophan hydroxylase 1 (Tph1, rs262731280) and serotonin receptor 3A (Htr3a, rs50670893) were associated with activity/anxiety behaviours, and microtubule-associated protein 2 (Map2, rs13475902) was associated with cognitive performance. All these genes except for Tph1 were expressed in the brain above the array median, and remained significantly associated with relevant behaviours after controlling for the family structure. Additionally, we found evidence for a correlation between Htr3a expression and activity. We discuss our findings in the light of the advantages and limitations of currently available mouse genetic tools, suggesting further directions for association studies in rodents. PMID:28145470

  15. Interplay between the key proteins of serotonin system in SSRI antidepressants efficacy.

    PubMed

    Kulikov, Alexander V; Gainetdinov, Raul R; Ponimaskin, Evgeni; Kalueff, Allan V; Naumenko, Vladimir S; Popova, Nina K

    2018-04-01

    Selective serotonin reuptake inhibitors (SSRIs) are the most effective and most used antidepressant drugs. Acting by inhibiting serotonin (5-HT) transporter, SSRIs display a typical 3-4-week delay in their therapeutic effects, with nearly 40% of depressed patients remaining treatment-resistant. Recent evidence suggests complex interplay between 5-HT receptors and key proteins of 5-HT metabolism in molecular mechanisms of such delay and resistance to SSRIs. Area covered: This paper concentrates on the interplay between 5-HT receptors in the delay of therapeutic effect of SSRIs, and the interaction between tryptophan hydroxylase 2 and 5-HT transporter in the SSRI resistance. Specifically, it discusses: (1) the data on the association between antidepressant drug efficacy and genetically defined characteristics of key proteins in the 5-HT signaling (TPH2, MAOA, SERT and 5-HT 1A receptor), (2) the effect of dimerization of 5-HT 7 and 5-HT 1A receptors on the internalization and functioning of 5-HT 1A presynaptic receptors, (3) the role of Tph2 deficiency in the resistance to SSRIs treatment. We shift the emphasis from individual proteins to their interactions in explaining antidepressant action of SSRI. Expert opinion: These interactions should be considered when developing more effective antidepressant drugs as well as for predicting and improving the efficacy of antidepressant therapies.

  16. Characteristics of petroleum-contaminated groundwater during natural attenuation: a case study in northeast China.

    PubMed

    Qian, Hong; Zhang, Yuling; Wang, Jiali; Si, Chaoqun; Chen, Zaixing

    2018-01-13

    The objective of this study was to investigate a petroleum-contaminated groundwater site in northeast China. We determined the physicochemical properties of groundwater that contained total petroleum hydrocarbons (TPH) with a view to developing a scientifically robust strategy for controlling and remediating pollution of groundwater already contaminated with petroleum. Samples were collected at regular intervals and were analyzed for dissolved oxygen (DO), iron (Fe 3+ ), sulfate (SO 4 2- ), electrical conductivity (Eh), pH, hydrogen carbonate (HCO 3 - ), and enzyme activities of catalase (CAT), peroxidase (HRP), catechol 1,2-dioxygenase (C12O), and catechol 2,3-dioxygenase (C23O). We used factor analysis in SPSS to determine the main environmental characteristics of the groundwater samples. The results confirmed that the study site was slightly contaminated and that TPH levels were decreasing slightly. Some of the physicochemical variables showed regular fluctuations; DO, Fe 3+ , and SO 4 2- contents decreased gradually, while the concentrations of one of the microbial degradation products, HCO 3 - , increased. Microorganism enzyme activities decreased gradually. The microbiological community deteriorated noticeably during the natural attenuation process, so microbiological degradation of pollutants receded gradually. The HCO 3 - content increased and the pH and Eh decreased gradually. The groundwater environment tended to be reducing.

  17. Evaluation of Plant- Compost -Microorganisms Synergy for the Remediation of Diesel contaminated Soil: Success Stories from the Field Station

    NASA Astrophysics Data System (ADS)

    Hussain, Imran; Wimmer, Bernhard; Soja, Gerhard; Sessitsch, Angela; Reichenauer, Thomas G.

    2016-04-01

    Total petroleum hydrocarbons (TPH) contain a mixture of crude oil, gasoline, creosote and diesel is one of the most common groups of persistent organic pollutants. TPH enters into the ecosystem (soil, water and air) through leakage of underground storage tanks (LUST), accidental oil spills, transportation losses and industrial processes. Pollution associated with diesel oil and its refined products is of great concern worldwide due to its threats/damages for human and ecosystem health, soil structure and ground water quality. Extensive soils pollution with petroleum hydrocarbons results in extreme harsh surroundings, produce hydrophobic conditions and infertile soils that ultimately lead towards less plant and microorganisms growth. Among biological methods, bioremediation and phytoremediation are promising technologies that have both technical and ecological benefits as compared to convention methods. Within phytoremediation, rhizoremediation based on stimulation of degrading microorganism's population influenced by plant rhizospheric effect is known as main mechanism for phytoremediation of petroleum polluted soils. Composting along with rhizodegradtion was used to remediate freshly spilled soils at Lysimeter station Siebersdof, Austria. Experiment was started in July 2013 and will be monitored up to September 2016. Field station has 12 Lysimeter in total; each has length, width and depth of 100 cm respectively. Each Lysimeter was filled with normal agricultural soil from Siebersdof (0-70 cm), sand (70-85 cm) and stones (85-100cm). Sand and stones were added to support the normal leaching and percolation of water as we collected leachate samples after regular intervals. After filling, commercial diesel oil (2% w/w of 0-70 cm soil) was spilled on top of each Lysimeter as accidental spill occurs in filed. Compost was added at 0-15 cm layer (5% w/w of soil) to stimulate plant as well as microorganisms growth. Whole Lysimeter station was divided into three treatments and four replicates; T1 was only planted with Lolium multiflorum and Lotus corniculatus, T2 was planted with both above mentioned plants inoculated with microbial consortium (mixture of strains: Pantoea sp. strains, ITSI10, BTRH79 and Pseudomonas sp. strain, MixRI75)and T3 was kept unplanted to support bioremediation. Germination percentage (GP) was monitored weekly until three weeks after seed sowing. Biometric parameters (plant height, fresh and dry weight of shoots) and leaf chlorophyll content were recorded in periodic intervals. Soil samples were taken in regular intervals (after every 6 month) and PHC content was measured by GC-FID. In the presentation we will report about the development of plants and the degradation of petroleum hydrocarbons in Lysimeter. The degradation of TPH will be reported for 7 layers inside each Lysimeter as well as in leachate samples.

  18. Lipopolysaccharide-induced inhibition of transcription of tlr4 in vitro is reversed by dexamethasone and correlates with presence of conserved NFκB binding sites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bonin, Camila P., E-mail: mila_bonin@yahoo.com.br; Baccarin, Raquel Y.A., E-mail: baccarin@usp.br; Nostell, Katarina, E-mail: katarina.nostell@slu.se

    2013-03-08

    Highlights: ► Chimpanzees, horses and humans have regions of similarity on TLR4 and MD2 promoters. ► Rodents have few regions of similarity on TLR4 promoter when compared to primates. ► Conserved NFkB binding sites were found in the promoters of TLR4 and MD2. ► LPS-induced inhibition of TLR4 transcription is reversed by dexamethasone. ► LPS-induced transcription of MD2 is inhibited by dexamethasone. -- Abstract: Engagement of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) is a master trigger of the deleterious effects of septic shock. Horses and humans are considered the most sensitive species to septic shock, but the mechanisms explainingmore » these phenomena remain elusive. Analysis of tlr4 promoters revealed high similarity among LPS-sensitive species (human, chimpanzee, and horse) and low similarity with LPS-resistant species (mouse and rat). Four conserved nuclear factor kappa B (NFκB) binding sites were found in the tlr4 promoter and two in the md2 promoter sequences that are likely to be targets for dexamethasone regulation. In vitro treatment of equine peripheral blood mononuclear cells (eqPBMC) with LPS decreased transcripts of tlr4 and increased transcription of md2 (myeloid differentiation factor 2) and cd14 (cluster of differentiation 14). Treatment with dexamethasone rescued transcription of tlr4 after LPS inhibition. LPS-induced transcription of md2 was inhibited in the presence of dexamethasone. Dexamethasone alone did not affect transcription of tlr4 and md2.« less

  19. Transcriptional Regulation of CYP2D6 Expression

    PubMed Central

    Pan, Xian; Ning, Miaoran

    2017-01-01

    CYP2D6-mediated drug metabolism exhibits large interindividual variability. Although genetic variations in the CYP2D6 gene are well known contributors to the variability, the sources of CYP2D6 variability in individuals of the same genotype remain unexplained. Accumulating data indicate that transcriptional regulation of CYP2D6 may account for part of CYP2D6 variability. Yet, our understanding of factors governing transcriptional regulation of CYP2D6 is limited. Recently, mechanistic studies of increased CYP2D6-mediated drug metabolism in pregnancy revealed two transcription factors, small heterodimer partner (SHP) and Krüppel-like factor 9, as a transcriptional repressor and an activator, respectively, of CYP2D6. Chemicals that increase SHP expression (e.g., retinoids and activators of farnesoid X receptor) were shown to downregulate CYP2D6 expression in the humanized mice as well as in human hepatocytes. This review summarizes the series of studies on the transcriptional regulation of CYP2D6 expression, potentially providing a basis to better understand the large interindividual variability in CYP2D6-mediated drug metabolism. PMID:27698228

  20. cDNA cloning and characterization of the human THRAP2 gene which maps to chromosome 12q24, and its mouse ortholog Thrap2.

    PubMed

    Musante, Luciana; Bartsch, Oliver; Ropers, Hans-Hilger; Kalscheuer, Vera M

    2004-05-12

    Characterization of a balanced t(2;12)(q37;q24) translocation in a patient with suspicion of Noonan syndrome revealed that the chromosome 12 breakpoint lies in the vicinity of a novel human gene, thyroid hormone receptor-associated protein 2 (THRAP2). We therefore characterized this gene and its mouse counterpart in more detail. Human and mouse THRAP2/Thrap2 span a genomic region of about 310 and >170 kilobases (kb), and both contain 31 exons. Corresponding transcripts are approximately 9.5 kb long. Their open reading frames code for proteins of 2210 and 2203 amino acids, which are 93% identical. By northern blot analysis, human and mouse THRAP2/Thrap2 genes showed ubiquitous expression. Transcripts were most abundant in human skeletal muscle and in mouse heart. THRAP2 protein is 56% identical to human TRAP240, which belongs to the thyroid hormone receptor associated protein (TRAP) complex and is evolutionary conserved up to yeast. This complex is involved in transcriptional regulation and is believed to serve as adapting interface between regulatory proteins bound to specific DNA sequences and RNA polymerase II.

  1. An evaluation of the urban stormwater pollutant removal efficiency of catch basin inserts.

    PubMed

    Morgan, Robert A; Edwards, Findlay G; Brye, Kristofor R; Burian, Stephen J

    2005-01-01

    In a storm sewer system, the catch basin is the interface between surface runoff and the sewer. Responding to the need to improve the quality of stormwater from urban areas and transportation facilities, and spurred by Phase I and II Stormwater Rules from the U.S. Environmental Protection Agency, several companies market catch basin inserts as best management practices for urban water quality management. However, little data have been collected under controlled tests that indicate the pollutant removal efficiency of these inserts when the inflow is near what can be expected to occur in the field. A stormwater simulator was constructed to test inserts under controlled and replicable conditions. The inserts were tested for removal efficiency of total suspended solids (TSS) and total petroleum hydrocarbons (TPH) at an inflow rate of 757 to 814 L/min, with influent pollutant concentrations of 225 mg/L TSS and 30 mg/L TPH. These conditions are similar to stormwater runoff from small commercial sites in the southeastern United States. Results from the tests indicate that at the test flowrate and pollutant concentration, average TSS removal efficiencies ranged from 11 to 42% and, for TPH, the removal efficiency ranged from 10 to 19%.

  2. Transcriptional Profiling of Human Endogenous Retrovirus Group HERV-K(HML-2) Loci in Melanoma

    PubMed Central

    Schmitt, Katja; Reichrath, Jörg; Roesch, Alexander; Meese, Eckart; Mayer, Jens

    2013-01-01

    Recent studies suggested a role for the human endogenous retrovirus (HERV) group HERV-K(HML-2) in melanoma because of upregulated transcription and expression of HERV-K(HML-2)-encoded proteins. Very little is known about which HML-2 loci are transcribed in melanoma. We assigned >1,400 HML-2 cDNA sequences generated from various melanoma and related samples to genomic HML-2 loci, identifying a total of 23 loci as transcribed. Transcription profiles of loci differed significantly between samples. One locus was found transcribed only in melanoma-derived samples but not in melanocytes and might represent a marker for melanoma. Several of the transcribed loci harbor ORFs for retroviral Gag and/or Env proteins. Env-encoding loci were transcribed only in melanoma. Specific investigation of rec and np9 transcripts indicated transcription of protein encoding loci in melanoma and melanocytes hinting at the relevance of Rec and Np9 in melanoma. UVB irradiation changed transcription profiles of loci and overall transcript levels decreased in melanoma and melanocytes. We further identified transcribed HML-2 loci formed by reverse transcription of spliced HML-2 transcripts by L1 machinery or in a retroviral fashion, with loci potentially encoding HML-2-like proteins. We reveal complex, sample-specific transcription of HML-2 loci in melanoma and related samples. Identified HML-2 loci and proteins encoded by those loci are particularly relevant for further studying the role of HML-2 in melanoma. Transcription of HERVs appears as a complex mechanism requiring specific studies to elucidate which HERV loci are transcribed and how transcribed HERVs may be involved in disease. PMID:23338945

  3. Molecular coevolution of mammalian ribosomal gene terminator sequences and the transcription termination factor TTF-I.

    PubMed Central

    Evers, R; Grummt, I

    1995-01-01

    Both the DNA elements and the nuclear factors that direct termination of ribosomal gene transcription exhibit species-specific differences. Even between mammals--e.g., human and mouse--the termination signals are not identical and the respective transcription termination factors (TTFs) which bind to the terminator sequence are not fully interchangeable. To elucidate the molecular basis for this species-specificity, we have cloned TTF-I from human and mouse cells and compared their structural and functional properties. Recombinant TTF-I exhibits species-specific DNA binding and terminates transcription both in cell-free transcription assays and in transfection experiments. Chimeric constructs of mouse TTF-I and human TTF-I reveal that the major determinant for species-specific DNA binding resides within the C terminus of TTF-I. Replacing 31 C-terminal amino acids of mouse TTF-I with the homologous human sequences relaxes the DNA-binding specificity and, as a consequence, allows the chimeric factor to bind the human terminator sequence and to specifically stop rDNA transcription. Images Fig. 2 Fig. 3 Fig. 4 PMID:7597036

  4. Effects of Oil Spillage on Vegetation, Land and Water Odu-Gboro Sagamu, Ogun State, South-Western Nigeria) Using Remote Sensing and GIS Techniques.

    NASA Astrophysics Data System (ADS)

    Oseni, O.

    2016-12-01

    This paper explores the impacts of oil spill on the physical environment with particular attention paid to the NNPC/PPMC pipeline system. It focuses on the environmental impacts of oil pollution in Nigeria, and discusses the increasing environmental contradictions of the area, and its influence on global warming. Nigeria's economy is highly dependent on earnings from the oil sector, which provides 20% of GDP, 95% of foreign exchange earnings, and about 65% of budgetary revenues. Since the discovery of oil in Nigeria in 1956, the country has been suffering the negative environmental consequences of oil exploration and exploitation. Between 1976 and 1996 a total of 4647 incidents resulted in the spill of approximately 2,369,470 barrels of oil into the environment. The study traces the effects of the oil spillage on the environment to determine whether oil spill is a major factor responsible for environmental pollution. By the use of remotely sensed data and other ancillary data, the major causes of oil spill in the region were identified; the presence of total petroleum hydrocarbon (TPH) in the environment, and it also determined the environmental impacts on land and water. Field observation and laboratory analysis of soil and water were used. Gas chromatography was used to determine the TPH concentration in soil extract and water extracts. Liquid-liquid extraction method was used for water and spectro-radiometer which is a very efficient process commonly used to determine spectral signature of various soil, water and plant samples obtained from the study area. Based largely on the GIS analysis, the findings showed that the main cause of oil spill is vandalism along the pipeline right of way; Vandalism which is an act of sabotage had the highest percentage compared to equipment failure, accident from oil tankers and accidental discharge during pipeline repairs. TPH were present at the site with soil samples having the high values, and the environmental impact on soil and water is due to poor resource management and control. Satellite imagery (Ikonos and Landsat series) helped in monitoring oil spill by providing the spill position. The petroleum industries should work with government agencies, universities and research centers to prevent oil spill incidents. Keywords: Environment, Pollution, GIS, TPH, Analysis, Vandalism.

  5. Two distinct promoters drive transcription of the human D1A dopamine receptor gene.

    PubMed

    Lee, S H; Minowa, M T; Mouradian, M M

    1996-10-11

    The human D1A dopamine receptor gene has a GC-rich, TATA-less promoter located upstream of a small, noncoding exon 1, which is separated from the coding exon 2 by a 116-base pair (bp)-long intron. Serial 3'-deletions of the 5'-noncoding region of this gene, including the intron and 5'-end of exon 2, resulted in 80 and 40% decrease in transcriptional activity of the upstream promoter in two D1A-expressing neuroblastoma cell lines, SK-N-MC and NS20Y, respectively. To investigate the function of this region, the intron and 245 bp at the 5'-end of exon 2 were investigated. Transient expression analyses using various chloramphenicol acetyltransferase constructs showed that the transcriptional activity of the intron is higher than that of the upstream promoter by 12-fold in SK-N-MC cells and by 5.5-fold in NS20Y cells in an orientation-dependent manner, indicating that the D1A intron is a strong promoter. Primer extension and ribonuclease protection assays revealed that transcription driven by the intron promoter is initiated at the junction of intron and exon 2 and at a cluster of nucleotides located 50 bp downstream from this junction. The same transcription start sites are utilized by the chloramphenicol acetyltransferase constructs employed in transfections as well as by the D1A gene expressed within the human caudate. The relative abundance of D1A transcripts originating from the upstream promoter compared with those transcribed from the intron promoter is 1.5-2.9 times in SK-N-MC cells and 2 times in the human caudate. Transcript stability studies in SK-N-MC cells revealed that longer D1A mRNA molecules containing exon 1 are degraded 1.8 times faster than shorter transcripts lacking exon 1. Although gel mobility shift assay could not detect DNA-protein interaction at the D1A intron, competitive co-transfection using the intron as competitor confirmed the presence of trans-acting factors at the intron. These data taken together indicate that the human D1A gene has two functional TATA-less promoters, both in D1A expressing cultured neuroblastoma cells and in the human striatum.

  6. Acute Fluoxetine Treatment Induces Slow Rolling of Leukocytes on Endothelium in Mice

    PubMed Central

    Herr, Nadine; Mauler, Maximilian; Witsch, Thilo; Stallmann, Daniela; Schmitt, Stefanie; Mezger, Julius; Bode, Christoph; Duerschmied, Daniel

    2014-01-01

    Objective Activated platelets release serotonin at sites of inflammation where it acts as inflammatory mediator and enhances recruitment of neutrophils. Chronic treatment with selective serotonin reuptake inhibitors (SSRI) depletes the serotonin storage pool in platelets, leading to reduced leukocyte recruitment in murine experiments. Here, we examined the direct and acute effects of SSRI on leukocyte recruitment in murine peritonitis. Methods C57Bl/6 and Tph1−/− (Tryptophan hydroxylase1) mice underwent acute treatment with the SSRI fluoxetine or vehicle. Serotonin concentrations were measured by ELISA. Leukocyte rolling and adhesion on endothelium was analyzed by intravital microscopy in mesentery venules with and without lipopolysaccharide challenge. Leukocyte extravasation in sterile peritonitis was measured by flow cytometry of abdominal lavage fluid. Results Plasma serotonin levels were elevated 2 hours after fluoxetine treatment (0.70±0.1 µg/ml versus 0.27±0.1, p = 0.03, n = 14), while serum serotonin did not change. Without further stimulation, acute fluoxetine treatment increased the number of rolling leukocytes (63±8 versus 165±17/0.04 mm2min−1) and decreased their velocity (61±6 versus 28±1 µm/s, both p<0.0001, n = 10). In Tph1−/− mice leukocyte rolling was not significantly influenced by acute fluoxetine treatment. Stimulation with lipopolysaccharide decreased rolling velocity and induced leukocyte adhesion, which was enhanced after fluoxetine pretreatment (27±3 versus 36±2/0.04 mm2, p = 0.008, n = 10). Leukocyte extravasation in sterile peritonitis, however, was not affected by acute fluoxetine treatment. Conclusions Acute fluoxetine treatment increased plasma serotonin concentrations and promoted leukocyte-endothelial interactions in-vivo, suggesting that serotonin is a promoter of acute inflammation. E-selectin was upregulated on endothelial cells in the presence of serotonin, possibly explaining the observed increase in leukocyte-endothelial interactions. However transmigration of neutrophils in sterile peritonitis was not affected by higher serotonin concentrations, indicating that the effect of fluoxetine was restricted to early steps in the leukocyte recruitment. Whether SSRI use in humans alters leukocyte recruitment remains to be investigated. PMID:24520366

  7. Cost-Effective, Ultra-Sensitive Groundwater Monitoring for Site Remediation and Management

    DTIC Science & Technology

    2015-05-01

    Example anion concentrations in groundwater used for feasibility studies. ................... 30 Table 5. Compounds screened in the laboratory for IS2...phase extraction ST storage tank SVOC semivolatile organic compound TCE trichloroethene TPH total petroleum hydrocarbon USEPA U.S. Environmental...Protection Agency UST underground storage tank V volt VOA volatile organic analysis VOC volatile organic compound Technical material

  8. Subsoil TPH contamination in two oil pipeline pumping stations and one pipeline right-of-way in north Mexico.

    PubMed

    Iturbe, R; Flores-Serrano, R M; Castro, A; Flores, C; Torres, L G

    2010-11-01

    This investigation deals with the characterization carried out in zones around two pipeline pumping stations and one pipeline right-of-way in the north of Mexico. In particular those areas where contamination was evaluated: (a) south area of the separation ditch in the Avalos station, (b) the area between the separation ditch at the Avalos station, (c) km 194+420 of the Moctuzma station, and (d) km 286+900 in the Candelaria station. Results of this investigation showed that only four samples showed TPH values higher than the Mexican limit for 2004: AVA 1B, with 21,191 mg kg(-1); AVA 1C, with 9348 mg kg(-1); AVA 2B, with 13,970 mg kg(-1); and MOC 2A, with 4108 mg kg(-1).None of the sampled points showed the presence of PAHs at values higher than those found in the Mexican or American legislations. PAH were detected in the range of 0.0004 and 13.05 mg kg(-1).It is suggested to implement surfactant soil washing as a remediation technique for the approximately 600 m(3) that need to be treated. Copyright 2010 Elsevier Ltd. All rights reserved.

  9. Use of sugarcane filter cake and nitrogen, phosphorus and potassium fertilization in the process of bioremediation of soil contaminated with diesel.

    PubMed

    Tellechea, Fernando Reynel Fundora; Martins, Marco Antônio; da Silva, Alexsandro Araujo; da Gama-Rodrigues, Emanuela Forestieri; Martins, Meire Lelis Leal

    2016-09-01

    This study evaluated the use of sugarcane filter cake and nitrogen, phosphorus and potassium (NPK) fertilization in the bioremediation of a soil contaminated with diesel fuel using a completely randomized design. Five treatments (uncontaminated soil, T1; soil contaminated with diesel, T2; soil contaminated with diesel and treated with 15 % (wt) filter cake, T3; soil contaminated with diesel and treated with NPK fertilizer, T4; and soil contaminated with diesel and treated with 15 % (wt) filter cake and NPK fertilizer, T5) and four evaluation periods (1, 60, 120, and 180 days after the beginning of the experiment) were used according to a 4 × 5 factorial design to analyze CO2 release. The variables total organic carbon (TOC) and total petroleum hydrocarbons (TPH) remaining in the soil were analyzed using a 5 × 2 factorial design, with the same treatments described above and two evaluation periods (1 and 180 days after the beginning of the experiment). In T3 and T5, CO2 release was significantly higher, compared with the other treatments. Significant TPH removal was observed on day 180, when percent removal values were 61.9, 70.1, 68.2, and 75.9 in treatments T2, T3, T4, and T5, respectively, compared with the initial value (T1).

  10. Influence of petroleum hydrocarbon contamination on microalgae and microbial activities in a long-term contaminated soil.

    PubMed

    Megharaj, M; Singleton, I; McClure, N C; Naidu, R

    2000-05-01

    Petroleum hydrocarbons are widespread environmental pollutants. Although biodegradation of petroleum hydrocarbons has been the subject of numerous investigations, information on their toxicity to microorganisms in soil is limited, with virtually no work conducted on soil algae. We carried out a screening experiment for total petroleum hydrocarbons (TPH) and their toxicity to soil algal populations, microbial biomass, and soil enzymes (dehydrogenase and urease) in a long-term TPH-polluted site with reference to an adjacent unpolluted site. Microbial biomass, soil enzyme activity, and microalgae declined in medium to high-level (5,200-21,430 mg kg(-1) soil) TPH-polluted soils, whereas low-level (<2,120 mg kg(-1) soil) pollution stimulated the algal populations and showed no effect on microbial biomass and enzymes. However, inhibition of all the tested parameters was more severe in soil considered to have medium-level pollution than in soils that were highly polluted. This result could not be explained by chemical analysis alone. Of particular interest was an observed shift in the species composition of algae in polluted soils with elimination of sensitive species in the medium to high polluted soils. Also, an algal growth inhibition test carried out using aqueous eluates prepared from polluted soils supported these results. Given the sensitivity of algae to synthetic pollutants, alteration in the algal species composition can serve as a useful bioindicator of pollution. The results of this experiment suggest that chemical analysis alone is not adequate for toxicological estimations and should be used in conjunction with bioassays. Furthermore, changes in species composition of algae proved to be more sensitive than microbial biomass and soil enzyme activity measurements.

  11. Elongin B-mediated epigenetic alteration of viral chromatin correlates with efficient human cytomegalovirus gene expression and replication.

    PubMed

    Hwang, Jiwon; Saffert, Ryan T; Kalejta, Robert F

    2011-01-01

    Elongins B and C are members of complexes that increase the efficiency of transcriptional elongation by RNA polymerase II (RNAPII) and enhance the monoubiquitination of histone H2B, an epigenetic mark of actively transcribed genes. Here we show that, in addition to its role in facilitating transcription of the cellular genome, elongin B also enhances gene expression from the double-stranded DNA genome of human cytomegalovirus (HCMV), a pathogenic herpesvirus. Reducing the level of elongin B by small interfering RNA- or short hairpin RNA-mediated knockdown decreased viral mRNA expression, viral protein accumulation, viral DNA replication, and infectious virion production. Chromatin immunoprecipitation analysis indicated viral genome occupancy of the elongating form of RNAPII, and monoubiquitinated histone H2B was reduced in elongin B-deficient cells. These data suggest that, in addition to the previously documented epigenetic regulation of transcriptional initiation, HCMV also subverts cellular elongin B-mediated epigenetic mechanisms for enhancing transcriptional elongation to enhance viral gene expression and virus replication. The genetic and epigenetic control of transcription initiation at both cellular and viral promoters is well documented. Recently, the epigenetic modification of histone H2B monoubiquitination throughout the bodies of cellular genes has been shown to enhance the elongation of RNA polymerase II-initiated transcripts. Mechanisms that might control the elongation of viral transcripts are less well studied. Here we show that, as with cellular genes, elongin B-mediated monoubiquitination of histone H2B also facilitates the transcriptional elongation of human cytomegalovirus genes. This and perhaps other epigenetic markings of actively transcribed regions may help in identifying viral genes expressed during in vitro latency or during natural infections of humans. Furthermore, this work identifies a novel, tractable model system to further study the regulation of transcriptional elongation in living cells.

  12. Roles of CDX2 and EOMES in human induced trophoblast progenitor cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chen, Ying, E-mail: ying.chen@hc.msu.edu; Wang, Kai; Gong, Yun Guo

    Highlights: ► CDX2 and EOMES play critical roles in human induced trophoblast progenitors (iTP). ► iTP cells directly transformed from fibroblasts. ► Differentiation of iTP cells into extravillous trophoblasts and syncytiotrophoblasts. -- Abstract: Abnormal trophoblast lineage proliferation and differentiation in early pregnancy have been associated with the pathogenesis of placenta diseases of pregnancy. However, there is still a gap in understanding the molecular mechanisms of early placental development due to the limited primary trophoblast cultures and fidelity of immortalized trophoblast lines. Trophoblasts stem (TS) cells, an in vitro model of trophectoderm that can differentiate into syncytiotrophoblasts and extravillous trophoblasts, canmore » be an attractive tool for early pregnancy research. TS cells are well established in mouse but not in humans due to insufficient knowledge of which trophoblast lineage-specific transcription factors are involved in human trophectoderm (TE) proliferation and differentiation. Here, we applied induced pluripotent stem cell technique to investigate the human trophoblast lineage-specific transcription factors. We established human induced trophoblast progenitor (iTP) cells by direct reprogramming the fibroblasts with a pool of mouse trophoblast lineage-specific transcription factors consisting of CDX2, EOMES, and ELF5. The human iTP cells exhibit epithelial morphology and can be maintained in vitro for more than 2 months. Gene expression profile of these cells was tightly clustered with human trophectoderm but not with human neuron progenitor cells, mesenchymal stem cells, or endoderm cells. These cells are capable of differentiating into cells with an invasive capacity, suggesting extravillous trophoblasts. They also form multi-nucleated cells which secrete human chorionic gonadotropin and estradiol, consistent with a syncytiotrophoblast phenotype. Our results provide the evidence that transcription factors CDX2 and EOMES may play critical roles in human iTP cell generation.« less

  13. Expression and regulation of estrogen-converting enzymes in ectopic human endometrial tissue.

    PubMed

    Fechner, Sabine; Husen, Bettina; Thole, Hubert; Schmidt, Markus; Gashaw, Isabella; Kimmig, Rainer; Winterhager, Elke; Grümmer, Ruth

    2007-10-01

    To investigate the regulation of estrogen-converting enzymes in human ectopic endometrial tissue. Animal study. Academic medical center. Sixty female nude mice with implanted human endometrial tissue. Twenty-two premenopausal women undergoing endometrial biopsy or hysterectomy. Human endometrial tissue was implanted into the peritoneal cavity of nude mice, and the effect of therapeutic drugs on transcription of steroid receptors and estrogen-converting enzymes was analyzed. Transcript levels of steroid hormone receptors, 17beta-hydroxysteroid dehydrogenase type 1 and 2, aromatase, and steroid sulfatase as well as proliferation rate were analyzed in the human ectopic endometrial tissue. Steroid receptors and estrogen-converting enzymes were expressed in the ectopic human endometrial fragments. Application of medroxyprogesterone acetate, dydrogesterone, danazol, and the aromatase inhibitor finrozole significantly inhibited aromatase transcription. In addition, danazol caused a significant decrease in transcription of steroid sulfatase, and finrozole, of 17beta-hydroxysteroid dehydrogenase type 1 in parallel to a decrease in proliferation rate in the ectopic human endometrial tissue. Pharmacological regulation of transcription of estrogen-converting enzymes in human endometrium cultured in nude mice may help to develop new therapeutic concepts based on local regulation of estrogen metabolism in endometriosis.

  14. Simultaneous determination of brominated and phosphate flame retardants in flame-retarded polyester curtains by a novel extraction method.

    PubMed

    Miyake, Yuichi; Tokumura, Masahiro; Nakayama, Hayato; Wang, Qi; Amagai, Takashi; Ogo, Sayaka; Kume, Kazunari; Kobayashi, Takeshi; Takasu, Shinji; Ogawa, Kumiko; Kannan, Kurunthachalam

    2017-12-01

    The use of novel brominated flame retardants (BFRs) and phosphate-based flame retardants (PFRs) has increased as substitutes for hexabromocyclododecane (HBCD) in many consumer products. To facilitate collection of data on chemicals used as flame retardants in textiles and fabrics, we developed an analytical method using liquid chromatography interfaced with tandem mass spectrometry (LC-MS/MS). We compared two extraction methods, one involving ultrasonic extraction (traditional method) using dichloromethane, toluene or acetone and the other encompassing complete dissolution of textile with 25% 1,1,1,3,3,3-hexafluoro-2-propanol/chloroform. The dissolution method extracted up to 204 times more BFRs and PFRs than the traditional ultrasonic extraction. Tris(2,3-dibromopropyl) isocyanurate (TDBP-TAZTO), triphenylphosphine oxide (TPhPO), tris(1,3-dichloro-2-propyl) phosphate (TDCPP), tricresyl phosphate (TCsP), and triphenyl phosphate (TPhP) were found in 40 flame-retarded curtain samples purchased from Japanese market in 2014. TDBP-TAZTO was detected in polyester curtains for the first time. Some of the flame-retarded curtain samples did not contain any of the known target analytes, which suggested the presence of other unknown flame retardants in those fabrics. Copyright © 2017. Published by Elsevier B.V.

  15. Expression of metastasis suppressor gene AES driven by a Yin Yang (YY) element in a CpG island promoter and transcription factor YY2.

    PubMed

    Kakizaki, Fumihiko; Sonoshita, Masahiro; Miyoshi, Hiroyuki; Itatani, Yoshiro; Ito, Shinji; Kawada, Kenji; Sakai, Yoshiharu; Taketo, M Mark

    2016-11-01

    We recently found that the product of the AES gene functions as a metastasis suppressor of colorectal cancer (CRC) in both humans and mice. Expression of amino-terminal enhancer of split (AES) protein is significantly decreased in liver metastatic lesions compared with primary colon tumors. To investigate its downregulation mechanism in metastases, we searched for transcriptional regulators of AES in human CRC and found that its expression is reduced mainly by transcriptional dysregulation and, in some cases, by additional haploidization of its coding gene. The AES promoter-enhancer is in a typical CpG island, and contains a Yin-Yang transcription factor recognition sequence (YY element). In human epithelial cells of normal colon and primary tumors, transcription factor YY2, a member of the YY family, binds directly to the YY element, and stimulates expression of AES. In a transplantation mouse model of liver metastases, however, expression of Yy2 (and therefore of Aes) is downregulated. In human CRC metastases to the liver, the levels of AES protein are correlated with those of YY2. In addition, we noticed copy-number reduction for the AES coding gene in chromosome 19p13.3 in 12% (5/42) of human CRC cell lines. We excluded other mechanisms such as point or indel mutations in the coding or regulatory regions of the AES gene, CpG methylation in the AES promoter enhancer, expression of microRNAs, and chromatin histone modifications. These results indicate that Aes may belong to a novel family of metastasis suppressors with a CpG-island promoter enhancer, and it is regulated transcriptionally. © 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

  16. Conserved and Divergent Features of Mesenchymal Progenitor Cell Types within the Cortical Nephrogenic Niche of the Human and Mouse Kidney.

    PubMed

    Lindström, Nils O; Guo, Jinjin; Kim, Albert D; Tran, Tracy; Guo, Qiuyu; De Sena Brandine, Guilherme; Ransick, Andrew; Parvez, Riana K; Thornton, Matthew E; Basking, Laurence; Grubbs, Brendan; McMahon, Jill A; Smith, Andrew D; McMahon, Andrew P

    2018-03-01

    Cellular interactions among nephron, interstitial, and collecting duct progenitors drive mammalian kidney development. In mice, Six2 + nephron progenitor cells (NPCs) and Foxd1 + interstitial progenitor cells (IPCs) form largely distinct lineage compartments at the onset of metanephric kidney development. Here, we used the method for analyzing RNA following intracellular sorting (MARIS) approach, single-cell transcriptional profiling, in situ hybridization, and immunolabeling to characterize the presumptive NPC and IPC compartments of the developing human kidney. As in mice, each progenitor population adopts a stereotypical arrangement in the human nephron-forming niche: NPCs capped outgrowing ureteric branch tips, whereas IPCs were sandwiched between the NPCs and the renal capsule. Unlike mouse NPCs, human NPCs displayed a transcriptional profile that overlapped substantially with the IPC transcriptional profile, and key IPC determinants, including FOXD1 , were readily detected within SIX2 + NPCs. Comparative gene expression profiling in human and mouse Six2/SIX2 + NPCs showed broad agreement between the species but also identified species-biased expression of some genes. Notably, some human NPC-enriched genes, including DAPL1 and COL9A2 , are linked to human renal disease. We further explored the cellular diversity of mesenchymal cell types in the human nephrogenic niche through single-cell transcriptional profiling. Data analysis stratified NPCs into two main subpopulations and identified a third group of differentiating cells. These findings were confirmed by section in situ hybridization with novel human NPC markers predicted through the single-cell studies. This study provides a benchmark for the mesenchymal progenitors in the human nephrogenic niche and highlights species-variability in kidney developmental programs. Copyright © 2018 by the American Society of Nephrology.

