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Sample records for active demethylation process

  1. AP endonucleases process 5-methylcytosine excision intermediates during active DNA demethylation in Arabidopsis

    PubMed Central

    Lee, Jiyoon; Jang, Hosung; Shin, Hosub; Choi, Woo Lee; Mok, Young Geun; Huh, Jin Hoe

    2014-01-01

    DNA methylation is a primary epigenetic modification regulating gene expression and chromatin structure in many eukaryotes. Plants have a unique DNA demethylation system in that 5-methylcytosine (5mC) is directly removed by DNA demethylases, such as DME/ROS1 family proteins, but little is known about the downstream events. During 5mC excision, DME produces 3′-phosphor-α, β-unsaturated aldehyde and 3′-phosphate by successive β- and δ-eliminations, respectively. The kinetic studies revealed that these 3′-blocking lesions persist for a significant amount of time and at least two different enzyme activities are required to immediately process them. We demonstrate that Arabidopsis AP endonucleases APE1L, APE2 and ARP have distinct functions to process such harmful lesions to allow nucleotide extension. DME expression is toxic to E. coli due to excessive 5mC excision, but expression of APE1L or ARP significantly reduces DME-induced cytotoxicity. Finally, we propose a model of base excision repair and DNA demethylation pathway unique to plants. PMID:25228464

  2. Active DNA Demethylation in Plants and Animals

    PubMed Central

    Zhang, H.; Zhu, J.-K.

    2013-01-01

    Active DNA demethylation regulates many vital biological processes, including early development and locus-specific gene expression in plants and animals. In Arabidopsis, bifunctional DNA glycosylases directly excise the 5-methylcytosine base and then cleave the DNA backbone at the abasic site. Recent evidence suggests that mammals utilize DNA glycosylases after 5-methylcytosine is oxidized and/or deaminated. In both cases, the resultant single-nucleotide gap is subsequently filled with an unmodified cytosine through the DNA base excision repair pathway. The enzymatic removal of 5-methylcytosine is tightly integrated with histone modifications and possibly noncoding RNAs. Future research will increase our understanding of the mechanisms and critical roles of active DNA demethylation in various cellular processes as well as inspire novel genetic and chemical therapies for epigenetic disorders. PMID:23197304

  3. Active DNA demethylation at enhancers during the vertebrate phylotypic period.

    PubMed

    Bogdanović, Ozren; Smits, Arne H; de la Calle Mustienes, Elisa; Tena, Juan J; Ford, Ethan; Williams, Ruth; Senanayake, Upeka; Schultz, Matthew D; Hontelez, Saartje; van Kruijsbergen, Ila; Rayon, Teresa; Gnerlich, Felix; Carell, Thomas; Veenstra, Gert Jan C; Manzanares, Miguel; Sauka-Spengler, Tatjana; Ecker, Joseph R; Vermeulen, Michiel; Gómez-Skarmeta, José Luis; Lister, Ryan

    2016-04-01

    The vertebrate body plan and organs are shaped during a conserved embryonic phase called the phylotypic stage. However, the mechanisms that guide the epigenome through this transition and their evolutionary conservation remain elusive. Here we report widespread DNA demethylation of enhancers during the phylotypic period in zebrafish, Xenopus tropicalis and mouse. These enhancers are linked to developmental genes that display coordinated transcriptional and epigenomic changes in the diverse vertebrates during embryogenesis. Binding of Tet proteins to (hydroxy)methylated DNA and enrichment of 5-hydroxymethylcytosine in these regions implicated active DNA demethylation in this process. Furthermore, loss of function of Tet1, Tet2 and Tet3 in zebrafish reduced chromatin accessibility and increased methylation levels specifically at these enhancers, indicative of DNA methylation being an upstream regulator of phylotypic enhancer function. Overall, our study highlights a regulatory module associated with the most conserved phase of vertebrate embryogenesis and suggests an ancient developmental role for Tet dioxygenases.

  4. Krüppel like factor 4 promoter undergoes active demethylation during monocyte/macrophage differentiation.

    PubMed

    Karpurapu, Manjula; Ranjan, Ravi; Deng, Jing; Chung, Sangwoon; Lee, Yong Gyu; Xiao, Lei; Nirujogi, Teja Srinivas; Jacobson, Jeffrey R; Park, Gye Young; Christman, John W

    2014-01-01

    The role of different lineage specific transcription factors in directing hematopoietic cell fate towards myeloid lineage is well established but the status of epigenetic modifications has not been defined during this important developmental process. We used non proliferating, PU.1 inducible myeloid progenitor cells and differentiating bone marrow derived macrophages to study the PU.1 dependent KLF4 transcriptional regulation and its promoter demethylation during monocyte/macrophage differentiation. Expression of KLF4 was regulated by active demethylation of its promoter and PU.1 specifically bound to KLF4 promoter oligo harboring the PU.1 consensus sequence. Methylation specific quantitative PCR and Bisulfite sequencing indicated demethylation of CpG residues most proximal to the transcription start site of KLF4 promoter. Cloned KLF4 promoter in pGL3 Luciferase and CpG free pcpgf-bas vectors showed accentuated reporter activity when co-transfected with the PU.1 expression vector. In vitro methylation of both KLF4 promoter oligo and cloned KLF4 promoter vectors showed attenuated in vitro DNA binding activity and Luciferase/mouse Alkaline phosphotase reporter activity indicating the negative influence of KLF4 promoter methylation on PU.1 binding. The Cytosine deaminase, Activation Induced Cytidine Deaminase (AICDA) was found to be critical for KLF4 promoter demethylation. More importantly, knock down of AICDA resulted in blockade of KLF4 promoter demethylation, decreased F4/80 expression and other phenotypic characters of macrophage differentiation. Our data proves that AICDA mediated active demethylation of the KLF4 promoter is necessary for transcriptional regulation of KLF4 by PU.1 during monocyte/macrophage differentiation.

  5. Parp Inhibition Prevents Ten-Eleven Translocase Enzyme Activation and Hyperglycemia-Induced DNA Demethylation

    PubMed Central

    Dhliwayo, Nyembezi; Sarras, Michael P.; Luczkowski, Ernest; Mason, Samantha M.

    2014-01-01

    Studies from human cells, rats, and zebrafish have documented that hyperglycemia (HG) induces the demethylation of specific cytosines throughout the genome. We previously documented that a subset of these changes become permanent and may provide, in part, a mechanism for the persistence of complications referred to as the metabolic memory phenomenon. In this report, we present studies aimed at elucidating the molecular machinery that is responsible for the HG-induced DNA demethylation observed. To this end, RNA expression and enzymatic activity assays indicate that the ten-eleven translocation (Tet) family of enzymes are activated by HG. Furthermore, through the detection of intermediates generated via conversion of 5-methyl-cytosine back to the unmethylated form, the data were consistent with the use of the Tet-dependent iterative oxidation pathway. In addition, evidence is provided that the activity of the poly(ADP-ribose) polymerase (Parp) enzyme is required for activation of Tet activity because the use of a Parp inhibitor prevented demethylation of specific loci and the accumulation of Tet-induced intermediates. Remarkably, this inhibition was accompanied by a complete restoration of the tissue regeneration deficit that is also induced by HG. The ultimate goal of this work is to provide potential new avenues for therapeutic discovery. PMID:24722243

  6. Thymine DNA Glycosylase Is Essential for Active DNA Demethylation by Linked Deamination-Base Excision Repair

    PubMed Central

    Cortellino, Salvatore; Xu, Jinfei; Sannai, Mara; Moore, Robert; Caretti, Elena; Cigliano, Antonio; Le Coz, Madeleine; Devarajan, Karthik; Wessels, Andy; Soprano, Dianne; Abramowitz, Lara K.; Bartolomei, Marisa S.; Rambow, Florian; Bassi, Maria Rosaria; Bruno, Tiziana; Fanciulli, Maurizio; Renner, Catherine; Klein-Szanto, Andres J.; Matsumoto, Yoshihiro; Kobi, Dominique; Davidson, Irwin; Alberti, Christophe; Larue, Lionel; Bellacosa, Alfonso

    2011-01-01

    Summary DNA methylation is a major epigenetic mechanism for gene silencing. While methyltransferases mediate cytosine methylation, it is less clear how unmethylated regions in mammalian genomes are protected from de novo methylation and whether an active demethylating activity is involved. Here we show that either knockout or catalytic inactivation of the DNA repair enzyme Thymine DNA Glycosylase (TDG) leads to embryonic lethality in mice. TDG is necessary for recruiting p300 to retinoic acid (RA)-regulated promoters, protection of CpG islands from hypermethylation, and active demethylation of tissue-specific, developmentally- and hormonally-regulated promoters and enhancers. TDG interacts with the deaminase AID and the damage-response protein GADD45a. These findings highlight a dual role for TDG in promoting proper epigenetic states during development and suggest a two-step mechanism for DNA demethylation in mammals, whereby 5-methylcytosine and 5-hydroxymethylcytosine are first deaminated by AID to thymine and 5-hydroxymethyluracil, respectively, followed by TDG-mediated thymine and 5-hydroxymethyluracil excision repair. PMID:21722948

  7. Impacts of Activated Carbon Amendment on Hg Methylation, Demethylation and Microbial Activity in Marsh Soils

    NASA Astrophysics Data System (ADS)

    Gilmour, C. C.; Ghosh, U.; Santillan, E. F. U.; Soren, A.; Bell, J. T.; Butera, D.; McBurney, A. W.; Brown, S.; Henry, E.; Vlassopoulos, D.