  17. Low-level expression of human ACAT2 gene in monocytic cells is regulated by the C/EBP transcription factors

    PubMed Central

    Guo, Dongqing; Lu, Ming; Hu, Xihan; Xu, Jiajia; Hu, Guangjing; Zhu, Ming; Zhang, Xiaowei; Li, Qin; Chang, Catherine C. Y.; Chang, Tayuan; Song, Baoliang; Xiong, Ying; Li, Boliang

    2016-01-01

    Acyl-coenzyme A:cholesterol acyltransferases (ACATs) are the exclusive intracellular enzymes that catalyze the formation of cholesteryl/steryl esters (CE/SE). In our previous work, we found that the high-level expression of human ACAT2 gene with the CpG hypomethylation of its whole promoter was synergistically regulated by two transcription factors Cdx2 and HNF1α in the intestine and fetal liver. Here, we first observed that the specific CpG-hypomethylated promoter was correlated with the low expression of human ACAT2 gene in monocytic cell line THP-1. Then, two CCAAT/enhancer binding protein (C/EBP) elements within the activation domain in the specific CpG-hypomethylation promoter region were identified, and the expression of ACAT2 in THP-1 cells was evidently decreased when the C/EBP transcription factors were knock-downed using RNAi technology. Furthermore, ChIP assay confirmed that C/EBPs directly bind to their elements for low-level expression of human ACAT2 gene in THP-1 cells. Significantly, the increased expressions of ACAT2 and C/EBPs were also found in macrophages differentiated from both ATRA-treated THP-1 cells and cultured human blood monocytes. These results demonstrate that the low-level expression of human ACAT2 gene with specific CpG-hypomethylated promoter is regulated by the C/EBP transcription factors in monocytic cells, and imply that the lowly expressed ACAT2 catalyzes the synthesis of certain CE/SE that are assembled into lipoproteins for the secretion. PMID:27688151

  18. Two populations of glutamatergic axons in the rat dorsal raphe nucleus defined by the vesicular glutamate transporters 1 and 2.

    PubMed

    Commons, Kathryn G; Beck, Sheryl G; Bey, Vincent W

    2005-03-01

    Most glutamatergic neurons in the brain express one of two vesicular glutamate transporters, vGlut1 or vGlut2. Cortical glutamatergic neurons highly express vGlut1, whereas vGlut2 predominates in subcortical areas. In this study immunohistochemical detection of vGlut1 or vGlut2 was used in combination with tryptophan hydroxylase (TPH) to characterize glutamatergic innervation of the dorsal raphe nucleus (DRN) of the rat. Immunofluorescence labeling of both vGlut1 and vGlut2 was punctate and homogenously distributed throughout the DRN. Puncta labeled for vGlut2 appeared more numerous then those labeled for vGlut1. Ultrastructural analysis revealed axon terminals containing vGlut1 and vGlut2 formed asymmetric-type synapses 80% and 95% of the time, respectively. Postsynaptic targets of vGlut1- and vGlut2-containing axons differed in morphology. vGlut1-labeled axon terminals synapsed predominantly on small-caliber (distal) dendrites (42%, 46/110) or dendritic spines (46%, 50/110). In contrast, vGlut2-containing axons synapsed on larger caliber (proximal) dendritic shafts (> 0.5 microm diameter; 48%, 78/161). A fraction of both vGlut1- or vGlut2-labeled axons synapsed onto TPH-containing dendrites (14% and 34%, respectively). These observations reveal that different populations of glutamate-containing axons innervate selective dendritic domains of serotonergic and non-serotonergic neurons, suggesting they play different functional roles in modulating excitation within the DRN.

  19. Human PIRH2 Enhances Androgen Receptor Signaling through Inhibition of Histone Deacetylase 1 and Is Overexpressed in Prostate Cancer

    PubMed Central

    Logan, Ian R.; Gaughan, Luke; McCracken, Stuart R. C.; Sapountzi, Vasileia; Leung, Hing Y.; Robson, Craig N.

    2006-01-01

    The androgen receptor (AR) is a hormone-dependent transcription factor critically involved in human prostate carcinogenesis. Optimal transcriptional control of androgen-responsive genes by AR may require complex interaction among multiple coregulatory proteins. We have previously shown that the AR coregulator TIP60 can interact with human PIRH2 (hPIRH2). In this study, we uncover important new functional role(s) for hPIRH2 in AR signaling: (i) hPIRH2 interacts with AR and enhances AR-mediated transcription with a dynamic pattern of recruitment to androgen response elements in the prostate-specific antigen (PSA) gene; (ii) hPIRH2 interacts with the AR corepressor HDAC1, leading to reduced HDAC1 protein levels and inhibition of transcriptional repression; (iii) hPIRH2 is required for optimal PSA expression; and (iv) hPIRH2 is involved in prostate cancer cell proliferation. In addition, overexpression of hPIRH2 protein was detected in 73 of 82 (89%) resected prostate cancers, with a strong correlation between increased hPIRH2 expression and aggressive disease, as signified by high Gleason sum scores and the presence of metastatic disease (P = <0.0001 and 0.0004, respectively). Collectively, our data establish hPIRH2 as a key modulator of AR function, opening a new direction for targeted therapy in aggressive human prostate cancer. PMID:16914734

  20. Human PIRH2 enhances androgen receptor signaling through inhibition of histone deacetylase 1 and is overexpressed in prostate cancer.

    PubMed

    Logan, Ian R; Gaughan, Luke; McCracken, Stuart R C; Sapountzi, Vasileia; Leung, Hing Y; Robson, Craig N

    2006-09-01

    The androgen receptor (AR) is a hormone-dependent transcription factor critically involved in human prostate carcinogenesis. Optimal transcriptional control of androgen-responsive genes by AR may require complex interaction among multiple coregulatory proteins. We have previously shown that the AR coregulator TIP60 can interact with human PIRH2 (hPIRH2). In this study, we uncover important new functional role(s) for hPIRH2 in AR signaling: (i) hPIRH2 interacts with AR and enhances AR-mediated transcription with a dynamic pattern of recruitment to androgen response elements in the prostate-specific antigen (PSA) gene; (ii) hPIRH2 interacts with the AR corepressor HDAC1, leading to reduced HDAC1 protein levels and inhibition of transcriptional repression; (iii) hPIRH2 is required for optimal PSA expression; and (iv) hPIRH2 is involved in prostate cancer cell proliferation. In addition, overexpression of hPIRH2 protein was detected in 73 of 82 (89%) resected prostate cancers, with a strong correlation between increased hPIRH2 expression and aggressive disease, as signified by high Gleason sum scores and the presence of metastatic disease (P = <0.0001 and 0.0004, respectively). Collectively, our data establish hPIRH2 as a key modulator of AR function, opening a new direction for targeted therapy in aggressive human prostate cancer.

  1. p21 as a Transcriptional Co-Repressor of S-Phase and Mitotic Control Genes

    PubMed Central

    Ferrándiz, Nuria; Caraballo, Juan M.; García-Gutierrez, Lucía; Devgan, Vikram; Rodriguez-Paredes, Manuel; Lafita, M. Carmen; Bretones, Gabriel; Quintanilla, Andrea; Muñoz-Alonso, M. Jose; Blanco, Rosa; Reyes, Jose C.; Agell, Neus; Delgado, M. Dolores; Dotto, G. Paolo; León, Javier

    2012-01-01

    It has been previously described that p21 functions not only as a CDK inhibitor but also as a transcriptional co-repressor in some systems. To investigate the roles of p21 in transcriptional control, we studied the gene expression changes in two human cell systems. Using a human leukemia cell line (K562) with inducible p21 expression and human primary keratinocytes with adenoviral-mediated p21 expression, we carried out microarray-based gene expression profiling. We found that p21 rapidly and strongly repressed the mRNA levels of a number of genes involved in cell cycle and mitosis. One of the most strongly down-regulated genes was CCNE2 (cyclin E2 gene). Mutational analysis in K562 cells showed that the N-terminal region of p21 is required for repression of gene expression of CCNE2 and other genes. Chromatin immunoprecipitation assays indicated that p21 was bound to human CCNE2 and other p21-repressed genes gene in the vicinity of the transcription start site. Moreover, p21 repressed human CCNE2 promoter-luciferase constructs in K562 cells. Bioinformatic analysis revealed that the CDE motif is present in most of the promoters of the p21-regulated genes. Altogether, the results suggest that p21 exerts a repressive effect on a relevant number of genes controlling S phase and mitosis. Thus, p21 activity as inhibitor of cell cycle progression would be mediated not only by the inhibition of CDKs but also by the transcriptional down-regulation of key genes. PMID:22662213

  2. Comprehensive Analysis of Human Endogenous Retrovirus Group HERV-W Locus Transcription in Multiple Sclerosis Brain Lesions by High-Throughput Amplicon Sequencing

    PubMed Central

    Schmitt, Katja; Richter, Christin; Backes, Christina; Meese, Eckart; Ruprecht, Klemens

    2013-01-01

    Human endogenous retroviruses (HERVs) of the HERV-W group comprise hundreds of loci in the human genome. Deregulated HERV-W expression and HERV-W locus ERVWE1-encoded Syncytin-1 protein have been implicated in the pathogenesis of multiple sclerosis (MS). However, the actual transcription of HERV-W loci in the MS context has not been comprehensively analyzed. We investigated transcription of HERV-W in MS brain lesions and white matter brain tissue from healthy controls by employing next-generation amplicon sequencing of HERV-W env-specific reverse transcriptase (RT) PCR products, thus revealing transcribed HERV-W loci and the relative transcript levels of those loci. We identified more than 100 HERV-W loci that were transcribed in the human brain, with a limited number of loci being predominantly transcribed. Importantly, relative transcript levels of HERV-W loci were very similar between MS and healthy brain tissue samples, refuting deregulated transcription of HERV-W env in MS brain lesions, including the high-level-transcribed ERVWE1 locus encoding Syncytin-1. Quantitative RT-PCR likewise did not reveal differences in MS regarding HERV-W env general transcript or ERVWE1- and ERVWE2-specific transcript levels. However, we obtained evidence for interindividual differences in HERV-W transcript levels. Reporter gene assays indicated promoter activity of many HERV-W long terminal repeats (LTRs), including structurally incomplete LTRs. Our comprehensive analysis of HERV-W transcription in the human brain thus provides important information on the biology of HERV-W in MS lesions and normal human brain, implications for study design, and mechanisms by which HERV-W may (or may not) be involved in MS. PMID:24109235

  3. Molecular cloning and functional characterization of the promoter region of the human uncoupling protein-2 gene.

    PubMed

    Tu, N; Chen, H; Winnikes, U; Reinert, I; Marmann, G; Pirke, K M; Lentes, K U

    1999-11-19

    As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5'-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt -141 to -66) a strong cis-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region. Copyright 1999 Academic Press.

  4. Molecular Mechanisms of Toxicity and Cell Damage by Chemicals in a Human Pancreatic Beta Cell Line, 1.1B4.

    PubMed

    Vasu, Srividya; McClenaghan, Neville H; Flatt, Peter R

    2016-10-01

    Mechanisms of toxicity and cell damage were investigated in novel clonal human pancreatic beta cell line, 1.1B4, after exposure to streptozotocin, alloxan, ninhydrin, and hydrogen peroxide. Viability, DNA damage, insulin secretion/content, [Ca]i, and glucokinase/hexokinase, mRNA expression were measured by MTT assay, comet assay, radioimmunoassay, fluorometric imaging plate reader, enzyme-coupled photometry, and real-time polymerase chain reaction, respectively. Chemicals significantly reduced 1.1B4 cell viability in a time/concentration-dependent manner. Chronic 18-hour exposure decreased cellular insulin, glucokinase, and hexokinase activities. Chemicals decreased transcription of INS, GCK, PCSK1, PCSK2, and GJA1 (involved in secretory function). Insulin release and [Ca]i responses to nutrients and membrane-depolarizing agents were impaired. Streptozotocin and alloxan up-regulated transcription of genes, SOD1 and SOD2 (antioxidant enzymes). Ninhydrin and hydrogen peroxide up-regulated SOD2 transcription, whereas alloxan and hydrogen peroxide increased CAT transcription. Chemicals induced DNA damage, apoptosis, and increased caspase 3/7 activity. Streptozotocin and alloxan decreased transcription of BCL2 while increasing transcription of BAX. Chemicals did not affect transcription of HSPA4 and HSPA5 and nitrite production. 1.1B4 cells represent a useful model of human beta cells. Chemicals impaired 1.1B4 cell secretory function and activated antioxidant defense and apoptotic pathways without activating endoplasmic reticulum stress response/nitrosative stress.

  5. Microbiological characteristics of multi-media PRB reactor in the bioremediation of groundwater contaminated by petroleum hydrocarbons.

    PubMed

    Liu, Hong; Zhang, Lanying; Deng, Haijing; Liu, Na; Liu, Cuizhu

    2011-10-01

    A multi-media bio-PRB reactor was designed to treat groundwater contaminated with petroleum hydrocarbons. After a 208-day bioremediation, combined with the total petroleum hydrocarbons content in the groundwater flowed through the reactor, microbiological characteristics of the PRB reactor including microbes immobilized and its dehydrogenase activity were investigated. TPH was significantly reduced by as much as 65% in the back of the second media layer, whereas in the third layer, the TPH content reached lower than 1 mg l⁻¹. For microbes immobilized on the media, the variations with depth in different media were significantly the same and the regularity was obvious in the forepart of the media, which increased with depth at first and then reduced gradually, while in the back-end, the microbes almost did not have any variations with depth but decreased with the distance. The dehydrogenase activity varied from 2.98 to 16.16 mg TF L⁻¹ h⁻¹ and its distribution illustrated a similar trend with numbers of microbial cell, therefore, the noticeable correlation was found between them.

  6. Cyclin E-Mediated Human Proopiomelanocortin Regulation as a Therapeutic Target for Cushing Disease.

    PubMed

    Liu, Ning-Ai; Araki, Takako; Cuevas-Ramos, Daniel; Hong, Jiang; Ben-Shlomo, Anat; Tone, Yukiko; Tone, Masahide; Melmed, Shlomo

    2015-07-01

    Cushing disease, due to pituitary corticotroph tumor ACTH hypersecretion, drives excess adrenal cortisol production with adverse morbidity and mortality. Loss of glucocorticoid negative feedback on the hypothalamic-pituitary-adrenal axis leads to autonomous transcription of the corticotroph precursor hormone proopiomelanocortin (POMC), consequent ACTH overproduction, and adrenal hypercortisolism. We previously reported that R-roscovitine (CYC202, seliciclib), a 2,6,9-trisubstituted purine analog, suppresses cyclin-dependent-kinase 2/cyclin E and inhibits ACTH in mice and zebrafish. We hypothesized that intrapituitary cyclin E signaling regulates corticotroph tumor POMC transcription independently of cell cycle progression. The aim was to investigate whether R-roscovitine inhibits human ACTH in corticotroph tumors by targeting the cyclin-dependent kinase 2/cyclin E signaling pathway. Primary cell cultures of surgically resected human corticotroph tumors were treated with or without R-roscovitine, ACTH measured by RIA and quantitative PCR, and/or Western blot analysis performed to investigate ACTH and lineage-specific transcription factors. Cyclin E and E2F transcription factor 1 (E2F1) small interfering RNA (siRNA) transfection was performed in murine corticotroph tumor AtT20 cells to elucidate mechanisms for drug action. POMC gene promoter activity in response to R-roscovitine treatment was analyzed using luciferase reporter and chromatin immunoprecipitation assays. R-roscovitine inhibits human corticotroph tumor POMC and Tpit/Tbx19 transcription with decreased ACTH expression. Cyclin E and E2F1 exhibit reciprocal positive regulation in corticotroph tumors. R-roscovitine disrupts E2F1 binding to the POMC gene promoter and suppresses Tpit/Tbx19 and other lineage-specific POMC transcription cofactors via E2F1-dependent and -independent pathways. R-roscovitine inhibits human pituitary corticotroph tumor ACTH by targeting the cyclin E/E2F1 pathway. Pituitary cyclin E/E2F1 signaling is a previously unappreciated molecular mechanism underlying neuroendocrine regulation of the hypothalamic-pituitary-adrenal axis, providing a subcellular therapeutic target for small molecule cyclin-dependent kinase 2 inhibitors of pituitary ACTH-dependent hypercortisolism, ie, Cushing disease.

  7. The Influence of Family Structure, the TPH2 G-703T and the 5-HTTLPR Serotonergic Genes upon Affective Problems in Children Aged 10-14 Years

    ERIC Educational Resources Information Center

    Nobile, Maria; Rusconi, Marianna; Bellina, Monica; Marino, Cecilia; Giorda, Roberto; Carlet, Ombretta; Vanzin, Laura; Molteni, Massimo; Battaglia, Marco

    2009-01-01

    Background: Both genetic and psychosocial risk factors influence the risk for depression in development. While the impacts of family structure and of serotonergic polymorphisms upon individual differences for affective problems have been investigated separately, they have never been considered together in a gene-environment interplay perspective.…

  8. Tissue-specific expression and regulation of the alternatively-spliced forms of lysyl hydroxylase 2 (LH2) in human kidney cells and skin fibroblasts.

    PubMed

    Walker, Linda C; Overstreet, Mayra A; Yeowell, Heather N

    2005-01-01

    Lysyl hydroxylases 1, 2, and 3 catalyse the hydroxylation of specific lysines in collagen. A small percentage of these hydroxylysine residues are precursors for the cross-link formation essential for the tensile strength of collagen. Lysyl hydroxylase 2 (LH2) exists as two alternatively-spliced forms; the long transcript (the major ubiquitously-expressed form) includes a 63 bp exon (13A) that is spliced out in the short form (expressed, together with the long form, in human kidney, spleen, liver, and placenta). This study shows that this alternative splicing event can be regulated by both cell density and cycloheximide (CHX). Although only the long form of LH2 is detected in untreated confluent human skin fibroblasts, after 24 h treatment with CHX the short LH2 transcript is also expressed. In kidney cells, in which both LH2 transcripts are equally expressed, the long LH2 transcript is significantly decreased after 24 h CHX treatment, whereas expression of the short transcript is slightly increased. This suggests that, in kidney cells, the splicing mechanism for the inclusion of exon 13A in LH2 requires a newly-synthesized protein factor that is suppressed by CHX, whereas, in skin fibroblasts in which levels of LH2 (long) are unaffected, CHX appears to suppress a factor that inhibits exclusion of exon 13A, thereby promoting expression of LH2 (short). As these alternate transcripts of LH2 may have specificity for hydroxylation of lysines in either telopeptide or helical collagen domains, their relative expression determines the type of cross-links formed, thereby affecting collagen strength. Therefore, any perturbation of the regulation of LH2 splicing could influence the stability of the extracellular matrix and contribute to specific connective tissue disorders.

  9. miRNA Enriched in Human Neuroblast Nuclei Bind the MAZ Transcription Factor and Their Precursors Contain the MAZ Consensus Motif.

    PubMed

    Goldie, Belinda J; Fitzsimmons, Chantel; Weidenhofer, Judith; Atkins, Joshua R; Wang, Dan O; Cairns, Murray J

    2017-01-01

    While the cytoplasmic function of microRNA (miRNA) as post-transcriptional regulators of mRNA has been the subject of significant research effort, their activity in the nucleus is less well characterized. Here we use a human neuronal cell model to show that some mature miRNA are preferentially enriched in the nucleus. These molecules were predominantly primate-specific and contained a sequence motif with homology to the consensus MAZ transcription factor binding element. Precursor miRNA containing this motif were shown to have affinity for MAZ protein in nuclear extract. We then used Ago1/2 RIP-Seq to explore nuclear miRNA-associated mRNA targets. Interestingly, the genes for Ago2-associated transcripts were also significantly enriched with MAZ binding sites and neural function, whereas Ago1-transcripts were associated with general metabolic processes and localized with SC35 spliceosomes. These findings suggest the MAZ transcription factor is associated with miRNA in the nucleus and may influence the regulation of neuronal development through Ago2-associated miRNA induced silencing complexes. The MAZ transcription factor may therefore be important for organizing higher order integration of transcriptional and post-transcriptional processes in primate neurons.

  10. Selective activation of human heat shock gene transcription by nitrosourea antitumor drugs mediated by isocyanate-induced damage and activation of heat shock transcription factor.

    PubMed Central

    Kroes, R A; Abravaya, K; Seidenfeld, J; Morimoto, R I

    1991-01-01

    Treatment of cultured human tumor cells with the chloroethylnitrosourea antitumor drug 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) selectively induces transcription and protein synthesis of a subset of the human heat shock or stress-induced genes (HSP90 and HSP70) with little effect on other stress genes or on expression of the c-fos, c-myc, or beta-actin genes. The active component of BCNU and related compounds appears to be the isocyanate moiety that causes carbamoylation of proteins and nucleic acids. Transcriptional activation of the human HSP70 gene by BCNU is dependent on the heat shock element and correlates with the level of heat shock transcription factor and its binding to the heat shock element in vivo. Unlike activation by heat or heavy metals, BCNU-mediated activation is strongly dependent upon new protein synthesis. This suggests that BCNU-induced, isocyanate-mediated damage to newly synthesized protein(s) may be responsible for activation of the heat shock transcription factor and increased transcription of the HSP90 and HSP70 genes. Images PMID:2052560

  11. Treadmill exercise alleviates depressive symptoms in rotenone-induced Parkinson disease rats

    PubMed Central

    Shin, Mal-Soon; Kim, Tae-Woon; Lee, Jae-Min; Sung, Yun-Hee; Lim, Baek-Vin

    2017-01-01

    Parkinson disease (PD) is characterized by selective loss of the dopaminergic neurons. The symptoms of depression following PD are closely associated with reduced activity of the serotonergic system in the dorsal raphe. We explored the antidepressive effect of exercise and its possible mechanism using the rotenone-induced PD rats. PD rats were induced by subcutaneously injection with rotenone for 14 days. The rats in the exercise groups were made to run on a treadmill for 30 min once a day during 14 consecutive days. Forced swimming test, immunohistochemistry for serotonin (5-hydroxytryptamine, 5-HT), tryptophan hydroxylase (TPH), and western blot for serotonin 1A (5-HT1A) receptor were conducted. Injection of rotenone induced PD rats. PD rats showed depressive state and treadmill exercise ameliorated this depressive state. 5-HT, TPH, and 5-HT1A receptor expressions in the dorsal raphe were suppressed by rotenone injection and treadmill exercise increased the expressions of 5-HT, TPH, and 5-HT1A receptor in the rotenone-injected rats. The present results show that treadmill exercise ameliorated depressive symptoms in the rotenone-induced PD rats. The antidepressive effect of treadmill exercise might be ascribed to the enhancement of serotonergic function through upregulation of 5-HT1A expression in the dorsal raphe. PMID:28503522

  12. Treadmill exercise alleviates depressive symptoms in rotenone-induced Parkinson disease rats.

    PubMed

    Shin, Mal-Soon; Kim, Tae-Woon; Lee, Jae-Min; Sung, Yun-Hee; Lim, Baek-Vin

    2017-04-01

    Parkinson disease (PD) is characterized by selective loss of the dopaminergic neurons. The symptoms of depression following PD are closely associated with reduced activity of the serotonergic system in the dorsal raphe. We explored the antidepressive effect of exercise and its possible mechanism using the rotenone-induced PD rats. PD rats were induced by subcutaneously injection with rotenone for 14 days. The rats in the exercise groups were made to run on a treadmill for 30 min once a day during 14 consecutive days. Forced swimming test, immunohistochemistry for serotonin (5-hydroxytryptamine, 5-HT), tryptophan hydroxylase (TPH), and western blot for serotonin 1A (5-HT1A) receptor were conducted. Injection of rotenone induced PD rats. PD rats showed depressive state and treadmill exercise ameliorated this depressive state. 5-HT, TPH, and 5-HT1A receptor expressions in the dorsal raphe were suppressed by rotenone injection and treadmill exercise increased the expressions of 5-HT, TPH, and 5-HT1A receptor in the rotenone-injected rats. The present results show that treadmill exercise ameliorated depressive symptoms in the rotenone-induced PD rats. The antidepressive effect of treadmill exercise might be ascribed to the enhancement of serotonergic function through upregulation of 5-HT1A expression in the dorsal raphe.

  13. A combined approach of physicochemical and biological methods for the characterization of petroleum hydrocarbon-contaminated soil.

    PubMed

    Masakorala, Kanaji; Yao, Jun; Chandankere, Radhika; Liu, Haijun; Liu, Wenjuan; Cai, Minmin; Choi, Martin M F

    2014-01-01

    Main physicochemical and microbiological parameters of collected petroleum-contaminated soils with different degrees of contamination from DaGang oil field (southeast of Tianjin, northeast China) were comparatively analyzed in order to assess the influence of petroleum contaminants on the physicochemical and microbiological properties of soil. An integration of microcalorimetric technique with urease enzyme analysis was used with the aim to assess a general status of soil metabolism and the potential availability of nitrogen nutrient in soils stressed by petroleum-derived contaminants. The total petroleum hydrocarbon (TPH) content of contaminated soils varied from 752.3 to 29,114 mg kg(−1). Although the studied physicochemical and biological parameters showed variations dependent on TPH content, the correlation matrix showed also highly significant correlation coefficients among parameters, suggesting their utility in describing a complex matrix such as soil even in the presence of a high level of contaminants. The microcalorimetric measures gave evidence of microbial adaptation under highest TPH concentration; this would help in assessing the potential of a polluted soil to promote self-degradation of oil-derived hydrocarbon under natural or assisted remediation. The results highlighted the importance of the application of combined approach in the study of those parameters driving the soil amelioration and bioremediation.

  14. Genetic analyses, phenotypic adaptability and stability in sugarcane genotypes for commercial cultivation in Pernambuco.

    PubMed

    Dutra Filho, J A; Junior, T C; Simões Neto, D E

    2015-10-05

    In the present study, we assessed the agro-industrial performance of 22 sugarcane genotypes adaptable to edaphoclimatic conditions in production microregions in the State of Pernambuco, Brazil, and we recommended the commercial cultivation of select genotypes. The variables analyzed were as follows: sucrose percentage in cane juice, tonnage of saccharose per hectare (TPH), sugarcane tonnage per hectare (TCH), fiber, solid soluble contents, total recoverable sugar tonnage (ATR), and total recoverable sugar tonnage per hectare (ATR t/ha). A randomized block design with 4 repeats was used. Combined variance of the experiments, genetic parameter estimates, and environment stratification were analyzed. Phenotypic adaptability and stability were analyzed using the Annicchiarico and Wricke methods and analysis of variance. Genetic gain was estimated using the classic index and sum of ranks. Genotype selection was efficient for TPH, TCH, and ATR t/ha. Genotypes presented a great potential for improvement and a similar response pattern in Litoral Norte and Mata Sul microregions for TPH and TCH and Litoral Norte and Litoral Sul microregions for ATR t/ha. Genotypes SP78-4764, RB813804, and SP79-101 showed better productivity and phenotypic adaptability and stability, according to the Wricke and Annicchiarico methods. These genotypes can be recommended for cultivation in the sugarcane belt in the State of Pernambuco.

  15. Effects of Low Temperature and Freeze-Thaw Cycles on Hydrocarbon Biodegradation in Arctic Tundra Soil

    PubMed Central

    Eriksson, Mikael; Ka, Jong-Ok; Mohn, William W.

    2001-01-01

    Degradation of petroleum hydrocarbons was monitored in microcosms with diesel fuel-contaminated Arctic tundra soil incubated for 48 days at low temperatures (−5, 0, and 7°C). An additional treatment was incubation for alternating 24-h periods at 7 and −5°C. Hydrocarbons were biodegraded at or above 0°C, and freeze-thaw cycles may have actually stimulated hydrocarbon biodegradation. Total petroleum hydrocarbon (TPH) removal over 48 days in the 7, 0, and 7 and −5°C treatments, respectively, was 450, 300, and 600 μg/g of soil. No TPH removal was observed at −5°C. Total carbon dioxide production suggested that TPH removal was due to biological mineralization. Bacterial metabolic activity, indicated by RNA/DNA ratios, was higher in the middle of the experiment (day 21) than at the start, in agreement with measured hydrocarbon removal and carbon dioxide production activities. The total numbers of culturable heterotrophs and of hydrocarbon degraders did not change significantly over the 48 days of incubation in any of the treatments. At the end of the experiment, bacterial community structure, evaluated by ribosomal intergenic spacer length analysis, was very similar in all of the treatments but the alternating 7 and −5°C treatment. PMID:11679333

  16. Effect of Piriformospora indica inoculation on root development and distribution of maize (Zea mays L.) in the presence of petroleum contaminated soil

    NASA Astrophysics Data System (ADS)

    Zamani, Javad; Hajabbasi, Mohammad Ali; Alaie, Ebrahim

    2014-05-01

    The root systems of most terrestrial plants are confronted to various abiotic and biotic stresses. One of these abiotic stresses is contamination of soil with petroleum hydrocarbon, which the efficiency of phytoremediation of petroleum hydrocarbons in soils is dependent on the ability of plant roots to development into the contaminated soils. Piriformospora indica represents a recently discovered fungus that transfers considerable beneficial impact to its host plants. A rhizotron experiment was conducted to study the effects of P. Indica inoculation on root distribution and root and shoot development of maize (Zea mays L.) in the presence of three patterns of petroleum contamination in the soil (subsurface contamination, continuous contamination and without contamination (control)). Root distribution and root and shoot development were monitored over time. The final root and shoot biomass and the final TPH concentration in the rhizosphere were determined. Analysis of digitized images which were prepared of the tracing of the appeared roots along the front rhizotrons showed the depth and total length of root network in the contamination treatments were significantly decreased. Although the degradation of TPH in the rhizosphere of maize was significant, but there were no significant differences between degradation of TPH in the rhizosphere of +P. indica plants in comparison to -P. indica plants.

  17. Bacterial community shift and hydrocarbon transformation during bioremediation of short-term petroleum-contaminated soil.

    PubMed

    Wu, Manli; Ye, Xiqiong; Chen, Kaili; Li, Wei; Yuan, Jing; Jiang, Xin

    2017-04-01

    A laboratory study was conducted to evaluate the impact of bioaugmentation plus biostimulation (BR, added both nutrients and bacterial consortia), and natural attenuation (NA) on hydrocarbon degradation efficiency and microflora characterization during remediation of a freshly contaminated soil. After 112 days of remediation, the initial level of total petroleum hydrocarbon (TPH) (61,000 mg/kg soil) was reduced by 4.5% and 5.0% in the NA and BR treatments, respectively. Bioremediation did not significantly enhance TPH biodegradation compared to natural attenuation. The degradation of the aliphatic fraction was the most active with the degradation rate of 30.3 and 28.7 mg/kg/day by the NA and BR treatments, respectively. Soil microbial activities and counts in soil were generally greater for bioremediation than for natural attenuation. MiSeq sequencing indicated that the diversity and structure of microbial communities were affected greatly by bioremediation. In response to bioremediation treatment, Promicromonospora, Pseudomonas, Microcella, Mycobacterium, Alkanibacter, and Altererythrobacter became dominant genera in the soil. The result indicated that combining bioaugmentation with biostimulation did not improve TPH degradation, but soil microbial activities and structure of microbial communities are sensitive to bioremediation in short-term and heavily oil-contaminated soil. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Assessment of three approaches of bioremediation (Natural Attenuation, Landfarming and Bioagumentation - Assistited Landfarming) for a petroleum hydrocarbons contaminated soil.

    PubMed

    Guarino, C; Spada, V; Sciarrillo, R

    2017-03-01

    Contamination with total petroleum hydrocarbons (TPH) subsequent to refining activities, is currently one of the major environmental problems. Among the biological remediation approaches, landfarming and in situ bioremediation strategies are of great interest. Purpose of this study was to verify the feasibility of a remediation process wholly based on biological degradation applied to contaminated soils from a decommissioned refinery. This study evaluated through a pot experiment three bioremediation strategies: a) Natural Attenuation (NA), b) Landfarming (L), c) Bioaugmentation-assisted Landfarming (LB) for the treatment of a contaminated soil with petroleum hydrocarbons (TPHs). After a 90-days trial, Bioagumentation - assistited Landfarming approach produced the best results and the greatest evident effect was shown with the most polluted samples reaching a reduction of about 86% of total petroleum hydrocarbons (TPH), followed by Landfarming (70%), and Natural Attenuation (57%). The results of this study demonstrated that the combined use of bioremediation strategies was the most advantageous option for the treatment of contaminated soil with petroleum hydrocarbons, as compared to natural attenuation, bioaugmentation or landfarming applied alone. Besides, our results indicate that incubation with an autochthonous bacterial consortium may be a promising method for bioremediation of TPH-contaminated soils. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Fate of hydrocarbon pollutants in source and non-source control sustainable drainage systems.

    PubMed

    Roinas, Georgios; Mant, Cath; Williams, John B

    2014-01-01

    Sustainable drainage (SuDs) is an established method for managing runoff from developments, and source control is part of accepted design philosophy. However, there are limited studies into the contribution source control makes to pollutant removal, especially for roads. This study examines organic pollutants, total petroleum hydrocarbons (TPH) and polycyclic aromatic hydrocarbons (PAHs), in paired source and non-source control full-scale SuDs systems. Sites were selected to cover local roads, trunk roads and housing developments, with a range of SuDs, including porous asphalt, swales, detention basins and ponds. Soil and water samples were taken bi-monthly over 12 months to assess pollutant loads. Results show first flush patterns in storm events for solids, but not for TPH. The patterns of removal for specific PAHs were also different, reflecting varying physico-chemical properties. The potential of trunk roads for pollution was illustrated by peak runoff for TPH of > 17,000 μg/l. Overall there was no significant difference between pollutant loads from source and non-source control systems, but the dynamic nature of runoff means that longer-term data are required. The outcomes of this project will increase understanding of organic pollutants behaviour in SuDs. This will provide design guidance about the most appropriate systems for treating these pollutants.

  20. Human Mitochondrial Transcription Factor B2 Is Required for Promoter Melting during Initiation of Transcription.

    PubMed

    Posse, Viktor; Gustafsson, Claes M

    2017-02-17

    The mitochondrial transcription initiation machinery in humans consists of three proteins: the RNA polymerase (POLRMT) and two accessory factors, transcription factors A and B2 (TFAM and TFB2M, respectively). This machinery is required for the expression of mitochondrial DNA and the biogenesis of the oxidative phosphorylation system. Previous experiments suggested that TFB2M is required for promoter melting, but conclusive experimental proof for this effect has not been presented. Moreover, the role of TFB2M in promoter unwinding has not been discriminated from that of TFAM. Here we used potassium permanganate footprinting, DNase I footprinting, and in vitro transcription from the mitochondrial light-strand promoter to study the role of TFB2M in transcription initiation. We demonstrate that a complex composed of TFAM and POLRMT was readily formed at the promoter but alone was insufficient for promoter melting, which only occurred when TFB2M joined the complex. We also show that mismatch bubble templates could circumvent the requirement of TFB2M, but TFAM was still required for efficient initiation. Our findings support a model in which TFAM first recruits POLRMT to the promoter, followed by TFB2M binding and induction of promoter melting. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Transcriptional activation of short interspersed elements by DNA-damaging agents.

    PubMed

    Rudin, C M; Thompson, C B

    2001-01-01

    Short interspersed elements (SINEs), typified by the human Alu repeat, are RNA polymerase III (pol III)-transcribed sequences that replicate within the genome through an RNA intermediate. Replication of SINEs has been extensive in mammalian evolution: an estimated 5% of the human genome consists of Alu repeats. The mechanisms regulating transcription, reverse transcription, and reinsertion of SINE elements in genomic DNA are poorly understood. Here we report that expression of murine SINE transcripts of both the B1 and B2 classes is strongly upregulated after prolonged exposure to cisplatin, etoposide, or gamma radiation. A similar induction of Alu transcripts in human cells occurs under these conditions. This induction is not due to a general upregulation of pol III activity in either species. Genotoxic treatment of murine cells containing an exogenous human Alu element induced Alu transcription. Concomitant with the increased expression of SINEs, an increase in cellular reverse transcriptase was observed after exposure to these same DNA-damaging agents. These findings suggest that genomic damage may be an important activator of SINEs, and that SINE mobility may contribute to secondary malignancy after exposure to DNA-damaging chemotherapy.

  2. Subsoil TPH and other petroleum fractions-contamination levels in an oil storage and distribution station in north-central Mexico.