    2015-12-01

    In-situ sorbent amendments are a low-impact approach for remediation of contaminants in sediments, particular in habitats like wetlands that provide important ecosystem services. Laboratory microcosm trials (Gilmour et al. 2013) and early field trials show that activated carbon (AC) can effectively increase partitioning of both inorganic Hg and methylmercury to the solid phase. Sediment-water partitioning can serve as a proxy for Hg and MeHg bioavailability in soils. One consideration in using AC in remediation is its potential impact on organisms. For mercury, a critical consideration is the potential impact on net MeHg accumulation and bioavailability. In this study, we specifically evaluated the impact of AC on rates of methylmercury production and degradation, and on overall microbial activity, in 4 different Hg-contaminated salt marsh soils. The study was done over 28 days in anaerobic, sulfate-reducing slurries. A double label of enriched mercury isotopes (Me199Hg and inorganic 201Hg) was used to separately follow de novo Me201Hg production and Me199Hg degradation. AC amendments decreased both methylation and demethylation rate constants relative to un-amended controls, but the impact on demethylation was stronger. The addition of 5% (dry weight) regenerated AC to soil slurries drove demethylation rate constants to nearly zero; i.e. MeHg sorption to AC almost totally blocked its degradation. The net impact was increased solid phase MeHg concentrations in some of the soil slurries with the highest methylation rate constants. However, the net impact of AC amendments was to increase MeHg (and inorganic Hg) partitioning to the soil phase and decrease concentrations in the aqueous phase. AC significantly decreased aqueous phase inorganic Hg and MeHg concentrations after 28 days. Overall, the efficacy of AC in reducing aqueous MeHg was highest in the soils with the highest MeHg concentrations. The AC addition did not significantly impact microbial activity, as

  8. An interplay of the base excision repair and mismatch repair pathways in active DNA demethylation

    PubMed Central

    Grin, Inga; Ishchenko, Alexander A.

    2016-01-01

    Active DNA demethylation (ADDM) in mammals occurs via hydroxylation of 5-methylcytosine (5mC) by TET and/or deamination by AID/APOBEC family enzymes. The resulting 5mC derivatives are removed through the base excision repair (BER) pathway. At present, it is unclear how the cell manages to eliminate closely spaced 5mC residues whilst avoiding generation of toxic BER intermediates and whether alternative DNA repair pathways participate in ADDM. It has been shown that non-canonical DNA mismatch repair (ncMMR) can remove both alkylated and oxidized nucleotides from DNA. Here, a phagemid DNA containing oxidative base lesions and methylated sites are used to examine the involvement of various DNA repair pathways in ADDM in murine and human cell-free extracts. We demonstrate that, in addition to short-patch BER, 5-hydroxymethyluracil and uracil mispaired with guanine can be processed by ncMMR and long-patch BER with concomitant removal of distant 5mC residues. Furthermore, the presence of multiple mispairs in the same MMR nick/mismatch recognition region together with BER-mediated nick formation promotes proficient ncMMR resulting in the reactivation of an epigenetically silenced reporter gene in murine cells. These findings suggest cooperation between BER and ncMMR in the removal of multiple mismatches that might occur in mammalian cells during ADDM. PMID:26843430

  9. Photochemical N-demethylation of alkaloids.

    PubMed

    Ripper, J A; Tiekink, E R; Scammells, P J

    2001-02-26

    Certain alkaloids were observed to undergo N-demethylation processes under photochemical conditions. Tropine, acetyltropine, tropinone, and atropine were cleanly N-demethylated upon treatment with tetraphenylporphin, oxygen, and light. Dextromethorphan also underwent a N-demethylation reaction, but reacted further to afford an imine. In contrast, 14-acyloxycodeinones underwent a photochemically induced tandem N-demethylation acyl migration.

  10. Role of the Arabidopsis DNA glycosylase/lyase ROS1 in active DNA demethylation

    PubMed Central

    Agius, Fernanda; Kapoor, Avnish; Zhu, Jian-Kang

    2006-01-01

    DNA methylation is a stable epigenetic mark for transcriptional gene silencing in diverse organisms including plants and many animals. In contrast to the well characterized mechanism of DNA methylation by methyltransferases, the mechanisms and function of active DNA demethylation have been controversial. Genetic evidence suggested that the DNA glycosylase domain-containing protein ROS1 of Arabidopsis is a putative DNA demethylase, because loss-of-function ros1 mutations cause DNA hypermethylation and enhance transcriptional gene silencing. We report here the biochemical characterization of ROS1 and the effect of its overexpression on the DNA methylation of target genes. Our data suggest that the DNA glycosylase activity of ROS1 removes 5-methylcytosine from the DNA backbone and then its lyase activity cleaves the DNA backbone at the site of 5-methylcytosine removal by successive β- and δ-elimination reactions. Overexpression of ROS1 in transgenic plants led to a reduced level of cytosine methylation and increased expression of a target gene. These results demonstrate that ROS1 is a 5-methylcytosine DNA glycosylase/lyase important for active DNA demethylation in Arabidopsis. PMID:16864782

  11. Enzyme-based electrochemical biosensor for sensitive detection of DNA demethylation and the activity of DNA demethylase.

    PubMed

    Zhou, Yunlei; Li, Bingchen; Wang, Mo; Yang, Zhiqing; Yin, Huanshun; Ai, Shiyun

    2014-08-20

    A novel electrochemical method is developed for detection of DNA demethylation and assay of DNA demethylase activity. This method is constructed by hybridizing the probe with biotin tagged hemi-methylated complementary DNA and further capturing streptavidin tagged alkaline phosphatase (SA-ALP) to catalyze the hydrolysis reaction of p-nitrophenyl phosphate. The hydrolysate of p-nitrophenol (PNP) is then used as electrochemical probe for detecting DNA demethylation and assaying the activity of DNA demethylase. Demethylation of target DNA initiates a degradation reaction of the double-stranded DNA (dsDNA) by restriction endonuclease of BstUI. It makes the failed immobilization of ALP, resulting in a decreased electrochemical oxidation signal of PNP. Through the change of this electrochemical signal, the DNA demethylation is identified and the activity of DNA demethylase is analyzed with low detection limit of 1.3 ng mL(-1). This method shows the advantages of simple operation, cheap and miniaturized instrument, high selectivity. Thus, it provides a useful platform for detecting DNA demethylation, analyzing demethylase activity and screening inhibited drug.

  12. Early steps of active DNA demethylation initiated by ROS1 glycosylase require three putative helix-invading residues.

    PubMed

    Parrilla-Doblas, Jara Teresa; Ponferrada-Marín, María Isabel; Roldán-Arjona, Teresa; Ariza, Rafael R

    2013-10-01

    Active DNA demethylation is crucial for epigenetic control, but the underlying enzymatic mechanisms are incompletely understood. REPRESSOR OF SILENCING 1 (ROS1) is a 5-methylcytosine (5-meC) DNA glycosylase/lyase that initiates DNA demethylation in plants through a base excision repair process. The enzyme binds DNA nonspecifically and slides along the substrate in search of 5-meC. In this work, we have used homology modelling and biochemical analysis to gain insight into the mechanism of target location and recognition by ROS1. We have found that three putative helix-intercalating residues (Q607, R903 and M905) are required for processing of 5-meC:G pairs, but dispensable for excision of mismatched 5-meC. Mutant proteins Q607A, R903A and M905G retain the capacity to process an abasic site opposite G, thus suggesting that all three residues play a critical role in early steps of the base extrusion process and likely contribute to destabilization of 5-meC:G pairs. While R903 and M905 are not essential for DNA binding, mutation of Q607 abrogates stable binding to both methylated and nonmethylated DNA. However, the mutant protein Q607A can form stable complexes with DNA substrates containing blocked ends, which suggests that Q607 intercalates into the helix and inhibits sliding. Altogether, our results suggest that ROS1 uses three predicted helix-invading residues to actively interrogate DNA in search for 5-meC.

  13. Active tissue-specific DNA demethylation conferred by somatic cell nuclei in stable heterokaryons

    PubMed Central

    Zhang, Fan; Pomerantz, Jason H.; Sen, George; Palermo, Adam T.; Blau, Helen M.

    2007-01-01

    DNA methylation is among the most stable epigenetic marks, ensuring tissue-specific gene expression in a heritable manner throughout development. Here we report that differentiated mesodermal somatic cells can confer tissue-specific changes in DNA methylation on epidermal progenitor cells after fusion in stable multinucleate heterokaryons. Myogenic factors alter regulatory regions of genes in keratinocyte cell nuclei, demethylating and activating a muscle-specific gene and methylating and silencing a keratinocyte-specific gene. Because these changes occur in the absence of DNA replication or cell division, they are mediated by an active mechanism. Thus, the capacity to transfer epigenetic changes to other nuclei is not limited to embryonic stem cells and oocytes but is also a property of highly specialized mammalian somatic cells. These results suggest the possibility of directing the reprogramming of readily available postnatal human progenitor cells toward specific tissue cell types. PMID:17360535

  14. DNA demethylation in PD-1 gene promoter induced by 5-azacytidine activates PD-1 expression on Molt-4 cells.

    PubMed

    Zhang, Min; Xiao, Xin Q; Jiang, Yong F; Liang, Yun S; Peng, Min Y; Xu, Yun; Gong, Guo Z

    2011-01-01

    The expression of the programmed death 1 (PD-1) gene is an indicator of exhausted T-cells with decreased activation and function. It remains unknown, however, whether the methylation status of the PD-1 gene promoter is associated with PD-1 expression level. This study shows the changes of PD-1 expression levels and the demethylation status of the PD-1 promoter region in Molt-4 cells under different concentrations of 5-azacytidine (5-Zac). The result demonstrated that DNA demethylation at PD-1 promoter may contribute to PD-1 overexpression.

  15. Isolation and characterization of a human intestinal bacterium, Eubacterium sp. ARC-2, capable of demethylating arctigenin, in the essential metabolic process to enterolactone.

    PubMed

    Jin, Jong-Sik; Zhao, Yu-Feng; Nakamura, Norio; Akao, Teruaki; Kakiuchi, Nobuko; Hattori, Masao

    2007-05-01

    Plant lignans, such as pinoresinol diglucoside, secoisolariciresinol diglucoside and arctiin, are metabolized to mammalian lignans, enterolactone or enterodiol, by human intestinal bacteria. Their metabolic processes include deglucosylation, ring cleavage, demethylation, dehydroxylation and oxidation. Here we isolated an intestinal bacterium capable of demethylating arctigenin, an aglycone of arctiin, to 2,3-bis(3,4-dihydroxybenzyl)butyrolactone (1) from human feces, and identified as an Eubacterium species (E. sp. ARC-2), which is similar to Eubacterium limosum on the basis of morphological and biochemical properties and 16S rRNA gene sequencing. By incubating with E. sp. ARC-2, arctigenin was converted to 1 through stepwise demethylation. Demethylation of arctigenin by E. sp. ARC-2 was tetrahydrofolate- and ATP-dependent, indicating that the reaction was catalyzed by methyltransferase. Moreover, E. sp. ARC-2 transformed secoisolariciresinol to 2,3-bis(3,4-dihydroxybenzyl)-1,4-butanediol by demethylation.

  16. Genistein mediated histone acetylation and demethylation activates tumor suppressor genes in prostate cancer cells.

    PubMed

    Kikuno, Nobuyuki; Shiina, Hiroaki; Urakami, Shinji; Kawamoto, Ken; Hirata, Hiroshi; Tanaka, Yuichiro; Majid, Shahana; Igawa, Mikio; Dahiya, Rajvir

    2008-08-01

    Genistein is a phytoestrogen that has been reported to suppress the AKT signaling pathway in several malignancies. However, the molecular mechanism of genistein action is not known. We tested the hypothesis that genistein activates expression of several aberrantly silenced tumor suppressor genes (TSGs) that have unmethylated promoters such as PTEN, CYLD, p53 and FOXO3a. We report here that genistein activates TSGs through remodeling of the heterochromatic domains at promoters in prostate cancer cells by modulating histone H3-Lysine 9 (H3-K9) methylation and deacetylation. Genistein activation involved demethylation and acetylation of H3-K9 at the PTEN and the CYLD promoter, while acetylation of H3-K9 at the p53 and the FOXO3a promoter occurred through reduction of endogenous SIRT1 activity. There was a decrease of SIRT1 expression and accumulation of SIRT1 in the cytoplasm from the nucleus. Increased expression of these TSGs was also reciprocally related to attenuation of phosphorylated-AKT and NF-kappaB binding activity in prostate cancer cells. This is the first report describing a novel epigenetic pathway that activates TSGs by modulating either histone H3-Lysine 9 (H3-K9) methylation or deacetylation at gene promoters leading to inhibition of the AKT signaling pathway. These findings strengthen the understanding of how genistein may be chemoprotective in prostate cancer.

  17. H(2)-CO(2)-Dependent Anaerobic O-Demethylation Activity in Subsurface Sediments and by an Isolated Bacterium.

    PubMed

    Liu, S; Suflita, J M

    1993-05-01

    The ability of microorganisms in sediments from the Atlantic Coastal Plain to biodegrade methoxylated aromatic compounds was examined. O-demethylation activity was detected in deep (121- and 406-m) sediments, as well as in the surface soil. A syringate-demethylating consortium, containing at least three types of bacteria, was enriched from a deep-sediment sample in a medium containing syringate as the sole organic carbon source and with a N(2)-CO(2) atmosphere. An isolate which demethylated syringate was obtained from the enrichment on an agar medium incubated under a H(2)-CO(2) but not a N(2)-CO(2) or N(2) atmosphere. O demethylation of syringate of this isolate was dependent on the presence of both H(2) and CO(2) in the gas phase. The metabolism of syringate occurred in a sequential manner: methylgallate accumulated transiently before it was converted to gallate. Mass balance analysis suggests that the stoichiometry of the reaction in this isolate proceeds in accordance with the following generalized equation: C(7)H(3)O(3)(OCH(3))(n) + nHCO(3) + nH(2) --> C(7)H(3)O(3)(OH)(n) + nCH(3)COO + nH(2)O.

  18. IL-2 and GM-CSF are regulated by DNA demethylation during activation of T cells, B cells and macrophages

    SciTech Connect

    Li, Yan; Ohms, Stephen J.; Shannon, Frances M.; Sun, Chao; Fan, Jun Y.

    2012-03-23

    Highlights: Black-Right-Pointing-Pointer DNA methylation is dynamic and flexible and changes rapidly upon cell activation. Black-Right-Pointing-Pointer DNA methylation controls the inducible gene expression in a given cell type. Black-Right-Pointing-Pointer Some enzymes are involved in maintaining the methylation profile of immune cells. -- Abstract: DNA demethylation has been found to occur at the promoters of a number of actively expressed cytokines and is believed to play a critical role in transcriptional regulation. While many DNA demethylation studies have focused on T cell activation, proliferation and differentiation, changes in DNA methylation in other types of immune cells are less well studied. We found that the expression of two cytokines (IL-2 and GM-CSF) responded differently to activation in three types of immune cells: EL4, A20 and RAW264.7 cells. Using the McrBC and MeDIP approaches, we observed decreases in DNA methylation at a genome-wide level and at the promoters of the genes of these cytokines. The expression of several potential enzymes/co-enzymes involved in the DNA demethylation pathways seemed to be associated with immune cell activation.

  19. An embryonic demethylation mechanism involving binding of transcription factors to replicating DNA.

    PubMed Central

    Matsuo, K; Silke, J; Georgiev, O; Marti, P; Giovannini, N; Rungger, D

    1998-01-01

    In vertebrates, transcriptionally active promoters are undermethylated. Since the transcription factor Sp1, and more recently NF-kappaB, have been implicated in the demethylation process, we examined the effect of transcription factors on demethylation by injecting in vitro methylated plasmid DNA into Xenopus fertilized eggs. We found that various transactivation domains, including a strong acidic activation domain from the viral protein VP16, can enhance demethylation of a promoter region when fused to a DNA binding domain which recognizes the promoter. Furthermore, demethylation occurs only after the midblastula transition, when the general transcription machinery of the host embryo becomes available. Nevertheless, transcription factor binding need not be followed by actual transcription, since demethylation is not blocked by alpha-amanitin treatment. Finally, replication of the target DNA is a prerequisite for efficient demethylation since only plasmids that carry the bovine papilloma virus sequences which support plasmid replication after the midblastula transition are demethylated. No demethylation is detectable in the oocyte system where DNA is not replicated. These results suggest that, in the Xenopus embryo, promoters for which transcription factors are available are demethylated by a replication-dependent, possibly passive mechanism. PMID:9482741

  20. The cytosolic Fe-S cluster assembly component MET18 is required for the full enzymatic activity of ROS1 in active DNA demethylation

    PubMed Central

    Wang, Xiaokang; Li, Qi; Yuan, Wei; Cao, Zhendong; Qi, Bei; Kumar, Suresh; Li, Yan; Qian, Weiqiang

    2016-01-01

    DNA methylation patterns in plants are dynamically regulated by DNA methylation and active DNA demethylation in response to both environmental changes and development of plant. Beginning with the removal of methylated cytosine by ROS1/DME family of 5-methylcytosine DNA glycosylases, active DNA demethylation in plants occurs through base excision repair. So far, many components involved in active DNA demethylation remain undiscovered. Through a forward genetic screening of Arabidopsis mutants showing DNA hypermethylation at the EPF2 promoter region, we identified the conserved iron-sulfur cluster assembly protein MET18. MET18 dysfunction caused DNA hypermethylation at more than 1000 loci as well as the silencing of reporter genes and some endogenous genes. MET18 can directly interact with ROS1 in vitro and in vivo. ROS1 activity was reduced in the met18 mutant plants and point mutation in the conserved Fe-S cluster binding motif of ROS1 disrupted its biological function. Interestingly, a large number of DNA hypomethylated loci, especially in the CHH context, were identified from the met18 mutants and most of the hypo-DMRs were from TE regions. Our results suggest that MET18 can regulate both active DNA demethylation and DNA methylation pathways in Arabidopsis. PMID:27193999

  1. Fungal demethylation of Kraft lignin.

    PubMed

    Zou, Linyou; Ross, Brian M; Hutchison, Leonard J; Christopher, Lew P; Dekker, Robert F H; Malek, Lada

    2015-06-01

    Demethylation of industrial lignin has been for long coveted as a pathway to the production of an abundant natural substitute for fossil-oil derived phenol. In an attempt to possibly identify a novel Kraft lignin-demethylating enzyme, we surveyed a collection of fungi by using selected ion flow tube-mass spectrometry (SIFT-MS). This method readily identifies methanol resulting from lignin demethylation activity. Absidia cylindrospora, and unidentified Cylindrocladium sp. and Aspergillus sp. were shown to metabolize lignin via different pathways, based on the HPLC analysis of lignin fragments. Of these three, Cylindrocladium and Aspergillus were shown to retain most of the lignin intact after 3 weeks in culture, while removing about 40% of the available methoxy groups. Our results demonstrate that after optimization of culture and lignin recovery methods, biological modification of Kraft lignin may be a feasible pathway to obtaining demethylated lignin for further industrial use.