    PubMed

    Iturbe, Rosario; Flores, Carlos; Flores, Rosa Ma; Torres, Luis G

    2005-12-01

    Many oil industry related sites have become contaminated due to the activities characteristic of this industry, such as oil exploration and production, refining, and petro-chemistry. In Mexico, reported hydrocarbon spills for the year 2000 amounted to 185203, equivalent to 6252 tons (PEMEX, 2000). The first step for the remediation of these polluted sites is to assess the size and intensity of the oil contamination affecting the subsoil and groundwater, followed by a health risk assessment to establish clean up levels. The aim of this work was to characterize the soil and water in a north-central Mexico Oil Storage and Distribution Station (ODSS), in terms of TPHs, gasoline and diesel fractions, BTEX, PAHs, MTBE, and some metals. Besides, measurements of the explosivity index along the ODSS were made and we describe and discuss the risk health assessment analysis performed at the ODSS, as well as the recommendations arising from it. Considering soils with TPH concentrations higher than 2000 mg kg(-1), the contaminated areas corresponding to the railway zone is about 12776.5 m2, to the south of the storage tanks is about 6558 m2, and to the south of the filling tanks is about 783 m2. Total area to be treated is about 20107 m2 (volume of 20107 m3), considering 1m depth.

  3. Human ISWI complexes are targeted by SMARCA5 ATPase and SLIDE domains to help resolve lesion-stalled transcription

    PubMed Central

    Aydin, Özge Z.; Marteijn, Jurgen A.; Ribeiro-Silva, Cristina; Rodríguez López, Aida; Wijgers, Nils; Smeenk, Godelieve; van Attikum, Haico; Poot, Raymond A.; Vermeulen, Wim; Lans, Hannes

    2014-01-01

    Chromatin compaction of deoxyribonucleic acid (DNA) presents a major challenge to the detection and removal of DNA damage. Helix-distorting DNA lesions that block transcription are specifically repaired by transcription-coupled nucleotide excision repair, which is initiated by binding of the CSB protein to lesion-stalled RNA polymerase II. Using live cell imaging, we identify a novel function for two distinct mammalian ISWI adenosine triphosphate (ATP)-dependent chromatin remodeling complexes in resolving lesion-stalled transcription. Human ISWI isoform SMARCA5/SNF2H and its binding partners ACF1 and WSTF are rapidly recruited to UV-C induced DNA damage to specifically facilitate CSB binding and to promote transcription recovery. SMARCA5 targeting to UV-C damage depends on transcription and histone modifications and requires functional SWI2/SNF2-ATPase and SLIDE domains. After initial recruitment to UV damage, SMARCA5 re-localizes away from the center of DNA damage, requiring its HAND domain. Our studies support a model in which SMARCA5 targeting to DNA damage-stalled transcription sites is controlled by an ATP-hydrolysis-dependent scanning and proofreading mechanism, highlighting how SWI2/SNF2 chromatin remodelers identify and bind nucleosomes containing damaged DNA. PMID:24990377

  4. Long-term Effects of Nutrient Addition and Phytoremediation on Diesel and Crude Oil Contaminated Soils in subarctic Alaska

    PubMed Central

    Leewis, Mary-Cathrine; Reynolds, Charles M.; Leigh, Mary Beth

    2014-01-01

    Phytoremediation is a potentially inexpensive method of detoxifying contaminated soils using plants and associated soil microorganisms. The remote locations and cold climate of Alaska provide unique challenges associated with phytoremediation such as finding effective plant species that can achieve successful site clean-up despite the extreme environmental conditions and with minimal site management. A long-term assessment of phytoremediation was performed which capitalized on a study established in Fairbanks in 1995. The original study sought to determine how the introduction of plants (Festuca rubra, Lolium multiflorum), nutrients (fertilizer), or their combination would affect degradation of petroleum hydrocarbon (TPH) contaminated soils (crude oil or diesel) over time. Within the year following initial treatments, the plots subjected to both planting and/or fertilization showed greater overall decreases in TPH concentrations in both the diesel and crude oil contaminated soils relative to untreated plots. We re-examined this field site after 15 years with no active site management to assess the long-term effects of phytoremediation on colonization by native and non-native plants, their rhizosphere microbial communities and on petroleum removal from soil. Native and non-native vegetation had extensively colonized the site, with more abundant vegetation found on the diesel contaminated soils than the more nutrient-poor, more coarse, and acidic crude oil contaminated soils. TPH concentrations achieved regulatory clean up levels in all treatment groups, with lower TPH concentrations correlating with higher amounts of woody vegetation (trees & shrubs). In addition, original treatment type has affected vegetation recruitment to each plot with woody vegetation and more native plants in unfertilized plots. Bacterial community structure also varies according to the originally applied treatments. This study suggests that initial treatment with native tree species in combination with grasses could be an effective means for phytoremediating petroleum contaminated soils and promoting ecological recovery in cold regions. PMID:24501438

  5. Characterization of EPA's 16 priority pollutant polycyclic aromatic hydrocarbons (PAHs) in tank bottom solids and associated contaminated soils at oil exploration and production sites in Texas.

    PubMed

    Bojes, Heidi K; Pope, Peter G

    2007-04-01

    The purpose of this study was to determine the concentration and types of polycyclic aromatic hydrocarbons (PAHs), a group of environmentally toxic and persistent chemicals, at contaminated oil exploration and production (E&P) sites located in environmentally sensitive and geographically distinct areas throughout Texas. Samples of tank bottom solids, the oily sediment that collects at the bottom of the tanks, were collected from inactive crude oil storage tanks at E&P sites and hydrocarbon contaminated soil samples were collected from the area surrounding each tank that was sampled. All samples were analyzed for the 16 PAH priority pollutant listed by US EPA and for total petroleum hydrocarbons (TPH). The results demonstrate that overall average PAH concentrations were significantly higher in tank bottom solids than in contaminated soils. Total PAH concentrations decreased predictably with diminishing hydrocarbon concentrations; but the percent fraction of carcinogenic PAHs per total measured PAH content increased from approximately 12% in tank bottom solids to about 46% in the contaminated soils. These results suggest that the PAH content found in tank bottom solids cannot reliably be used to predict the PAH content in associated contaminated soil. Comparison of PAHs to conservative risk-based screening levels for direct exposure to soil and leaching from soil to groundwater indicate that PAHs are not likely to exceed default risk-based thresholds in soils containing TPH of 1% (10,000mg/kg) or less. These results show that the magnitude of TPH concentration may be a useful indicator of potential risk from PAHs in crude oil-contaminated soils. The results also provide credibility to the 1% (10,000mg/kg) TPH cleanup level, used in Texas as a default management level at E&P sites located in non-sensitive areas, with respect to PAH toxicity.

  6. DOR/Tp53inp2 and Tp53inp1 constitute a metazoan gene family encoding dual regulators of autophagy and transcription.

    PubMed

    Sancho, Ana; Duran, Jordi; García-España, Antonio; Mauvezin, Caroline; Alemu, Endalkachew A; Lamark, Trond; Macias, Maria J; DeSalle, Rob; Royo, Miriam; Sala, David; Chicote, Javier U; Palacín, Manuel; Johansen, Terje; Zorzano, Antonio

    2012-01-01

    Human DOR/TP53INP2 displays a unique bifunctional role as a modulator of autophagy and gene transcription. However, the domains or regions of DOR that participate in those functions have not been identified. Here we have performed structure/function analyses of DOR guided by identification of conserved regions in the DOR gene family by phylogenetic reconstructions. We show that DOR is present in metazoan species. Invertebrates harbor only one gene, DOR/Tp53inp2, and in the common ancestor of vertebrates Tp53inp1 may have arisen by gene duplication. In keeping with these data, we show that human TP53INP1 regulates autophagy and that different DOR/TP53INP2 and TP53INP1 proteins display transcriptional activity. The use of molecular evolutionary information has been instrumental to determine the regions that participate in DOR functions. DOR and TP53INP1 proteins share two highly conserved regions (region 1, aa residues 28-42; region 2, 66-112 in human DOR). Mutation of conserved hydrophobic residues in region 1 of DOR (that are part of a nuclear export signal, NES) reduces transcriptional activity, and blocks nuclear exit and autophagic activity under autophagy-activated conditions. We also identify a functional and conserved LC3-interacting motif (LIR) in region 1 of DOR and TP53INP1 proteins. Mutation of conserved acidic residues in region 2 of DOR reduces transcriptional activity, impairs nuclear exit in response to autophagy activation, and disrupts autophagy. Taken together, our data reveal DOR and TP53INP1 as dual regulators of transcription and autophagy, and identify two conserved regions in the DOR family that concentrate multiple functions crucial for autophagy and transcription.

  7. DOR/Tp53inp2 and Tp53inp1 Constitute a Metazoan Gene Family Encoding Dual Regulators of Autophagy and Transcription

    PubMed Central

    Sancho, Ana; Duran, Jordi; García-España, Antonio; Mauvezin, Caroline; Alemu, Endalkachew A.; Lamark, Trond; Macias, Maria J.; DeSalle, Rob; Royo, Miriam; Sala, David; Chicote, Javier U.; Palacín, Manuel; Johansen, Terje; Zorzano, Antonio

    2012-01-01

    Human DOR/TP53INP2 displays a unique bifunctional role as a modulator of autophagy and gene transcription. However, the domains or regions of DOR that participate in those functions have not been identified. Here we have performed structure/function analyses of DOR guided by identification of conserved regions in the DOR gene family by phylogenetic reconstructions. We show that DOR is present in metazoan species. Invertebrates harbor only one gene, DOR/Tp53inp2, and in the common ancestor of vertebrates Tp53inp1 may have arisen by gene duplication. In keeping with these data, we show that human TP53INP1 regulates autophagy and that different DOR/TP53INP2 and TP53INP1 proteins display transcriptional activity. The use of molecular evolutionary information has been instrumental to determine the regions that participate in DOR functions. DOR and TP53INP1 proteins share two highly conserved regions (region 1, aa residues 28–42; region 2, 66–112 in human DOR). Mutation of conserved hydrophobic residues in region 1 of DOR (that are part of a nuclear export signal, NES) reduces transcriptional activity, and blocks nuclear exit and autophagic activity under autophagy-activated conditions. We also identify a functional and conserved LC3-interacting motif (LIR) in region 1 of DOR and TP53INP1 proteins. Mutation of conserved acidic residues in region 2 of DOR reduces transcriptional activity, impairs nuclear exit in response to autophagy activation, and disrupts autophagy. Taken together, our data reveal DOR and TP53INP1 as dual regulators of transcription and autophagy, and identify two conserved regions in the DOR family that concentrate multiple functions crucial for autophagy and transcription. PMID:22470510

  8. Anthropogenic pollution indicators in marine environment of the Eastern Part of the Gulf of Finland

    NASA Astrophysics Data System (ADS)

    Zhakovskaya, Zoya; Nikiforov, Vladimir; Mamontova, Varvara; Khoroshko, Larisa; Chernova, Ekaterina; Russkikh, Iana

    2014-05-01

    Pollution involving hazardous substances is considered one of the major problems affecting the state of the Baltic marine environment. However, assessment of the vast majority of the hazardous substances (including accepted as pollution indicators) in the environment have not been monitored in Russian Federation yet. Moreover there are no official guideline values for their presence or release in environment. For our investigation we have selected the organotin biocides and widespread pharmaceutical diclofenac. The study is focused on surface marine water and bottom sediments, collected from the eastern part of the Gulf of Finland during the navigation seasons of 2012-2013. Organotin compounds belong to a large group of key marine contaminants. They had been widely used in the world industry as antifouling paints, fungicides and biocides until the middle of 1980s. Tributyltin (TBT) and triphenyltin (TPhT) are the most hazardous of all organotin compounds, causing such biological effects as shell deformation, endocrine disruption, imposex and intersex phenomena at the concentration of 2 ng/L. The use of TBT in antifouling paints was banned within EU in 2003 and within Russian Federation in 2008. Monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT) and triphenyltin (TPhT) were analysed as ethyl derivatives using electron impact gas chromatography-mass spectrometry (GC-MS-EI) in single ion monitoring mode (SIM). TBT and TPhT were frequently found above MAC of 1.5 ng/L and 2 ng/g dw respectively in both water and bottom sediment samples collected from the Gulf of Finland water basin. The highest detected concentration detected mainly in coastal areas with dense ship traffic were 670 ng/L (TBT) in water samples, 440 ng/g dw (TBT), 160 ng/g dw (TPhT) in sediment samples. Potential risks from the environmental presence of pharmaceuticals and personal care products (PPCP), such as medicine, hormones, means of personal hygiene, etc. reveal in abnormal physiological processes and reproductive impairment, increasing number of cancer incidences and increasing of bacterial antibiotic resistance. Diclofenac one of anthropogenic markers, was analyzed by the method of liquid chromatography high-resolution mass-spectrometry, using LTQ Orbitrap (Thermo Finnigan) in natural water and sediment samples. Mass spectra were recorded in several modes: full scan, SIM and MRM using positive and negative ionization. Resolution was 30000. Diclofenac were detected in several water samples (in the range of 3,9-270,0 ng/L). The obtained results are using for "Biota spatial distribution/Geological diversity/Pollution" model validation. This study was supported by projects TOPCONS («Transboundary tool for spatial planning and conservation of the Gulf of Finland»), HELCOM projects BALTHAZAR Phase II and BASE.

  9. Chorionic gonadotropin regulates the transcript level of VHL, p53, and HIF-2alpha in human granulosa lutein cells.

    PubMed

    Herr, D; Keck, C; Tempfer, C; Pietrowski, Detlef

    2004-12-01

    The ovarian corpus luteum plays a critical role in reproduction being the primary source of circulating progesterone. After ovulation the corpus luteum is build by avascular granulosa lutein cells through rapid vascularization regulated by gonadotropic hormones. The present study was performed to investigate whether this process might be influenced by the human chorionic gonadotropin (hCG)-dependent expression of different tumor suppressor genes and hypoxia dependent transcription factors. RNA was isolated from cultured granulosa lutein cells, transcribed into cDNA, and the transcript level of following genes were determined: RB-1, VHL, NF-1, NF-2, Wt-1, p53, APC, and hypoxia inducible factor-1 (HIF-1), -2, and -3alpha. Additionally, the influence of hCG on the expression of VHL, p53, and HIf2alpha were investigated. We demonstrate that in human granulosa lutein cells the tumor suppressor genes RB-1, VHL, NF-1, NF-2, Wt-1, p53, and APC and the hypoxia dependent transcription factors HIF-1alpha, -2alpha, and -3alpha are expressed. In addition, we showed that hCG regulates the expression of p53, VHL, and HIF-2alpha. Our results indicate that hCG may determine the growth and development of the corpus luteum by mediating hypoxic and apoptotic pathways in human granulosa lutein cells. Copyright 2004 Wiley-Liss, Inc.

  10. Transcriptional profiling of human bronchial epithelial cell BEAS-2B exposed to diesel and biomass ultrafine particles.

    PubMed

    Grilli, Andrea; Bengalli, Rossella; Longhin, Eleonora; Capasso, Laura; Proverbio, Maria Carla; Forcato, Mattia; Bicciato, Silvio; Gualtieri, Maurizio; Battaglia, Cristina; Camatini, Marina

    2018-04-27

    Emissions from diesel vehicles and biomass burning are the principal sources of primary ultrafine particles (UFP). The exposure to UFP has been associated to cardiovascular and pulmonary diseases, including lung cancer. Although many aspects of the toxicology of ambient particulate matter (PM) have been unraveled, the molecular mechanisms activated in human cells by the exposure to UFP are still poorly understood. Here, we present an RNA-seq time-course experiment (five time point after single dose exposure) used to investigate the differential and temporal changes induced in the gene expression of human bronchial epithelial cells (BEAS-2B) by the exposure to UFP generated from diesel and biomass combustion. A combination of different bioinformatics tools (EdgeR, next-maSigPro and reactome FI app-Cytoscape and prioritization strategies) facilitated the analyses the temporal transcriptional pattern, functional gene set enrichment and gene networks related to cellular response to UFP particles. The bioinformatics analysis of transcriptional data reveals that the two different UFP induce, since the earliest time points, different transcriptional dynamics resulting in the activation of specific genes. The functional enrichment of differentially expressed genes indicates that the exposure to diesel UFP induces the activation of genes involved in TNFα signaling via NF-kB and inflammatory response, and hypoxia. Conversely, the exposure to ultrafine particles from biomass determines less distinct modifications of the gene expression profiles. Diesel UFP exposure induces the secretion of biomarkers associated to inflammation (CCXL2, EPGN, GREM1, IL1A, IL1B, IL6, IL24, EREG, VEGF) and transcription factors (as NFE2L2, MAFF, HES1, FOSL1, TGIF1) relevant for cardiovascular and lung disease. By means of network reconstruction, four genes (STAT3, HIF1a, NFKB1, KRAS) have emerged as major regulators of transcriptional response of bronchial epithelial cells exposed to diesel exhaust. Overall, this work highlights modifications of the transcriptional landscape in human bronchial cells exposed to UFP and sheds new lights on possible mechanisms by means of which UFP acts as a carcinogen and harmful factor for human health.

  11. Contributions of in vitro transcription to the understanding of human RNA polymerase III transcription

    PubMed Central

    Dumay-Odelot, Hélène; Durrieu-Gaillard, Stéphanie; El Ayoubi, Leyla; Parrot, Camila; Teichmann, Martin

    2014-01-01

    Human RNA polymerase III transcribes small untranslated RNAs that contribute to the regulation of essential cellular processes, including transcription, RNA processing and translation. Analysis of this transcription system by in vitro transcription techniques has largely contributed to the discovery of its transcription factors and to the understanding of the regulation of human RNA polymerase III transcription. Here we review some of the key steps that led to the identification of transcription factors and to the definition of minimal promoter sequences for human RNA polymerase III transcription. PMID:25764111

  12. Transcriptional regulation of the human Na{sup +}/H{sup +} exchanger NHE3 by serotonin in intestinal epithelial cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Amin, Md Ruhul; Ghannad, Leda; Othman, Ahmad

    2009-05-08

    Serotonin (5-HT) decreases NHE2 and NHE3 activities under acute conditions in human intestinal epithelial cells. Here, we have investigated the effects of 5-HT on expression of the human NHE3 gene and the mechanisms underlying its transcriptional regulation in differentiated C2BBe1 cells. Treatment of the human intestinal epithelial cell line, C2BBe1, with 5-HT (20 {mu}M) resulted in a significant decrease in NHE3 mRNA and protein expression. In transient transfection studies, 5-HT repressed the NHE3 promoter activity by {approx}55%. The repression of the NHE3 promoter activity in response to 5-HT was accompanied by reduced DNA-binding activity of transcription factors Sp1 and Sp3more » to the NHE3 promoter without alteration in their nuclear levels. Pharmacological inhibitors of protein kinase C reversed the inhibitory effect of 5-HT on the promoter activity. Our data indicate that 5-HT suppresses the transcriptional activity of the NHE3 promoter and this effect may be mediated by PKC{alpha} and modulation of DNA-binding affinities of Sp1 and Sp3.« less

  13. Discovery of a Regulatory Motif for Human Satellite DNA Transcription in Response to BATF2 Overexpression.

    PubMed

    Bai, Xuejia; Huang, Wenqiu; Zhang, Chenguang; Niu, Jing; Ding, Wei

    2016-03-01

    One of the basic leucine zipper transcription factors, BATF2, has been found to suppress cancer growth and migration. However, little is known about the genes downstream of BATF2. HeLa cells were stably transfected with BATF2, then chromatin immunoprecipitation-sequencing was employed to identify the DNA motifs responsive to BATF2. Comprehensive bioinformatics analyses indicated that the most significant motif discovered as TTCCATT[CT]GATTCCATTC[AG]AT was primarily distributed among the chromosome centromere regions and mostly within human type II satellite DNA. Such motifs were able to prime the transcription of type II satellite DNA in a directional and asymmetrical manner. Consistently, satellite II transcription was up-regulated in BATF2-overexpressing cells. The present study provides insight into understanding the role of BATF2 in tumours and the importance of satellite DNA in the maintenance of genomic stability. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  14. Structural Basis of Mitochondrial Transcription Initiation.

    PubMed

    Hillen, Hauke S; Morozov, Yaroslav I; Sarfallah, Azadeh; Temiakov, Dmitry; Cramer, Patrick

    2017-11-16

    Transcription in human mitochondria is driven by a single-subunit, factor-dependent RNA polymerase (mtRNAP). Despite its critical role in both expression and replication of the mitochondrial genome, transcription initiation by mtRNAP remains poorly understood. Here, we report crystal structures of human mitochondrial transcription initiation complexes assembled on both light and heavy strand promoters. The structures reveal how transcription factors TFAM and TFB2M assist mtRNAP to achieve promoter-dependent initiation. TFAM tethers the N-terminal region of mtRNAP to recruit the polymerase to the promoter whereas TFB2M induces structural changes in mtRNAP to enable promoter opening and trapping of the DNA non-template strand. Structural comparisons demonstrate that the initiation mechanism in mitochondria is distinct from that in the well-studied nuclear, bacterial, or bacteriophage transcription systems but that similarities are found on the topological and conceptual level. These results provide a framework for studying the regulation of gene expression and DNA replication in mitochondria. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Treatment of real coal gasification wastewater using a novel integrated system of anoxic hybrid two stage aerobic processes: performance and the role of pure oxygen microbubble.

    PubMed

    Zhuang, Haifeng; Han, Hongjun; Shan, Shengdao

    2016-06-01

    A novel integrated system of anoxic-pure oxygen microbubble-activated sludge reactor-moving bed biofilm reactor was employed in treatment of real coal gasification wastewater. The results showed the integrated system had efficient performance of pollutants removal in short hydraulic retention time. While pure oxygen microbubble with the flow rate of 1.5 L/h and NaHCO3 dosage ratio of 2:1 (amount NaHCO3 to NH4 (+)-N ratio, mol: mol) were used, the removal efficiencies of COD, total phenols (TPh) and NH4 (+)-N reached 90, 95, and 95 %, respectively, with the influent loading rates of 3.4 kg COD/(m(3) d), 0.81 kg TPh/(m(3) d), and 0.28 kg NH4 (+)-N/(m(3) d). With the recycle ratio of 300 %, the concentrations of NO2 (-)-N and NO3 (-)-N in effluent decreased to 12 and 59 mg/L, respectively. Meanwhile, pure oxygen microbubble significantly improved the enzymatic activities and affected the effluent organic compositions and reduced the foam expansion. Thus, the novel integrated system with efficient, stable, and economical advantages was suitable for engineering application.

  16. Regulation of MET by FOXP2, genes implicated in higher cognitive dysfunction and autism risk.

    PubMed

    Mukamel, Zohar; Konopka, Genevieve; Wexler, Eric; Osborn, Gregory E; Dong, Hongmei; Bergman, Mica Y; Levitt, Pat; Geschwind, Daniel H

    2011-08-10

    Autism spectrum disorder (ASD) is a highly heritable, behaviorally defined, heterogeneous disorder of unknown pathogenesis. Several genetic risk genes have been identified, including the gene encoding the receptor tyrosine kinase MET, which regulates neuronal differentiation and growth. An ASD-associated polymorphism disrupts MET gene transcription, and there are reduced levels of MET protein expression in the mature temporal cortex of subjects with ASD. To address the possible neurodevelopmental contribution of MET to ASD pathogenesis, we examined the expression and transcriptional regulation of MET by a transcription factor, FOXP2, which is implicated in regulation of cognition and language, two functions altered in ASD. MET mRNA expression in the midgestation human fetal cerebral cortex is strikingly restricted, localized to portions of the temporal and occipital lobes. Within the cortical plate of the temporal lobe, the pattern of MET expression is highly complementary to the expression pattern of FOXP2, suggesting the latter may play a role in repression of gene expression. Consistent with this, MET and FOXP2 also are reciprocally expressed by differentiating normal human neuronal progenitor cells (NHNPs) in vitro, leading us to assess whether FOXP2 transcriptionally regulates MET. Indeed, FOXP2 binds directly to the 5' regulatory region of MET, and overexpression of FOXP2 results in transcriptional repression of MET. The expression of MET in restricted human neocortical regions, and its regulation in part by FOXP2, is consistent with genetic evidence for MET contributing to ASD risk.

  17. CREB-1 and AP-1 transcription factors JunD and Fra-2 regulate bone sialoprotein gene expression in human breast cancer cells.

    PubMed

    Detry, C; Lamour, V; Castronovo, V; Bellahcène, A

    2008-02-01

    Bone sialoprotein (BSP) expression is detected in a variety of human osteotropic cancers. High expression of BSP in breast and prostate primary carcinomas is associated with progression and bone metastases development. In this study, we examined the transcriptional regulation of BSP gene expression in MDA-MB-231 and MCF-7 human breast cancer cells compared with Saos-2 human osteoblast-like cells. BSP human promoter deletion analyses delineated a -56/-84 region, which comprises a cAMP response element (CRE) that was sufficient for maximal promoter activity in breast cancer cell lines. We found that the basic fibroblast growth factor response element (FRE) also located in the proximal promoter was a crucial regulator of human BSP promoter activity in Saos-2 but not in breast cancer cells. Promoter activity experiments in combination with DNA mobility shift assays demonstrated that BSP promoter activity is under the control of the CRE element, through CREB-1, JunD and Fra-2 binding, in MDA-MB-231, MCF-7 and in Saos-2 cells. Forskolin, a protein kinase A pathway activator, failed to enhance BSP transcriptional activity suggesting that CRE site behaves as a constitutive rather than an inducible element in these cell lines. Over-expression of JunD and Fra-2 increased BSP promoter activity and upregulated endogenous BSP protein expression in MCF-7 and Saos-2 cells while siRNA-mediated inhibition of both factors expression significantly reduced BSP protein level in MDA-MB-231. Collectively, these data provide with new transcriptional mechanisms, implicating CREB and AP-1 factors, that control BSP gene expression in breast cancer cells.

  18. Genome-Wide Reprogramming of Transcript Architecture by Temperature Specifies the Developmental States of the Human Pathogen Histoplasma

    PubMed Central

    Gilmore, Sarah A.; Voorhies, Mark; Gebhart, Dana; Sil, Anita

    2015-01-01

    Eukaryotic cells integrate layers of gene regulation to coordinate complex cellular processes; however, mechanisms of post-transcriptional gene regulation remain poorly studied. The human fungal pathogen Histoplasma capsulatum (Hc) responds to environmental or host temperature by initiating unique transcriptional programs to specify multicellular (hyphae) or unicellular (yeast) developmental states that function in infectivity or pathogenesis, respectively. Here we used recent advances in next-generation sequencing to uncover a novel re-programming of transcript length between Hc developmental cell types. We found that ~2% percent of Hc transcripts exhibit 5’ leader sequences that differ markedly in length between morphogenetic states. Ribosome density and mRNA abundance measurements of differential leader transcripts revealed nuanced transcriptional and translational regulation. One such class of regulated longer leader transcripts exhibited tight transcriptional and translational repression. Further examination of these dually repressed genes revealed that some control Hc morphology and that their strict regulation is necessary for the pathogen to make appropriate developmental decisions in response to temperature. PMID:26177267

  19. Genome-Wide Reprogramming of Transcript Architecture by Temperature Specifies the Developmental States of the Human Pathogen Histoplasma.

    PubMed

    Gilmore, Sarah A; Voorhies, Mark; Gebhart, Dana; Sil, Anita

    2015-07-01

    Eukaryotic cells integrate layers of gene regulation to coordinate complex cellular processes; however, mechanisms of post-transcriptional gene regulation remain poorly studied. The human fungal pathogen Histoplasma capsulatum (Hc) responds to environmental or host temperature by initiating unique transcriptional programs to specify multicellular (hyphae) or unicellular (yeast) developmental states that function in infectivity or pathogenesis, respectively. Here we used recent advances in next-generation sequencing to uncover a novel re-programming of transcript length between Hc developmental cell types. We found that ~2% percent of Hc transcripts exhibit 5' leader sequences that differ markedly in length between morphogenetic states. Ribosome density and mRNA abundance measurements of differential leader transcripts revealed nuanced transcriptional and translational regulation. One such class of regulated longer leader transcripts exhibited tight transcriptional and translational repression. Further examination of these dually repressed genes revealed that some control Hc morphology and that their strict regulation is necessary for the pathogen to make appropriate developmental decisions in response to temperature.

  20. Alternative splicing of the tyrosinase gene transcript in normal human melanocytes and lymphocytes.

    PubMed

    Fryer, J P; Oetting, W S; Brott, M J; King, R A

    2001-11-01

    We have identified and isolated ectopically expressed tyrosinase transcripts in normal human melanocytes and lymphocytes and in a human melanoma (MNT-1) cell line to establish a baseline for the expression pattern of this gene in normal tissue. Tyrosinase mRNA from human lymphoblastoid cell lines was reverse transcribed and amplified using specific "nested" primers. This amplification yielded eight identifiable transcripts; five that resulted from alternative splicing patterns arising from the utilization of normal and alternative splice sequences. Identical splicing patterns were found in transcripts from human primary melanocytes in culture and a melanoma cell line, indicating that lymphoblastoid cell lines provide an accurate reflection of transcript processing in melanocytes. Similar splicing patterns have also been found with murine melanocyte tyrosinase transcripts. Our results demonstrate that alternative splicing of human tyrosinase gene transcript produces a number of predictable and identifiable transcripts, and that human lymphoblastoid cell lines provide a source of ectopically expressed transcripts that can be used to study the biology of tyrosinase gene expression in humans.

  1. ZEB2 drives immature T-cell lymphoblastic leukaemia development via enhanced tumour-initiating potential and IL-7 receptor signalling

    PubMed Central

    Goossens, Steven; Radaelli, Enrico; Blanchet, Odile; Durinck, Kaat; Van der Meulen, Joni; Peirs, Sofie; Taghon, Tom; Tremblay, Cedric S.; Costa, Magdaline; Ghahremani, Morvarid Farhang; De Medts, Jelle; Bartunkova, Sonia; Haigh, Katharina; Schwab, Claire; Farla, Natalie; Pieters, Tim; Matthijssens, Filip; Van Roy, Nadine; Best, J. Adam; Deswarte, Kim; Bogaert, Pieter; Carmichael, Catherine; Rickard, Adam; Suryani, Santi; Bracken, Lauryn S.; Alserihi, Raed; Canté-Barrett, Kirsten; Haenebalcke, Lieven; Clappier, Emmanuelle; Rondou, Pieter; Slowicka, Karolina; Huylebroeck, Danny; Goldrath, Ananda W.; Janzen, Viktor; McCormack, Matthew P.; Lock, Richard B.; Curtis, David J.; Harrison, Christine; Berx, Geert; Speleman, Frank; Meijerink, Jules P. P.; Soulier, Jean; Van Vlierberghe, Pieter; Haigh, Jody J.

    2015-01-01

    Early T-cell precursor leukaemia (ETP-ALL) is a high-risk subtype of human leukaemia that is poorly understood at the molecular level. Here we report translocations targeting the zinc finger E-box-binding transcription factor ZEB2 as a recurrent genetic lesion in immature/ETP-ALL. Using a conditional gain-of-function mouse model, we demonstrate that sustained Zeb2 expression initiates T-cell leukaemia. Moreover, Zeb2-driven mouse leukaemia exhibit some features of the human immature/ETP-ALL gene expression signature, as well as an enhanced leukaemia-initiation potential and activated Janus kinase (JAK)/signal transducers and activators of transcription (STAT) signalling through transcriptional activation of IL7R. This study reveals ZEB2 as an oncogene in the biology of immature/ETP-ALL and paves the way towards pre-clinical studies of novel compounds for the treatment of this aggressive subtype of human T-ALL using our Zeb2-driven mouse model. PMID:25565005

  2. NKX2-5 regulates human cardiomyogenesis via a HEY2 dependent transcriptional network.

    PubMed

    Anderson, David J; Kaplan, David I; Bell, Katrina M; Koutsis, Katerina; Haynes, John M; Mills, Richard J; Phelan, Dean G; Qian, Elizabeth L; Leitoguinho, Ana Rita; Arasaratnam, Deevina; Labonne, Tanya; Ng, Elizabeth S; Davis, Richard P; Casini, Simona; Passier, Robert; Hudson, James E; Porrello, Enzo R; Costa, Mauro W; Rafii, Arash; Curl, Clare L; Delbridge, Lea M; Harvey, Richard P; Oshlack, Alicia; Cheung, Michael M; Mummery, Christine L; Petrou, Stephen; Elefanty, Andrew G; Stanley, Edouard G; Elliott, David A

    2018-04-10

    Congenital heart defects can be caused by mutations in genes that guide cardiac lineage formation. Here, we show deletion of NKX2-5, a critical component of the cardiac gene regulatory network, in human embryonic stem cells (hESCs), results in impaired cardiomyogenesis, failure to activate VCAM1 and to downregulate the progenitor marker PDGFRα. Furthermore, NKX2-5 null cardiomyocytes have abnormal physiology, with asynchronous contractions and altered action potentials. Molecular profiling and genetic rescue experiments demonstrate that the bHLH protein HEY2 is a key mediator of NKX2-5 function during human cardiomyogenesis. These findings identify HEY2 as a novel component of the NKX2-5 cardiac transcriptional network, providing tangible evidence that hESC models can decipher the complex pathways that regulate early stage human heart development. These data provide a human context for the evaluation of pathogenic mutations in congenital heart disease.

  3. Multilayered epithelium in a rat model and human Barrett's esophagus: Similar expression patterns of transcription factors and differentiation markers

    PubMed Central

    Chen, Xiaoxin; Qin, Rong; Liu, Ba; Ma, Yan; Su, Yinghao; Yang, Chung S; Glickman, Jonathan N; Odze, Robert D; Shaheen, Nicholas J

    2008-01-01

    Background In rats, esophagogastroduodenal anastomosis (EGDA) without concomitant chemical carcinogen treatment leads to gastroesophageal reflux disease, multilayered epithelium (MLE, a presumed precursor in intestinal metaplasia), columnar-lined esophagus, dysplasia, and esophageal adenocarcinoma. Previously we have shown that columnar-lined esophagus in EGDA rats resembled human Barrett's esophagus (BE) in its morphology, mucin features and expression of differentiation markers (Lab. Invest. 2004;84:753–765). The purpose of this study was to compare the phenotype of rat MLE with human MLE, in order to gain insight into the nature of MLE and its potential role in the development of BE. Methods Serial sectioning was performed on tissue samples from 32 EGDA rats and 13 patients with established BE. Tissue sections were immunohistochemically stained for a variety of transcription factors and differentiation markers of esophageal squamous epithelium and intestinal columnar epithelium. Results We detected MLE in 56.3% (18/32) of EGDA rats, and in all human samples. As expected, both rat and human squamous epithelium, but not intestinal metaplasia, expressed squamous transcription factors and differentiation markers (p63, Sox2, CK14 and CK4) in all cases. Both rat and human intestinal metaplasia, but not squamous epithelium, expressed intestinal transcription factors and differentiation markers (Cdx2, GATA4, HNF1α, villin and Muc2) in all cases. Rat MLE shared expression patterns of Sox2, CK4, Cdx2, GATA4, villin and Muc2 with human MLE. However, p63 and CK14 were expressed in a higher proportion of rat MLE compared to humans. Conclusion These data indicate that rat MLE shares similar properties to human MLE in its expression pattern of these markers, not withstanding small differences, and support the concept that MLE may be a transitional stage in the metaplastic conversion of squamous to columnar epithelium in BE. PMID:18190713

  4. Multilayered epithelium in a rat model and human Barrett's esophagus: similar expression patterns of transcription factors and differentiation markers.

    PubMed

    Chen, Xiaoxin; Qin, Rong; Liu, Ba; Ma, Yan; Su, Yinghao; Yang, Chung S; Glickman, Jonathan N; Odze, Robert D; Shaheen, Nicholas J

    2008-01-11

    In rats, esophagogastroduodenal anastomosis (EGDA) without concomitant chemical carcinogen treatment leads to gastroesophageal reflux disease, multilayered epithelium (MLE, a presumed precursor in intestinal metaplasia), columnar-lined esophagus, dysplasia, and esophageal adenocarcinoma. Previously we have shown that columnar-lined esophagus in EGDA rats resembled human Barrett's esophagus (BE) in its morphology, mucin features and expression of differentiation markers (Lab. Invest. 2004;84:753-765). The purpose of this study was to compare the phenotype of rat MLE with human MLE, in order to gain insight into the nature of MLE and its potential role in the development of BE. Serial sectioning was performed on tissue samples from 32 EGDA rats and 13 patients with established BE. Tissue sections were immunohistochemically stained for a variety of transcription factors and differentiation markers of esophageal squamous epithelium and intestinal columnar epithelium. We detected MLE in 56.3% (18/32) of EGDA rats, and in all human samples. As expected, both rat and human squamous epithelium, but not intestinal metaplasia, expressed squamous transcription factors and differentiation markers (p63, Sox2, CK14 and CK4) in all cases. Both rat and human intestinal metaplasia, but not squamous epithelium, expressed intestinal transcription factors and differentiation markers (Cdx2, GATA4, HNF1alpha, villin and Muc2) in all cases. Rat MLE shared expression patterns of Sox2, CK4, Cdx2, GATA4, villin and Muc2 with human MLE. However, p63 and CK14 were expressed in a higher proportion of rat MLE compared to humans. These data indicate that rat MLE shares similar properties to human MLE in its expression pattern of these markers, not withstanding small differences, and support the concept that MLE may be a transitional stage in the metaplastic conversion of squamous to columnar epithelium in BE.

  5. Effects of gonadectomy and serotonin depletion on inter-individual differences in anxiety-like behaviour in male Wistar rats.

    PubMed

    Näslund, Jakob; Studer, Erik; Johansson, Elin; Eriksson, Elias

    2016-07-15

    Previous studies in Wistar rats suggest inter-individual differences in anxiety-like behaviour as assessed using the elevated plus maze (EPM), both between sexes and among males, to be abolished by serotonin depletion. To shed further light on the influence of sex steroids and serotonin - and on the interplay between the two - on proneness for EPM-assessed anxiety in males, outbred Wistar rats were divided into those with high and low anxiety, respectively, and exposed to gonadectomy or sham operation followed by administration of a serotonin synthesis inhibitor, para-chlorophenylalanine, or saline. Whereas gonadectomy enhanced anxiety-like behaviour in low anxiety rats so that these no longer differed in this regard from the high anxiety group, serotonin depletion reversed this effect, and also reduced anxiety in the low anxiety group regardless of gonadal state. A previously observed association between high anxiety-like behaviour and high expression of the serotonin-synthesizing enzyme tryptophan hydroxylase 2 (Tph2) in the raphe was confirmed in sham-operated animals but absent in gonadectomised rats, an ANCOVA revealing a significant interactive effect of baseline anxiety and gonadal state on Tph2 expression. It is suggested that androgens may contribute to upholding inter-individual differences in anxiety-like behaviour in male rats by interacting with serotonergic neurotransmission. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  6. Transcriptional and functional studies of Human Endogenous Retrovirus envelope EnvP(b) and EnvV genes in human trophoblasts

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Vargas, Amandine, E-mail: amandine.vargas@voila.fr; Thiery, Maxime, E-mail: thiery.maxime@courrier.uqam.ca; Lafond, Julie, E-mail: lafond.julie@uqam.ca

    2012-03-30

    HERV (Human Endogenous Retrovirus)-encoded envelope proteins are implicated in the development of the placenta. Indeed, Syncytin-1 and -2 play a crucial role in the fusion of human trophoblasts, a key step in placentation. Other studies have identified two other HERV env proteins, namely EnvP(b) and EnvV, both expressed in the placenta. In this study, we have fully characterized both env transcripts and their expression pattern and have assessed their implication in trophoblast fusion. Through RACE analyses, standard spliced transcripts were detected, while EnvV transcripts demonstrated alternative splicing at its 3 Prime end. Promoter activity and expression of both genes weremore » induced in forskolin-stimulated BeWo cells and in primary trophoblasts. Although we have confirmed the fusogenic activity of EnvP(b), overexpression or silencing experiments revealed no impact of this protein on trophoblast fusion. Our results demonstrate that both env genes are expressed in human trophoblasts but are not required for syncytialization.« less

  7. Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: role of serotonin.

    PubMed

    Guzzetti, Sara; Calcagno, Eleonora; Canetta, Alessandro; Sacchetti, Giuseppina; Fracasso, Claudia; Caccia, Silvio; Cervo, Luigi; Invernizzi, Roberto W

    2008-10-10

    We studied the antidepressant-like effect of paroxetine in strains of mice carrying different isoforms of tryptophan hydroxylase-2 (TPH-2), the enzyme responsible for the synthesis of brain serotonin (5-HT). The effect of paroxetine alone and in combination with pharmacological treatments enhancing or lowering 5-HT synthesis or melatonin was assessed in the forced swimming test in mice carrying allelic variants of TPH-2 (1473C in C57BL/6 and 1473G in DBA/2 and BALB/c). Changes in brain 5-hydroxytryptophan (5-HTP) accumulation and melatonin levels were measured by high-performance liquid chromatography. Paroxetine (2.5 and 5 mg/kg) reduced immobility time in C57BL/6J and C57BL/6N mice but had no such effect in DBA/2J, DBA/2N and BALB/c mice, even at 10 mg/kg. Enhancing 5-HT synthesis with tryptophan reinstated the antidepressant-like effect of paroxetine in DBA/2J, DBA/2N and BALB/c mice whereas inhibition of 5-HT synthesis prevented the effect of paroxetine in C57BL/6N mice. The response to paroxetine was not associated with changes in locomotor activity, brain melatonin or brain levels of the drug measured at the end of the behavioral test. These results support the importance of 5-HT synthesis in the response to SSRIs and suggest that melatonin does not contribute to the ability of tryptophan to rescue the antidepressant-like effect of paroxetine.

  8. Large scale treatment of total petroleum-hydrocarbon contaminated groundwater using bioaugmentation.

    PubMed

    Poi, Gregory; Shahsavari, Esmaeil; Aburto-Medina, Arturo; Mok, Puah Chum; Ball, Andrew S

    2018-05-15

    Bioaugmentation or the addition of microbes to contaminated sites has been widely used to treat contaminated soil or water; however this approach is often limited to laboratory based studies. In the present study, large scale bioaugmentation has been applied to total petroleum hydrocarbons (TPH)-contaminated groundwater at a petroleum facility. Initial TPH concentrations of 1564 mg L -1 in the field were reduced to 89 mg L -1 over 32 days. This reduction was accompanied by improved ecotoxicity, as shown by Brassica rapa germination numbers that increased from 52 at day 0 to 82% by the end of the treatment. Metagenomic analysis indicated that there was a shift in the microbial community when compared to the beginning of the treatment. The microbial community was dominated by Proteobacteria and Bacteroidetes from day 0 to day 32, although differences at the genus level were observed. The predominant genera at the beginning of the treatment (day 0 just after inoculation) were Cloacibacterium, Sediminibacterium and Brevundimonas while at the end of the treatment members of Flavobacterium dominated, reaching almost half the population (41%), followed by Pseudomonas (6%) and Limnobacter (5.8%). To the author's knowledge, this is among the first studies to report the successful large scale biodegradation of TPH-contaminated groundwater (18,000 L per treatment session) at an offshore petrochemical facility. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. Contrast-enhanced transrectal ultrasound for prediction of prostate cancer aggressiveness: The role of normal peripheral zone time-intensity curves.

    PubMed

    Huang, Hui; Zhu, Zheng-Qiu; Zhou, Zheng-Guo; Chen, Ling-Shan; Zhao, Ming; Zhang, Yang; Li, Hong-Bo; Yin, Li-Ping

    2016-12-08

    To assess the role of time-intensity curves (TICs) of the normal peripheral zone (PZ) in the identification of biopsy-proven prostate nodules using contrast-enhanced transrectal ultrasound (CETRUS). This study included 132 patients with 134 prostate PZ nodules. Arrival time (AT), peak intensity (PI), mean transit time (MTT), area under the curve (AUC), time from peak to one half (TPH), wash in slope (WIS) and time to peak (TTP) were analyzed using multivariate linear logistic regression and receiver operating characteristic (ROC) curves to assess whether combining nodule TICs with normal PZ TICs improved the prediction of prostate cancer (PCa) aggressiveness. The PI, AUC (p < 0.001 for both), MTT and TPH (p = 0.011 and 0.040 respectively) values of the malignant nodules were significantly higher than those of the benign nodules. Incorporating the PI and AUC values (both, p < 0.001) of the normal PZ TIC, but not the MTT and TPH values (p = 0.076 and 0.159 respectively), significantly improved the AUC for prediction of malignancy (PI: 0.784-0.923; AUC: 0.758-0.891) and assessment of cancer aggressiveness (p < 0.001). Thus, all these findings indicate that incorporating normal PZ TICs with nodule TICs in CETRUS readings can improve the diagnostic accuracy for PCa and cancer aggressiveness assessment.

  10. Comparative study of rhizobacterial community structure of plant species in oil-contaminated soil.

    PubMed

    Lee, Eun-Hee; Cho, Kyong-Suk; Kim, Jaisoo

    2010-09-01

    In this study, the identity and distribution of plants and the structure of their associated rhizobacterial communities were examined in an oil-contaminated site. The number of plant species that formed a community or were scattered was 24. The species living in soil highly contaminated with total petroleum hydrocarbon (TPH) (9,000-4,5000 mg/g-soil) were Cynodon dactylon, Persicaria lapathifolia, and Calystegia soldanella (a halophytic species). Among the 24 plant species, the following have been known to be effective for oil removal: C. dactylon, Digitaria sanguinalis, and Cyperus orthostachyus. Denaturing gradient gel electrophoresis (DGGE) profile analysis showed that the following pairs of plant species had highly similar (above 70%) rhizobacterial community structures: Artemisia princeps and Hemistepta lyrata; C. dactylon and P. lapathifolia; Carex kobomugi and Cardamine flexuosa; and Equisetum arvense and D. sanguinalis. The major groups of rhizobacteria were Betaproteobacteria, Gamma-proteobacteria, Chloroflexi, Actinobacteria, and unknown. Based on DGGE analysis, P. lapathifolia, found for the first time in this study growing in the presence of high TPH, may be a good species for phytoremediation of oil-contaminated soils and in particular, C. soldanella may be useful for soils with high TPH and salt concentrations. Overall, this study suggests that the plant roots, regardless of plant species, may have a similar influence on the bacterial community structure in oil-contaminated soil.

  11. Does fertilizer (N15P15K15) amendment enhance phytoremediation of petroleum-polluted aquatic ecosystem in the presence of water hyacinth (Eichhornia crassipes [Mart.] Solms)?

    PubMed

    Ndimele, Prince Emeka; Jenyo-Oni, Adetola; Chukwuka, Kanayo S; Ndimele, Chinatu Charity; Ayodele, Ibukunoluwa Augustine

    2015-01-01

    This study investigated the effects of inorganic fertilizer (N15P15K15) amendments on crude oil uptake by water hyacinth. Experimental units (water hyacinth grown in fresh water) were spiked with 0, 20, 40 and 60 mg/L crude oil. After 24 h, they were randomly assigned fertilizer (N15P15K15) at three different concentrations; 0, 6 and 10 mg/L. Crude oil degradation and absorption were determined by measuring total petroleum hydrocarbon (TPH) in the water column and water hyacinth, respectively. The measurements were taken monthly for six months (February-August 2010). The results showed that TPH concentration in the water column in the treatment amended at 6 mg/L (0.30 ± 0.01 mg/L) was significantly lower (p < .05) than the treatment amended at 10 mg/L (0.76 ± 0.15 mg/L) but was similar to the control (0.33 ± 0.03 mg/L). The water hyacinth in the control (phytoremediation) absorbed significantly higher (p < .05) TPH than the fertilizer-amended treatments. The first-order kinetic model gave a better description of the degradation of petroleum hydrocarbons. The study showed that phytoremediation of crude oil by water hyacinth and biostimulation with fertilizer (N15P15K15) is possible.

  12. Differential behavioral sensitivity to carbon dioxide (CO2) inhalation in rats

    PubMed Central

    Winter, Andrew; Ahlbrand, Rebecca; Naik, Devanshi; Sah, Renu

    2017-01-01

    Inhalation of carbon dioxide (CO2) is frequently employed as a biological challenge to evoke intense fear and anxiety. In individuals with panic disorder, CO2 reliably evokes panic attacks. Sensitivity to CO2 is highly heterogeneous among individuals, and although a genetic component is implicated, underlying mechanisms are not clear. Preclinical models that can simulate differential responsivity to CO2 are therefore relevant. In the current study we investigated CO2-evoked behavioral responses in four different rat strains: Sprague-Dawley (SD), Wistar (W), Long Evans (LE) and Wistar-Kyoto, (WK) rats. We also assessed tryptophan hydroxylase 2 (TPH-2)-positive serotonergic neurons in anxiety/panic regulatory subdivisions of the dorsal raphe nucleus (DR), as well as dopamine β hydroxylase (DβH)-positive noradrenergic neurons in the locus coeruleus, implicated in central CO2-chemosensitivity. Behavioral responsivity to CO2 inhalation varied between strains. CO2-evoked immobility was significantly higher in LE and WK rats as compared with W and SD cohorts. Differences were also observed in CO2-evoked rearing and grooming behaviors. Exposure to CO2 did not produce conditioned behavioral responses upon re-exposure to CO2 context in any strain. Reduced TPH-2 positive cell counts were observed specifically in the panic-regulatory dorsal raphe ventrolateral (DRVL)-ventrolateral periaqueductal grey (VLPAG) subdivision in CO2-sensitive strains. Conversely, DβH positive cell counts within the LC were significantly higher in CO2-sensitive strains. Collectively, our data provide evidence for strain dependent, differential CO2-sensitivity and potential differences in monoaminergic systems regulating panic and anxiety. Comparative studies between CO2-vulnerable and resistant strains may facilitate the mechanistic understanding of differential CO2-sensitivity in the development of panic and anxiety disorders. PMID:28087339

  13. MicroRNA-30b-Mediated Regulation of Catalase Expression in Human ARPE-19 Cells

    PubMed Central

    Haque, Rashidul; Chun, Eugene; Howell, Jennifer C.; Sengupta, Trisha; Chen, Dan; Kim, Hana

    2012-01-01

    Background Oxidative injury to retinal pigment epithelium (RPE) and retinal photoreceptors has been linked to a number of retinal diseases, including age-related macular degeneration (AMD). Reactive oxygen species (ROS)-mediated gene expression has been extensively studied at transcriptional levels. Also, the post-transcriptional control of gene expression at the level of translational regulation has been recently reported. However, the microRNA (miRNA/miR)-mediated post-transcriptional regulation in human RPE cells has not been thoroughly looked at. Increasing evidence points to a potential role of miRNAs in diverse physiological processes. Methodology/Principal Findings We demonstrated for the first time in a human retinal pigment epithelial cell line (ARPE-19) that the post-transcriptional control of gene expression via miRNA modulation regulates human catalase, an important and potent component of cell's antioxidant defensive network, which detoxifies hydrogen peroxide (H2O2) radicals. Exposure to several stress-inducing agents including H2O2 has been reported to alter miRNA expression profile. Here, we demonstrated that a sublethal dose of H2O2 (200 µM) up-regulated the expression of miR-30b, a member of the miR-30 family, which inhibited the expression of endogenous catalase both at the transcript and protein levels. However, antisense (antagomirs) of miR-30b was not only found to suppress the miR-30b mimics-mediated inhibitions, but also to dramatically increase the expression of catalase even under an oxidant environment. Conclusions/Significance We propose that a microRNA antisense approach could enhance cytoprotective mechanisms against oxidative stress by increasing the antioxidant defense system. PMID:22880027

  14. Molecular analysis of the human SLC13A4 sulfate transporter gene promoter

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jefferis, J.; Rakoczy, J.; School of Biomedical Sciences, University of Queensland, St. Lucia, Queensland

    2013-03-29

    Highlights: ► Basal promoter activity of SLC13A4 −57 to −192 nt upstream of transcription initiation site. ► Human SLC13A4 5′-flanking region has conserved motifs with other placental species. ► Putative NFY, SP1 and KLF7 motifs in SLC13A4 5′-flanking region enhance transcription. -- Abstract: The human solute linked carrier (SLC) 13A4 gene is primarily expressed in the placenta where it is proposed to mediate the transport of nutrient sulfate from mother to fetus. The molecular mechanisms involved in the regulation of SLC13A4 expression remain unknown. To investigate the regulation of SLC13A4 gene expression, we analysed the transcriptional activity of the humanmore » SLC13A4 5′-flanking region in the JEG-3 placental cell line using luciferase reporter assays. Basal transcriptional activity was identified in the region −57 to −192 nucleotides upstream of the SLC13A4 transcription initiation site. Mutational analysis of the minimal promoter region identified Nuclear factor Y (NFY), Specificity protein 1 (SP1) and Krüppel like factor 7 (KLF7) motifs which conferred positive transcriptional activity, as well as Zinc finger protein of the cerebellum 2 (ZIC2) and helix–loop–helix protein 1 (HEN1) motifs that repressed transcription. The conserved NFY, SP1, KLF7, ZIC2 and HEN1 motifs in the SLC13A4 promoter of placental species but not in non-placental species, suggests a potential role for these putative transcriptional factor binding motifs in the physiological control of SLC13A4 mRNA expression.« less

  15. Sustainable Horizontal Bioventing and Vertical Biosparging Implementation (Invited)

    NASA Astrophysics Data System (ADS)

    Leu, J.; Lin, J.; Ferris, S.

    2013-12-01

    A former natural gas processing site with total petroleum hydrocarbons (TPH) and benzene, toluene, ethylbenzene, and xylene (BTEX) impacts in both soil and groundwater was partially excavated to remove 2,400 cubic yards of impacted soil. However, due to active natural gas pipelines within the impacted footprint, excavation was discontinued and an area of impacted soil containing maximum concentrations of 5,000 mg/kg gasoline-range organics (GRO), 8,600 mg/kg diesel-range organics (DRO), and 130 mg/kg motor oil-range organics (ORO). Groundwater was impacted with concentrations up to 2,300 μg/L GRO and 4,200 μg/L DRO remained in place. Taking advantage of the open excavation, horizontal-screened piping was placed in the backfill to deliver air for bioventing, which resulted in successful remediation of soil in a physically inaccessible area. The combined use of excavation of the source area, bioventing of surrounding inaccessible soil, and biosparging of the groundwater and smear zone resulted in nearing a no-further-action status at the site. The sustainable bioventing system consisted of one 3-HP blower and eight horizontal air injection wells. Five dual-depth nested vapor monitoring points (VMPs) were installed at 5 feet and 10 feet below ground surface as part of the monitoring system for human health and system performance. The bioventing system operated for one year followed by a three-month rebound test. During the one-year operation, air flow was periodically adjusted to maximize removal of volatile organic compounds (VOCs) from the vent wells with elevated photo-ionization detector readings. After the bioventing successfully remediated the inaccessible impacted soil, the biosparging system incorporated the pre-existing bioventing unit with an upgraded 5-HP blower and three vertical biosparging wells to biodegrade dissolved phase impacts in the groundwater. The subsequent monitoring system includes the VMPs, the air injection wells, and four groundwater monitoring wells including three existing wells. The system is scheduled to operate for at least one year followed by a three-month rebound test. The flow rate was adjusted between 5 and 10 scfm during operations to focus the biosparging in the impacted area of the site. After the bioventing system was operated and optimized for a year, average VOC concentrations were reduced from approximately 120 to 5 ppmv in the vadose zone. TPH gasoline and BTEX concentrations experienced reductions up to 99%. Fugitive VOCs were not detected outside the property boundary or at possible fugitive gas monitoring points. During the rebound test, no significant rebound of VOC concentrations was observed. The average hydrocarbon biodegradation rate was estimated to be approximately 2.5 mg TPH/kg soil/day. During biosparging, the migration of injected air also stimulated biodegradation in the vadose zone. Within six months of operation, the groundwater GRO and DRO concentrations decreased approximately 70% and 50%, respectively, at the monitoring well within the excavation/backfill area. Bioventing followed by biosparging has proven to be successful in decreasing soil vapor chemicals of concern in the native soil of the inaccessible area and in groundwater of the excavation/backfill area.

  16. Paradoxical Regulation of Human FGF21 by Both Fasting and Feeding Signals: Is FGF21 a Nutritional Adaptation Factor?

    PubMed Central

    Uebanso, Takashi; Taketani, Yutaka; Yamamoto, Hironori; Amo, Kikuko; Ominami, Hirokazu; Arai, Hidekazu; Takei, Yuichiro; Masuda, Masashi; Tanimura, Ayako; Harada, Nagakatsu; Yamanaka-Okumura, Hisami; Takeda, Eiji

    2011-01-01

    Fibroblast growth factor 21 (FGF21) has recently emerged as a metabolic hormone involved in regulating glucose and lipid metabolism in mouse, but the regulatory mechanisms and actions of FGF21 in humans remain unclear. Here we have investigated the regulatory mechanisms of the human FGF21 gene at the transcriptional level. A deletion study of the human FGF21 promoter (−1672 to +230 bp) revealed two fasting signals, including peroxisome proliferator-activated receptor α (PPARα) and glucagon signals, that independently induced human FGF21 gene transcription in mouse primary hepatocytes. In addition, two feeding signals, glucose and xylitol, also dose-dependently induced human FGF21 gene transcription and mRNA expression in both human HepG2 cells and mouse primary hepatocytes. FGF21 protein expression and secretion were also induced by high glucose stimulation. The human FGF21 promoter (−1672 to +230 bp) was found to have a carbohydrate-responsive element at −380 to −366 bp, which is distinct from the PPAR response element (PPRE). Knock-down of the carbohydrate response element binding protein by RNAi diminished glucose-induced human FGF21 transcription. Moreover, we found that a region from −555 to −443 bp of the human FGF21 promoter region exerts an important role in the activation of basic transcription. In conclusion, human FGF21 gene expression is paradoxically and independently regulated by both fasting and feeding signals. These regulatory mechanisms suggest that human FGF21 is increased with nutritional crisis, including starvation and overfeeding. PMID:21829679

  17. Vasohibin 2 promotes human luminal breast cancer angiogenesis in a non-paracrine manner via transcriptional activation of fibroblast growth factor 2.

    PubMed

    Tu, Min; Lu, Cheng; Lv, Nan; Wei, Jishu; Lu, Zipeng; Xi, Chunhua; Chen, Jianmin; Guo, Feng; Jiang, Kuirong; Li, Qiang; Wu, Junli; Song, Guoxin; Wang, Shui; Gao, Wentao; Miao, Yi

    2016-12-28

    Vasohibin 2 (VASH2) is an angiogenic factor and cancer-related protein that acts via paracrine mechanisms. Here, we investigated the angiogenic function and mechanism of action of VASH2 in 200 human breast cancer tissues by performing immunohistochemical staining, western blot, indirect sandwich enzyme-linked immunosorbent assay (ELISA), and a semi-quantitative sandwich-based antibody array. Breast cancer cells stably overexpressing VASH2 or with knocked-down VASH2 were established and used for in vivo and in vitro models. In human luminal tissue, but not in HER2-positive or basal-like breast cancer tissues, VASH2 was positively correlated with CD31-positive microvascular density, induced angiogenesis in xenograft tumors, and promoted human umbilical vein endothelial cell tube formation in vitro. VASH2 expression was absent in the concentrated conditioned medium collected from knocked-down VASH2 and VASH2-overexpressing luminal breast cancer cells. Further, VASH2 regulated the expression of fibroblast growth factor 2 (FGF2) in human luminal breast cancer cells, and the pro-angiogenic effect induced by VASH2 overexpression was blocked by FGF2 neutralization in vitro. Additionally, dual luciferase reporter assay and Chromatin immunoprecipitation analysis results showed that FGF2 promoter was transcriptionally activated by VASH2 via histone modifications. In conclusion, VASH2 expression is positively correlated with FGF2 expression and promotes angiogenesis in human luminal breast cancer by transcriptional activation of fibroblast growth factor 2 through non-paracrine mechanisms. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  18. Alternate promoter selection within a human cytomegalovirus immediate-early and early transcription unit (UL119-115) defines true late transcripts containing open reading frames for putative viral glycoproteins.

    PubMed Central

    Leatham, M P; Witte, P R; Stinski, M F

    1991-01-01

    The human cytomegalovirus open reading frames (ORFs) UL119 through UL115 (UL119-115) are located downstream of the immediate-early 1 and 2 transcription units. The promoter upstream of UL119 is active at all times after infection and drives the synthesis of a spliced 3.1-kb mRNA. The viral mRNA initiates in UL119, contains UL119-117 and UL116, and terminates just downstream of UL115. True late transcripts that are detected only after viral DNA synthesis originate from this transcription unit. True late mRNAs of 2.1 kb, containing ORFs UL116 and UL115, and 1.2 kb, containing ORF UL115 only, are synthesized. The true late viral mRNAs are 3' coterminal with the 3.1-kb mRNA. This transcription unit is an example of late promoters nested within an immediate-early-early transcription unit. The gene products of UL119-117, UL116, and UL115 are predicted to be glycoproteins. Efficient expression of the downstream ORFs at late times after infection may be related to alternate promoter usage and downstream cap site selection. Images PMID:1717716

  19. IMMUNOCYTOCHEMICAL IDENTIFICATION OF ELECTRONEUTRAL NA+-COUPLED HCO3− TRANSPORTERS IN FRESHLY DISSOCIATED MOUSE MEDULLARY RAPHÉ NEURONS

    PubMed Central

    COLEY, A. A.; RUFFIN, V. A.; MOSS, F. J.; HOPFER, U.; BORON, W. F.

    2013-01-01

    The medullary raphé (MR) of the medulla oblongata contains chemosensitive neurons that respond to increases in arterial [CO2], by altering firing rate, with increases being associated with serotonergic (5-hydroxytryptamine [5HT]) neurons and decreases, with GABAergic neurons. Both types of neurons contribute to increased alveolar ventilation. Decreases in intracellular pH are thought to link the rise in [CO2] to increased ventilation. Because electroneutral Na+-coupled HCO3− transporters (nNCBTs), which help protect cells from intracellular acidosis, are expressed robustly in the neurons of the central nervous system, a key question is whether these transporters are present in chemosensitive neurons. Therefore, we used an immunocytochemistry approach to identify neurons (using a microtubule associated protein-2 monoclonal antibody) and specifically 5HT neurons (TPH monoclonal antibody) or GABAergic neurons (GAD2 monoclonal antibody) in freshly dissociated cells from the mouse MR. We also co-labeled with polyclonal antibodies against the three nNCBTs: NBCn1, NDCBE, and NBCn2. We exploited ePet-EYFP (enhanced yellow fluorescent protein) mice (with EYFP-labeled 5HT neurons) as well as mice genetically deficient in each of the three nNCBTs. Quantitative image analysis distinguished positively stained cells from background signals. We found that >80% of GAD2+ cells also were positive for NDCBE, and >90% of the TPH+ and GAD2+ cells were positive for the other nNCBTs. Assuming that the transporters are independently distributed among neurons, we can conclude that virtually all chemosensitive MR neurons contain at least one nNCBT. PMID:23500099

  20. Remediation of aged diesel contaminated soil by alkaline activated persulfate.

    PubMed

    Lominchar, M A; Santos, A; de Miguel, E; Romero, A

    2018-05-01

    The present work studies the efficiency of alkaline activated persulfate (PS) to remediate an aged diesel fuel contaminated soil from a train maintenance facility. The Total Petroleum Hydrocarbon (TPH) concentration in soil was approximately 5000mgkg -1 with a ratio of aliphatic:aromatic compounds of 70:30. Aromatic compounds were mainly naphtalenes and phenanthrenes. The experiments were performed in batch mode where different initial concentrations of persulfate (105mM, 210mM and 420mM) and activator:persulfate ratios (2 and 4) were evaluated, with NaOH used as activator. Runs were carried out during 56days. Complete TPH conversion was obtained with the highest concentration of PS and activator, whereas in the other runs the elimination of fuel ranged between 60 and 77%. Besides, the abatement of napthalenes and phenantrenes was faster than aliphatic reduction (i. e. after 4days of treatment, the conversions of the aromatic compounds were around 0.8 meanwhile the aliphatic abatements were 0.55) and no aromatic oxidation intermediates from naphtalenes or phenantrenes were detected. These results show that this technology is effective for the remediation of aged diesel in soil with alkaline pH. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Determination of hydrocarbons transported by urban runoff in sediments of São Gonçalo Channel (Pelotas - RS, Brazil).

    PubMed

    Sanches Filho, Pedro J; Böhm, Emerson M; Böhm, Giani M B; Montenegro, Gissele O; Silveira, Lucas A; Betemps, Glauco R

    2017-01-30

    A high concentration of hydrocarbons in the environment is indicative of pollution. To evaluate the effect of hydrocarbons transported by urban runoff, the present study analyzed total petroleum hydrocarbons (TPHs), aliphatic hydrocarbons (AHs), unresolved complex mixture (UCM), and n-alkanes of the sediments of the canal that cross the urban area of Pelotas, Rio Grande do Sul, Brazil. The carbon preference index (CPI), terrigenous/aquatic ratio (TAR), and pristane/phytane ratio were determined. The TPH content ranged from 177,043.7μg·kg -1 ±13.4% to 5,892,667.0μg·kg -1 ±5.9%. The total aliphatic content ranged from 116,268.8μg·kg -1 ±11.1% to 2,393,592.6μg·kg -1 ±7.7%, indicating chronic contamination of n-alkanes petrogenic and biogenic sources. The levels of hydrocarbons (TPH, AHs, and n-alkanes) were considered relatively high, confirming the effect of urban runoff on the drainage system of cities and their consequent effect on the estuarine region of Patos Lagoon and other water resources. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Microcosm assays and Taguchi experimental design for treatment of oil sludge containing high concentration of hydrocarbons.

    PubMed

    Castorena-Cortés, G; Roldán-Carrillo, T; Zapata-Peñasco, I; Reyes-Avila, J; Quej-Aké, L; Marín-Cruz, J; Olguín-Lora, P

    2009-12-01

    Microcosm assays and Taguchi experimental design was used to assess the biodegradation of an oil sludge produced by a gas processing unit. The study showed that the biodegradation of the sludge sample is feasible despite the high level of pollutants and complexity involved in the sludge. The physicochemical and microbiological characterization of the sludge revealed a high concentration of hydrocarbons (334,766+/-7001 mg kg(-1) dry matter, d.m.) containing a variety of compounds between 6 and 73 carbon atoms in their structure, whereas the concentration of Fe was 60,000 mg kg(-1) d.m. and 26,800 mg kg(-1) d.m. of sulfide. A Taguchi L(9) experimental design comprising 4 variables and 3 levels moisture, nitrogen source, surfactant concentration and oxidant agent was performed, proving that moisture and nitrogen source are the major variables that affect CO(2) production and total petroleum hydrocarbons (TPH) degradation. The best experimental treatment yielded a TPH removal of 56,092 mg kg(-1) d.m. The treatment was carried out under the following conditions: 70% moisture, no oxidant agent, 0.5% of surfactant and NH(4)Cl as nitrogen source.

  3. Phytoremediation of fuel oil and lead co-contaminated soil by Chromolaena odorata in association with Micrococcus luteus.

    PubMed

    Jampasri, Kongkeat; Pokethitiyook, Prayad; Kruatrachue, Maleeya; Ounjai, Puey; Kumsopa, Acharaporn

    2016-10-02

    Phytoremediation is widely promoted as a cost-effective technology for treating heavy metal and total petroleum hydrocarbon (TPH) co-contaminated soil. This study investigated the concurrent removal of TPHs and Pb in co-contaminated soil (27,000 mg kg(-1) TPHs, 780 mg kg(-1) Pb) by growing Siam weed (Chromolaena odorata) in a pot experiment for 90 days. There were four treatments: co-contaminated soil; co-contaminated soil with C. odorata only; co-contaminated soil with C. odorata and Micrococcus luteus inoculum; and co-contaminated soil with M. luteus only. C. odorata survived and grew well in the co-contaminated soil. C. odorata with M. luteus showed the highest Pb accumulation (513.7 mg kg(-1)) and uptake (7.7 mg plant(-1)), and the highest reduction percentage of TPHs (52.2%). The higher TPH degradation in vegetated soils indicated the interaction between the rhizosphere microorganisms and plants. The results suggested that C. odorata together with M. luteus and other rhizosphere microorganisms is a promising candidate for the removal of Pb and TPHs in co-contaminated soils.

  4. TPH and PAH concentrations in the subsoil of polyduct segments, oil pipeline pumping stations, and right-of-way pipelines from Central Mexico

    NASA Astrophysics Data System (ADS)

    Iturbe, Rosario; Castro, Alejandrina; Perez, Guillermina; Flores, Carlos; Torres, Luis G.

    2008-10-01

    For the year 1996, 366 incidents related with clandestine poaching of oil-products were reported in Mexico, 159 in 1997, and 240 in 1998. For the year 2003 (the most recently reported figure), there were 136 events. Petroleos Mexicanos (PEMEX), very concerned with the environmental agenda, has developed programs oriented to diminish contamination levels in all of its oil facilities. This work was aimed at characterizing zones around polyduct segments, pipelines, pumping stations, and right-of-way pipelines located in the center of Mexico. The TPH contaminated sites were, in decreasing order, polyduct km 39 + 150 > polyduct km 25 + 020 > Zoquital > Tepetitlan > Catalina > Venta Prieta > Ceiba. Most of the sampled points showed the presence of more than one of the 16 PAHs considered by USEPA as priority pollutants. Except point TEPE 2A, where no PAHs were detected, all the sampled points showed values from low to medium concentrations; however, values found at the sites did not exceed the limits according to the Mexican or the American legislation. The place with the largest contaminated area corresponded to the polyduct km 39 + 150, with 130 m2 and 260 m3 to be treated. The least contaminated area was that around the JUAN 4 point at Juandho station, with 20 m2 and 22 m3 of contaminated soil. The total area to be treated is about 230 m2 and 497 m3.

  5. Factors of transforming growth factor beta signalling are co-regulated in human hepatocellular carcinoma.

    PubMed

    Longerich, Thomas; Breuhahn, Kai; Odenthal, Margarete; Petmecky, Katharina; Schirmacher, Peter

    2004-12-01

    Transforming growth factor beta (TGFbeta) is a central mitoinhibitory factor for epithelial cells, and alterations of TGFbeta signalling have been demonstrated in many different human cancers. We have analysed human hepatocellular carcinomas (HCCs) for potential pro-tumourigenic alterations in regard to expression of Smad4 and mutations and expression changes of the pro-oncogenic transcriptional co-repressors Ski and SnoN, as well as mRNA levels of matrix metalloproteinase-2 (MMP2), which is transcriptionally regulated by TGFbeta. Smad4 mRNA was detected in all HCCs; while, using immunohistology, loss of Smad4 expression was found in 10% of HCCs. Neither mutations in the transformation-relevant sequences nor significant pro-tumourigenic expression changes of the Ski and SnoN genes were detected. In HCC cell lines, expression of both genes was regulated, potentially involving phosphorylation. Ski showed a distinct nuclear speckled pattern, indicating recruitment to active transcription complexes. MMP2 mRNA levels were increased in 19% of HCCs, whereas MMP2 mRNA was not detectable in HCC cell lines, suggesting that MMP2 was derived only from tumour stroma cells. Transcript levels of Smad4, Ski, SnoN and MMP2 correlated well. These data argue against a significant role of Ski and SnoN in human hepatocarcinogenesis and suggest that, in the majority of HCCs, the analysed factors are co-regulated by an upstream mechanism, potentially by TGFbeta itself.

  6. Physical structure and chromosomal localization of a gene encoding human p58[sup clk-1], a cell division control related protein kinase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Eipers, P.G.

    1992-01-01

    The gene for the human p58[sup clk[minus]1] protein kinase, a cell division control-related gene, has been mapped by somatic cell hybrid analyses, in situ localization with the chromosomal gene, and nested polymerase chain reaction amplification of microdissected chromosomes. These studies indicate that the expressed p58[sup clk[minus]1] chromosomal gene maps to 1p36, while a highly related p58[sup clk[minus]1] sequence of unknown nature maps to chromosome 15. Assignment of a p34[sup cdc2]-related gene to 1p36 region, including neuroblastoma, ductal carcinoma of the breast, malignant melanoma, Merkel cell carcinoma and endocrine neoplasia among others. Aberrant expression of this protein kinase negatively regulates normalmore » cellular growth. The p58[sup clk[minus]1] protein contains a central domain of 299 amino acids that is 46% identical to human p34[sup cdc2], the master mitotic protein kinase. This dissertation details the complete structure of the p58[sup clk[minus]1] chromosomal gene, including its putative promoter region, transcriptional start sites, exonic sequences, and intron/exon boundary sequences. The gene is 10 kb in size and contains 12 exons and 11 introns. Interestingly, the rather large 2.0 kb 3[prime] untranslated region is interrupted by an intron that separates a region containing numerous AUUUA destabilization motifs from the coding region. Furthermore, the expression of this gene in normal human tissues, as well as several human tumor cell samples and lines, is examined. The origin of multiple human transcripts from the same chromosomal gene, and the possible differential stability of these various transcripts, is discussed with regard to the transcriptional and post-transcriptional regulation of this gene. This is the first report of the chromosomal gene structure of a member of the p34[sup cdc2] supergene family.« less

  7. Regulatory Nexus of Synthesis and Degradation Deciphers Cellular Nrf2 Expression Levels

    PubMed Central

    Suzuki, Takafumi; Shibata, Tatsuhiro; Takaya, Kai; Shiraishi, Kouya; Kohno, Takashi; Kunitoh, Hideo; Tsuta, Koji; Furuta, Koh; Goto, Koichi; Hosoda, Fumie; Sakamoto, Hiromi; Motohashi, Hozumi

    2013-01-01

    Transcription factor Nrf2 (NF-E2-related factor 2) is essential for oxidative and electrophilic stress responses. While it has been well characterized that Nrf2 activity is tightly regulated at the protein level through proteasomal degradation via Keap1 (Kelch-like ECH-associated protein 1)-mediated ubiquitination, not much attention has been paid to the supply side of Nrf2, especially regulation of Nrf2 gene transcription. Here we report that manipulation of Nrf2 transcription is effective in changing the final Nrf2 protein level and activity of cellular defense against oxidative stress even in the presence of Keap1 and under efficient Nrf2 degradation, determined using genetically engineered mouse models. In excellent agreement with this finding, we found that minor A/A homozygotes of a single nucleotide polymorphism (SNP) in the human NRF2 upstream promoter region (rs6721961) exhibited significantly diminished NRF2 gene expression and, consequently, an increased risk of lung cancer, especially those who had ever smoked. Our results support the notion that in addition to control over proteasomal degradation and derepression from degradation/repression, the transcriptional level of the Nrf2 gene acts as another important regulatory point to define cellular Nrf2 levels. These results thus verify the critical importance of human SNPs that influence the levels of transcription of the NRF2 gene for future personalized medicine. PMID:23572560

  8. P-TEFb regulation of transcription termination factor Xrn2 revealed by a chemical genetic screen for Cdk9 substrates

    PubMed Central

    Sansó, Miriam; Levin, Rebecca S.; Lipp, Jesse J.; Wang, Vivien Ya-Fan; Greifenberg, Ann Katrin; Quezada, Elizabeth M.; Ali, Akbar; Ghosh, Animesh; Larochelle, Stéphane; Rana, Tariq M.; Geyer, Matthias; Tong, Liang; Shokat, Kevan M.; Fisher, Robert P.