  2. Integrated genomic analysis of colorectal cancer progression reveals activation of EGFR through demethylation of the EREG promoter

    PubMed Central

    Qu, X; Sandmann, T; Frierson, H; Fu, L; Fuentes, E; Walter, K; Okrah, K; Rumpel, C; Moskaluk, C; Lu, S; Wang, Y; Bourgon, R; Penuel, E; Pirzkall, A; Amler, L; Lackner, M R; Tabernero, J; Hampton, G M; Kabbarah, O

    2016-01-01

    Key molecular drivers that underlie transformation of colonic epithelium into colorectal adenocarcinoma (CRC) are well described. However, the mechanisms through which clinically targeted pathways are activated during CRC progression have yet to be elucidated. Here, we used an integrative genomics approach to examine CRC progression. We used laser capture microdissection to isolate colonic crypt cells, differentiated surface epithelium, adenomas, carcinomas and metastases, and used gene expression profiling to identify pathways that were differentially expressed between the different cell types. We identified a number of potentially important transcriptional changes in developmental and oncogenic pathways, and noted a marked upregulation of EREG in primary and metastatic cancer cells. We confirmed this pattern of gene expression by in situ hybridization and observed staining consistent with autocrine expression in the tumor cells. Upregulation of EREG during the adenoma–carcinoma transition was associated with demethylation of two key sites within its promoter, and this was accompanied by an increase in the levels of epidermal growth factor receptor (EGFR) phosphorylation, as assessed by reverse-phase protein analysis. In CRC cell lines, we demonstrated that EREG demethylation led to its transcriptional upregulation, higher levels of EGFR phosphorylation, and sensitization to EGFR inhibitors. Low levels of EREG methylation in patients who received cetuximab as part of a phase II study were associated with high expression of the ligand and a favorable response to therapy. Conversely, high levels of promoter methylation and low levels of EREG expression were observed in tumors that progressed after treatment. We also noted an inverse correlation between EREG methylation and expression levels in several other cancers, including those of the head and neck, lung and bladder. Therefore, we propose that upregulation of EREG expression through promoter demethylation

  3. The methylcytosine dioxygenase Tet2 promotes DNA demethylation and activation of cytokine gene expression in T cells.

    PubMed

    Ichiyama, Kenji; Chen, Tingting; Wang, Xiaohu; Yan, Xiaowei; Kim, Byung-Seok; Tanaka, Shinya; Ndiaye-Lobry, Delphine; Deng, Yuhua; Zou, Yanli; Zheng, Pan; Tian, Qiang; Aifantis, Iannis; Wei, Lai; Dong, Chen

    2015-04-21

    Epigenetic regulation of lineage-specific genes is important for the differentiation and function of T cells. Ten-eleven translocation (Tet) proteins catalyze 5-methylcytosine (5 mC) conversion to 5-hydroxymethylcytosine (5 hmC) to mediate DNA demethylation. However, the roles of Tet proteins in the immune response are unknown. Here, we characterized the genome-wide distribution of 5 hmC in CD4(+) T cells and found that 5 hmC marks putative regulatory elements in signature genes associated with effector cell differentiation. Moreover, Tet2 protein was recruited to 5 hmC-containing regions, dependent on lineage-specific transcription factors. Deletion of Tet2 in T cells decreased their cytokine expression, associated with reduced p300 recruitment. In vivo, Tet2 plays a critical role in the control of cytokine gene expression in autoimmune disease. Collectively, our findings suggest that Tet2 promotes DNA demethylation and activation of cytokine gene expression in T cells.

  4. Evidence that active demethylation mechanisms maintain the genome of carcinoma in situ cells hypomethylated in the adult testis

    PubMed Central

    Kristensen, D G; Nielsen, J E; Jørgensen, A; Skakkebæk, N E; Rajpert-De Meyts, E; Almstrup, K

    2014-01-01

    Background: Developmental arrest of fetal germ cells may lead to neoplastic transformation and formation of germ cell tumours via carcinoma in situ (CIS) cells. Normal fetal germ cell development requires complete erasure and re-establishment of DNA methylation. In contrast to normal spermatogonia, the genome of CIS cells remains unmethylated in the adult testis. We here investigated the possible active and passive pathways that can sustain the CIS genome hypomethylated in the adult testis. Methods: The levels of 5-methyl-cytosine (5mC) and 5-hydroxy-methyl-cytosine (5hmC) in DNA from micro-dissected CIS cells were assessed by quantitative measurements. The expression of TET1, TET2, APOBEC1, MBD4, APEX1, PARP1, DNMT1, DNMT3A, DNMT3B and DNMT3L in adult testis specimens with CIS and in human fetal testis was investigated by immunohistochemistry and immunofluorescence. Results: DNA from micro-dissected CIS cells contained very low levels of 5hmC produced by ten eleven translocation (TET) enzymes. CIS cells and fetal germ cells expressed the suggested initiator of active demethylation, APOBEC1, and the base excision repair proteins MBD4, APEX1 and PARP1, whereas TETs – the alternative initiators were absent. Both maintenance and de novo methyltransferases were detected in CIS cells. Conclusion: The data are consistent with the presence of an active DNA de-methylation pathway in CIS cells. The hypomethylated genome of CIS cells may contribute to phenotypic plasticity and invasive capabilities of this testicular cancer precursor. PMID:24292451

  5. Dynamics of 5-carboxylcytosine during hepatic differentiation: potential general role for active demethylation by DNA repair in lineage specification.

    PubMed

    Lewis, Lara C; Lo, Peggy Cho Kiu; Foster, Jeremy M; Dai, Nan; Corrêa, Ivan R; Durczak, Paulina M; Duncan, Gary; Ramsawhook, Ashley; Aithal, Guruprasad Padur; Denning, Chris; Hannan, Nicholas R F; Ruzov, Alexey

    2017-03-07

    Patterns of DNA methylation (5-methylcytosine, 5mC) are rearranged during differentiation contributing to the regulation of cell type-specific gene expression. TET proteins oxidize 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). Both 5fC and 5caC can be recognized and excised from DNA by thymine-DNA glycosylase (TDG) followed by the subsequent incorporation of unmodified cytosine into the abasic site via the base excision repair (BER) pathway. We previously demonstrated that 5caC accumulates during lineage specification of neural stem cells (NSCs) suggesting that such active demethylation pathway is operative in this system; however, it is still unknown if TDG/BER-dependent demethylation is utilized during other types of cellular differentiation. Here we analyze dynamics of the global levels of 5hmC and 5caC during differentiation of human pluripotent stem cells towards hepatic endoderm. We show that, similar to differentiating NSCs, 5caC transiently accumulates during hepatic differentiation. The levels of 5caC increase during specification of foregut, peak at the stage of hepatic endoderm commitment, and drop in differentiating cells concurrently with the onset of expression of alpha fetoprotein, a marker of committed hepatic progenitors. Moreover, we show that 5caC accumulates at promoter regions of several genes expressed during hepatic specification at differentiation stages corresponding to the beginning of their expression. Our data indicate that transient 5caC accumulation is a common feature of two different types (neural/glial and endoderm/hepatic) of cellular differentiation. This suggests that oxidation of 5mC may represent a general mechanism of rearrangement of 5mC profiles during lineage specification of somatic cells in mammals.

  6. Interleukin-4-Mediated 15-Lipoxygenase-1 Trans-Activation Requires UTX Recruitment and H3K27me3 Demethylation at the Promoter in A549 Cells

    PubMed Central

    Han, Hongya; Xu, Dawei; Liu, Cheng; Claesson, Hans-Erik; Björkholm, Magnus; Sjöberg, Jan

    2014-01-01

    Arachidonate 15-lipoxygenase-1 (ALOX15) oxygenates polyunsaturated fatty acids and bio-membranes, generating multiple lipid signalling mediators involved in inflammation. Several lines of evidence indicate that ALOX15 activation in the respiratory tract contributes to asthma progression. Recent experimental data reveals that histone modification at the promoter plays a critical role in ALOX15 gene transcription. In the present study, we examined the status of histone H3 trimethyl-lysine 27 (H3K27me3) at the ALOX15 promoter by chromatin immunoprecipitation assay in human lung epithelial carcinoma A549 cells incubated with or without interleukin (IL)-4. We identified demethylation of H3K27me3 at the ALOX15 promoter after IL-4 treatment. Furthermore, we found that the H3K27me2/3-specific demethylase, ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX), mediates the H3K27me3 demethylation during ALOX15 transcriptional activation. When UTX expression was knocked down using siRNA, IL-4-mediated H3K27me3 demethylation and ALOX15 induction were significantly attenuated. The critical role of UTX in ALOX15 expression was confirmed in human monocytes and the Hodgkin lymphoma (HL) cell line L1236, but was in these cells not related to H3K27me3-demethylase activity. These results demonstrate that UTX is implicated in IL-4 mediated transcriptional activation of the ALOX15 gene. PMID:24465480

  7. Anaerobic O-demethylation of phenylmethylethers

    SciTech Connect

    Frazer, A.C.; Young, L.Y.

    1990-01-01

    Anaerobic O-demethylation (AOD) of phenylmethylethers is a process of both basic and applied significance. The aryl-O-methyl ethers are abundant in natural products, particularly as components of lignin. They are present as methoxylated lignin monomers in anaerobic environments and can be completely degraded there by mixed microbial populations. AOD is an essential early step in this process, and it is also a key reaction in the utilization of the O-methyl substituent as a C-one substrate by acetogens. An understanding of the AOD reaction mechanism might suggest new ways in which chemicals could be derived from lignocellulosic materials. The biochemical mechanism for the anaerobic cleavage of the aryl-O-methyl ether bond is an intriguing, but relatively unexplored process. In contrast to aerobic O-demethylating enzymes, AOD appears to involve methyl group transfer. Thus, novel biochemical information on an important biotransformation reaction will be gained from the research proposed. Recently, we have shown that AOD activity is inducible and have developed an assay for detecting AOD activity in cell-free extracts of Acetobacterium woodii. AOD activity is stimulated in vitro by the addition of ATP (1mM) and pyruvate (30 mM), the K{sub M} for vanillate being 0.4 mM. In collaboration with protein purification experts, we proposed to purify the AOD enzyme and characterize the protein(s) and the enzymatic reaction involved. 8 figs., 5 tabs.