    2016-01-01

    The transcription cycle of RNA polymerase II (Pol II) is regulated at discrete transition points by cyclin-dependent kinases (CDKs). Positive transcription elongation factor b (P-TEFb), a complex of Cdk9 and cyclin T1, promotes release of paused Pol II into elongation, but the precise mechanisms and targets of Cdk9 action remain largely unknown. Here, by a chemical genetic strategy, we identified ∼100 putative substrates of human P-TEFb, which were enriched for proteins implicated in transcription and RNA catabolism. Among the RNA processing factors phosphorylated by Cdk9 was the 5′-to-3′ “torpedo” exoribonuclease Xrn2, required in transcription termination by Pol II, which we validated as a bona fide P-TEFb substrate in vivo and in vitro. Phosphorylation by Cdk9 or phosphomimetic substitution of its target residue, Thr439, enhanced enzymatic activity of Xrn2 on synthetic substrates in vitro. Conversely, inhibition or depletion of Cdk9 or mutation of Xrn2-Thr439 to a nonphosphorylatable Ala residue caused phenotypes consistent with inefficient termination in human cells: impaired Xrn2 chromatin localization and increased readthrough transcription of endogenous genes. Therefore, in addition to its role in elongation, P-TEFb regulates termination by promoting chromatin recruitment and activation of a cotranscriptional RNA processing enzyme, Xrn2. PMID:26728557

  9. Ecosystem effects and the management of petroleum-contaminated soils on subantarctic islands.

    PubMed

    Errington, Ingrid; King, Catherine K; Wilkins, Daniel; Spedding, Tim; Hose, Grant C

    2018-03-01

    Human activity in the Polar Regions has resulted in petroleum contamination of soils. In this context, subantarctic islands are a unique management challenge for climatic, biological and logistical reasons. In this review we identify the main abiotic factors affecting petroleum-contaminated soils in the subantarctic environment, the primary effects of such contamination on biota, and lessons learned with regards to remediation techniques in this region. The sensitivity of biota to contamination depends on organism life stage, on soil properties, and on the degree of contaminant weathering. Initial studies using species endemic to subantarctic islands suggest that for fresh diesel fuel, sensitivities may range between 103 and 20 000 mg total petroleum hydrocarbons (TPH) kg -1 soil. Diesel that has undergone a short period of weathering is generally more toxic, with sensitivities ranging between 52 and 13 000 mg TPH kg -1 soil for an earthworm and a grass respectively (based on EC 20 and IC 50 values). A sufficient body of data from which to develop remediation targets for existing spills in the region does not yet exist for the region, but there has been a recent increase in research attention to address this data gap. A range of remediation methods have also now been trialled, and techniques such as in-ground aeration and nutrient addition have achieved some success. Passive management techniques such as permeable reactive barriers and phytoremediation are in preliminary stages of investigation for the region and show promise, not least because they cause less collateral disturbance than other methods. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  10. A Minimal Chimera of Human Cyclin T1 and Tat Binds TAR and Activates Human Immunodeficiency Virus Transcription in Murine Cells

    PubMed Central

    Fujinaga, Koh; Irwin, Dan; Taube, Ran; Zhang, Fan; Geyer, Matthias; Peterlin, B. Matija

    2002-01-01

    The transcriptional elongation of human immunodeficiency virus type 1 (HIV-1) is mediated by the virally encoded transactivator Tat and its cellular cofactor, positive transcription elongation factor b (P-TEFb). The human cyclin T1 (hCycT1) subunit of P-TEFb forms a stable complex with Tat and the transactivation response element (TAR) RNA located at the 5′ end of all viral transcripts. Previous studies have demonstrated that hCycT1 binds Tat in a Zn2+-dependent manner via the cysteine at position 261, which is a tyrosine in murine cyclin T1. In the present study, we mutated all other cysteines and histidines that could be involved in this Zn2+-dependent interaction. Because all of these mutant proteins except hCycT1(C261Y) activated viral transcription in murine cells, no other cysteine or histidine in hCycT1 is responsible for this interaction. Next, we fused the N-terminal 280 residues in hCycT1 with Tat. Not only the full-length chimera but also the mutant hCycT1 with an N-terminal deletion to position 249, which retained the Tat-TAR recognition motif, activated HIV-1 transcription in murine cells. This minimal hybrid mutant hCycT1-Tat protein bound TAR RNA as well as human and murine P-TEFb in vitro. We conclude that this minimal chimera not only reproduces the high-affinity binding among P-TEFb, Tat, and TAR but also will be invaluable for determining the three-dimensional structure of this RNA-protein complex. PMID:12438619

  11. Fibroblast growth factor 2 regulates bone sialoprotein gene transcription in human breast cancer cells.

    PubMed

    Li, Zhengyang; Wang, Zhitao; Yang, Li; Li, Xinyue; Sasaki, Yoko; Wang, Shuang; Araki, Shouta; Mezawa, Masaru; Takai, Hideki; Nakayama, Youhei; Ogata, Yorimasa

    2010-03-01

    Bone sialoprotein (BSP) is a major non-collagenous, extracellular matrix glycoprotein associated with mineralized tissues. Fibroblast growth factor 2 (FGF2) is recognized as a potent mitogen for a variety of mesenchymal cells. FGF2 produced by osteoblasts accumulates in the bone matrix and acts as an autocrine/paracrine regulator of osteoblasts. We previously reported that FGF2 regulates BSP gene transcription through the FGF2 response element (FRE) and activator protein 1 (AP1) binding site overlapping with the glucocorticoid response element in the rat BSP gene promoter. In the present study, FGF2 (10 ng/ml) increased BSP and Runx2 mRNA levels at 6 h in MCF7 human breast cancer cells. Transient transfection analyses were performed using chimeric constructs of the human BSP gene promoter linked to a luciferase reporter gene. Treatment of MCF7 cells with FGF2 (10 ng/ml) increased the luciferase activity of the constructs between -84LUC and -927LUC. Gel mobility shift analyses showed that FGF2 increased the binding of AP1 and CRE2. The CRE2- and AP1-protein complexes were disrupted by antibodies against CREB1, c-Fos, c-Jun, Fra2, p300 and Runx2. These studies demonstrate that FGF2 stimulates BSP transcription in MCF7 human breast cancer cells by targeting the AP1 and CRE2 elements in the human BSP gene promoter.

  12. RSR-2, the Caenorhabditis elegans Ortholog of Human Spliceosomal Component SRm300/SRRM2, Regulates Development by Influencing the Transcriptional Machinery

    PubMed Central

    Fontrodona, Laura; Porta-de-la-Riva, Montserrat; Morán, Tomás; Niu, Wei; Díaz, Mònica; Aristizábal-Corrales, David; Villanueva, Alberto; Schwartz, Simó; Reinke, Valerie; Cerón, Julián

    2013-01-01

    Protein components of the spliceosome are highly conserved in eukaryotes and can influence several steps of the gene expression process. RSR-2, the Caenorhabditis elegans ortholog of the human spliceosomal protein SRm300/SRRM2, is essential for viability, in contrast to the yeast ortholog Cwc21p. We took advantage of mutants and RNA interference (RNAi) to study rsr-2 functions in C. elegans, and through genetic epistasis analysis found that rsr-2 is within the germline sex determination pathway. Intriguingly, transcriptome analyses of rsr-2(RNAi) animals did not reveal appreciable splicing defects but instead a slight global decrease in transcript levels. We further investigated this effect in transcription and observed that RSR-2 colocalizes with DNA in germline nuclei and coprecipitates with chromatin, displaying a ChIP-Seq profile similar to that obtained for the RNA Polymerase II (RNAPII). Consistent with a novel transcription function we demonstrate that the recruitment of RSR-2 to chromatin is splicing-independent and that RSR-2 interacts with RNAPII and affects RNAPII phosphorylation states. Proteomic analyses identified proteins associated with RSR-2 that are involved in different gene expression steps, including RNA metabolism and transcription with PRP-8 and PRP-19 being the strongest interacting partners. PRP-8 is a core component of the spliceosome and PRP-19 is the core component of the PRP19 complex, which interacts with RNAPII and is necessary for full transcriptional activity. Taken together, our study proposes that RSR-2 is a multifunctional protein whose role in transcription influences C. elegans development. PMID:23754964

  13. A human Polycomb isoform lacking the Pc box does not participate to PRC1 complexes but forms protein assemblies and represses transcription.

    PubMed

    Völkel, Pamela; Le Faou, Perrine; Vandamme, Julien; Pira, Dorcas; Angrand, Pierre-Olivier

    2012-05-01

    Polycomb repression controls the expression of hundreds of genes involved in development and is mediated by essentially two classes of chromatin-associated protein complexes. The Polycomb repressive complex 2 (PRC2) trimethylates histone H3 at lysine 27, an epigenetic mark that serves as a docking site for the PRC1 protein complex. Drosophila core PRC1 is composed of four subunits: Polycomb (Pc), Posterior sex combs (Psc), Polyhomeotic (Ph) and Sex combs extra (Sce). Each of these proteins has multiple orthologs in vertebrates, thus generating an enormous scope for potential combinatorial diversity. In particular, mammalian genomes encode five Pc family members: CBX2, CBX4, CBX6, CBX7 and CBX8. To complicate matters further, distinct isoforms might arise from single genes. Here, we address the functional role of the two human CBX2 isoforms. Owing to different polyadenylation sites and alternative splicing events, the human CBX2 locus produces two transcripts: a 5-exon transcript that encodes the 532-amino acid CBX2-1 isoform that contains the conserved chromodomain and Pc box and a 4-exon transcript encoding a shorter isoform, CBX2-2, lacking the Pc box but still possessing a chromodomain. Using biochemical approaches and a novel in vivo imaging assay, we show that the short CBX2-2 isoform lacking the Pc box, does not participate in PRC1 protein complexes, but self-associates in vivo and forms complexes of high molecular weight. Furthermore, the CBX2 short isoform is still able to repress transcription, suggesting that Polycomb repression might occur in the absence of PRC1 formation.

  14. Role of the POZ zinc finger transcription factor FBI-1 in human and murine adipogenesis.

    PubMed

    Laudes, Matthias; Christodoulides, Constantinos; Sewter, Ciaran; Rochford, Justin J; Considine, Robert V; Sethi, Jaswinder K; Vidal-Puig, Antonio; O'Rahilly, Stephen

    2004-03-19

    Poxvirus zinc finger (POZ) zinc finger domain transcription factors have been shown to play a role in the control of growth arrest and differentiation in several types of mesenchymal cells but not, as yet, adipocytes. We found that a POZ domain protein, factor that binds to inducer of short transcripts-1 (FBI-1), was induced during both murine and human preadipocyte differentiation with maximal expression levels seen at days 2-4. FBI-1 mRNA was expressed in human adipose tissue with the highest levels found in samples from morbidly obese subjects. Murine cell lines constitutively expressing FBI-1 showed evidence for accelerated adipogenesis with earlier induction of markers of differentiation and enhanced lipid accumulation, suggesting that FBI-1 may be an active participant in the differentiation process. Consistent with the properties of this family of proteins in other cell systems, 3T3L1 cells stably overexpressing FBI-1 showed reduced DNA synthesis and reduced expression of cyclin A, cyclin-dependent kinase 2, and p107, proteins known to be involved in the regulation of mitotic clonal expansion. In addition, FBI-1 reduced the transcriptional activity of the cyclin A promoter. Thus, FBI-1, a POZ zinc finger transcription factor, is induced during the early phases of human and murine preadipocyte differentiation where it may contribute to adipogenesis through influencing the switch from cellular proliferation to terminal differentiation.

  15. A screen for nuclear transcripts identifies two linked noncoding RNAs associated with SC35 splicing domains

    PubMed Central

    Hutchinson, John N; Ensminger, Alexander W; Clemson, Christine M; Lynch, Christopher R; Lawrence, Jeanne B; Chess, Andrew

    2007-01-01

    Background Noncoding RNA species play a diverse set of roles in the eukaryotic cell. While much recent attention has focused on smaller RNA species, larger noncoding transcripts are also thought to be highly abundant in mammalian cells. To search for large noncoding RNAs that might control gene expression or mRNA metabolism, we used Affymetrix expression arrays to identify polyadenylated RNA transcripts displaying nuclear enrichment. Results This screen identified no more than three transcripts; XIST, and two unique noncoding nuclear enriched abundant transcripts (NEAT) RNAs strikingly located less than 70 kb apart on human chromosome 11: NEAT1, a noncoding RNA from the locus encoding for TncRNA, and NEAT2 (also known as MALAT-1). While the two NEAT transcripts share no significant homology with each other, each is conserved within the mammalian lineage, suggesting significant function for these noncoding RNAs. NEAT2 is extraordinarily well conserved for a noncoding RNA, more so than even XIST. Bioinformatic analyses of publicly available mouse transcriptome data support our findings from human cells as they confirm that the murine homologs of these noncoding RNAs are also nuclear enriched. RNA FISH analyses suggest that these noncoding RNAs function in mRNA metabolism as they demonstrate an intimate association of these RNA species with SC35 nuclear speckles in both human and mouse cells. These studies show that one of these transcripts, NEAT1 localizes to the periphery of such domains, whereas the neighboring transcript, NEAT2, is part of the long-sought polyadenylated component of nuclear speckles. Conclusion Our genome-wide screens in two mammalian species reveal no more than three abundant large non-coding polyadenylated RNAs in the nucleus; the canonical large noncoding RNA XIST and NEAT1 and NEAT2. The function of these noncoding RNAs in mRNA metabolism is suggested by their high levels of conservation and their intimate association with SC35 splicing domains in multiple mammalian species. PMID:17270048

  16. Ovarian-specific expression of a new gene regulated by the goat PIS region and transcribed by a FOXL2 bidirectional promoter.

    PubMed

    Pannetier, Maëlle; Renault, Lauriane; Jolivet, Geneviève; Cotinot, Corinne; Pailhoux, Eric

    2005-06-01

    Studies on XX sex reversal in polled goats (PIS mutation: polled intersex syndrome) have led to the discovery of a female-specific locus crucial for ovarian differentiation. This genomic region is composed of at least two genes, FOXL2 and PISRT1, sharing a common transcriptional regulatory region, PIS. In this paper, we describe a third gene, PFOXic (promoter FOXL2 inverse complementary), located near FOXL2 in the opposite orientation. This gene composed of five exons encodes a 1723-bp cDNA, enclosing two repetitive elements in its 3' end. PFOXic mRNA encodes a putative protein of 163 amino acids with no homologies in any of the databases tested. The transcriptional expression of PFOXic is driven by a bidirectional promoter also enhancing FOXL2 transcription. In goats, PFOXic is expressed in developing ovaries, from 36 days postcoitum until adulthood. Ovarian-specific expression of PFOXic is regulated by the PIS region. PFOXic is found conserved only in Bovidae. But, a human gene located in the opposite orientation relative to FOXL2 can be considered a human PFOXic. Finally, we discuss evidence arguing for regulation of the level of FOXL2 transcription via the bidirectional promoter and the level of transcription of PFOXic.

  17. Effect of exposure intensity and post-cure temperature storage on hardness of contemporary photo-activated composites.

    PubMed

    Quance, S C; Shortall, A C; Harrington, E; Lumley, P J

    2001-11-01

    The effect of variation in post-exposure storage temperature (18 vs. 37 degrees C) and light intensity (200 vs. 500mW/cm(2)) on micro-hardness of seven light-activated resin composite materials, cured with a Prismetics Mk II (Dentsply) light activation unit, were studied. Hardness values at the upper and lower surfaces of 2mm thick disc shaped specimens of seven light-cured resin composite materials (Herculite XRV and Prodigy/Kerr, Z100 and Silux Plus/3M, TPH/Dentsply, Pertac-Hybrid/Espe, and Charisma/Kulzer), which had been stored dry, were determined 24h after irradiation with a Prismetics Mk II (Dentsply) light activation unit. Hardness values varied with product, surface, storage temperature, and curing light intensity. In no case did the hardness at the lower surface equal that of the upper surface, and the combination of 500mW/cm(2) intensity and 37 degrees C storage produced the best hardness results at the lower surface. Material composition had a significant influence on surface hardness. Only one of the seven products (TPH) produced a mean hardness values at the lower surface >80% of the maximum mean upper surface hardness obtained for the corresponding product at 500mW/cm(2) intensity/37 degrees C storage temperature when subjected to all four test regimes. Despite optimum post-cure storage conditions, 200mW/cm(2) intensity curing for 40s will not produce acceptable hardness at the lower surface of 2mm increments of the majority of products tested.

  18. An integrated bioremediation process for petroleum hydrocarbons removal and odor mitigation from contaminated marine sediment.

    PubMed

    Zhang, Zhen; Lo, Irene M C; Yan, Dickson Y S

    2015-10-15

    This study developed a novel integrated bioremediation process for the removal of petroleum hydrocarbons and the mitigation of odor induced by reduced sulfur from contaminated marine sediment. The bioremediation process consisted of two phases. In Phase I, acetate was dosed into the sediment as co-substrate to facilitate the sulfate reduction process. Meanwhile, akaganeite (β-FeOOH) was dosed in the surface layer of the sediment to prevent S(2-) release into the overlying seawater. In Phase II, NO3(-) was injected into the sediment as an electron acceptor to facilitate the denitrification process. After 20 weeks of treatment, the sequential integration of the sulfate reduction and denitrification processes led to effective biodegradation of total petroleum hydrocarbons (TPH), in which about 72% of TPH was removed. In Phase I, the release of S(2-) was effectively controlled by the addition of akaganeite. The oxidation of S(2-) by Fe(3+) and the precipitation of S(2-) by Fe(2+) were the main mechanisms for S(2-) removal. In Phase II, the injection of NO3(-) completely inhibited the sulfate reduction process. Most of residual AVS and S(0) were removed within 4 weeks after NO3(-) injection. The 16S rRNA clone library-based analysis revealed a distinct shift of bacterial community structure in the sediment over different treatment phases. The clones affiliated with Desulfobacterales and Desulfuromonadales were the most abundant in Phase I, while the clones related to Thioalkalivibrio sulfidophilus, Thiohalomonas nitratireducens and Sulfurimonas denitrificans predominated in Phase II. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Profiling of m6A RNA modifications identified an age-associated regulation of AGO2 mRNA stability.

    PubMed

    Min, Kyung-Won; Zealy, Richard W; Davila, Sylvia; Fomin, Mikhail; Cummings, James C; Makowsky, Daniel; Mcdowell, Catherine H; Thigpen, Haley; Hafner, Markus; Kwon, Sang-Ho; Georgescu, Constantin; Wren, Jonathan D; Yoon, Je-Hyun

    2018-06-01

    Gene expression is dynamically regulated in a variety of mammalian physiologies. During mammalian aging, there are changes that occur in protein expression that are highly controlled by the regulatory steps in transcription, post-transcription, and post-translation. Although there are global profiles of human transcripts during the aging processes available, the mechanism(s) by which transcripts are differentially expressed between young and old cohorts remains unclear. Here, we report on N6-methyladenosine (m6A) RNA modification profiles of human peripheral blood mononuclear cells (PBMCs) from young and old cohorts. An m6A RNA profile identified a decrease in overall RNA methylation during the aging process as well as the predominant modification on proteincoding mRNAs. The m6A-modified transcripts tend to be more highly expressed than nonmodified ones. Among the many methylated mRNAs, those of DROSHA and AGO2 were heavily methylated in young PBMCs which coincided with a decreased steady-state level of AGO2 mRNA in the old PBMC cohort. Similarly, downregulation of AGO2 in proliferating human diploid fibroblasts (HDFs) also correlated with a decrease in AGO2 mRNA modifications and steady-state levels. In addition, the overexpression of RNA methyltransferases stabilized AGO2 mRNA but not DROSHA and DICER1 mRNA in HDFs. Moreover, the abundance of miRNAs also changed in the young and old PBMCs which are possibly due to a correlation with AGO2 expression as observed in AGO2-depleted HDFs. Taken together, we uncovered the role of mRNA methylation on the abundance of AGO2 mRNA resulting in the repression of miRNA expression during the process of human aging. © 2018 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

  20. Transcriptional consequences of XPA disruption in human cell lines

    PubMed Central

    Manandhar, Mandira; Lowery, Megan G.; Boulware, Karen S.; Lin, Kevin H.; Lu, Yue; Wood, Richard D.

    2017-01-01

    Nucleotide excision repair (NER) in mammalian cells requires the xeroderma pigmentosum group A protein (XPA) as a core factor. Remarkably, XPA and other NER proteins have been detected by chromatin immunoprecipitation at some active promoters, and NER deficiency is reported to influence the activated transcription of selected genes. However, the global influence of XPA on transcription in human cells has not been determined. We analyzed the human transcriptome by RNA sequencing (RNA-Seq). We first confirmed that XPA is confined to the cell nucleus even in the absence of external DNA damage, in contrast to previous reports that XPA is normally resident in the cytoplasm and is imported following DNA damage. We then analyzed four genetically matched human cell line pairs deficient or proficient in XPA. Of the ∼14,000 genes transcribed in each cell line, 325 genes (2%) had a significant XPA-dependent directional change in gene expression that was common to all four pairs (with a false discovery rate of 0.05). These genes were enriched in pathways for the maintenance of mitochondria. Only 27 common genes were different by more than 1.5-fold. The most significant hits were AKR1C1 and AKR1C2, involved in steroid hormone metabolism. AKR1C2 protein was lower in all of the immortalized XPA-deficient cells. Retinoic acid treatment led to modest XPA-dependent activation of some genes with transcription-related functions. We conclude that XPA status does not globally influence human gene transcription. However, XPA significantly influences expression of a small subset of genes important for mitochondrial functions and steroid hormone metabolism. The results may help explain defects in neurological function and sterility in individuals with xeroderma pigmentosum. PMID:28704716

  1. Effectiveness and potential ecological effects of offshore surface dispersant use during the Deepwater Horizon oil spill: a retrospective analysis of monitoring data.

    PubMed

    Bejarano, Adriana C; Levine, Edwin; Mearns, Alan J

    2013-12-01

    The Special Monitoring of Applied Response Technologies (SMART) program was used during the Deepwater Horizon oil spill as a strategy to monitor the effectiveness of sea surface dispersant use. Although SMART was implemented during aerial and vessel dispersant applications, this analysis centers on the effort of a special dispersant missions onboard the M/V International Peace, which evaluated the effectiveness of surface dispersant applications by vessel only. Water samples (n = 120) were collected from background sites, and under naturally and chemically dispersed oil slicks, and were analyzed for polycyclic aromatic hydrocarbons (TPAHs), total petroleum hydrocarbons (TPH), and a chemical marker of Corexit (dipropylene glycol n-butyl ether, DPnB). Water chemistry results were analyzed relative to SMART field assessments of dispersant effectiveness ("not effective," "effective," and "very effective"), based on in situ fluorometry. Chemistry data were also used to indirectly determine if the use of dispersants increased the risk of acute effects to water column biota, by comparison to toxicity benchmarks. TPAH and TPH concentrations in background, and naturally and chemically dispersed samples were extremely variable, and differences were not statistically detected across sample types. Ratios of TPAH and TPH between chemically and naturally dispersed samples provided a quantitative measure of dispersant effectiveness over natural oil dispersion alone, and were in reasonable agreement with SMART field assessments of dispersant effectiveness. Samples from "effective" and "very effective" dispersant applications had ratios of TPAH and TPH up to 35 and 64, respectively. In two samples from an "effective" dispersant application, TPHs and TPAHs exceeded acute benchmarks (0.81 mg/L and 8 μg/L, respectively), while none exceeded DPnB's chronic value (1,000 μg/L). Although the primary goal of the SMART program is to provide near real-time effectiveness data to the response, and not to address concerns regarding acute biological effects, the analyses presented here demonstrate that SMART can generate information of value to a larger scientific audience. A series of recommendations for future SMART planning are also provided.

  2. Intergenic disease-associated regions are abundant in novel transcripts.

    PubMed

    Bartonicek, N; Clark, M B; Quek, X C; Torpy, J R; Pritchard, A L; Maag, J L V; Gloss, B S; Crawford, J; Taft, R J; Hayward, N K; Montgomery, G W; Mattick, J S; Mercer, T R; Dinger, M E

    2017-12-28

    Genotyping of large populations through genome-wide association studies (GWAS) has successfully identified many genomic variants associated with traits or disease risk. Unexpectedly, a large proportion of GWAS single nucleotide polymorphisms (SNPs) and associated haplotype blocks are in intronic and intergenic regions, hindering their functional evaluation. While some of these risk-susceptibility regions encompass cis-regulatory sites, their transcriptional potential has never been systematically explored. To detect rare tissue-specific expression, we employed the transcript-enrichment method CaptureSeq on 21 human tissues to identify 1775 multi-exonic transcripts from 561 intronic and intergenic haploblocks associated with 392 traits and diseases, covering 73.9 Mb (2.2%) of the human genome. We show that a large proportion (85%) of disease-associated haploblocks express novel multi-exonic non-coding transcripts that are tissue-specific and enriched for GWAS SNPs as well as epigenetic markers of active transcription and enhancer activity. Similarly, we captured transcriptomes from 13 melanomas, targeting nine melanoma-associated haploblocks, and characterized 31 novel melanoma-specific transcripts that include fusion proteins, novel exons and non-coding RNAs, one-third of which showed allelically imbalanced expression. This resource of previously unreported transcripts in disease-associated regions ( http://gwas-captureseq.dingerlab.org ) should provide an important starting point for the translational community in search of novel biomarkers, disease mechanisms, and drug targets.

  3. Parathyroid hormone regulation of the human bone sialoprotein gene transcription is mediated through two cAMP response elements.

    PubMed

    Araki, Shouta; Mezawa, Masaru; Sasaki, Yoko; Yang, Li; Li, Zhengyang; Takai, Hideki; Nakayama, Youhei; Ogata, Yorimasa

    2009-03-01

    Parathyroid hormone (PTH) regulates serum calcium and inorganic phosphate levels through its actions on kidney and bone. Bone sialoprotein (BSP) is an early marker of osteoblast differentiation and bone metabolism. We here report that two cAMP response elements (CRE) in the human BSP gene promoter are target of PTH. In human osteoblast-like Saos2 cells, PTH (human 1-34 PTH, 10 nM) increased BSP mRNA and protein levels at 3 h. From transient transfection assays, 2- to 2.5-fold increase in transcription by PTH was observed at 3 and 6 h in -184, -211, -428, -868, and -927 luciferase constructs that included the human BSP gene promoter. Effect of PTH was abrogated by 2 bp mutations in either the CRE1 (-79 to -72) or CRE2 (-674 to -667). Luciferase activities induced by PTH were blocked by protein kinase A inhibitor H89 and tyrosine kinase inhibitor herbimycin A. Gel shift analyses showed that PTH increased binding of nuclear proteins to the CRE1 and CRE2 elements. The CRE1-protein and CRE2-protein complexes were disrupted by CRE binding protein 1 (CREB1) antibodies and supershifted by phospho-CREB1 antibody. ChIP assays detected binding of CREB1 and phospho-CREB1 to a chromatin fragment containing CRE1 and CRE2, and increased binding of phospho-CREB1 to the both sites. These studies demonstrate that PTH stimulates human BSP gene transcription by targeting the two CREs in the promoter of the human BSP gene.

  4. Distinct organization of the candidate tumor suppressor gene RFP2 in human and mouse: multiple mRNA isoforms in both species- and human-specific antisense transcript RFP2OS.

    PubMed

    Baranova, Ancha; Hammarsund, Marianne; Ivanov, Dmitry; Skoblov, Mikhail; Sangfelt, Olle; Corcoran, Martin; Borodina, Tatiana; Makeeva, Natalia; Pestova, Anna; Tyazhelova, Tatiana; Nazarenko, Svetlana; Gorreta, Francesco; Alsheddi, Tariq; Schlauch, Karen; Nikitin, Eugene; Kapanadze, Bagrat; Shagin, Dmitry; Poltaraus, Andrey; Ivanovich Vorobiev, Andrey; Zabarovsky, Eugene; Lukianov, Sergey; Chandhoke, Vikas; Ibbotson, Rachel; Oscier, David; Einhorn, Stefan; Grander, Dan; Yankovsky, Nick

    2003-12-04

    In the present study, we describe the human and mouse RFP2 gene structure, multiple RFP2 mRNA isoforms in the two species that have different 5' UTRs and a human-specific antisense transcript RFP2OS. Since the human RFP2 5' UTR is not conserved in mouse, these findings might indicate a different regulation of RFP2 in the two species. The predicted human and mouse RFP2 proteins are shown to contain a tripartite RING finger-B-box-coiled-coil domain (RBCC), also known as a TRIM domain, and therefore belong to a subgroup of RING finger proteins that are often involved in developmental and tumorigenic processes. Because homozygous deletions of chromosomal region 13q14.3 are found in a number of malignancies, including chronic lymphocytic leukemia (CLL) and multiple myeloma (MM), we suggest that RFP2 might be involved in tumor development. This study provides necessary information for evaluation of the role of RFP2 in malignant transformation and other biological processes.

  5. Base Realignment and Closure (BRAC) Cleanup Plan, Ford Ord, Monterey, California

    DTIC Science & Technology

    1994-03-25

    2,036.39 1 10 Maria Antonia Field 563.19 1 11 Stephen Joseph Field 1,018.02 1 April 1944 Key: I = Undocumented o45.sj Fort Ord, California - 25 March 1994...geophysical anomalies Further investigation of canal containoc .etroleum hydrocarbons discharge area which depending on and vanous organic compounds...detected at various areas. Concentration below TPH cleanup standard. Canal discharge area soil contained Pb, Sb, and Cr at concentration of concern. For

  6. Inactivation of the FoxO3a transcription factor is associated with the production of reactive oxygen species during protein kinase CK2 downregulation-mediated senescence in human colon cancer and breast cancer cells.

    PubMed

    Park, Seong-Yeol; Bae, Young-Seuk

    2016-09-09

    We previously showed that protein kinase CK2 downregulation mediates senescence through the reactive oxygen species (ROS)-p53-p21(Cip1/WAF1) pathway in various human cells. In the present study, we investigated whether the FoxO3a transcription factor is associated with ROS production during CK2 downregulation-induced senescence in human colon cancer HCT116 and breast cancer MCF-7 cells. FoxO3a overexpression suppressed ROS production and p53 stabilization induced by a CK2α knockdown. CK2α downregulation induced nuclear export of FoxO3a through stimulation of AKT-mediated phosphorylation of FoxO3a and decreased transcription of its target genes (Cu/ZnSOD, MnSOD, and catalase). In contrast, CK2α overexpression inhibited AKT-mediated FoxO3a phosphorylation. This resulted in nuclear accumulation of FoxO3a, and elevated expression of its target genes. Therefore, these data indicate for the first time that CK2 downregulation stimulates ROS generation by inhibiting FoxO3a during premature senescence in human colon and breast cancer cells. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Organotin and Irgarol-1051 contamination in Singapore coastal waters.

    PubMed

    Basheer, C; Tan, K S; Lee, H K

    2002-07-01

    The seas surrounding Singapore are principally utilized by the shipping industry but are now also increasingly used for a variety of other purposes, including desalination for supplies of drinking water and intensive aquaculture of food fish. While stringent environmental pollution standards are in place for industrial effluents, there is currently no legislative control over pollution from anti-fouling paints in Singapore. In this study, the concentrations of toxic antifouling agents tributyltin (TBT), triphenyltin (TPhT) and Irgarol-1051 (2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine) were determined from seawater obtained from 26 locations along and off the coast of Singapore in October and November 2000. These compounds were isolated by liquid-liquid extraction derivatized under controlled microwave heating and quantified by gas chromatography-mass spectrometry. TBT concentrations in seawater ranged between 0.43 and 3.20 microg 1(-1) with a mean value of 1.40 +/- 0.60 micro 1(-1). The mean values of DBT and MBT were 1.07 +/- 0.80 microg 11(-1) and 0.34 +/- 0.50 microg 1(-1) respectively, while TPhT concentrations of up to 0.40 microg 1(-1) were found. Monophenyltin and diphenyltin were not detected in all samples analysed. Irgarol-1051 was found to be present at concentrations of between 3.02 microg 1(-1) and 4.20 microg 1(-1) in seawater with a mean value of 2.00 +/- 1.20 microg 1(-1).

  8. Effects of adding bulking agent, inorganic nutrient and microbial inocula on biopile treatment for oil-field drilling waste.

    PubMed

    Ma, Jie; Yang, Yongqi; Dai, Xiaoli; Chen, Yetong; Deng, Hanmei; Zhou, Huijun; Guo, Shaohui; Yan, Guangxu

    2016-05-01

    Contamination from oil-field drilling waste is a worldwide environmental problem. This study investigated the performance of four bench-scale biopiles in treating drilling waste: 1) direct biopile (DW), 2) biopile plus oil-degrading microbial consortium (DW + M), 3) biopile plus microbial consortium and bulking agents (saw dust) (DW + M + BA), 4) biopile plus microbial consortium, bulking agents, and inorganic nutrients (Urea and K2HPO4) (DW + M + BA + N). Ninety days of biopiling removed 41.0%, 44.0%, 55.7% and 87.4% of total petroleum hydrocarbon (TPH) in the pile "DW", "DW + M", "DW + M + BA", and "DW + M + BA + N" respectively. Addition of inorganic nutrient and bulking agents resulted in a 56.9% and 26.6% increase in TPH removal efficiency respectively. In contrast, inoculation of hydrocarbon-degrading microorganisms only slightly enhanced the contaminant removal (increased 7.3%). The biopile with stronger contaminant removal also had higher pile temperature and lower pile pH (e.g., in "DW + M + BA + N"). GC-MS analysis shows that biopiling significantly reduced the total number of detected contaminants and changed the chemical composition. Overall, this study shows that biopiling is an effective remediation technology for drilling waste. Adding inorganic nutrients and bulking agents can significantly improve biopile performance while addition of microbial inocula had minimal positive impacts on contaminant removal. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. A High Proportion of Chromosome 21 Promoter Polymorphisms Influence Transcriptional Activity

    PubMed Central

    Buckland, Paul R.; Coleman, Sharol L.; Hoogendoorn, Bastiaan; Guy, Carol; Smith, S. Kaye; O’Donovan, Michael C.