  8. Tissue-specific DNA demethylation is required for proper B-cell differentiation and function

    PubMed Central

    Orlanski, Shari; Labi, Verena; Reizel, Yitzhak; Spiro, Adam; Lichtenstein, Michal; Levin-Klein, Rena; Koralov, Sergei B.; Skversky, Yael; Rajewsky, Klaus; Cedar, Howard; Bergman, Yehudit

    2016-01-01

    There is ample evidence that somatic cell differentiation during development is accompanied by extensive DNA demethylation of specific sites that vary between cell types. Although the mechanism of this process has not yet been elucidated, it is likely to involve the conversion of 5mC to 5hmC by Tet enzymes. We show that a Tet2/Tet3 conditional knockout at early stages of B-cell development largely prevents lineage-specific programmed demethylation events. This lack of demethylation affects the expression of nearby B-cell lineage genes by impairing enhancer activity, thus causing defects in B-cell differentiation and function. Thus, tissue-specific DNA demethylation appears to be necessary for proper somatic cell development in vivo. PMID:27091986

  9. Fluconazole Binding and Sterol Demethylation in Three CYP51 Isoforms Indicate Differences in Active Site Topology

    SciTech Connect

    Bellamine, A.; Lepesheva, Galina I.; Waterman, Mike

    2010-11-16

    14{alpha}-Demethylase (CYP51) is a key enzyme in all sterol biosynthetic pathways (animals, fungi, plants, protists, and some bacteria), catalyzing the removal of the C-14 methyl group following cyclization of squalene. Based on mutations found in CYP51 genes from Candida albicans azole-resistant isolates obtained after fluconazole treatment of fungal infections, and using site-directed mutagenesis, we have found that fluconazole binding and substrate metabolism vary among three different CYP51 isoforms: human, fungal, and mycobacterial. In C. albicans, the Y132H mutant from isolates shows no effect on fluconazole binding, whereas the F145L mutant results in a 5-fold increase in its IC{sub 50} for fluconazole, suggesting that F145 (conserved only in fungal 14{alpha}-demethylases) interacts with this azole. In C. albicans, F145L accounts, in part, for the difference in fluconazole sensitivity reported between mammals and fungi, providing a basis for treatment of fungal infections. The C. albicans Y132H and human Y145H CYP51 mutants show essentially no effect on substrate metabolism, but the Mycobacterium tuberculosis F89H CYP51 mutant loses both its substrate binding and metabolism. Because these three residues align in the three isoforms, the results indicate that their active sites contain important structural differences, and further emphasize that fluconazole and substrate binding are uncoupled properties.

  10. Demethylation of methylarsonic acid by a microbial community.

    PubMed

    Yoshinaga, Masafumi; Cai, Yong; Rosen, Barry P

    2011-05-01

    Arsenic is one of the most widespread environmental carcinogens and has created devastating human health problems worldwide, yet little is known about mechanisms of biotransformation in contaminated regions. Methylarsonic acid [MAs(V)], extensively utilized as an herbicide, is largely demethylated to more toxic inorganic arsenite, which causes environmental problems. To understand the process of demethylation of methylarsenicals, soil samples commonly used on Florida golf courses were studied. Several soil extracts were found to demethylate MAs(V) to inorganic arsenite [As(III)]. From these extracts, a bacterial isolate was capable of reducing MAs(V) to MAs(III) but not of demethylating to As(III). A second bacterial isolate was capable of demethylating MAs(III) to As(III) but not of reducing MAs(V). A mixed culture could carry out the complete process of reduction and demethylation, demonstrating that demethylation of MAs(V) to As(III) is a two-step process. Analysis of the 16S ribosomal DNA sequences of the two organisms identified the MAs(V)-reducing and the MAs(III)-demethylating isolates as belong to Burkholderia and Streptomyces species respectively. This is the first report of a novel pathway of degradation of a methylarsenical herbicide by sequential reduction and demethylation in a microbial soil community, which we propose plays a significant role in the arsenic biogeocycle.

  11. Demethylation of methylarsonic acid by a microbial community

    PubMed Central

    Yoshinaga, Masafumi; Cai, Yong; Rosen, Barry P.

    2013-01-01

    Summary Arsenic is one of the most widespread environmental carcinogens and has created devastating human health problems worldwide, yet little is known about mechanisms of biotransformation in contaminated regions. Methylarsonic acid [MAs(V)], extensively utilized as an herbicide, is largely demethylated to more toxic inorganic arsenite, which causes environmental problems. To understand the process of demethylation of methylarsenicals, soil samples commonly used on Florida golf courses were studied. Several soil extracts were found to demethylate MAs(V) to inorganic arsenite [As(III)]. From these extracts, a bacterial isolate was capable of reducing MAs(V) to MAs(III) but not of demethylating to As(III). A second bacterial isolate was capable of demethylating MAs(III) to As(III) but not of reducing MAs(V). A mixed culture could carry out the complete process of reduction and demethylation, demonstrating that demethylation of MAs(V) to As(III) is a two-step process. Analysis of the 16S ribosomal DNA sequences of the two organisms identified the MAs(V)-reducing and the MAs(III)-demethylating isolates as belong to Burkholderia and Streptomyces species respectively. This is the first report of a novel pathway of degradation of a methylarsenical herbicide by sequential reduction and demethylation in a microbial soil community, which we propose plays a significant role in the arsenic biogeocycle. PMID:21272184

  12. Protein arginine methylation/demethylation and cancer

    PubMed Central

    Poulard, Coralie; Corbo, Laura; Le Romancer, Muriel

    2016-01-01

    Protein arginine methylation is a common post-translational modification involved in numerous cellular processes including transcription, DNA repair, mRNA splicing and signal transduction. Currently, there are nine known members of the protein arginine methyltransferase (PRMT) family, but only one arginine demethylase has been identified, namely the Jumonji domain-containing 6 (JMJD6). Although its demethylase activity was initially challenged, its dual activity as an arginine demethylase and a lysine hydroxylase is now recognized. Interestingly, a growing number of substrates for arginine methylation and demethylation play key roles in tumorigenesis. Though alterations in the sequence of these enzymes have not been identified in cancer, their overexpression is associated with various cancers, suggesting that they could constitute targets for therapeutic strategies. In this review, we present the recent knowledge of the involvement of PRMTs and JMJD6 in tumorigenesis. PMID:27556302

  13. CNS germinomas are characterized by global demethylation, chromosomal instability and mutational activation of the Kit-, Ras/Raf/Erk- and Akt-pathways

    PubMed Central

    Schulte, Simone Laura; Waha, Andreas; Steiger, Barbara; Denkhaus, Dorota; Dörner, Evelyn; Calaminus, Gabriele; Leuschner, Ivo; Pietsch, Torsten

    2016-01-01

    CNS germinomas represent a unique germ cell tumor entity characterized by undifferentiated tumor cells and a high response rate to current treatment protocols. Limited information is available on their underlying genomic, epigenetic and biological alterations. We performed a genome-wide analysis of genomic copy number alterations in 49 CNS germinomas by molecular inversion profiling. In addition, CpG dinucleotide methylation was studied by immunohistochemistry for methylated cytosine residues. Mutational analysis was performed by resequencing of candidate genes including KIT and RAS family members. Ras/Erk and Akt pathway activation was analyzed by immunostaining with antibodies against phospho-Erk, phosho-Akt, phospho-mTOR and phospho-S6. All germinomas coexpressed Oct4 and Kit but showed an extensive global DNA demethylation compared to other tumors and normal tissues. Molecular inversion profiling showed predominant genomic instability in all tumors with a high frequency of regional gains and losses including high level gene amplifications. Activating mutations of KIT exons 11, 13, and 17 as well as a case with genomic KIT amplification and activating mutations or amplifications of RAS gene family members including KRAS, NRAS and RRAS2 indicated mutational activation of crucial signaling pathways. Co-activation of Ras/Erk and Akt pathways was present in 83% of germinomas. These data suggest that CNS germinoma cells display a demethylated nuclear DNA similar to primordial germ cells in early development. This finding has a striking coincidence with extensive genomic instability. In addition, mutational activation of Kit-, Ras/Raf/Erk- and Akt- pathways indicate the biological importance of these pathways and their components as potential targets for therapy. PMID:27391150

  14. Production of Hydrolysable Tannin-Like Structures During the Microbial Demethylation of lignin: An Assessment Using13C-Labeled Tetramethylammonium Hydroxide Thermochemolysis.

    NASA Astrophysics Data System (ADS)

    Filley, T.; Blanchette, R.; Nierop, K.; Gamblin, D.