    2004-01-01

    We have sought to obtain an unbiased estimate of the proportion of polymorphisms in promoters of human genes that have functional effects. We carried out polymorphism discovery on a randomly selected group of 51 gene promoters mapping to human chromosome 21 and successfully analyzed the effect on transcription of 38 of the sequence variants. To achieve this, a total of 53 different haplotypes from 20 promoters were cloned into a modified pGL3 luciferase reporter gene vector and were tested for their abilities to promote transcription in HEK293t and JEG-3 cells. Up to seven (18%) of the 38 tested variants altered transcription by 1.5-fold, confirming that a surprisingly high proportion of promoter region polymorphisms are likely to be functionally important. The functional variants were distributed across the promoters of CRYAA, IFNAR1, KCNJ15, NCAM2, IGSF5, and B3GALT5. Three of the genes (NCAM2, IFNAR1, and CRYAA) have been previously associated with human phenotypes and the polymorphisms we describe here may therefore play a role in those phenotypes. PMID:15200235

  10. Small RNAs Targeting Transcription Start Site Induce Heparanase Silencing through Interference with Transcription Initiation in Human Cancer Cells

    PubMed Central

    Pu, Jiarui; Mei, Hong; Zhao, Jun; Huang, Kai; Zeng, Fuqing; Tong, Qiangsong

    2012-01-01

    Heparanase (HPA), an endo-h-D-glucuronidase that cleaves the heparan sulfate chain of heparan sulfate proteoglycans, is overexpressed in majority of human cancers. Recent evidence suggests that small interfering RNA (siRNA) induces transcriptional gene silencing (TGS) in human cells. In this study, transfection of siRNA against −9/+10 bp (siH3), but not −174/−155 bp (siH1) or −134/−115 bp (siH2) region relative to transcription start site (TSS) locating at 101 bp upstream of the translation start site, resulted in TGS of heparanase in human prostate cancer, bladder cancer, and gastric cancer cells in a sequence-specific manner. Methylation-specific PCR and bisulfite sequencing revealed no DNA methylation of CpG islands within heparanase promoter in siH3-transfected cells. The TGS of heparanase did not involve changes of epigenetic markers histone H3 lysine 9 dimethylation (H3K9me2), histone H3 lysine 27 trimethylation (H3K27me3) or active chromatin marker acetylated histone H3 (AcH3). The regulation of alternative splicing was not involved in siH3-mediated TGS. Instead, siH3 interfered with transcription initiation via decreasing the binding of both RNA polymerase II and transcription factor II B (TFIIB), but not the binding of transcription factors Sp1 or early growth response 1, on the heparanase promoter. Moreover, Argonaute 1 and Argonaute 2 facilitated the decreased binding of RNA polymerase II and TFIIB on heparanase promoter, and were necessary in siH3-induced TGS of heparanase. Stable transfection of the short hairpin RNA construct targeting heparanase TSS (−9/+10 bp) into cancer cells, resulted in decreased proliferation, invasion, metastasis and angiogenesis of cancer cells in vitro and in athymic mice models. These results suggest that small RNAs targeting TSS can induce TGS of heparanase via interference with transcription initiation, and significantly suppress the tumor growth, invasion, metastasis and angiogenesis of cancer cells. PMID:22363633

  11. Erythroid activator NF-E2, TAL1 and KLF1 play roles in forming the LCR HSs in the human adult β-globin locus.

    PubMed

    Kim, Yea Woon; Yun, Won Ju; Kim, AeRi

    2016-06-01

    The β-like globin genes are developmental stage specifically transcribed in erythroid cells. The transcription of the β-like globin genes requires erythroid specific activators such as GATA-1, NF-E2, TAL1 and KLF1. However, the roles of these activators have not fully elucidated in transcription of the human adult β-globin gene. Here we employed hybrid MEL cells (MEL/ch11) where a human chromosome containing the β-globin locus is present and the adult β-globin gene is highly transcribed by induction. The roles of erythroid specific activators were analyzed by inhibiting the expression of NF-E2, TAL1 or KLF1 in MEL/ch11 cells. The loss of each activator decreased the transcription of human β-globin gene, locus wide histone hyperacetylation and the binding of other erythroid specific activators including GATA-1, even though not affecting the expression of other activators. Notably, sensitivity to DNase I was reduced in the locus control region (LCR) hypersensitive sites (HSs) with the depletion of activators. These results indicate that NF-E2, TAL1 and KLF1, all activators play a primary role in HSs formation in the LCR. It might contribute to the transcription of human adult β-globin gene by allowing the access of activators and cofactors. The roles of activators in the adult β-globin locus appear to be different from the roles in the early fetal locus. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Effects of Citalopram on Serotonin and CRF Systems in the Midbrain of Primates with Differences in Stress Sensitivity

    PubMed Central

    Bethea, Cynthia L.; Lima, Fernanda B.; Centeno, Maria L.; Weissheimer, Karin V.; Senashova, Olga; Reddy, Arubala P.; Cameron, Judy L.

    2011-01-01

    This chapter reviews the neurobiological effects of stress sensitivity and CIT treatment observed in our nonhuman primate model of Functional Hypothalamic Amenorrhea (FHA). This type of infertility, also known as stress-induced amenorrhea, is exhibited by cynomolgus macaques. In small populations, some individuals are stress sensitive (SS) and others are highly stress resilient (HSR). The SS macaques have suboptimal secretion of estrogen and progesterone during normal menstrual cycles. SS monkeys also have decreased serotonin gene expression and increased CRF expression compared to HSR monkeys. Recently, we found that s-citalopram (CIT) treatment improved ovarian steroid secretion in SS monkeys, but had no effect in HSR monkeys. Examination of the serotonin system revealed that SS monkeys had significantly lower Fev (fifth Ewing variant, rodent Pet1), TPH2 (tryptophan hydroxylase 2), 5HT1A autoreceptor and SERT (serotonin reuptake transporter) expression in the dorsal raphe than SR monkeys. However, CIT did not alter the expression of either Fev, TPH2, SERT or 5HT1A mRNAs. In contrast, SS monkeys tended to a higher density of CRF fiber innervation of the dorsal raphe than HSR monkeys, and CIT significantly decreased the CRF fiber density in SS animals. In addition, CIT increased CRF-R2 gene expression in the dorsal raphe. We speculate that in a 15-week time frame, the therapeutic effect of S-citalopram may be achieved through a mechanism involving extracellular serotonin inhibition of CRF and stimulation of CRF-R2, rather than alteration of serotonin-related gene expression. PMID:21683135

  13. IQCJ-SCHIP1, a novel fusion transcript encoding a calmodulin-binding IQ motif protein

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kwasnicka-Crawford, Dorota A.; Carson, Andrew R.; Scherer, Stephen W.

    The existence of transcripts that span two adjacent, independent genes is considered rare in the human genome. This study characterizes a novel human fusion gene named IQCJ-SCHIP1. IQCJ-SCHIP1 is the longest isoform of a complex transcriptional unit that bridges two separate genes that encode distinct proteins, IQCJ, a novel IQ motif containing protein and SCHIP1, a schwannomin interacting protein that has been previously shown to interact with the Neurofibromatosis type 2 (NF2) protein. IQCJ-SCHIP1 is located on the chromosome 3q25 and comprises a 1692-bp transcript encompassing 11 exons spanning 828 kb of the genomic DNA. We show that IQCJ-SCHIP1 mRNAmore » is highly expressed in the brain. Protein encoded by the IQCJ-SCHIP1 gene was localized to cytoplasm and actin-rich regions and in differentiated PC12 cells was also seen in neurite extensions.« less

  14. Pseudogap Behavior of the Nuclear Spin-Lattice Relaxation Rate in FeSe Probed by 77Se-NMR

    NASA Astrophysics Data System (ADS)

    Shi, Anlu; Arai, Takeshi; Kitagawa, Shunsaku; Yamanaka, Takayoshi; Ishida, Kenji; Böhmer, Anna E.; Meingast, Christoph; Wolf, Thomas; Hirata, Michihiro; Sasaki, Takahiko

    2018-01-01

    We conducted 77Se-nuclear magnetic resonance studies of the iron-based superconductor FeSe in magnetic fields of 0.6 to 19 T to investigate the superconducting and normal-state properties. The nuclear spin-lattice relaxation rate divided by the temperature (T1T)-1 increases below the structural transition temperature Ts but starts to be suppressed below T*, well above the superconducting transition temperature Tc(H), resulting in a broad maximum of (T1T)-1 at Tp(H). This is similar to the pseudogap behavior in optimally doped cuprate superconductors. Because T* and Tp(H) decrease in the same manner as Tc(H) with increasing H, the pseudogap behavior in FeSe is ascribed to superconducting fluctuations, which presumably originate from the theoretically predicted preformed pair above Tc(H).

  15. Remediation of soil co-contaminated with petroleum and heavy metals by the integration of electrokinetics and biostimulation.

    PubMed

    Dong, Zhi-Yong; Huang, Wen-Hui; Xing, Ding-Feng; Zhang, Hong-Feng

    2013-09-15

    Successful remediation of soil co-contaminated with high levels of organics and heavy metals is a challenging task, because that metal pollutants in soil can partially or completely suppress normal heterotrophic microbial activity and thus hamper biodegradation of organics. In this study, the benefits of integrating electrokinetic (EK) remediation with biodegradation for decontaminating soil co-contaminated with crude oil and Pb were evaluated in laboratory-scale experiments lasting for 30 days. The treated soil contained 12,500 mg/kg of total petroleum hydrocarbons (TPH) and 450 mg/kg Pb. The amendments of EDTA and Tween 80, together with a regular refreshing of electrolyte showed the best performance to remediate this contaminated soil. An important function of EDTA-enhanced EK treatment was to eliminate heavy metal toxicity from the soil, thus activating microbial degradation of oil. Although Tween 80 reduced current, it could serve as a second substrate for enhancing microbial growth and biodegradation. It was found that oil biodegradation degree and microbial numbers increased toward the anode and cathode. Microbial metabolism was found to be beneficial to metal release from the soil matrix. Under the optimum conditions, the soil Pb and TPH removal percentages after 30 days of running reached 81.7% and 88.3%, respectively. After treatment, both the residual soil Pb and TPH concentrations met the requirement of the Chinese soil environmental quality standards. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Bioremediation of oil-based drill cuttings by a halophilic consortium isolated from oil-contaminated saline soil.

    PubMed

    Rezaei Somee, Maryam; Shavandi, Mahmoud; Dastgheib, Seyed Mohammad Mehdi; Amoozegar, Mohammad Ali

    2018-05-01

    Oil-based drill cuttings are hazardous wastes containing complex hydrocarbons, heavy metals, and brine. Their remediation is a crucial step before release to the environment. In this work, we enriched a halophilic consortium, from oil-polluted saline soil, which is capable of degrading diesel as the main pollutant of oil-based drill cuttings. The degradation ability of the consortium was evaluated in microcosms using two different diluting agents (fine sand and biologically active soil). During the bioremediation process, the bacterial community dynamics of the microcosms was surveyed using PCR amplification of a fragment of 16S rRNA gene followed by denaturing gradient gel electrophoresis (DGGE). The diesel degradation rates were monitored by total petroleum hydrocarbon (TPH) measurement and the total count of heterotrophic and diesel-degrading bacteria. After 3 months, the microcosm containing fine sand and drill cuttings with the ratio of 1:1 (initial TPH of 36,000 mg/kg) showed the highest TPH removal (40%) and its dominant bacterial isolates belonged to the genera Dietzia, Arthrobacter , and Halomonas . DGGE results also confirmed the role of these genera in drill cuttings remediation. DGGE analysis of the bacterial diversity showed that Propionibacterium, Salinimicrobium, Marinobacter , and Dietzia are dominant in active soil microcosm; whereas Bacillus, Salinibacillus , and Marinobacter are abundant in sand microcosm. Our results suggest that the bioaugmentation strategy would be more successful if the diluting agent does not contain a complex microbial community.

  17. Occurrence of organotin compounds in river sediments under the dynamic water level conditions in the Three Gorges Reservoir Area, China.

    PubMed

    Gao, Jun-Min; Zhang, Ke; Chen, You-Peng; Guo, Jin-Song; Wei, Yun-Mei; Jiang, Wen-Chao; Zhou, Bin; Qiu, Hui

    2015-06-01

    The Three Gorges Project is the largest hydro project in the world, and the water level of the Three Gorges Reservoir (TGR) is dynamic and adjustable with the aim of flood control and electrical power generation. It is necessary to investigate the pollutants and their underlying contamination processes under dynamic water levels to determine their environmental behaviors in the Three Gorges Reservoir Area (TGRA). Here, we report the assessment of organotin compounds (OTs) pollution in the river sediments of the TGRA. Surface sediment samples were collected in the TGRA at low and high water levels. Tributyltin (TBT), triphenyltin (TPhT), and their degradation products in sediments were quantified by gas chromatography-mass spectrometry. Butyltins (BTs) and phenyltins (PhTs) were detected in sediments, and BTs predominated over PhTs in the whole study area under dynamic water level conditions. The concentrations of OTs in sediments varied markedly among locations, and significant concentrations were found in river areas with high levels of boat traffic and wastewater discharge. Sediments at all stations except Cuntan were lightly contaminated with TBT, and total organic carbon (TOC) was a significant factor affecting the fate of TBT in the TGRA. The butyltin and phenyltin degradation indices showed no recent inputs of TBT or TPhT into this region, with the exception of fresh TPhT input at Xiakou Town. Shipping activity, wastewater discharge, and agriculture are the most likely sources of OTs in the TGRA.

  18. Assessment of the effectiveness of onsite exsitu remediation by enhanced natural attenuation in the Niger Delta region, Nigeria.

    PubMed

    Okparanma, Reuben N; Azuazu, Ikeabiama; Ayotamuno, Josiah M

    2017-12-15

    This study was conducted to quantify and rank the effectiveness of onsite exsitu remediation by enhanced natural attenuation using soil quality index. The investigation was conducted at three oil spill sites in the Niger Delta (5.317°N, 6.467°E), Nigeria with a predominance of Oxisols. Baseline assessment and a two-step post-remediation monitoring of the sites were conducted. Target contaminants including total petroleum hydrocarbon (TPH) and BTEX (benzene, toluene, ethylbenzene, and xylene) were analyzed by gas chromatography-mass spectrometry. Results of the baseline assessment showed that TPH concentrations across the study sites averaged between 5113 and 7640 mg/kg at 0- to 1-m depth, which was higher than the local regulatory value of 5000 mg/kg. The soil quality index across the sites ranged between 68 and 45, suggesting medium to high potential ecological health risks with medium to high priority for remediation. BTEX concentrations followed a similar trend. However, after remediation TPH degraded rapidly initially and then slowly but asymptotically during the post-remediation monitoring period. Then, soil quality index across the study sites ranged between 100 and 58, indicating very low to medium potential ecological health risks. This demonstrates the effectiveness of onsite exsitu remediation by enhanced natural attenuation as a remediation strategy for petroleum-contaminated soils, which holds great promise for the Niger Delta province. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Application of a continuously stirred tank bioreactor (CSTR) for bioremediation of hydrocarbon-rich industrial wastewater effluents.

    PubMed

    Gargouri, Boutheina; Karray, Fatma; Mhiri, Najla; Aloui, Fathi; Sayadi, Sami

    2011-05-15

    A continuously stirred tank bioreactor (CSTR) was used to optimize feasible and reliable bioprocess system in order to treat hydrocarbon-rich industrial wastewaters. A successful bioremediation was developed by an efficient acclimatized microbial consortium. After an experimental period of 225 days, the process was shown to be highly efficient in decontaminating the wastewater. The performance of the bioaugmented reactor was demonstrated by the reduction of COD rates up to 95%. The residual total petroleum hydrocarbon (TPH) decreased from 320 mg TPH l(-1) to 8 mg TPH l(-1). Analysis using gas chromatography-mass spectrometry (GC-MS) identified 26 hydrocarbons. The use of the mixed cultures demonstrated high degradation performance for hydrocarbons range n-alkanes (C10-C35). Six microbial isolates from the CSTR were characterized and species identification was confirmed by sequencing the 16S rRNA genes. The partial 16S rRNA gene sequences demonstrated that 5 strains were closely related to Aeromonas punctata (Aeromonas caviae), Bacillus cereus, Ochrobactrum intermedium, Stenotrophomonas maltophilia and Rhodococcus sp. The 6th isolate was affiliated to genera Achromobacter. Besides, the treated wastewater could be considered as non toxic according to the phytotoxicity test since the germination index of Lepidium sativum ranged between 57 and 95%. The treatment provided satisfactory results and presents a feasible technology for the treatment of hydrocarbon-rich wastewater from petrochemical industries and petroleum refineries. Copyright © 2011 Elsevier B.V. All rights reserved.

  20. Phosphorus flame retardants: properties, production, environmental occurrence, toxicity and analysis.

    PubMed

    van der Veen, Ike; de Boer, Jacob

    2012-08-01

    Since the ban on some brominated flame retardants (BFRs), phosphorus flame retardants (PFRs), which were responsible for 20% of the flame retardant (FR) consumption in 2006 in Europe, are often proposed as alternatives for BFRs. PFRs can be divided in three main groups, inorganic, organic and halogen containing PFRs. Most of the PFRs have a mechanism of action in the solid phase of burning materials (char formation), but some may also be active in the gas phase. Some PFRs are reactive FRs, which means they are chemically bound to a polymer, whereas others are additive and mixed into the polymer. The focus of this report is limited to the PFRs mentioned in the literature as potential substitutes for BFRs. The physico-chemical properties, applications and production volumes of PFRs are given. Non-halogenated PFRs are often used as plasticisers as well. Limited information is available on the occurrence of PFRs in the environment. For triphenyl phosphate (TPhP), tricresylphosphate (TCP), tris(2-chloroethyl)phosphate (TCEP), tris(chloropropyl)phosphate (TCPP), tris(1,3-dichloro-2-propyl)phosphate (TDCPP), and tetrekis(2-chlorethyl)dichloroisopentyldiphosphate (V6) a number of studies have been performed on their occurrence in air, water and sediment, but limited data were found on their occurrence in biota. Concentrations found for these PFRs in air were up to 47 μg m(-3), in sediment levels up to 24 mg kg(-1) were found, and in surface water concentrations up to 379 ng L(-1). In all these matrices TCPP was dominant. Concentrations found in dust were up to 67 mg kg(-1), with TDCPP being the dominant PFR. PFR concentrations reported were often higher than polybrominated diphenylether (PBDE) concentrations, and the human exposure due to PFR concentrations in indoor air appears to be higher than exposure due to PBDE concentrations in indoor air. Only the Cl-containing PFRs are carcinogenic. Other negative human health effects were found for Cl-containing PFRs as well as for TCP, which suggest that those PFRs would not be suitable alternatives for BFRs. TPhP, diphenylcresylphosphate (DCP) and TCP would not be suitable alternatives either, because they are considered to be toxic to (aquatic) organisms. Diethylphosphinic acid is, just like TCEP, considered to be very persistent. From an environmental perspective, resorcinol-bis(diphenylphosphate) (RDP), bisphenol-A diphenyl phosphate (BADP) and melamine polyphosphate, may be suitable good substitutes for BFRs. Information on PFR analysis in air, water and sediment is limited to TCEP, TCPP, TPhP, TCP and some other organophosphate esters. For air sampling passive samplers have been used as well as solid phase extraction (SPE) membranes, SPE cartridges, and solid phase micro-extraction (SPME). For extraction of PFRs from water SPE is recommended, because this method gives good recoveries (67-105%) and acceptable relative standard deviations (RSDs) (<20%), and offers the option of on-line coupling with a detection system. For the extraction of PFRs from sediment microwave-assisted extraction (MAE) is recommended. The recoveries (78-105%) and RSDs (3-8%) are good and the method is faster and requires less solvent compared to other methods. For the final instrumental analysis of PFRs, gas chromatography-flame photometric detection (GC-FPD), GC-nitrogen-phosphorus detection (NPD), GC-atomic emission detection (AED), GC-mass spectrometry (MS) as well as liquid chromatography (LC)-MS/MS and GC-Inductively-coupled plasma-MS (ICP-MS) are used. GC-ICP-MS is a promising method, because it provides much less complex chromatograms while offering the same recoveries and limits of detection (LOD) (instrumental LOD is 5-10 ng mL(-1)) compared to GC-NPD and GC-MS, which are frequently used methods for PFR analysis. GC-MS offers a higher selectivity than GC-NPD and the possibility of using isotopically labeled compounds for quantification. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. 25 years monitoring of PAHs and petroleum hydrocarbons biodegradation in soil.

    PubMed

    Harmsen, Joop; Rietra, René P J J

    2018-05-10

    Biodegradation of polycyclic aromatic hydrocarbons (PAHs) and total petroleum hydrocarbons (TPH) in sediment and soil has been monitored on seven experimental fields during periods up to 25 years. With this unique dataset, we investigated long-term very slow biodegradation under field conditions. . The data show that three biodegradation rates can be distinguished for PAHs: 1) rapid degradation during the first year, 2) slow degradation during the following 6 years and 3), subject of this paper, a very slow degradation after 7 years until at least 25 years. Beside 2-, 3- and 4-ring PAHs, also 5- and 6-ring PAHs (aromatic rings) were degraded, all at the same rate during very slow degradation. In the period of very slow degradation, 6% yr -1 of the PAHs present were removed in five fields and 2% yr -1 in two other fields, while in the same period no very slow degradation of TPH could be observed. The remaining petroleum hydrocarbons were high boiling and non-toxic. Using the calculated degradation rates and the independently measured bioavailability of the PAHs (Tenax-method), the PAHs degradation curves of all seven monitored fields could be modelled. Applying the model and data obtained with the Tenax-method for fresh contaminated material, results of long-term biodegradation can be predicted, which can support the use of bioremediation in order to obtain a legally acceptable residual concentration. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. Sequence Elements Upstream of the Core Promoter Are Necessary for Full Transcription of the Capsule Gene Operon in Streptococcus pneumoniae Strain D39

    PubMed Central

    Wen, Zhensong; Sertil, Odeniel; Cheng, Yongxin; Zhang, Shanshan; Liu, Xue; Wang, Wen-Ching

    2015-01-01

    Streptococcus pneumoniae is a major bacterial pathogen in humans. Its polysaccharide capsule is a key virulence factor that promotes bacterial evasion of human phagocytic killing. While S. pneumoniae produces at least 94 antigenically different types of capsule, the genes for biosynthesis of almost all capsular types are arranged in the same locus. The transcription of the capsular polysaccharide (cps) locus is not well understood. This study determined the transcriptional features of the cps locus in the type 2 virulent strain D39. The initial analysis revealed that the cps genes are cotranscribed from a major transcription start site at the −25 nucleotide (G) upstream of cps2A, the first gene in the locus. Using unmarked chromosomal truncations and a luciferase-based transcriptional reporter, we showed that the full transcription of the cps genes not only depends on the core promoter immediately upstream of cps2A, but also requires additional elements upstream of the core promoter, particularly a 59-bp sequence immediately upstream of the core promoter. Unmarked deletions of these promoter elements in the D39 genome also led to significant reduction in CPS production and virulence in mice. Lastly, common cps gene (cps2ABCD) mutants did not show significant abnormality in cps transcription, although they produced significantly less CPS, indicating that the CpsABCD proteins are involved in the encapsulation of S. pneumoniae in a posttranscriptional manner. This study has yielded important information on the transcriptional characteristics of the cps locus in S. pneumoniae. PMID:25733517

  3. Transcriptional specificity in various p53-mutant cells.

    PubMed

    Okaichi, Kumio; Izumi, Nanaka; Takamura, Yuma; Fukui, Shoichi; Kudo, Takashi

    2013-03-01

    Mutation of the tumor suppressor gene p53 is the most common genetic alteration observed in human tumors. However, the relationship between the mutation point of p53 and the transcriptional specificity is not so obvious. We prepared Saos-2 cells with various mutations of p53 that are found in human tumors, and examined the resulting transcriptional alterations in the cells. Loss of function and gain of function were observed in all p53 mutants. Hot-spot mutations of p53 are frequently found in tumor cells. We compared hot-spot mutations and other mutations of p53 and found that a more than 2-fold transcription of CADPS2, PIWIL4 and TRIM9 was induced by hot spot mutations, but not by other mutations. As PIWIL4 suppresses the p16(INK4A) and ARF pathway, restraining cell growth and genomic instability, induction of PIWIL4 expression may be one reason why hot-spot mutations are frequently found in tumor cells.

  4. Cloning of Novel Isoforms of the Human Gli2 Oncogene and Their Activities To Enhance Tax-Dependent Transcription of the Human T-Cell Leukemia Virus Type 1 Genome

    PubMed Central

    Tanimura, Akira; Dan, Shingo; Yoshida, Mitsuaki

    1998-01-01

    The expression of human T-cell leukemia virus type 1 (HTLV-1) is activated by interaction of a viral transactivator protein, Tax, and cellular transcription factor, CREB (cyclic AMP response element binding protein), which bind to a 21-bp enhancer in the long terminal repeats (LTR). THP (Tax-helping protein) was previously determined to enhance the transactivation by Tax protein. Here we report novel forms of the human homolog of a member of the Gli oncogene family, Gli2 (also termed Gli2/THP), an extended form of a zinc finger protein, THP, which was described previously. Four possible isoforms (hGli2 α, β, γ, and δ) are formed by combinations of two independent alternative splicings, and all the isoforms could bind to a DNA motif, TRE2S, in the LTR. The longer isoforms, α and β, were abundantly expressed in various cell lines including HTLV-1-infected T-cell lines. Fusion proteins of the hGli2 isoforms with the DNA-binding domain of Gal4 activated transcription when the reporter contained a Gal4-binding site and one copy of the 21-bp sequence, to which CREB binds. This activation was observed only in the presence of Tax. The 21-bp sequence in the reporter was also essential for the activation. These results suggest that simultaneous binding of hGli2 and CREB to the respective sites in the reporter seems to be critical for Tax protein to activate transcription. Consequently, it is probable that the LTR can be regulated by two independent signals through hGli2 and CREB, since the LTR contains the 21-bp and TRE2S sequences in the vicinity. PMID:9557682

  5. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Monte, D.; Coutte, L.; Dewitte, F.

    The ERM protein belongs to the family of Ets transcription factors. We show here that the human ERM gene is organized into 14 exons distributed along 65 kb of genomic DNA on chromosome 3. The two main functional domains of ERM, the acidic domain and the DNA-binding ETS domain, are overlapped by three different exons each. The 3{prime}-untranslated region of ERM is 2.1 kb, whereas the 5{prime}-untranslated region is about 0.3 kb; this allows the transcription of ERM transcripts of approximately 4 kb. The human ERM gene is localized to the q27-q29 region of chromosome 3. 17 refs., 3 figs.

  6. Differential HFE Gene Expression Is Regulated by Alternative Splicing in Human Tissues

    PubMed Central

    Proença, Daniela; Faustino, Paula

    2011-01-01

    Background The pathophysiology of HFE-derived Hereditary Hemochromatosis and the function of HFE protein in iron homeostasis remain uncertain. Also, the role of alternative splicing in HFE gene expression regulation and the possible function of the corresponding protein isoforms are still unknown. The aim of this study was to gain insights into the physiological significance of these alternative HFE variants. Methodology/Principal Findings Alternatively spliced HFE transcripts in diverse human tissues were identified by RT-PCR, cloning and sequencing. Total HFE transcripts, as well as two alternative splicing transcripts were quantified using a real-time PCR methodology. Intracellular localization, trafficking and protein association of GFP-tagged HFE protein variants were analysed in transiently transfected HepG2 cells by immunoprecipitation and immunofluorescence assays. Alternatively spliced HFE transcripts present both level- and tissue-specificity. Concerning the exon 2 skipping and intron 4 inclusion transcripts, the liver presents the lowest relative level, while duodenum presents one of the highest amounts. The protein resulting from exon 2 skipping transcript is unable to associate with β2M and TfR1 and reveals an ER retention. Conversely, the intron 4 inclusion transcript gives rise to a truncated, soluble protein (sHFE) that is mostly secreted by cells to the medium in association with β2M. Conclusions/Significance HFE gene post-transcriptional regulation is clearly affected by a tissue-dependent alternative splicing mechanism. Among the corresponding proteins, a sHFE isoform stands out, which upon being secreted into the bloodstream, may act in remote tissues. It could be either an agonist or antagonist of the full length HFE, through hepcidin expression regulation in the liver or by controlling dietary iron absorption in the duodenum. PMID:21407826

  7. Differential HFE gene expression is regulated by alternative splicing in human tissues.

    PubMed

    Martins, Rute; Silva, Bruno; Proença, Daniela; Faustino, Paula

    2011-03-03

    The pathophysiology of HFE-derived Hereditary Hemochromatosis and the function of HFE protein in iron homeostasis remain uncertain. Also, the role of alternative splicing in HFE gene expression regulation and the possible function of the corresponding protein isoforms are still unknown. The aim of this study was to gain insights into the physiological significance of these alternative HFE variants. Alternatively spliced HFE transcripts in diverse human tissues were identified by RT-PCR, cloning and sequencing. Total HFE transcripts, as well as two alternative splicing transcripts were quantified using a real-time PCR methodology. Intracellular localization, trafficking and protein association of GFP-tagged HFE protein variants were analysed in transiently transfected HepG2 cells by immunoprecipitation and immunofluorescence assays. Alternatively spliced HFE transcripts present both level- and tissue-specificity. Concerning the exon 2 skipping and intron 4 inclusion transcripts, the liver presents the lowest relative level, while duodenum presents one of the highest amounts. The protein resulting from exon 2 skipping transcript is unable to associate with β2M and TfR1 and reveals an ER retention. Conversely, the intron 4 inclusion transcript gives rise to a truncated, soluble protein (sHFE) that is mostly secreted by cells to the medium in association with β2M. HFE gene post-transcriptional regulation is clearly affected by a tissue-dependent alternative splicing mechanism. Among the corresponding proteins, a sHFE isoform stands out, which upon being secreted into the bloodstream, may act in remote tissues. It could be either an agonist or antagonist of the full length HFE, through hepcidin expression regulation in the liver or by controlling dietary iron absorption in the duodenum.

  8. Facilitated recycling protects human RNA polymerase III from repression by Maf1 in vitro.

    PubMed

    Cabart, Pavel; Lee, JaeHoon; Willis, Ian M

    2008-12-26

    Yeast cells synthesize approximately 3-6 million molecules of tRNA every cell cycle at a rate of approximately 2-4 transcripts/gene/s. This high rate of transcription is achieved through many rounds of reinitiation by RNA polymerase (pol) III on stable DNA-bound complexes of the initiation factor TFIIIB. Studies in yeast have shown that the rate of reinitiation is increased by facilitated recycling, a process that involves the repeated reloading of the polymerase on the same transcription unit. However, when nutrients become limiting or stress conditions are encountered, RNA pol III transcription is rapidly repressed through the action of the conserved Maf1 protein. Here we examine the relationship between Maf1-mediated repression and facilitated recycling in a human RNA pol III in vitro system. Using an immobilized template transcription assay, we demonstrate that facilitated recycling is conserved from yeast to humans. We assessed the ability of recombinant human Maf1 to inhibit different steps in transcription before and after preinitiation complex assembly. We show that recombinant Maf1 can inhibit the recruitment of TFIIIB and RNA pol III to immobilized templates. However, RNA pol III bound to preinitiation complexes or in elongation complexes is protected from repression by Maf1 and can undergo several rounds of initiation. This indicates that recombinant Maf1 is unable to inhibit facilitated recycling. The data suggest that additional biochemical steps may be necessary for rapid Maf1-dependent repression of RNA pol III transcription.

  9. LIM Domain Only 2 Regulates Endothelial Proliferation, Angiogenesis, and Tissue Regeneration.

    PubMed

    Meng, Shu; Matrone, Gianfranco; Lv, Jie; Chen, Kaifu; Wong, Wing Tak; Cooke, John P

    2016-10-06

    LIM domain only 2 (LMO2, human gene) is a key transcription factor that regulates hematopoiesis and vascular development. However, its role in adult endothelial function has been incompletely characterized. In vitro loss- and gain-of-function studies on LMO2 were performed in human umbilical vein endothelial cells with lentiviral overexpression or short hairpin RNA knockdown (KD) of LMO2, respectively. LMO2 KD significantly impaired endothelial proliferation. LMO2 controls endothelial G1/S transition through transcriptional regulation of cyclin-dependent kinase 2 and 4 as determined by reverse transcription polymerase chain reaction (PCR), western blot, and chromatin immunoprecipitation, and also influences the expression of Cyclin D1 and Cyclin A1. LMO2 KD also impaired angiogenesis by reducing transforming growth factor-β (TGF-β) expression, whereas supplementation of exogenous TGF-β restored defective network formation in LMO2 KD human umbilical vein endothelial cells. In a zebrafish model of caudal fin regeneration, RT-PCR revealed that the lmo2 (zebrafish gene) gene was upregulated at day 5 postresection. The KD of lmo2 by vivo-morpholino injections in adult Tg(fli1:egfp) y1 zebrafish reduced 5-bromo-2'-deoxyuridine incorporation in endothelial cells, impaired neoangiogenesis in the resected caudal fin, and substantially delayed fin regeneration. The transcriptional factor LMO2 regulates endothelial proliferation and angiogenesis in vitro. Furthermore, LMO2 is required for angiogenesis and tissue healing in vivo. Thus, LMO2 is a critical determinant of vascular and tissue regeneration. © 2016 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.

  10. Mesenchymal stem cells derived from human exocrine pancreas express transcription factors implicated in beta-cell development.

    PubMed

    Baertschiger, Reto M; Bosco, Domenico; Morel, Philippe; Serre-Beinier, Veronique; Berney, Thierry; Buhler, Leo H; Gonelle-Gispert, Carmen

    2008-07-01

    Transplantation of in vitro generated islets or insulin-producing cells represents an attractive option to overcome organ shortage. The aim of this study was to isolate, expand, and characterize cells from human exocrine pancreas and analyze their potential to differentiate into beta cells. Fibroblast-like cells growing out of human exocrine tissue were characterized by flow cytometry and by their capacity to differentiate into mesenchymal cell lineages. During cell expansion and after differentiation toward beta cells, expression of transcription factors of endocrine pancreatic progenitors was analyzed by reverse transcription polymerase chain reaction. Cells emerged from 14/18 human pancreatic exocrine fractions and were expanded up to 40 population doublings. These cells displayed surface antigens similar to mesenchymal stem cells from bone marrow. A culture of these cells in adipogenic and chondrogenic differentiation media allowed differentiation into adipocyte- and chondrocyte-like cells. During expansion, cells expressed transcription factors implicated in islet development such as Isl1, Nkx2.2, Nkx6.1, nestin, Ngn3, Pdx1, and NeuroD. Activin A and hepatocyte growth factor induced an expression of insulin, glucagon, and glucokinase. Proliferating cells with characteristics of mesenchymal stem cells and endocrine progenitors were isolated from exocrine tissue. Under specific conditions, these cells expressed little insulin. Human pancreatic exocrine tissue might thus be a source of endocrine cell progenitors.

  11. A fast-evolving human NPAS3 enhancer gained reporter expression in the developing forebrain of transgenic mice

    PubMed Central

    Kamm, Gretel B.; López-Leal, Rodrigo; Lorenzo, Juan R.; Franchini, Lucía F.

    2013-01-01

    The developmental brain gene NPAS3 stands out as a hot spot in human evolution because it contains the largest number of human-specific, fast-evolving, conserved, non-coding elements. In this paper we studied 2xHAR142, one of these elements that is located in the fifth intron of NPAS3. Using transgenic mice, we show that the mouse and chimp 2xHAR142 orthologues behave as transcriptional enhancers driving expression of the reporter gene lacZ to a similar NPAS3 expression subdomain in the mouse central nervous system. Interestingly, the human 2xHAR142 orthologue drives lacZ expression to an extended expression pattern in the nervous system. Thus, molecular evolution of 2xHAR142 provides the first documented example of human-specific heterotopy in the forebrain promoted by a transcriptional enhancer and suggests that it may have contributed to assemble the unique properties of the human brain. PMID:24218632

  12. Suppression of human fibrosarcoma cell growth by transcription factor, Egr-1, involves down-regulation of Bcl-2.

    PubMed

    Huang, R P; Fan, Y; Peng, A; Zeng, Z L; Reed, J C; Adamson, E D; Boynton, A L

    1998-09-11

    Previously, we showed that the transcription factor Egr-1 suppressed the proliferation of v-sis transformed NIH3T3 cells and also a number of human tumor cells. Here, we investigate the possible mechanisms responsible for this function. We show that transfected Egr-1 in human fibrosarcoma cells HT1080 leads to down-regulation of Bcl-2. Transient CAT transfection assays reveal that expression of Egr-1 suppresses Bcl-2 promoter activity in a dose-dependent manner. Furthermore, overexpression of Bcl-2 in Egr-1-expressing HT1080 cells enhanced cell proliferation in monolayer culture and increased anchorage-independent growth. Our results suggest that suppression of tumor cell proliferation by Egr-1 may be at least partially mediated through the down-regulation of Bcl-2.

  13. Total Petroleum Hydrocarbons (TPH): ToxFAQs

    MedlinePlus

    ... Favorites Del.icio.us Digg Facebook Google Bookmarks Yahoo MyWeb Page last reviewed: February 4, 2014 Page ... Favorites Del.icio.us Digg Facebook Google Bookmarks Yahoo MyWeb Contact Us: Agency for Toxic Substances and ...