    2003-12-01

    Phenolic compounds in soils are important mediators of microbial activity, metal mobility, soil redox, and soil organic matter building processes. Direct tannin input and the microbial decomposition of lignin in litter and soil are important contributors to this pool of phenols. The ability to accurately assess the relative differences in lignin decay (which are initiated by demethylation and side chain oxidation) among synapyl, coniferyl, and p-coumaryl components of detrital lignin requires the ability to determine microbial demethylation within the complex soil residues. Differentiating between hydrolysable tannins and contributions from advanced lignin decay can be problematic for many of the most common molecular techniques such as alkaline CuO oxidation, pyrolysis GC, and tetramethylammonium hydroxide thermochemolysis because of either the masking effects of derivatizing agents, oxidative damage to ortho-phenols or low volatility of lignin monomers. In this study we investigate lignin demethylation and polyhydroxyl-aromatic production in BC and C horizons of sandy forest soils dominated by oak, the A horizon from a red spruce forest, and controlled microbial inoculation studies of woody tissue using in-line 13C-labeled tetramethylammonium hydroxide thermochemolysis. Both white-rot and brown-rot decay resulted in syringyl demethylation, with the latter exhibiting more aggressive demethylation chemistry, while coniferyl monomer demethylation was essentially restricted to brown-rot decay. In a typical brown-rot sequence demethylation of syringyl components occurs more rapidly than coniferyl units within the same tissue and lower molecular weight fragments are likewise more demethylated than lignin monomers containing the full glycerol side chain. Demethylation of both methoxyl groups in the syringyl monomer is evident in soil horizons as well as laboratory inoculations. The latter may suggest demethylation after lignin depolymerization. Low molecular weight

  15. DNA-demethylating and anti-tumor activity of synthetic miR-29b mimics in multiple myeloma.

    PubMed

    Amodio, Nicola; Leotta, Marzia; Bellizzi, Dina; Di Martino, Maria Teresa; D'Aquila, Patrizia; Lionetti, Marta; Fabiani, Fernanda; Leone, Emanuela; Gullà, Anna Maria; Passarino, Giuseppe; Caraglia, Michele; Negrini, Massimo; Neri, Antonino; Giordano, Antonio; Tagliaferri, Pierosandro; Tassone, Pierfrancesco

    2012-10-01

    Aberrant DNA methylation plays a relevant role in multiple myeloma (MM) pathogenesis. MicroRNAs (miRNAs) are a class of small non-coding RNAs that recently emerged as master regulator of gene expression by targeting protein-coding mRNAs. However, miRNAs involvement in the regulation of the epigenetic machinery and their potential use as therapeutics in MM remain to be investigated. Here, we provide evidence that the expression of de novo DNA methyltransferases (DNMTs) is deregulated in MM cells. Moreover, we show that miR-29b targets DNMT3A and DNMT3B mRNAs and reduces global DNA methylation in MM cells. In vitro transfection of MM cells with synthetic miR-29b mimics significantly impairs cell cycle progression and also potentiates the growth-inhibitory effects induced by the demethylating agent 5-azacitidine. Most importantly, in vivo intratumor or systemic delivery of synthetic miR-29b mimics, in two clinically relevant murine models of human MM, including the SCID-synth-hu system, induces significant anti-tumor effects. All together, our findings demonstrate that aberrant DNMTs expression is efficiently modulated by tumor suppressive synthetic miR-29b mimics, indicating that methyloma modulation is a novel matter of investigation in miRNA-based therapy of MM.

  16. DNA-demethylating and anti-tumor activity of synthetic miR-29b mimics in multiple myeloma

    PubMed Central

    Amodio, Nicola; Leotta, Marzia; Bellizzi, Dina; Di Martino, Maria Teresa; D'Aquila, Patrizia; Lionetti, Marta; Fabiani, Fernanda; Leone, Emanuela; Gullà, Anna Maria; Passarino, Giuseppe; Caraglia, Michele; Negrini, Massimo; Neri, Antonino; Giordano, Antonio; Tagliaferri, Pierosandro; Tassone, Pierfrancesco

    2012-01-01

    Aberrant DNA methylation plays a relevant role in multiple myeloma (MM) pathogenesis. MicroRNAs (miRNAs) are a class of small non-coding RNAs that recently emerged as master regulator of gene expression by targeting protein-coding mRNAs. However, miRNAs involvement in the regulation of the epigenetic machinery and their potential use as therapeutics in MM remain to be investigated. Here, we provide evidence that the expression of de novo DNA methyltransferases (DNMTs) is deregulated in MM cells. Moreover, we show that miR-29b targets DNMT3A and DNMT3B mRNAs and reduces global DNA methylation in MM cells. In vitro transfection of MM cells with synthetic miR-29b mimics significantly impairs cell cycle progression and also potentiates the growth-inhibitory effects induced by the demethylating agent 5-azacitidine. Most importantly, in vivo intratumor or systemic delivery of synthetic miR-29b mimics, in two clinically relevant murine models of human MM, including the SCID-synth-hu system, induces significant anti-tumor effects. All together, our findings demonstrate that aberrant DNMTs expression is efficiently modulated by tumor suppressive synthetic miR-29b mimics, indicating that methyloma modulation is a novel matter of investigation in miRNA-based therapy of MM. PMID:23100393

  17. A pair of transposon-derived proteins function in a histone acetyltransferase complex for active DNA demethylation

    PubMed Central

    Duan, Cheng-Guo; Wang, Xingang; Xie, Shaojun; Pan, Li; Miki, Daisuke; Tang, Kai; Hsu, Chuan-Chih; Lei, Mingguang; Zhong, Yingli; Hou, Yueh-Ju; Wang, Zhijuan; Zhang, Zhengjing; Mangrauthia, Satendra K; Xu, Huawei; Zhang, Heng; Dilkes, Brian; Tao, W Andy; Zhu, Jian-Kang

    2017-01-01

    Transposons are generally kept silent by epigenetic mechanisms including DNA methylation. Here, we identified a pair of Harbinger transposon-derived proteins (HDPs), HDP1 and HDP2, as anti-silencing factors in Arabidopsis. hdp1 and hdp2 mutants displayed an enhanced silencing of transgenes and some transposons. Phylogenetic analyses revealed that HDP1 and HDP2 were co-domesticated from the Harbinger transposon-encoded transposase and DNA-binding protein, respectively. HDP1 interacts with HDP2 in the nucleus, analogous to their transposon counterparts. Moreover, HDP1 and HDP2 are associated with IDM1, IDM2, IDM3 and MBD7 that constitute a histone acetyltransferase complex functioning in DNA demethylation. HDP2 and the methyl-DNA-binding protein MBD7 share a large set of common genomic binding sites, indicating that they jointly determine the target specificity of the histone acetyltransferase complex. Thus, our data revealed that HDP1 and HDP2 constitute a functional module that has been recruited to a histone acetyltransferase complex to prevent DNA hypermethylation and epigenetic silencing. PMID:27934869

  18. [Effects of phospholipids on oxidative demethylation of dimethylcyclohexylamine by cumene hydroperoxide involving methemoglobin].

    PubMed

    Kiseleva, S N; Khatyleva, S Iu; Kisel', M A; Kiselev, P A; Akhrem, A A

    1981-12-01

    The reaction of oxidative demethylation of N-dimethylcyclohexylamine by cumene hydroperoxide involving methemoglobin was studied. Data from differential spectroscopy and kinetic analysis revealed the formation of a methemoglobin--N-dimethylcyclohexylamine--cumene hydroperoxide complex. The inhibiting analysis revealed the radical stages in the process of demethylation. An addition to the reaction mixture of phosphatidyl serine, phosphatidyl inositol and lysophosphatidyl choline at a ratio of 50 divided by 500 molecules per 1 molecule of protein increased the rate of the reaction product accumulation 2--3-fold. Phosphatidyl choline and the ionic detergent sodium cholate did not practically affect the reaction rate under the given experimental conditions. The nature of the activating effect of some phospholipids on oxidative demethylation is discussed.

  19. Regioselective demethylation of quinoline derivatives. A DFT rationalization

    NASA Astrophysics Data System (ADS)

    Belferdi, Fatiha; Merabet, Naima; Belkhiri, Lotfi; Douara, Bachir

    2016-08-01

    Demethylation of compound 2,7-dimethoxyquinoline-3-carbaldehyde 1, is carried out using BBr3. However, all attempts led, either to the starting material or to the regioselective demethylation at position 2 affording the product 4a. The nature (donor or acceptor) and the position of the R (CHO or CN) group is likely to play a role in the preventing the demethylation at position 7. To address this phenomena, the demethylation of 2-chloro-7-methoxyquinoline-3-carbaldehyde 2 and 2,7-dimethoxyquinoline-3-carbaldehyde 3 has been carried out. To support the results obtained, theoretical computations at DFT level (vide infra) have been carried out upon compound 1. The exploration of how the gas-phase demethylation process on Quinoline can be affected at a position 7 center by stepwise substation effects using different electro-donor and attractor groups, show that demethylation process seems to be more favorable when substituent is an electro-donor. This is sustained by bond energy and thermodynamic analyses (vide infra).

  20. NF-κB mediates Gadd45β expression and DNA demethylation in the hippocampus during fear memory formation

    PubMed Central

    Jarome, Timothy J.; Butler, Anderson A.; Nichols, Jessica N.; Pacheco, Natasha L.; Lubin, Farah D.

    2015-01-01

    Gadd45-mediated DNA demethylation mechanisms have been implicated in the process of memory formation. However, the transcriptional mechanisms involved in the regulation of Gadd45 gene expression during memory formation remain unexplored. NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) controls transcription of genes in neurons and is a critical regulator of synaptic plasticity and memory formation. In silico analysis revealed several NF-κB (p65/RelA and cRel) consensus sequences within the Gadd45β gene promoter. Whether NF-κB activity regulates Gadd45 expression and associated DNA demethylation in neurons during memory formation is unknown. Here, we found that learning in a fear conditioning paradigm increased Gadd45β gene expression and brain-derivedneurotrophic factor (BDNF) DNA demethylation in area CA1 of the hippocampus, both of which were prevented with pharmacological inhibition of NF-κB activity. Further experiments found that conditional mutations in p65/RelA impaired fear memory formation but did not alter changes in Gadd45β expression. The learning-induced increases in Gadd45β mRNA levels, Gadd45β binding at the BDNF gene and BDNF DNA demethylation were blocked in area CA1 of the c-rel knockout mice. Additionally, local siRNA-mediated knockdown of c-rel in area CA1 prevented fear conditioning-induced increases in Gadd45β expression and BDNF DNA demethylation, suggesting that c-Rel containing NF-κB transcription factor complex is responsible for Gadd45β regulation during memory formation. Together, these results support a novel transcriptional role for NF-κB in regulation of Gadd45β expression and DNA demethylation in hippocampal neurons during fear memory. PMID:26441517