  14. Role of the POZ Zinc Finger Transcription Factor FBI-1 in Human and Murine Adipogenesis

    PubMed Central

    Laudes, Matthias; Christodoulides, Constantinos; Sewter, Ciaran; Rochford, Justin J.; Considine, Robert V.; Sethi, Jaswinder K.; Vidal-Puig, Antonio; O’Rahilly, Stephen

    2015-01-01

    Poxvirus zinc finger (POZ) zinc finger domain transcription factors have been shown to play a role in the control of growth arrest and differentiation in several types of mesenchymal cells but not, as yet, adipocytes. We found that a POZ domain protein, factor that binds to inducer of short transcripts-1 (FBI-1), was induced during both murine and human preadipocyte differentiation with maximal expression levels seen at days 2–4. FBI-1 mRNA was expressed in human adipose tissue with the highest levels found in samples from morbidly obese subjects. Murine cell lines constitutively expressing FBI-1 showed evidence for accelerated adipogenesis with earlier induction of markers of differentiation and enhanced lipid accumulation, suggesting that FBI-1 may be an active participant in the differentiation process. Consistent with the properties of this family of proteins in other cell systems, 3T3L1 cells stably overexpressing FBI-1 showed reduced DNA synthesis and reduced expression of cyclin A, cyclin-dependent kinase 2, and p107, proteins known to be involved in the regulation of mitotic clonal expansion. In addition, FBI-1 reduced the transcriptional activity of the cyclin A promoter. Thus, FBI-1, a POZ zinc finger transcription factor, is induced during the early phases of human and murine preadipocyte differentiation where it may contribute to adipogenesis through influencing the switch from cellular proliferation to terminal differentiation. PMID:14701838

  15. Utility of next-generation RNA-sequencing in identifying chimeric transcription involving human endogenous retroviruses.

    PubMed

    Sokol, Martin; Jessen, Karen Margrethe; Pedersen, Finn Skou

    2016-01-01

    Several studies have shown that human endogenous retroviruses and endogenous retrovirus-like repeats (here collectively HERVs) impose direct regulation on human genes through enhancer and promoter motifs present in their long terminal repeats (LTRs). Although chimeric transcription in which novel gene isoforms containing retroviral and human sequence are transcribed from viral promoters are commonly associated with disease, regulation by HERVs is beneficial in other settings; for example, in human testis chimeric isoforms of TP63 induced by an ERV9 LTR protect the male germ line upon DNA damage by inducing apoptosis, whereas in the human globin locus the γ- and β-globin switch during normal hematopoiesis is mediated by complex interactions of an ERV9 LTR and surrounding human sequence. The advent of deep sequencing or next-generation sequencing (NGS) has revolutionized the way researchers solve important scientific questions and develop novel hypotheses in relation to human genome regulation. We recently applied next-generation paired-end RNA-sequencing (RNA-seq) together with chromatin immunoprecipitation with sequencing (ChIP-seq) to examine ERV9 chimeric transcription in human reference cell lines from Encyclopedia of DNA Elements (ENCODE). This led to the discovery of advanced regulation mechanisms by ERV9s and other HERVs across numerous human loci including transcription of large gene-unannotated genomic regions, as well as cooperative regulation by multiple HERVs and non-LTR repeats such as Alu elements. In this article, well-established examples of human gene regulation by HERVs are reviewed followed by a description of paired-end RNA-seq, and its application in identifying chimeric transcription genome-widely. Based on integrative analyses of RNA-seq and ChIP-seq, data we then present novel examples of regulation by ERV9s of tumor suppressor genes CADM2 and SEMA3A, as well as transcription of an unannotated region. Taken together, this article highlights the high suitability of contemporary sequencing methods in future analyses of human biology in relation to evolutionary acquired retroviruses in the human genome. © 2016 APMIS. Published by John Wiley & Sons Ltd.

  16. Effect of brain-derived neurotrophic factor on behavior and key members of the brain serotonin system in genetically predisposed to behavioral disorders mouse strains.

    PubMed

    Naumenko, V S; Kondaurova, E M; Bazovkina, D V; Tsybko, A S; Tikhonova, M A; Kulikov, A V; Popova, N K

    2012-07-12

    The effect of brain-derived neurotrophic factor (BDNF) on depressive-like behavior and serotonin (5-HT) system in the brain of antidepressant sensitive cataleptics (ASC)/Icg mouse strain, characterized by depressive-like behavior, in comparison with the parental nondepressive CBA/Lac mouse strain was examined. Significant decrease of catalepsy and tail suspension test (TST) immobility was shown 17days after acute central BDNF administration (300ng i.c.v.) in ASC mice. In CBA mouse strain, BDNF moderately decreased catalepsy without any effect on TST immobility time. Significant difference between ASC and CBA mice in the effect of BDNF on 5-HT system was revealed. It was shown that central administration of BDNF led to increase of 5-HT(1A) receptor gene expression but not 5-HT(1A) functional activity in ASC mice. Increased tryptophan hydroxylase-2 (Tph-2) and 5-HT(2A) receptor genes expression accompanied by 5-HT(2A) receptor sensitization was shown in BDNF-treated ASC but not in CBA mouse strain, suggesting BDNF-induced increase of the brain 5-HT system functional activity and activation of neurogenesis in "depressive" ASC mice. There were no changes found in the 5-HT transporter mRNA level in BDNF-treated ASC and CBA mice. In conclusion, central administration of BDNF produced prolonged ameliorative effect on depressive-like behavior accompanied by increase of the Tph-2, 5-HT(1A) and 5-HT(2A) genes expression and 5-HT(2A) receptor functional activity in animal model of hereditary behavior disorders. Copyright © 2012 IBRO. Published by Elsevier Ltd. All rights reserved.

  17. Effect of long-term actual spaceflight on the expression of key genes encoding serotonin and dopamine system

    NASA Astrophysics Data System (ADS)

    Popova, Nina; Shenkman, Boris; Naumenko, Vladimir; Kulikov, Alexander; Kondaurova, Elena; Tsybko, Anton; Kulikova, Elisabeth; Krasnov, I. B.; Bazhenova, Ekaterina; Sinyakova, Nadezhda

    The effect of long-term spaceflight on the central nervous system represents important but yet undeveloped problem. The aim of our work was to study the effect of 30-days spaceflight of mice on Russian biosatellite BION-M1 on the expression in the brain regions of key genes of a) serotonin (5-HT) system (main enzymes in 5-HT metabolism - tryptophan hydroxylase-2 (TPH-2), monoamine oxydase A (MAO A), 5-HT1A, 5-HT2A and 5-HT3 receptors); b) pivotal enzymes in DA metabolism (tyrosine hydroxylase, COMT, MAO A, MAO B) and D1, D2 receptors. Decreased expression of genes encoding the 5-HT catabolism (MAO A) and 5-HT2A receptor in some brain regions was shown. There were no differences between “spaceflight” and control mice in the expression of TPH-2 and 5-HT1A, 5-HT3 receptor genes. Significant changes were found in genetic control of DA system. Long-term spaceflight decreased the expression of genes encoding the enzyme in DA synthesis (tyrosine hydroxylase in s.nigra), DA metabolism (MAO B in the midbrain and COMT in the striatum), and D1 receptor in hypothalamus. These data suggested that 1) microgravity affected genetic control of 5-HT and especially the nigrostriatal DA system implicated in the central regulation of muscular tonus and movement, 2) the decrease in the expression of genes encoding key enzyme in DA synthesis, DA degradation and D1 receptor contributes to the movement impairment and dyskinesia produced by the spaceflight. The study was supported by Russian Foundation for Basic Research grant No. 14-04-00173.

  18. Tryptophan Hydroxylase 2 Gene Polymorphism in Anxiety and Depressive Disorder in Kashmiri Population

    PubMed Central

    Shoib, Sheikh; Shah, Tabindah; Mushtaq, Sahil

    2014-01-01

    Background: The gene of tryptophan hydroxylase is widely recognized as a major candidate gene in many psychiatric disorders. However, no study has been done which investigates tryptophan hydroxylase 2 gene polymorphism in anxiety and depressive disorders in Kashmiri population (India). Objectives: To study tryptophan hydroxylase 2 (TPH2) C 11993 A gene polymorphism in anxiety and depressive disorders. Method: Sixty patients of depression disorder, 60 patients of anxiety disorder and 40 unrelated healthy volunteers (control) were studied in a case control design. Polymorphism was determined using polymerase chain reaction (PCR) and agarose gel electrophoresis after digestion with HAP II enzyme. Genotypes and allele frequencies were compared using Chi-square tests, Fischer’s exact test, odds ratio, 95% confidence interval (C.I) and p-value of <0.05 was considered to be statistical significant. Results: The mean age ± SD of anxiety, depression and control group was 32.73±10.99, 32.20±10 and 29.75±10.12 respectively and the difference was found to be statistically non significant (p=0.349).The mean HAM-A (Hamilton rating scale for anxiety) score and HAM-D (Hamilton rating scale for depression) score was high in both groups (anxiety and depression) and found to be statistically significant (p=0.001).Depression group had AA genotype (55.2%) than control (37.5%) and was found to be statistically non significant (p=0.890).Comparison of allelic frequency revealed no association of A allele in anxiety group (76.67%) compared with control (75.5%) and was found to be statistically non significant (p= 0.866), OR 1.09 (0.56-2.11). Conclusion: TPH2C 11993 A gene was not found to be associated with major depressive disorder (MDD) and anxiety disorder in Kashmiri population. PMID:25121048

  19. Novel noncoding RNA from human Y distal heterochromatic block (Yq12) generates testis-specific chimeric CDC2L2

    PubMed Central

    Jehan, Zeenath; Vallinayagam, Sambandam; Tiwari, Shrish; Pradhan, Suman; Singh, Lalji; Suresh, Amritha; Reddy, Hemakumar M.; Ahuja, Y.R.; Jesudasan, Rachel A.

    2007-01-01

    The human Y chromosome, because it is enriched in repetitive DNA, has been very intractable to genetic and molecular analyses. There is no previous evidence for developmental stage- and testis-specific transcription from the male-specific region of the Y (MSY). Here, we present evidence for the first time for a developmental stage- and testis-specific transcription from MSY distal heterochromatic block. We isolated two novel RNAs, which localize to Yq12 in multiple copies, show testis-specific expression, and lack active X-homologs. Experimental evidence shows that one of the above Yq12 noncoding RNAs (ncRNAs) trans-splices with CDC2L2 mRNA from chromosome 1p36.3 locus to generate a testis-specific chimeric β sv13 isoform. This 67-nt 5′UTR provided by the Yq12 transcript contains within it a Y box protein-binding CCAAT motif, indicating translational regulation of the β sv13 isoform in testis. This is also the first report of trans-splicing between a Y chromosomal and an autosomal transcript. PMID:17095710

  20. Non-coding RNA derived from the region adjacent to the human HO-1 E2 enhancer selectively regulates HO-1 gene induction by modulating Pol II binding

    PubMed Central

    Maruyama, Atsushi; Mimura, Junsei; Itoh, Ken

    2014-01-01

    Recent studies have disclosed the function of enhancer RNAs (eRNAs), which are long non-coding RNAs transcribed from gene enhancer regions, in transcriptional regulation. However, it remains unclear whether eRNAs are involved in the regulation of human heme oxygenase-1 gene (HO-1) induction. Here, we report that multiple nuclear-enriched eRNAs are transcribed from the regions adjacent to two human HO-1 enhancers (i.e. the distal E2 and proximal E1 enhancers), and some of these eRNAs are induced by the oxidative stress-causing reagent diethyl maleate (DEM). We demonstrated that the expression of one forward direction (5′ to 3′) eRNA transcribed from the human HO-1 E2 enhancer region (named human HO-1enhancer RNA E2-3; hereafter called eRNA E2-3) was induced by DEM in an NRF2-dependent manner in HeLa cells. Conversely, knockdown of BACH1, a repressor of HO-1 transcription, further increased DEM-inducible eRNA E2-3 transcription as well as HO-1 expression. In addition, we showed that knockdown of eRNA E2-3 selectively down-regulated DEM-induced HO-1 expression. Furthermore, eRNA E2-3 knockdown attenuated DEM-induced Pol II binding to the promoter and E2 enhancer regions of HO-1 without affecting NRF2 recruitment to the E2 enhancer. These findings indicate that eRNAE2-3 is functional and is required for HO-1 induction. PMID:25404134

  1. Comparative analysis of A-to-I editing in human and non-human primate brains reveals conserved patterns and context-dependent regulation of RNA editing.

    PubMed

    O'Neil, Richard T; Wang, Xiaojing; Morabito, Michael V; Emeson, Ronald B

    2017-04-06

    A-to-I RNA editing is an important process for generating molecular diversity in the brain through modification of transcripts encoding several proteins important for neuronal signaling. We investigated the relationships between the extent of editing at multiple substrate transcripts (5HT2C, MGLUR4, CADPS, GLUR2, GLUR4, and GABRA3) in brain tissue obtained from adult humans and rhesus macaques. Several patterns emerged from these studies revealing conservation of editing across primate species. Additionally, variability in the human population allows us to make novel inferences about the co-regulation of editing at different editing sites and even across different brain regions.

  2. Type I human T cell leukemia virus tax protein transforms rat fibroblasts through the cyclic adenosine monophosphate response element binding protein/activating transcription factor pathway.

    PubMed Central

    Smith, M R; Greene, W C

    1991-01-01

    The Tax oncoprotein of the type I human T cell leukemia virus (HTLV-I) activates transcription of cellular and viral genes through at least two different transcription factor pathways. Tax activates transcription of the c-fos proto-oncogene by a mechanism that appears to involve members of the cAMP response element binding protein (CREB) and activating transcription factor (ATF) family of DNA-binding proteins. Tax also induces the nuclear expression of the NF-kappa B family of rel oncogene-related enhancer-binding proteins. We have investigated the potential role of these CREB/ATF and NF-kappa B/Rel transcription factors in Tax-mediated transformation by analyzing the oncogenic potential of Tax mutants that functionally segregate these two pathways of transactivation. Rat fibroblasts (Rat2) stably expressing either the wild-type Tax protein or a Tax mutant selectively deficient in the ability to induce NF-kappa B/Rel demonstrated marked changes in morphology and growth characteristics including the ability to form tumors in athymic mice. In contrast, Rat2 cells stably expressing a Tax mutant selectively deficient in the ability to activate transcription through CREB/ATF demonstrated no detectable changes in morphology or growth characteristics. These results suggest that transcriptional activation through the CREB/ATF pathway may play an important role in Tax-mediated cellular transformation. Images PMID:1832173

  3. Identification of a mouse synaptic glycoprotein gene in cultured neurons.

    PubMed

    Yu, Albert Cheung-Hoi; Sun, Chun Xiao; Li, Qiang; Liu, Hua Dong; Wang, Chen Ran; Zhao, Guo Ping; Jin, Meilei; Lau, Lok Ting; Fung, Yin-Wan Wendy; Liu, Shuang

    2005-10-01

    Neuronal differentiation and aging are known to involve many genes, which may also be differentially expressed during these developmental processes. From primary cultured cerebral cortical neurons, we have previously identified various differentially expressed gene transcripts from cultured cortical neurons using the technique of arbitrarily primed PCR (RAP-PCR). Among these transcripts, clone 0-2 was found to have high homology to rat and human synaptic glycoprotein. By in silico analysis using an EST database and the FACTURA software, the full-length sequence of 0-2 was assembled and the clone was named as mouse synaptic glycoprotein homolog 2 (mSC2). DNA sequencing revealed transcript size of mSC2 being smaller than the human and rat homologs. RT-PCR indicated that mSC2 was expressed differentially at various culture days. The mSC2 gene was located in various tissues with higher expression in brain, lung, and liver. Functions of mSC2 in neurons and other tissues remain elusive and will require more investigation.

  4. Identification of Viral MicroRNAs Expressed in Human Sacral Ganglia Latently Infected with Herpes Simplex Virus 2▿

    PubMed Central

    Umbach, Jennifer L.; Wang, Kening; Tang, Shuang; Krause, Philip R.; Mont, Erik K.; Cohen, Jeffrey I.; Cullen, Bryan R.

    2010-01-01

    Deep sequencing of small RNAs isolated from human sacral ganglia latently infected with herpes simplex virus 2 (HSV-2) was used to identify HSV-2 microRNAs (miRNAs) expressed during latent infection. This effort resulted in the identification of five distinct HSV-2 miRNA species, two of which, miR-H3/miR-I and miR-H4/miR-II, have been previously reported. Three novel HSV-2 miRNAs were also identified, and two of these, miR-H7 and miR-H9, are derived from the latency-associated transcript (LAT) and are located antisense to the viral transcript encoding transactivator ICP0. A third novel HSV-2 miRNA, miR-H10, is encoded within the unique long (UL) region of the genome, 3′ to the UL15 open reading frame, and is presumably excised from a novel, latent HSV-2 transcript distinct from LAT. PMID:19889786

  5. Identification of viral microRNAs expressed in human sacral ganglia latently infected with herpes simplex virus 2.

    PubMed

    Umbach, Jennifer L; Wang, Kening; Tang, Shuang; Krause, Philip R; Mont, Erik K; Cohen, Jeffrey I; Cullen, Bryan R

    2010-01-01

    Deep sequencing of small RNAs isolated from human sacral ganglia latently infected with herpes simplex virus 2 (HSV-2) was used to identify HSV-2 microRNAs (miRNAs) expressed during latent infection. This effort resulted in the identification of five distinct HSV-2 miRNA species, two of which, miR-H3/miR-I and miR-H4/miR-II, have been previously reported. Three novel HSV-2 miRNAs were also identified, and two of these, miR-H7 and miR-H9, are derived from the latency-associated transcript (LAT) and are located antisense to the viral transcript encoding transactivator ICP0. A third novel HSV-2 miRNA, miR-H10, is encoded within the unique long (U(L)) region of the genome, 3' to the U(L)15 open reading frame, and is presumably excised from a novel, latent HSV-2 transcript distinct from LAT.

  6. Influence of ethacrynic acid on glutathione S-transferase pi transcript and protein half-lives in human colon cancer cells.

    PubMed

    Shen, H; Ranganathan, S; Kuzmich, S; Tew, K D

    1995-10-12

    Ethacrynic acid (EA) is a plant phenolic acid that is both an inhibitor and an inducer of glutathione S-transferase (GST) activity. To determine contributory factors in the increased GST activity caused by EA treatment, human colon carcinoma HT29 cells were compared with a cloned EA-resistant population (HT6-8) maintained in medium containing 72 microM EA. Several factors are involved in the increased expression of GST pi in HT6-8. For example, nuclear run-on experiments showed an approximately 2-fold increase in the rate of transcription of GST pi. In addition, the half-life of GST pi transcript was increased from 4.1 (wild type, HT29, HT4-1) to 8.4 hr. The half-life of GST pi protein was 1-2 hr in HT4-1 cells versus 8-9 hr in HT6-8 cells. When either human ovarian carcinoma cells (SKOV3) or human prostatic carcinoma cells (DU145) were treated with EA, the half-life of the GST pi transcript was also increased. The transcript half-lives of another thiol-metabolism enzyme, gamma-glutamylcysteine synthetase (gamma-GCS), and a phase II detoxification enzyme, dihydrodiol dehydrogenase (DDH), were also increased in HT6-8, SKOV3 and DU145 cells treated with EA. However, the half-lives of transcripts from "housekeeping genes," such as glyceraldehyde 3-phosphate dehydrogenase (G3PDH), beta-actin and beta-tubulin, were not changed in these cell lines following EA. Apparently, a number of coordinated factors are involved in EA-enhanced expression of GST pi and other detoxification enzymes.

  7. Characterization of docosahexaenoic acid (DHA)-induced heme oxygenase-1 (HO-1) expression in human cancer cells: the importance of enhanced BTB and CNC homology 1 (Bach1) degradation.

    PubMed

    Wang, Shuai; Hannafon, Bethany N; Wolf, Roman F; Zhou, Jundong; Avery, Jori E; Wu, Jinchang; Lind, Stuart E; Ding, Wei-Qun

    2014-05-01

    The effect of docosahexaenoic acid (DHA) on heme oxygenase-1 (HO-1) expression in cancer cells has never been characterized. This study examines DHA-induced HO-1 expression in human cancer cell model systems. DHA enhanced HO-1 gene expression in a time- and concentration-dependent manner, with maximal induction at 21 h of treatment. This induction of HO-1 expression was confirmed in vivo using a xenograft nude mouse model fed a fish-oil-enriched diet. The increase in HO-1 gene transcription induced by DHA was significantly attenuated by the antioxidant N-acetyl cysteine, suggesting the involvement of oxidative stress. This was supported by direct measurement of lipid peroxide levels after DHA treatment. Using a human HO-1 gene promoter reporter construct, we identified two antioxidant response elements (AREs) that mediate the DHA-induced increase in HO-1 gene transcription. Knockdown of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) expression compromised the DHA-induced increase in HO-1 gene transcription, indicating the importance of the Nrf2 pathway in this event. However, the nuclear protein levels of Nrf2 remained unchanged upon DHA treatment. Further studies demonstrated that DHA reduces nuclear Bach1 protein expression by promoting its degradation and attenuates Bach1 binding to the AREs in the HO-1 gene promoter. In contrast, DHA enhanced Nrf2 binding to the AREs without affecting nuclear Nrf2 expression levels, indicating a new cellular mechanism that mediates DHA's induction of HO-1 gene transcription. To our knowledge, this is the first characterization of DHA-induced HO-1 expression in human malignant cells. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Cell-type-dependent activities of regulatory regions and E2 proteins derived from carcinogenic and non-carcinogenic human alphapapillomaviruses.

    PubMed

    Schenker, Astrid; Straub, Elke; Iftner, Thomas; Stubenrauch, Frank

    2013-06-01

    A large number of studies have revealed that persistent infections with certain human papillomavirus (HPV) types are necessary for the development of invasive cancer of the cervix. Recent studies have shown that not only do the major carcinogenic HPV types 16 and 18 encode E6 and E7 oncoproteins with immortalizing activity but also the very weakly or non-carcinogenic types 53, 66, 70 and 82. Currently, it is unknown whether transcriptional differences exist between these viruses that account for carcinogenicity in vivo. Therefore, we compared for the first time the activities of the upstream regulatory regions (URRs) that drive E6 and E7 expression derived from HPV16, -18, -31, -53, -66, -70 and -82 in the absence and presence of the viral E2 transcriptional regulator. URR activities in the absence of E2 varied widely and were further modulated by the cellular background. The co-expression of homologous E2 proteins resulted in repression of the URRs of only some HPV types and this varied with cell type. Activation by E2 proteins was less cell-type dependent but differed in an HPV-type-dependent manner. However, basal URR activity, repression of the URR by E2 and transcriptional activation by E2 did not correlate with HPV carcinogenicity in vivo. In summary, our data do not support the model that the transcriptional activity of human alphapapillomavirus types correlates with epidemiological risk classification.

  9. Epstein-Barr virus latency switch in human B-cells: a physico-chemical model.

    PubMed

    Werner, Maria; Ernberg, Ingemar; Zou, Jiezhi; Almqvist, Jenny; Aurell, Erik

    2007-08-31

    The Epstein-Barr virus is widespread in all human populations and is strongly associated with human disease, ranging from infectious mononucleosis to cancer. In infected cells the virus can adopt several different latency programs, affecting the cells' behaviour. Experimental results indicate that a specific genetic switch between viral latency programs, reprograms human B-cells between proliferative and resting states. Each of these two latency programs makes use of a different viral promoter, Cp and Qp, respectively. The hypothesis tested in this study is that this genetic switch is controlled by both human and viral transcription factors; Oct-2 and EBNA-1. We build a physico-chemical model to investigate quantitatively the dynamical properties of the promoter regulation and experimentally examine protein level variations between the two latency programs. Our experimental results display significant differences in EBNA-1 and Oct-2 levels between resting and proliferating programs. With the model we identify two stable latency programs, corresponding to a resting and proliferating cell. The two programs differ in robustness and transcriptional activity. The proliferating state is markedly more stable, with a very high transcriptional activity from its viral promoter. We predict the promoter activities to be mutually exclusive in the two different programs, and our relative promoter activities correlate well with experimental data. Transitions between programs can be induced, by affecting the protein levels of our transcription factors. Simulated time scales are in line with experimental results. We show that fundamental properties of the Epstein-Barr virus involvement in latent infection, with implications for tumor biology, can be modelled and understood mathematically. We conclude that EBNA-1 and Oct-2 regulation of Cp and Qp is sufficient to establish mutually exclusive expression patterns. Moreover, the modelled genetic control predict both mono- and bistable behavior and a considerable difference in transition dynamics, based on program stability and promoter activities. Both these phenomena we hope can be further investigated experimentally, to increase the understanding of this important switch. Our results also stress the importance of the little known regulation of human transcription factor Oct-2.

  10. Inhibition of human T cell leukemia virus type 2 replication by the suppressive action of class II transactivator and nuclear factor Y.

    PubMed

    Tosi, Giovanna; Pilotti, Elisabetta; Mortara, Lorenzo; De Lerma Barbaro, Andrea; Casoli, Claudio; Accolla, Roberto S

    2006-08-22

    The master regulator of MHC-II gene transcription, class II transactivator (CIITA), acts as a potent inhibitor of human T cell leukemia virus type 2 (HTLV-2) replication by blocking the activity of the viral Tax-2 transactivator. Here, we show that this inhibitory effect takes place at the nuclear level and maps to the N-terminal 1-321 region of CIITA, where we identified a minimal domain, from positions 64-144, that is strictly required to suppress Tax-2 function. Furthermore, we show that Tax-2 specifically cooperates with cAMP response element binding protein-binding protein (CBP) and p300, but not with p300/CBP-associated factor, to enhance transcription from the viral promoter. This finding represents a unique difference with respect to Tax-1, which uses all three coactivators to transactivate the human T cell leukemia virus type 1 LTR. Direct sequestering of CBP or p300 is not the primary mechanism by which CIITA causes suppression of Tax-2. Interestingly, we found that the transcription factor nuclear factor Y, which interacts with CIITA to increase transcription of MHC-II genes, exerts a negative regulatory action on the Tax-2-mediated HTLV-2 LTR transactivation. Thus, CIITA may inhibit Tax-2 function, at least in part, through nuclear factor Y. These findings demonstrate the dual defensive role of CIITA against pathogens: it increases the antigen-presenting function for viral determinants and suppresses HTLV-2 replication in infected cells.

  11. A common FADS2 promoter polymorphism increases promoter activity and facilitates binding of transcription factor ELK1

    PubMed Central

    Lattka, E.; Eggers, S.; Moeller, G.; Heim, K.; Weber, M.; Mehta, D.; Prokisch, H.; Illig, T.; Adamski, J.

    2010-01-01

    Fatty acid desaturases (FADS) play an important role in the formation of omega-6 and omega-3 highly unsaturated fatty acids (HUFAs). The composition of HUFAs in the human metabolome is important for membrane fluidity and for the modulation of essential physiological functions such as inflammation processes and brain development. Several recent studies reported significant associations of single nucleotide polymorphisms (SNPs) in the human FADS gene cluster with HUFA levels and composition. The presence of the minor allele correlated with a decrease of desaturase reaction products and an accumulation of substrates. We performed functional studies with two of the associated polymorphisms (rs3834458 and rs968567) and showed an influence of polymorphism rs968567 on FADS2 promoter activity by luciferase reporter gene assays. Electrophoretic mobility shift assays proved allele-dependent DNA-binding ability of at least two protein complexes to the region containing SNP rs968567. One of the proteins binding to this region in an allele-specific manner was shown to be the transcription factor ELK1 (a member of ETS domain transcription factor family). These results indicate that rs968567 influences FADS2 transcription and offer first insights into the modulation of complex regulation mechanisms of FADS2 gene transcription by SNPs. PMID:19546342

  12. A common FADS2 promoter polymorphism increases promoter activity and facilitates binding of transcription factor ELK1.

    PubMed

    Lattka, E; Eggers, S; Moeller, G; Heim, K; Weber, M; Mehta, D; Prokisch, H; Illig, T; Adamski, J

    2010-01-01

    Fatty acid desaturases (FADS) play an important role in the formation of omega-6 and omega-3 highly unsaturated fatty acids (HUFAs). The composition of HUFAs in the human metabolome is important for membrane fluidity and for the modulation of essential physiological functions such as inflammation processes and brain development. Several recent studies reported significant associations of single nucleotide polymorphisms (SNPs) in the human FADS gene cluster with HUFA levels and composition. The presence of the minor allele correlated with a decrease of desaturase reaction products and an accumulation of substrates. We performed functional studies with two of the associated polymorphisms (rs3834458 and rs968567) and showed an influence of polymorphism rs968567 on FADS2 promoter activity by luciferase reporter gene assays. Electrophoretic mobility shift assays proved allele-dependent DNA-binding ability of at least two protein complexes to the region containing SNP rs968567. One of the proteins binding to this region in an allele-specific manner was shown to be the transcription factor ELK1 (a member of ETS domain transcription factor family). These results indicate that rs968567 influences FADS2 transcription and offer first insights into the modulation of complex regulation mechanisms of FADS2 gene transcription by SNPs.

  13. Chemometric assessment of enhanced bioremediation of oil contaminated soils.

    PubMed

    Soleimani, Mohsen; Farhoudi, Majid; Christensen, Jan H

    2013-06-15

    Bioremediation is a promising technique for reclamation of oil polluted soils. In this study, six methods for enhancing bioremediation were tested on oil contaminated soils from three refinery areas in Iran (Isfahan, Arak, and Tehran). The methods included bacterial enrichment, planting, and addition of nitrogen and phosphorous, molasses, hydrogen peroxide, and a surfactant (Tween 80). Total petroleum hydrocarbon (TPH) concentrations and CHEMometric analysis of Selected Ion Chromatograms (SIC) termed CHEMSIC method of petroleum biomarkers including terpanes, regular, diaromatic and triaromatic steranes were used for determining the level and type of hydrocarbon contamination. The same methods were used to study oil weathering of 2 to 6 ring polycyclic aromatic compounds (PACs). Results demonstrated that bacterial enrichment and addition of nutrients were most efficient with 50% to 62% removal of TPH. Furthermore, the CHEMSIC results demonstrated that the bacterial enrichment was more efficient in degradation of n-alkanes and low molecular weight PACs as well as alkylated PACs (e.g. C₃-C₄ naphthalenes, C₂ phenanthrenes and C₂-C₃ dibenzothiophenes), while nutrient addition led to a larger relative removal of isoprenoids (e.g. norpristane, pristane and phytane). It is concluded that the CHEMSIC method is a valuable tool for assessing bioremediation efficiency. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Improvement of phytoremediation of an aged petroleum hydrocarbon-contaminated soil by Rhodococcus erythropolis CD 106 strain.

    PubMed

    Płociniczak, Tomasz; Fic, Ewa; Pacwa-Płociniczak, Magdalena; Pawlik, Małgorzata; Piotrowska-Seget, Zofia

    2017-07-03

    The aim of this study was to assess the impact of soil inoculation with the Rhodococcus erythropolis CD 106 strain on the effectiveness of the phytoremediation of an aged hydrocarbon-contaminated [approx. 1% total petroleum hydrocarbon (TPH)] soil using ryegrass (Lolium perenne). The introduction of CD 106 into the soil significantly increased the biomass of ryegrass and the removal of hydrocarbons in planted soil. The fresh weight of the shoots and roots of plants inoculated with CD 106 increased by 49% and 30%, respectively. After 210 days of the experiment, the concentration of TPH was reduced by 31.2%, whereas in the planted, non-inoculated soil, it was reduced by 16.8%. By contrast, the concentration of petroleum hydrocarbon decreased by 18.7% in non-planted soil bioaugmented with the CD 106 strain. The rifampicin-resistant CD 106 strain survived after inoculation into soil and was detected in the soil during the entire experimental period, but the number of CD 106 cells decreased constantly during the enhanced phytoremediation and bioaugmentation experiments. The plant growth-promoting and hydrocarbon-degrading properties of CD 106, which are connected with its long-term survival and limited impact on autochthonous microflora, make this strain a good candidate for improving the phytoremediation efficiency of soil contaminated with hydrocarbons.

  15. Investigation of lab-scale horizontal subsurface flow constructed wetlands treating industrial cork boiling wastewater.

    PubMed

    Gomes, Arlindo C; Silva, Lúcia; Albuquerque, António; Simões, Rogério; Stefanakis, Alexandros I

    2018-09-01

    The feasibility and treatment efficiency of horizontal subsurface flow constructed wetlands (HSFCW) was assessed for the first time for cork boiling wastewater (CBW) through laboratory experiments. CBW is known for its high content of phenolic compounds, complex composition of biorecalcitrant and toxic nature. Two lab-scale units, one planted with Phragmites australis (CWP) and one unplanted (CWC), were used to evaluate the removals of COD, BOD, total phenolic compounds (TPh) and decolourization over a 2.5-years monitoring period under Mediterranean climatic conditions. Seven organic and hydraulic loading rates ranging from 2.6 to 11.5 g COD/m 2 /d and 5.7-9.1 L/m 2 /d were tested under average hydraulic retention time (HRT) of 5 ± 1 days required due to the CWB limited biodegradability (i.e., BOD 5 /COD of 0.19). Average removals of the CWP exceeded those of the CWC and reached 74.6%, 91.7% and 69.1% for COD, BOD 5 and TPh, respectively, with respective mass removals rates up to 7.0, 1.7 and 0.5 (in g/m 2 /d). Decolourization was limited to 35%, since it mainly depends on physical processes rather than biodegradation. CBW concentration of nine phenolic compounds ranged from 1.2 to 38.4 mg/L (for the syringic and ellagic acids, respectively) in the raw CBW, with respective removals in the CWP unit ranging from 41.8 to 76.3%, higher than those in the control unit. Despite CBW high concentration of TPhs (average of 116.3 mg/L), the HSFCW reached organic load removals higher than those of conventional biological treatment methods. Copyright © 2018 Elsevier Ltd. All rights reserved.

  16. Human Oncoprotein MDM2 Up-regulates Expression of NF-κB2 Precursor p100 Conferring a Survival Advantage to Lung Cells

    PubMed Central

    Vaughan, Catherine; Mohanraj, Lathika; Singh, Shilpa; Dumur, Catherine I.; Ramamoorthy, Mahesh; Garrett, Carleton T.; Windle, Brad; Yeudall, W. Andrew; Deb, Sumitra

    2011-01-01

    The current model predicts that MDM2 is primarily overexpressed in cancers with wild-type (WT) p53 and contributes to oncogenesis by degrading p53. Following a correlated expression of MDM2 and NF-κB2 transcripts in human lung tumors, we have identified a novel transactivation function of MDM2. Here, we report that in human lung tumors, overexpression of MDM2 was found in approximately 30% of cases irrespective of their p53 status, and expression of MDM2 and NF-κB2 transcripts showed a highly significant statistical correlation in tumors with WT p53. We investigated the significance of this correlated expression in terms of mechanism and biological function. Increase in MDM2 expression from its own promoter in transgenic mice remarkably enhanced expression of NF-κB2 compared with its non-transgenic littermates. Knockdown or elimination of endogenous MDM2 expression in cultured non-transformed or lung tumor cells drastically reduced expression of NF-κB2 transcripts, suggesting a normal physiological role of MDM2 in regulating NF-κB2 transcription. MDM2 could up-regulate expression of NF-κB2 transcripts when its p53-interaction domain was blocked with Nutlin-3, indicating that the MDM2-p53 interaction is dispensable for up-regulation of NF-κB2 expression. Consistently, analysis of functional domains of MDM2 indicated that although the p53-interaction domain of MDM2 contributes to the up-regulation of the NFκB2 promoter, MDM2 does not require direct interactions with p53 for this function. Accordingly, MDM2 overexpression in non-transformed or lung cancer cells devoid of p53 also generated a significant increase in the expression of NF-κB2 transcript and its targets CXCL-1 and CXCL-10, whereas elimination of MDM2 expression had the opposite effects. MDM2-mediated increase in p100/NF-κB2 expression reduced cell death mediated by paclitaxel. Furthermore, knockdown of NF-κB2 expression retarded cell proliferation. Based on these data, we propose that MDM2-mediated NF-κB2 up-regulation is a combined effect of p53-dependent and independent mechanisms and that it confers a survival advantage to lung cancer cells. PMID:22701761

  17. Dampening DNA binding: a common mechanism of transcriptional repression for both ncRNAs and protein domains.

    PubMed

    Goodrich, James A; Kugel, Jennifer F

    2010-01-01

    With eukaryotic non-coding RNAs (ncRNAs) now established as critical regulators of cellular transcription, the true diversity with which they can elicit biological effects is beginning to be appreciated. Two ncRNAs, mouse B2 RNA and human Alu RNA, have been found to repress mRNA transcription in response to heat shock. They do so by binding directly to RNA polymerase II, assembling into complexes on promoter DNA, and disrupting contacts between the polymerase and the DNA. Such a mechanism of repression had not previously been observed for a eukaryotic ncRNA; however, there are examples of eukaryotic protein domains that repress transcription by blocking essential protein-DNA interactions. Comparing the mechanism of transcriptional repression utilized by these protein domains to that used by B2 and Alu RNAs raises intriguing questions regarding transcriptional control, and how B2 and Alu RNAs might themselves be regulated.