  1. Minimal role of base excision repair in TET-induced global DNA demethylation in HEK293T cells

    PubMed Central

    Jin, Chunlei; Qin, Taichun; Barton, Michelle Craig; Jelinek, Jaroslav; Issa, Jean-Pierre J

    2015-01-01

    Oxidation of 5-methylcytosine by TET family proteins can induce DNA replication-dependent (passive) DNA demethylation and base excision repair (BER)-based (active) DNA demethylation. The balance of active vs. passive TET-induced demethylation remains incompletely determined. In the context of large scale DNA demethylation, active demethylation may require massive induction of the DNA repair machinery and thus compromise genome stability. To study this issue, we constructed a tetracycline-controlled TET-induced global DNA demethylation system in HEK293T cells. Upon TET overexpression, we observed induction of DNA damage and activation of a DNA damage response; however, BER genes are not upregulated to promote DNA repair. Depletion of TDG (thymine DNA glycosylase) or APEX1 (apurinic/apyrimidinic endonuclease 1), two key BER enzymes, enhances rather than impairs global DNA demethylation, which can be explained by stimulated proliferation. By contrast, growth arrest dramatically blocks TET-induced global DNA demethylation. Thus, in the context of TET-induction in HEK293T cells, the DNA replication-dependent passive mechanism functions as the predominant pathway for global DNA demethylation. In the same context, BER-based active demethylation is markedly restricted by limited BER upregulation, thus potentially preventing a disastrous DNA damage response to extensive active DNA demethylation. PMID:26440216

  2. Mercury demethylation in waterbird livers: Dose-response thresholds and differences among species

    USGS Publications Warehouse

    Eagles-Smith, C. A.; Ackerman, J.T.; Julie, Y.E.E.; Adelsbach, T.L.

    2009-01-01

    We assessed methylmercury (MeHg) demethylation in the livers of adults and chicks of four waterbird species that commonly breed in San Francisco Bay: American avocets, black-necked stilts, Caspian terns, and Forster's terns. In adults (all species combined), we found strong evidence for a threshold, model where MeHg demethylation occurred above a hepatic total mercury concentration threshold of 8.51 ?? 0.93 ??g/g dry weight, and there was a strong decline in %MeHg values as total mercury (THg) concentrations increased above 8.51 ??g/g dry weight. Conversely, there was no evidence for a demethylation threshold in chicks, and we found that %MeHg values declined linearly with increasing THg concentrations. For adults, we also found taxonomie differences in the demethylation responses, with avocets and stilts showing a higher demethylation rate than that of terns when concentrations exceeded the threshold, whereas terns had a lower demethylation threshold (7.48 ?? 1.48 ??g/g dry wt) than that of avocets and stilts (9.91 ?? 1.29 ??g/g dry wt). Finally, we assessed the role of selenium (Se) in the demethylation process. Selenium concentrations were positively correlated with inorganic Hg in livers of birds above the demethylation threshold but not below. This suggests that Se may act as a binding site for demethylated Hg and may reduce the potential for secondary toxicity. Our findings indicate that waterbirds demethylate mercury in their livers if exposure exceeds a threshold value and suggest that taxonomie differences in demethylation ability may be an important factor in evaluating species-specific risk to MeHg exposure. Further, we provide strong evidence for a threshold of approximately 8.5 ??g/g dry weight of THg in the liver where demethylation is initiated. ?? 2009 SETAC.

  3. All-trans retinoic acid enhances the effect of 5-aza-2'-deoxycytidine on p16INK4a demethylation, and the two drugs synergistically activate retinoic acid receptor β gene expression in the human erythroleukemia K562 cell line.

    PubMed

    Xiang, Lili; Dong, Weimin; Wang, Rong; Wei, Jiang; Qiu, Guoqiang; Cen, Jiannong; Chen, Zixing; Zheng, Xiao; Hu, Shaoyan; Xie, Xiaobao; Cao, Xiangshan; Gu, Weiying

    2014-07-01

    The aim of the current study was to investigate the antineoplastic activities of 5-aza-2'-deoxycytidine (also known as decitabine; DAC) and all-trans retinoic acid (ATRA), administered alone or in combination, in K562 cells in vitro, as well as the effects on the expression of the tumor suppressor genes, p16INK4a (p16) and retinoic acid receptor β (RAR-β). Cell growth inhibition, differentiation and apoptosis in K562 cells treated with DAC and/or ATRA were detected. The methylation of the p16 and RAR-β genes in the K562 cells was detected using the methylation-specific polymerase chain reaction (PCR) method. Quantitative PCR was used for the detection of the mRNA expression of the p16 and RAR-β genes, and western blot analysis was used to detect protein expression. DAC and ATRA, alone or in combination, had no effect on the growth inhibition, differentiation and apoptosis of the K562 cells. DAC alone induced the demethylation of the p16 gene, and combination of DAC and ATRA demonstrated more evident demethylation of the p16 gene, however, ATRA alone had no effect on methylation. The RAR-β promoter region was not methylated in the K562 cells. DAC in combination with ATRA appeared to produce a greater activation of the RAR-β gene, which led to the upregulation of the RAR-β expression level. ATRA enhanced the effect of DAC on p16 demethylation, and the combination of the two drugs was found to activate RAR-β expression, which indicated that DAC used in combination with ATRA has clinical potential in the treatment of human erythroleukemia.

  4. Anaerobic Mercury Methylation and Demethylation by Geobacter bemidjiensis Bem

    DOE PAGES

    Lu, Xia; Liu, Yurong; Johs, Alexander; ...

    2016-03-28

    Two competing processes controlling the net production and bioaccumulation of neurotoxic methylmercury (MeHg) in natural ecosystems are microbial methylation and demethylation. Though mercury (Hg) methylation by anaerobic microorganisms and demethylation by aerobic Hg-resistant bacteria have both been extensively studied, little attention has been given to MeHg degradation by anaerobic bacteria, particularly the iron-reducing bacterium Geobacter bemidjensis Bem. Here we report, for the first time, that the strain G. bemidjensis Bem can methylate inorganic Hg and degrade MeHg concurrently under anoxic conditions. Our results suggest that G. bemidjensis cells utilize a reductive demethylation pathway to degrade MeHg, with elemental Hg(0) asmore » the major reaction product, possibly due to the presence of homologs encoding both organo-mercurial lyase (MerB) and mercuric reductase (MerA) in this organism. In addition, the cells can mediate multiple reactions including Hg/MeHg sorption, Hg reduction and oxidation, resulting in both time and concentration dependent Hg species transformations. Moderate concentrations (10 500 M) of Hg-binding ligands such as cysteine enhance Hg(II) methylation but inhibit MeHg degradation. These findings indicate a cycle of methylation and demethylation among anaerobic bacteria and suggest that mer-mediated demethylation may play a role in the net balance of MeHg production in anoxic water and sediments.« less

  5. Biochemistry and Occurrence of O-Demethylation in Plant Metabolism

    PubMed Central

    Hagel, Jillian M.; Facchini, Peter J.

    2010-01-01

    Demethylases play a pivitol role in numerous biological processes from covalent histone modification and DNA repair to specialized metabolism in plants and microorganisms. Enzymes that catalyze O- and N-demethylation include 2-oxoglutarate (2OG)/Fe(II)-dependent dioxygenases, cytochromes P450, Rieske-domain proteins and flavin adenine dinucleotide (FAD)-dependent oxidases. Proposed mechanisms for demethylation by 2OG/Fe(II)-dependent enzymes involve hydroxylation at the O- or N-linked methyl group followed by formaldehyde elimination. Members of this enzyme family catalyze a wide variety of reactions in diverse plant metabolic pathways. Recently, we showed that 2OG/Fe(II)-dependent dioxygenases catalyze the unique O-demethylation steps of morphine biosynthesis in opium poppy, which provides a rational basis for the widespread occurrence of demethylases in benzylisoquinoline alkaloid metabolism. PMID:21423357

  6. In vitro identification of the cytochrome P450 isozymes involved in the N-demethylation of the active opioid metabolite nortilidine to bisnortilidine.

    PubMed

    Wustrow, Isabel; Riedel, Klaus-Dieter; Mikus, Gerd; Weiss, Johanna

    2012-06-01

    Tilidine exhibits the highest consumption of opioids in Germany. The prodrug is hepatically metabolised in a sequential N-demethylation reaction. Its primary metabolite nortilidine is a selective μ-opioid receptor agonist which can penetrate the blood-brain barrier. Cytochrome P450 isozymes (CYP) 3A4 and CYP2C19 were previously identified as isozymes mediating the formation of nortilidine. This study was set up to identify the enzymes and kinetics of the subsequent N-demethylation to bisnortilidine, thus being able to understand clinical interactions. Human liver microsomes and recombinant CYPs were used to investigate the metabolism of nortilidine to bisnortilidine. Nortilidine and bisnortilidine were quantified using liquid chromatography tandem mass spectrometry. Inhibitor screening kits were used to quantify the inhibition of CYP3A4, CYP2C19, CYP2B6 and CYP2D6 by bisnortilidine. Nortilidine metabolism to bisnortilidine followed the Michaelis-Menten kinetics with K (m) = 141.6 ± 15 μM and V (max) = 46.2 ± 3 nmol/mg/h. Inhibitors of CYP3A4, CYP2C19 and CYP2B6 inhibited this reaction. Assays with recombinant CYPs verified that the N-demethylation is catalysed by CYP3A4, CYP2C19 and CYP2B6. Our results also demonstrated that the metabolism from tilidine to nortilidine is not only mediated by CYP3A4 and CYP2C19, but also by CYP2B6. Moreover, bisnortilidine is a weak inhibitor of CYP3A4 and CYP2B6, a strong inhibitor of CYP2D6, but not an inhibitor of CYP2C19. Our study demonstrated that nortilidine is metabolised via the same CYP isozymes as the prodrug tilidine, whereas the formation of bisnortilidine appears to be the rate-limiting step in the metabolism of tilidine. Pharmacokinetic interactions can be expected with inhibitors or inducers of CYP3A4, CYP2C19 or CYP2B6.