  18. Studies on crude oil removal from pebbles by the application of biodiesel.

    PubMed

    Xia, Wen-xiang; Xia, Yan; Li, Jin-cheng; Zhang, Dan-feng; Zhou, Qing; Wang, Xin-ping

    2015-02-15

    Oil residues along shorelines are hard to remove after an oil spill. The effect of biodiesel to eliminate crude oil from pebbles alone and in combination with petroleum degrading bacteria was investigated in simulated systems. Adding biodiesel made oil detach from pebbles and formed oil-biodiesel mixtures, most of which remained on top of seawater. The total petroleum hydrocarbon (TPH) removal efficiency increased with biodiesel quantities but the magnitude of augment decreased gradually. When used with petroleum degrading bacteria, the addition of biodiesel (BD), nutrients (NUT) and BD+NUT increased the dehydrogenase activity and decreased the biodegradation half lives. When BD and NUT were replenished at the same time, the TPH removal efficiency was 7.4% higher compared to the total improvement of efficiency when BD and NUT was added separately, indicating an additive effect of biodiesel and nutrients on oil biodegradation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. In vivo expression of Salmonella enterica serotype Typhi genes in the blood of patients with typhoid fever in Bangladesh.

    PubMed

    Sheikh, Alaullah; Charles, Richelle C; Sharmeen, Nusrat; Rollins, Sean M; Harris, Jason B; Bhuiyan, Md Saruar; Arifuzzaman, Mohammad; Khanam, Farhana; Bukka, Archana; Kalsy, Anuj; Porwollik, Steffen; Leung, Daniel T; Brooks, W Abdullah; LaRocque, Regina C; Hohmann, Elizabeth L; Cravioto, Alejandro; Logvinenko, Tanya; Calderwood, Stephen B; McClelland, Michael; Graham, James E; Qadri, Firdausi; Ryan, Edward T

    2011-12-01

    Salmonella enterica serotype Typhi is the cause of typhoid fever. It is a human-restricted pathogen, and few data exist on S. Typhi gene expression in humans. We applied an RNA capture and amplification technique, Selective Capture of Transcribed Sequences (SCOTS), and microarray hybridization to identify S. Typhi transcripts expressed in the blood of five humans infected with S. Typhi in Bangladesh. In total, we detected the expression of mRNAs for 2,046 S. Typhi genes (44% of the S. Typhi genome) in human blood; expression of 912 genes was detected in all 5 patients, and expression of 1,100 genes was detected in 4 or more patients. Identified transcripts were associated with the virulence-associated PhoP regulon, Salmonella pathogenicity islands, the use of alternative carbon and energy sources, synthesis and transport of iron, thiamine, and biotin, and resistance to antimicrobial peptides and oxidative stress. The most highly represented group were genes currently annotated as encoding proteins designated as hypothetical, unknown, or unclassified. Of the 2,046 detected transcripts, 1,320 (29% of the S. Typhi genome) had significantly different levels of detection in human blood compared to in vitro cultures; detection of 141 transcripts was significantly different in all 5 patients, and detection of 331 transcripts varied in at least 4 patients. These mRNAs encode proteins of unknown function, those involved in energy metabolism, transport and binding, cell envelope, cellular processes, and pathogenesis. We confirmed increased expression of a subset of identified mRNAs by quantitative-PCR. We report the first characterization of bacterial transcriptional profiles in the blood of patients with typhoid fever. S. Typhi is an important global pathogen whose restricted host range has greatly inhibited laboratory studies. Our results suggest that S. Typhi uses a largely uncharacterized genetic repertoire to survive within cells and utilize alternate energy sources during infection.

  20. Pigment Production Analysis in Human Melanoma Cells.

    PubMed

    Hopkin, Amelia Soto; Paterson, Elyse K; Ruiz, Rolando; Ganesan, Anand K

    2016-05-25

    The human epidermal melanocyte is a highly specialized pigmented cell that serves to protect the epidermis from ultraviolet (UV) damage through the production of melanin, or melanogenesis. Misregulation in melanogenesis leading to either hyper- or hypo-pigmentation is found in human diseases such as malasma and vitiligo. Current therapies for these diseases are largely unsuccessful and the need for new therapies is necessary. In order to identify genes and or compounds that can alter melanogenesis, methods are required that can detect changes in pigment production as well as expression of key melanogenesis transcription factors and enzymes. Here we describe methods to detect changes in melanogenesis in a human melanoma cell line, MNT-1, by (1) analyzing pigment production by measuring the absorbance of melanin present by spectrophotometry, (2) analyzing transcript expression of potent regulators of melanogenesis by qunatitative reverse-transcription (RT)PCR and (3) analyzing protein expression of potent regulators of melanogenesis by Western blot (WB).

  1. Preclinical evaluation of transcriptional targeting strategy for human hepatocellular carcinoma in an orthotopic xenograft mouse model.

    PubMed

    Sia, Kian Chuan; Huynh, Hung; Chung, Alexander Yaw Fui; Ooi, London Lucien Peng Jin; Lim, Kiat Hon; Hui, Kam Man; Lam, Paula Yeng Po

    2013-08-01

    Gene regulation of many key cell-cycle players in S-, G(2) phase, and mitosis results from transcriptional repression in their respective promoter regions during the G(0) and G(1) phases of cell cycle. Within these promoter regions are phylogenetically conserved sequences known as the cell-cycle-dependent element (CDE) and cell-cycle genes homology regions (CHR) sites. Thus, we hypothesize that transcriptional regulation of cell-cycle regulation via the CDE/CHR region together with liver-specific apolipoprotein E (apoE)-hAAT promoter could bring about a selective transgene expression in proliferating human hepatocellular carcinoma. We show that the newly generated vector AH-6CC-L2C could mediate hepatocyte-targeted luciferase gene expression in tumor cells and freshly isolated short-term hepatocellular carcinoma cultures from patient biopsy. In contrast, normal murine and human hepatocytes infected with AH-6CC-L2C expressed minimal or low luciferase activities. In the presence of prodrug 5-fluorocytosine (5-FC), AH-6CC-L2C effectively suppressed the growth of orthotopic hepatocellular carcinoma patient-derived xenograft mouse model via the expression of yeast cytosine deaminase (yCD) that converts 5-FC to anticancer metabolite 5-fluoruracil. More importantly, we show that combination treatment of AH-6CC-L2C with an EZH2 inhibitor, DZNep, that targets EpCAM-positive hepatocellular carcinoma, can bring about a greater therapeutic efficacy compared with a single treatment of virus or inhibitor. Our study showed that targeting proliferating human hepatocellular carcinoma cells through the transcriptional control of therapeutic gene could represent a feasible approach against hepatocellular carcinoma.

  2. Cloning and Biochemical Characterization of TAF-172, a Human Homolog of Yeast Mot1

    PubMed Central

    Chicca, John J.; Auble, David T.; Pugh, B. Franklin

    1998-01-01

    The TATA binding protein (TBP) is a central component of the eukaryotic transcriptional machinery and is the target of positive and negative transcriptional regulators. Here we describe the cloning and biochemical characterization of an abundant human TBP-associated factor (TAF-172) which is homologous to the yeast Mot1 protein and a member of the larger Snf2/Swi2 family of DNA-targeted ATPases. Like Mot1, TAF-172 binds to the conserved core of TBP and uses the energy of ATP hydrolysis to dissociate TBP from DNA (ADI activity). Interestingly, ATP also causes TAF-172 to dissociate from TBP, which has not been previously observed with Mot1. Unlike Mot1, TAF-172 requires both TBP and DNA for maximal (∼100-fold) ATPase activation. TAF-172 inhibits TBP-driven RNA polymerase II and III transcription but does not appear to affect transcription driven by TBP-TAF complexes. As it does with Mot1, TFIIA reverses TAF-172-mediated repression of TBP. Together, these findings suggest that human TAF-172 is the functional homolog of yeast Mot1 and uses the energy of ATP hydrolysis to remove TBP (but apparently not TBP-TAF complexes) from DNA. PMID:9488487

  3. Effects Of Oil Spillage On Vegetation, Land And Water(Odu-Gboro, Sagamu,Ogun State, South-Western, Nigeria) Using Remote Sensing And Gis Techniques

    NASA Astrophysics Data System (ADS)

    Oseni, O.

    2017-12-01

    This paper explores the impacts of oil spill on the physical environment (soil, water and plants) with particular attention paid to the NNPC/PPMC pipeline system. It focuses on the environmental impacts of oil pollution in Nigeria, and discusses the increasing environmental contradictions of the area, and its influence on global warming. The discovery of oil in Nigeria in 1956, the country has been suffering the negative environmental consequences of oil exploration and exploitation. Between 1976 and 1996 a total of 4647 incidents resulted in the spill of approximately 2,369,470 barrels of oil into the environment. In addition, between 1997 and 2001, Nigeria also recorded a total number of 2,097 oil spill incidents. The study traces the effects of the oil spillage on the environment in order to determine whether oil spill is a major factor responsible for environmental pollution. By the use of remotely sensed data and other ancillary data, it identified the major causes of oil spill in the region; the presence of total petroleum hydrocarbon (TPH) in the environment, and it also determined the environmental impacts on land and water. Personal interview, field observation and laboratory analysis of soil and water were used. Gas chromatography was used to determine the TPH concentration in soil extract and water extracts. Liquid-liquid extraction method was used for water and spectro-radiometer which is a very efficient process commonly used to determine spectral signature of various soil, water and plant samples obtained from the study area.Values of analyzed soil and water samples in the oil impacted area were compared to the control area (region with no spill). Based largely onthe GISanalysis, the findings showed that the main cause of oil spill is vandalism along the pipeline right of way; Vandalism which is an act of sabotage had the highest percentage compared to equipment failure, accident from oil tankers and accidental discharge during pipeline repairs.TPH were present at the site with soil samples having the high values, and the environmental impact onsoil and water is due to poor resource management and control. Satellite imagery (Ikonos and Landsat series)helped in monitoring oil spill by providing the spill position.

  4. A comprehensive transcript index of the human genome generated using microarrays and computational approaches

    PubMed Central

    Schadt, Eric E; Edwards, Stephen W; GuhaThakurta, Debraj; Holder, Dan; Ying, Lisa; Svetnik, Vladimir; Leonardson, Amy; Hart, Kyle W; Russell, Archie; Li, Guoya; Cavet, Guy; Castle, John; McDonagh, Paul; Kan, Zhengyan; Chen, Ronghua; Kasarskis, Andrew; Margarint, Mihai; Caceres, Ramon M; Johnson, Jason M; Armour, Christopher D; Garrett-Engele, Philip W; Tsinoremas, Nicholas F; Shoemaker, Daniel D

    2004-01-01

    Background Computational and microarray-based experimental approaches were used to generate a comprehensive transcript index for the human genome. Oligonucleotide probes designed from approximately 50,000 known and predicted transcript sequences from the human genome were used to survey transcription from a diverse set of 60 tissues and cell lines using ink-jet microarrays. Further, expression activity over at least six conditions was more generally assessed using genomic tiling arrays consisting of probes tiled through a repeat-masked version of the genomic sequence making up chromosomes 20 and 22. Results The combination of microarray data with extensive genome annotations resulted in a set of 28,456 experimentally supported transcripts. This set of high-confidence transcripts represents the first experimentally driven annotation of the human genome. In addition, the results from genomic tiling suggest that a large amount of transcription exists outside of annotated regions of the genome and serves as an example of how this activity could be measured on a genome-wide scale. Conclusions These data represent one of the most comprehensive assessments of transcriptional activity in the human genome and provide an atlas of human gene expression over a unique set of gene predictions. Before the annotation of the human genome is considered complete, however, the previously unannotated transcriptional activity throughout the genome must be fully characterized. PMID:15461792

  5. Oil refinery wastewater treatment using coupled electrocoagulation and fixed film biological processes

    NASA Astrophysics Data System (ADS)

    Pérez, Laura S.; Rodriguez, Oscar M.; Reyna, Silvia; Sánchez-Salas, José Luis; Lozada, J. Daniel; Quiroz, Marco A.; Bandala, Erick R.

    2016-02-01

    Oil refinery wastewater was treated using a coupled treatment process including electrocoagulation (EC) and a fixed film aerobic bioreactor. Different variables were tested to identify the best conditions using this procedure. After EC, the effluent was treated in an aerobic biofilter. EC was capable to remove over 88% of the overall chemical oxygen demand (COD) in the wastewater under the best working conditions (6.5 V, 0.1 M NaCl, 4 electrodes without initial pH adjustment) with total petroleum hydrocarbon (TPH) removal slightly higher than 80%. Aluminum release from the electrodes to the wastewater was found an important factor for the EC efficiency and closely related with several operational factors. Application of EC allowed to increase the biodegradability of the sample from 0.015, rated as non-biodegradable, up to 0.5 widely considered as biodegradable. The effluent was further treated using an aerobic biofilter inoculated with a bacterial consortium including gram positive and gram negative strains and tested for COD and TPH removal from the EC treated effluent during 30 days. Cell count showed the typical bacteria growth starting at day three and increasing up to a maximum after eight days. After day eight, cell growth showed a plateau which agreed with the highest decrease on contaminant concentration. Final TPHs concentration was found about 600 mgL-1 after 30 days whereas COD concentration after biological treatment was as low as 933 mgL-1. The coupled EC-aerobic biofilter was capable to remove up to 98% of the total TPH amount and over 95% of the COD load in the oil refinery wastewater.

  6. A novel luciferase knock-in reporter system for studying transcriptional regulation of the human Sox2 gene.

    PubMed

    Xiao, Dan; Zhang, Weifeng; Li, Yan; Liu, Kuan; Zhao, Junli; Sun, Xiaohong; Shan, Linlin; Mao, Qinwen; Xia, Haibin

    2016-02-10

    Sox2 is an important transcriptional factor that has multiple functions in stem cell maintenance and tumorigenesis. To investigate the transcriptional regulation of the Sox2 gene, a luciferase knock-in reporter system was established in HEK293 cells by placing the luciferase gene in the genome under the control of the Sox2 gene promoter using a transcription activator-like effector nuclease (TALEN)-mediated genome editing technique. PCR and Southern blot results confirmed the site-specific integration of a single copy of the exogenous luciferase gene into the genome. To prove the reliability and sensitivity of this novel luciferase knock-in system, a CRISPR/Cas transcription activation system for the Sox2 gene was constructed and applied to the knock-in system. The results indicated that luciferase activity was directly correlated with the activity of the Sox2 endogenous promoter. This novel system will be a useful tool to study the transcriptional regulation of Sox2, and has great potential in medical and industrial applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Involvement of atypical transcription factor E2F8 in the polyploidization during mouse and human decidualization.

    PubMed

    Qi, Qian-Rong; Zhao, Xu-Yu; Zuo, Ru-Juan; Wang, Tong-Song; Gu, Xiao-Wei; Liu, Ji-Long; Yang, Zeng-Ming

    2015-01-01

    Polyploid decidual cells are specifically differentiated cells during mouse uterine decidualization. However, little is known about the regulatory mechanism and physiological significance of polyploidization in pregnancy. Here we report a novel role of E2F8 in the polyploidization of decidual cells in mice. E2F8 is highly expressed in decidual cells and regulated by progesterone through HB-EGF/EGFR/ERK/STAT3 signaling pathway. E2F8 transcriptionally suppresses CDK1, thus triggering the polyploidization of decidual cells. E2F8-mediated polyploidization is a response to stresses which are accompanied by decidualization. Interestingly, polyploidization is not detected during human decidualization with the down-regulation of E2F8, indicating differential expression of E2F8 may lead to the difference of decidual cell polyploidization between mice and humans.

  8. Involvement of atypical transcription factor E2F8 in the polyploidization during mouse and human decidualization

    PubMed Central

    Qi, Qian-Rong; Zhao, Xu-Yu; Zuo, Ru-Juan; Wang, Tong-Song; Gu, Xiao-Wei; Liu, Ji-Long; Yang, Zeng-Ming

    2015-01-01

    Polyploid decidual cells are specifically differentiated cells during mouse uterine decidualization. However, little is known about the regulatory mechanism and physiological significance of polyploidization in pregnancy. Here we report a novel role of E2F8 in the polyploidization of decidual cells in mice. E2F8 is highly expressed in decidual cells and regulated by progesterone through HB-EGF/EGFR/ERK/STAT3 signaling pathway. E2F8 transcriptionally suppresses CDK1, thus triggering the polyploidization of decidual cells. E2F8-mediated polyploidization is a response to stresses which are accompanied by decidualization. Interestingly, polyploidization is not detected during human decidualization with the down-regulation of E2F8, indicating differential expression of E2F8 may lead to the difference of decidual cell polyploidization between mice and humans. PMID:25892397

  9. Differential splicing of human androgen receptor pre-mRNA in X-linked reifenstein syndrome, because of a deletion involving a putative branch site

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ris-Stalpers, C.; Verleun-Mooijman, M.C.T.; Blaeij, T.J.P. de

    1994-04-01

    The analysis of the androgen receptor (AR) gene, mRNA, and protein in a subject with X-linked Reifenstein syndrome (partial androgen insensitivity) is reported. The presence of two mature AR transcripts in genital skin fibroblasts of the patient is established, and, by reverse transcriptase-PCR and RNase transcription analysis, the wild-type transcript and a transcript in which exon 3 sequences are absent without disruption of the translational reading frame are identified. Sequencing and hybridization analysis show a deletion of >6 kb in intron 2 of the human AR gene, starting 18 bp upstream of exon 3. The deletion includes the putative branch-pointmore » sequence (BPS) but not the acceptor splice site on the intron 2/exon 3 boundary. The deletion of the putative intron 2 BPS results in 90% inhibition of wild-type splicing. The mutant transcript encodes an AR protein lacking the second zinc finger of the DNA-binding domain. Western/immunoblotting analysis is used to show that the mutant AR protein is expressed in genital skin fibroblasts of the patient. The residual 10% wild-type transcript can be the result of the use of a cryptic BPS located 63 bp upstream of the intron 2/exon 3 boundary of the mutant AR gene. The mutated AR protein has no transcription-activating potential and does not influence the transactivating properties of the wild-type AR, as tested in cotransfection studies. It is concluded that the partial androgen-insensitivity syndrome of this patient is the consequence of the limited amount of wild-type AR protein expressed in androgen target cells, resulting from the deletion of the intron 2 putative BPS. 42 refs., 6 figs., 1 tab.« less

  10. 78 FR 34421 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-07

    ... affiliated with a single TPH organization proves attractive to market participants on other exchanges, such... Commission summarily may temporarily suspend such rule change if it appears to the Commission that such...

  11. Inactivation of the FoxO3a transcription factor is associated with the production of reactive oxygen species during protein kinase CK2 downregulation-mediated senescence in human colon cancer and breast cancer cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Park, Seong-Yeol; Bae, Young-Seuk, E-mail: ysbae@knu.ac.kr

    We previously showed that protein kinase CK2 downregulation mediates senescence through the reactive oxygen species (ROS)–p53–p21{sup Cip1/WAF1} pathway in various human cells. In the present study, we investigated whether the FoxO3a transcription factor is associated with ROS production during CK2 downregulation-induced senescence in human colon cancer HCT116 and breast cancer MCF-7 cells. FoxO3a overexpression suppressed ROS production and p53 stabilization induced by a CK2α knockdown. CK2α downregulation induced nuclear export of FoxO3a through stimulation of AKT-mediated phosphorylation of FoxO3a and decreased transcription of its target genes (Cu/ZnSOD, MnSOD, and catalase). In contrast, CK2α overexpression inhibited AKT-mediated FoxO3a phosphorylation. This resulted inmore » nuclear accumulation of FoxO3a, and elevated expression of its target genes. Therefore, these data indicate for the first time that CK2 downregulation stimulates ROS generation by inhibiting FoxO3a during premature senescence in human colon and breast cancer cells. - Highlights: • FoxO3a overexpression inhibited ROS production mediated by CK2α knockdown. • CK2α downregulation induced nuclear export of FoxO3a via AKT activation. • CK2α downregulation reduced transcription of FoxO3a target genes including SOD. • CK2α upregulation elevated nuclear import and target gene expression of FoxO3a. • This study indicates that CK2 can modulate the intracellular ROS level via FoxO3a.« less

  12. Growth factors FGF8 and FGF2 and their receptor FGFR1, transcriptional factors Msx-1 and MSX-2, and apoptotic factors p19 and RIP5 participate in the early human limb development.

    PubMed

    Becic, Tina; Kero, Darko; Vukojevic, Katarina; Mardesic, Snjezana; Saraga-Babic, Mirna

    2018-04-01

    The expression pattern of fibroblast growth factors FGF8 and FGF2 and their receptor FGFR1, transcription factors MSX-1 and MSX-2, as well as cell proliferation (Ki-67) and cell death associated caspase-3, p19 and RIP5 factors were analyzed in histological sections of eight 4th-9th-weeks developing human limbs by immunohistochemistry and semi-thin sectioning. Increasing expression of all analyzed factors (except FGF8) characterized both the multilayered human apical ectodermal ridge (AER), sub-ridge mesenchyme (progress zone) and chondrocytes in developing human limbs. While cytoplasmic co-expression of MSX-1 and MSX-2 was observed in both limb epithelium and mesenchyme, p19 displayed strong cytoplasmic expression in non-proliferating cells. Nuclear expression of Ki-67 proliferating cells, and partly of MSX-1 and MSX-2 was detected in the whole limb primordium. Strong expression of factors p19 and RIP5, both in the AER and mesenchyme of human developing limbs indicates their possible involvement in control of cell senescence and cell death. In contrast to animal studies, expression of FGFR1 in the surface ectoderm and p19 in the whole limb primordium might reflect interspecies differences in limb morphology. Expression of FGF2 and downstream RIP5 gene, and transcription factors Msx-1 and MSX-2 did not show human-specific changes in expression pattern. Based on their spatio-temporal expression during human limb development, our study indicates role of FGFs and Msx genes in stimulation of cell proliferation, limb outgrowth, digit elongation and separation, and additionally MSX-2 in control of vasculogenesis. The cascade of orchestrated gene expressions, including the analyzed developmental factors, jointly contribute to the complex human limb development. Copyright © 2018 Elsevier GmbH. All rights reserved.

  13. Managing Exposure to Benzene and Total Petroleum Hydrocarbons at Two Oil Refineries 1977-2014.

    PubMed

    Tuomi, Tapani; Veijalainen, Henna; Santonen, Tiina

    2018-01-24

    Air concentrations of and inhalation exposure to total petroleum hydrocarbons (TPH) and benzene was monitored separately at two oil refineries from 1977 to 2014. Prevention policies and control measures that may explain changes were surveyed. The aim was to evaluate how the application of of Occupational Health and Safety Assessment Series OHSAS 18001.04 principles as well as Environmental protection Agency EPA and European Oil Company Organisation for Environment, Health and Safety CONCAWE practices have influenced air concentrations. Benzene air concentrations declined in 11 of 17 units, six of which were associated with declining exposures. Benzene air concentrations declined across all units on average by 46%. This amounts to an average yearly decline of 1.7%. TPH air concentrations declined in 10 of 17 units, seven of which were associated with declining exposures. The average decline in TPH air concentrations was 49%, corresponding to 1.3% per year. As a result, average working day exposure in 10 of 17 units have declined significantly and today, benzene and TPH exposure in most units are well below 10% of the current Occupational Exposure Limit (OEL 8h :s). A decline in air concentrations have coincided with consistent implementation of control measures. Such measures include on-line monitoring of leaks; benzene recovery; floating container roofs; improved valves and seals; hermetic pumps; recovery of loading gases and instalment of torches in terminals; cutback in coke combustion; a new production line spanning directly from the dock to aromatics production; and recovery of loading gases in the doc. Other tools in exposure management include personal leak monitors, on-line measurements, monitoring campaigns, risk assessment, and availability and user training of protective equipment. However, improvements are still needed. Hydrocarbon or benzene air concentrations have not declined in 8 of 17 units, in some of which concentrations exceed 10% of the relevant OEL 8h value. In addition, for benzene even 10% of the current OEL, 0.1 ppm, might still possess a risk. With this in mind, methods to estimate exposure at the refineries need to be improved to enable measuring benzene concentrations <0.1 ppm. Shut downs of the refinery have been associated with peaks in exposure concentrations. Consequently, effort should be placed on safe working methods pertaining to shutdowns. Also, the connection and detachment of hoses continues to be problematic from the point of view of controlling exposure.

  14. ETO2-GLIS2 Hijacks Transcriptional Complexes to Drive Cellular Identity and Self-Renewal in Pediatric Acute Megakaryoblastic Leukemia.

    PubMed

    Thirant, Cécile; Ignacimouttou, Cathy; Lopez, Cécile K; Diop, M'Boyba; Le Mouël, Lou; Thiollier, Clarisse; Siret, Aurélie; Dessen, Phillipe; Aid, Zakia; Rivière, Julie; Rameau, Philippe; Lefebvre, Céline; Khaled, Mehdi; Leverger, Guy; Ballerini, Paola; Petit, Arnaud; Raslova, Hana; Carmichael, Catherine L; Kile, Benjamin T; Soler, Eric; Crispino, John D; Wichmann, Christian; Pflumio, Françoise; Schwaller, Jürg; Vainchenker, William; Lobry, Camille; Droin, Nathalie; Bernard, Olivier A; Malinge, Sébastien; Mercher, Thomas

    2017-03-13

    Chimeric transcription factors are a hallmark of human leukemia, but the molecular mechanisms by which they block differentiation and promote aberrant self-renewal remain unclear. Here, we demonstrate that the ETO2-GLIS2 fusion oncoprotein, which is found in aggressive acute megakaryoblastic leukemia, confers megakaryocytic identity via the GLIS2 moiety while both ETO2 and GLIS2 domains are required to drive increased self-renewal properties. ETO2-GLIS2 directly binds DNA to control transcription of associated genes by upregulation of expression and interaction with the ETS-related ERG protein at enhancer elements. Importantly, specific interference with ETO2-GLIS2 oligomerization reverses the transcriptional activation at enhancers and promotes megakaryocytic differentiation, providing a relevant interface to target in this poor-prognosis pediatric leukemia. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Assessment of soil-gas and groundwater contamination at the Gibson Road landfill, Fort Gordon, Georgia, 2011

    USGS Publications Warehouse

    Falls, W. Fred; Caldwell, Andral W.; Guimaraes, Wladmir G.; Ratliff, W. Hagan; Wellborn, John B.; Landmeyer, James E.

    2012-01-01

    Soil-gas and groundwater assessments were conducted at the Gibson Road landfill in 201 to provide screening-level environmental contamination data to supplement the data collected during previous environmental studies at the landfill. Passive samplers were used in both assessments to detect volatile and semivolatile organic compounds and polycyclic aromatic hydrocarbons in soil gas and groundwater. A total of 56 passive samplers were deployed in the soil in late July and early August for the soil-gas assessment. Total petroleum hydrocarbons (TPH) were detected at masses greater than the method detection level of 0.02 microgram in all samplers and masses greater than 2.0 micrograms in 13 samplers. Three samplers located between the landfill and a nearby wetland had TPH masses greater than 20 micrograms. Diesel was detected in 28 of the 56 soil-gas samplers. Undecane, tridecane, and pentadecane were detected, but undecane was the most common diesel compound with 23 detections. Only five detections exceeded a combined diesel mass of 0.10 microgram, including the highest mass of 0.27 microgram near the wetland. Toluene was detected in only five passive samplers, including masses of 0.65 microgram near the wetland and 0.85 microgram on the southwestern side of the landfill. The only other gasoline-related compound detected was octane in two samplers. Naphthalene was detected in two samplers in the gully near the landfill and two samplers along the southwestern side of the landfill, but had masses less than or equal to 0.02 microgram. Six samplers located southeast of the landfill had detections of chlorinated compounds, including one perchloroethene detections (0.04 microgram) and five chloroform detections (0.05 to0.08 microgram). Passive samplers were deployed and recovered on August 8, 2011, in nine monitoring wells along the southwestern, southeastern and northeastern sides of the landfill and down gradient from the eastern corner of the landfill. Six of the nine samplers had TPH concentrations greater than 100 micrograms per liter. TPH concentrations declined from 320 micrograms per liter in a sampler near the landfill to 18 micrograms in a sampler near the wetland. Five of the samplers had detections of one or more diesel compounds but detections of individual diesel compounds had concentrations below a method detection level of 0.01 microgram per liter. Benzene was detected in three samplers and exceeded the national primary drinking-water standard of 5 micrograms per liter set by the U.S. Environmental Protection Agency. The concentrations of benzene, and therefore BTEX, were 6.1 micrograms per liter in the sampler near the eastern corner of the landfill, 27 micrograms per liter in the sampler near the wetland, and 37 micrograms per liter in the sampler at the southern corner of the landfill. Nonfuel-related compounds were detected in the four wells that are aligned between the eastern corner of the landfill and the wetland. The sampler deployed nearest the eastern corner of the landfill had the greatest number of detected organic compounds and had the only detections of two trimethylbenzene compounds, naphthalene, 2-methyl naphthalene, and 1,4-dichlorobenzene. The two up gradient samplers had the greatest number of chlorinated compounds with five compounds each, compared to detections of four compounds and one compound in the two down gradient samplers. All four samplers had detections of 1,1-dichloroethane which ranged from 42 to 1,300 micrograms per liter. Other detections of chlorinated compounds included trichloroethene, perchloroethene, cis-1,2-dichloroethene, 1,1,1-trichloroethane and chloroform.

  16. Bio-treatment of oily sludge: the contribution of amendment material to the content of target contaminants, and the biodegradation dynamics.

    PubMed

    Kriipsalu, Mait; Marques, Marcia; Nammari, Diauddin R; Hogland, William

    2007-09-30

    The objective was to investigate the aerobic biodegradation of oily sludge generated by a flotation-flocculation unit (FFU) of an oil refinery wastewater treatment plant. Four 1m(3) pilot bioreactors with controlled air-flow were filled with FFU sludge mixed with one of the following amendments: sand (M1); matured oil compost (M2); kitchen waste compost (M3) and shredded waste wood (M4). The variables monitored were: pH, total petroleum hydrocarbons (TPHs), polycyclic aromatic hydrocarbons (PAHs), total carbon (C(tot)), total nitrogen (N(tot)) and total phosphorus (P(tot)). The reduction of TPH based on mass balance in M1, M2, M3 and M4 after 373 days of treatment was 62, 51, 74 and 49%; the reduction of PAHs was 97%, +13% (increase), 92 and 88%, respectively. The following mechanisms alone or in combination might explain the results: (i) most organics added with amendments biodegrade faster than most petroleum hydrocarbons, resulting in a relative increase in concentration of these recalcitrant contaminants; (ii) some amendments result in increased amounts of TPH and PAHs to be degraded in the mixture; (iii) sorption-desorption mechanisms involving hydrophobic compounds in the organic matrix reduce bioavailability, biodegradability and eventually extractability; (iv) mixture heterogeneity affecting sampling. Total contaminant mass reduction seems to be a better parameter than concentration to assess degradation efficiency in mixtures with high content of biodegradable amendments.

  17. Maternal vitamin B6 deficient or supplemented diets on expression of genes related to GABAergic, serotonergic, or glutamatergic pathways in hippocampus of rat dams and their offspring.

    PubMed

    Almeida, Mara Ribeiro; Mabasa, Lawrence; Crane, Courtney; Park, Chung S; Venâncio, Vinícius Paula; Bianchi, Maria Lourdes Pires; Antunes, Lusânia Maria Greggi

    2016-07-01

    Vitamin B6 plays crucial roles on brain development and its maternal deficiency impacts the gamma-aminobutyric acid (GABA)ergic, serotonergic, glutamatergic, and dopaminergic systems in offspring. However, the molecular mechanisms underlying these neurological changes are not well understood. Thus, we aimed at evaluating which components of those neurotransmitter metabolism and signaling pathways can be modulated by maternal vitamin B6 -deficient or B6 -supplementated diets in the hippocampus of rat dams and their offspring. Female Wistar rats were fed three different diets: control (6 mg vitamin B6 /kg), supplemented (30 mg vitamin B6 /kg) or deficient diet (0 mg vitamin B6 /kg), from 4 weeks before pregnancy through lactation. Newborn pups (10 days old) from rat dams fed vitamin B6 -deficient diet presented hyperhomocysteinemia and had a significant increase in mRNA levels of glutamate decarboxylase 1 (Gad1), fibroblast growth factor 2 (Fgf2), and glutamate-ammonia ligase (Glul), while glutaminase (Gls) and tryptophan hydroxylase 1 (Tph1) mRNAs were downregulated. Vitamin B6 supplementation or deficiency did not change hippocampal global DNA methylation. A maternal vitamin B6 -deficient diet affects the expression of genes related to GABA, glutamate, and serotonin metabolisms in offspring by regulating Gad1, Glul, Gls, and Tph1 mRNA expression. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Rapid identification of oil-contaminated soils using visible near-infrared diffuse reflectance spectroscopy.

    PubMed

    Chakraborty, Somsubhra; Weindorf, David C; Morgan, Cristine L S; Ge, Yufeng; Galbraith, John M; Li, Bin; Kahlon, Charanjit S

    2010-01-01

    In the United States, petroleum extraction, refinement, and transportation present countless opportunities for spillage mishaps. A method for rapid field appraisal and mapping of petroleum hydrocarbon-contaminated soils for environmental cleanup purposes would be useful. Visible near-infrared (VisNIR, 350-2500 nm) diffuse reflectance spectroscopy (DRS) is a rapid, nondestructive, proximal-sensing technique that has proven adept at quantifying soil properties in situ. The objective of this study was to determine the prediction accuracy of VisNIR DRS in quantifying petroleum hydrocarbons in contaminated soils. Forty-six soil samples (including both contaminated and reference samples) were collected from six different parishes in Louisiana. Each soil sample was scanned using VisNIR DRS at three combinations of moisture content and pretreatment: (i) field-moist intact aggregates, (ii) air-dried intact aggregates, (iii) and air-dried ground soil (sieved through a 2-mm sieve). The VisNIR spectra of soil samples were used to predict total petroleum hydrocarbon (TPH) content in the soil using partial least squares (PLS) regression and boosted regression tree (BRT) models. Each model was validated with 30% of the samples that were randomly selected and not used in the calibration model. The field-moist intact scan proved best for predicting TPH content with a validation r2 of 0.64 and relative percent difference (RPD) of 1.70. Because VisNIR DRS was promising for rapidly predicting soil petroleum hydrocarbon content, future research is warranted to evaluate the methodology for identifying petroleum contaminated soils.

  19. Nrf2 pathway modulates Substance P-induced human mast cell activation and degranulation in the hair follicle.

    PubMed

    Jadkauskaite, Laura; Bahri, Rajia; Farjo, Nilofer; Farjo, Bessam; Jenkins, Gail; Bhogal, Ranjit; Haslam, Iain; Bulfone-Paus, Silvia; Paus, Ralf

    2018-05-30

    Activation of Nrf2 in primary human mast cells exposed to oxidative stress induced by substance P suppresses pro-inflammatory gene transcription, activation and degranulation. Copyright © 2018. Published by Elsevier Inc.

  20. ASGR1 and ASGR2, the Genes that Encode the Asialoglycoprotein Receptor (Ashwell Receptor), Are Expressed in Peripheral Blood Monocytes and Show Interindividual Differences in Transcript Profile

    PubMed Central

    Harris, Rebecca Louise; van den Berg, Carmen Wilma; Bowen, Derrick John

    2012-01-01

    Background. The asialoglycoprotein receptor (ASGPR) is a hepatic receptor that mediates removal of potentially hazardous glycoconjugates from blood in health and disease. The receptor comprises two proteins, asialoglycoprotein receptor 1 and 2 (ASGR1 and ASGR2), encoded by the genes ASGR1 and ASGR2. Design and Methods. Using reverse transcription amplification (RT-PCR), expression of ASGR1 and ASGR2 was investigated in human peripheral blood monocytes. Results. Monocytes were found to express ASGR1 and ASGR2 transcripts. Correctly spliced transcript variants encoding different isoforms of ASGR1 and ASGR2 were present in monocytes. The profile of transcript variants from both ASGR1 and ASGR2 differed among individuals. Transcript expression levels were compared with the hepatocyte cell line HepG2 which produces high levels of ASGPR. Monocyte transcripts were 4 to 6 orders of magnitude less than in HepG2 but nonetheless readily detectable using standard RT-PCR. The monocyte cell line THP1 gave similar results to monocytes harvested from peripheral blood, indicating it may provide a suitable model system for studying ASGPR function in this cell type. Conclusions. Monocytes transcribe and correctly process transcripts encoding the constituent proteins of the ASGPR. Monocytes may therefore represent a mobile pool of the receptor, capable of reaching sites remote from the liver. PMID:22919488

Top