  7. TET1 promotes cisplatin-resistance via demethylating the vimentin promoter in ovarian cancer.

    PubMed

    Han, Xi; Zhou, Yuanyuan; You, Yuanyi; Lu, Jiaojiao; Wang, Lijie; Hou, Huilian; Li, Jing; Chen, Wei; Zhao, Le; Li, Xu

    2017-04-01

    The development of chemo-resistance impairs the outcome of the first line platinum-based chemotherapies for ovarian cancer. Deregulation of DNA methylation/demethylation provides a critical mechanism for the occurrence of chemo-resistance. The ten-eleven translocation (TET) family of dioxygenases including TET1/2/3 plays an important part in DNA demethylation, but their roles in cisplatin resistance have not been elucidated. Using cisplatin-sensitive and cisplatin-resistant ovarian cancer cell models, we found that TET1 was significantly upregulated in cisplatin-resistant CP70 cells compared with that in cisplatin-sensitive A2780 cells. Ectopic expression of TET1 in A2780 cells promoted cisplatin resistance and decreased cytotoxicity induced by cisplatin, while inhibition of TET1 by siRNA transfection in CP70 cells attenuated cisplatin resistance and enhanced cytotoxicity of cisplatin. Increased TET1 induced re-expression of vimentin through active DNA demethylation, and cause partial epithelial-to-mesenchymal (EMT) in A2780 cells. Contrarily, knocking down of TET1 in CP70 cells reduced vimentin expression and reversed EMT process. Immunohistochemical analysis of TET1 in human ovarian cancer tissues revealed that TET1 existed in nucleus and cytoplasm in ovarian cancer tissues. And the expression of nuclear TET1 was positively correlated with residual tumor and chemotherapeutic response. Thus, TET1 expression causes resistance to cisplatin and one of the targets of TET1 action is vimentin in ovarian cancer.

  8. Redistribution of demethylated RNA helicase A during foot-and-mouth disease virus infection: Role of Jumonji C-domain containing protein 6 in RHA demethylation

    SciTech Connect

    Lawrence, Paul; Conderino, Joseph S.; Rieder, Elizabeth

    2014-03-15

    Previously, RNA helicase A (RHA) re-localization from the nucleus to the cytoplasm in foot-and-mouth disease virus (FMDV) infected cells was shown to coincide with loss of RHA methylated arginine residues at its C-terminus. The potential interaction between RHA and Jumonji C-domain (JmjC) protein 6 (JMJD6) arginine demethylase in infected cells was investigated. Treatment with N-oxalylglycine (NOG) inhibitor of JmjC demethylases prevented FMDV-induced RHA demethylation and re-localization, and also decreased viral protein synthesis and virus titers. Physical interaction between JMJD6 and RHA was demonstrated via reciprocal co-immunoprecipitation, where RHA preferentially bound JMJD6 monomers. Nuclear efflux of demethylated RHA (DM-RHA) coincided with nuclear influx of JMJD6, which was not observed using another picornavirus. A modified biochemical assay demonstrated JMJD6 induced dose-dependent demethylation of RHA and two RHA-derived isoforms, which could be inhibited by NOG. We propose a role for JMJD6 in RHA demethylation stimulated by FMDV, that appears to facilitate virus replication. - Highlights: • We examined the role of JMJD6 in FMDV-induced RHA demethylation process. • Using an arginine demethylation assay showed that JMJD6 is involved in RHA demethylation. • A demethylases inhibitor reduced cytoplasmic accumulation of RHA and FMDV titers.

  9. Non-small-cell lung cancer-induced immunosuppression by increased human regulatory T cells via Foxp3 promoter demethylation.

    PubMed

    Ke, Xing; Zhang, Shuping; Xu, Jian; Liu, Genyan; Zhang, Lixia; Xie, Erfu; Gao, Li; Li, Daqian; Sun, Ruihong; Wang, Fang; Pan, Shiyang

    2016-05-01

    Patients with non-small-cell lung cancer (NSCLC) have immune defects that are poorly understood. Forkhead box protein P3 (Foxp3) is crucial for immunosuppression by CD4(+) regulatory T cells (Tregs). It is not well known how NSCLC induces Foxp3 expression and causes immunosuppression in tumor-bearing patients. Our study found a higher percentage of CD4(+) Tregs in the peripheral blood of NSCLC compared with healthy donors. NSCLC patients showed demethylation of eight CpG sites within the Foxp3 promoter with methylation ratios negatively correlated with CD4(+)CD25(+)Foxp3(+) T levels. Foxp3 expression in CD4(+) Tregs was directly regulated by Foxp3 promoter demethylation and was involved in immunosuppression by NSCLC. To verify the effect of tumor cells on the phenotype and function of CD4(+) Tregs, we established a coculture system using NSCLC cell line and healthy CD4(+) T cells and showed that SPC-A1 induced IL-10 and TGF-β1 secretion by affecting the function of CD4(+) Tregs. The activity of DNA methyltransferases from CD4(+) T was decreased during this process. Furthermore, eight CpG sites within the Foxp3 promoter also appeared to have undergone demethylation. Foxp3 is highly expressed in CD4(+) T cells, and this may be caused by gene promoter demethylation. These induced Tregs are highly immunosuppressive and dramatically inhibit the proliferative activity of naïve CD4(+) T cells. Our study provides one possible mechanism describing Foxp3 promoter demethylation changes by which NSCLC down-regulates immune responses and contributes to tumor progression. Foxp3 represents an important target for NSCLC anti-tumor immunotherapy.

  10. A promoter DNA demethylation landscape of human hematopoietic differentiation

    PubMed Central

    Calvanese, Vincenzo; Fernández, Agustín F.; Urdinguio, Rocío G.; Suárez-Alvarez, Beatriz; Mangas, Cristina; Pérez-García, Vicente; Bueno, Clara; Montes, Rosa; Ramos-Mejía, Verónica; Martínez-Camblor, Pablo; Ferrero, Cecilia; Assenov, Yassen; Bock, Christoph; Menendez, Pablo; Carrera, Ana Clara; Lopez-Larrea, Carlos; Fraga, Mario F.

    2012-01-01

    Global mechanisms defining the gene expression programs specific for hematopoiesis are still not fully understood. Here, we show that promoter DNA demethylation is associated with the activation of hematopoietic-specific genes. Using genome-wide promoter methylation arrays, we identified 694 hematopoietic-specific genes repressed by promoter DNA methylation in human embryonic stem cells and whose loss of methylation in hematopoietic can be associated with gene expression. The association between promoter methylation and gene expression was studied for many hematopoietic-specific genes including CD45, CD34, CD28, CD19, the T cell receptor (TCR), the MHC class II gene HLA-DR, perforin 1 and the phosphoinositide 3-kinase (PI3K) and results indicated that DNA demethylation was not always sufficient for gene activation. Promoter demethylation occurred either early during embryonic development or later on during hematopoietic differentiation. Analysis of the genome-wide promoter methylation status of induced pluripotent stem cells (iPSCs) generated from somatic CD34+ HSPCs and differentiated derivatives from CD34+ HSPCs confirmed the role of DNA methylation in regulating the expression of genes of the hemato-immune system, and indicated that promoter methylation of these genes may be associated to stemness. Together, these data suggest that promoter DNA demethylation might play a role in the tissue/cell-specific genome-wide gene regulation within the hematopoietic compartment. PMID:21911366

  11. Anaerobic Mercury Methylation and Demethylation by Geobacter bemidjiensis Bem.

    PubMed

    Lu, Xia; Liu, Yurong; Johs, Alexander; Zhao, Linduo; Wang, Tieshan; Yang, Ziming; Lin, Hui; Elias, Dwayne A; Pierce, Eric M; Liang, Liyuan; Barkay, Tamar; Gu, Baohua

    2016-04-19

    Microbial methylation and demethylation are two competing processes controlling the net production and bioaccumulation of neurotoxic methylmercury (MeHg) in natural ecosystems. Although mercury (Hg) methylation by anaerobic microorganisms and demethylation by aerobic Hg-resistant bacteria have both been extensively studied, little attention has been given to MeHg degradation by anaerobic bacteria, particularly the iron-reducing bacterium Geobacter bemidjiensis Bem. Here we report, for the first time, that the strain G. bemidjiensis Bem can mediate a suite of Hg transformations, including Hg(II) reduction, Hg(0) oxidation, MeHg production and degradation under anoxic conditions. Results suggest that G. bemidjiensis utilizes a reductive demethylation pathway to degrade MeHg, with elemental Hg(0) as the major reaction product, possibly due to the presence of genes encoding homologues of an organomercurial lyase (MerB) and a mercuric reductase (MerA). In addition, the cells can strongly sorb Hg(II) and MeHg, reduce or oxidize Hg, resulting in both time and concentration-dependent Hg species transformations. Moderate concentrations (10-500 μM) of Hg-binding ligands such as cysteine enhance Hg(II) methylation but inhibit MeHg degradation. These findings indicate a cycle of Hg methylation and demethylation among anaerobic bacteria, thereby influencing net MeHg production in anoxic water and sediments.

  12. Methylmercury decomposition in sediments and bacterial cultures: Involvement of methanogens and sulfate reducers in oxidative demethylation

    USGS Publications Warehouse

    Oremland, R.S.; Culbertson, C.W.; Winfrey, M.R.

    1991-01-01

    Demethylation of monomethylmercury in freshwater and estuarine sediments and in bacterial cultures was investigated with 14CH3HgI. Under anaerobiosis, results with inhibitors indicated partial involvement of both sulfate reducers and methanogens, the former dominating estuarine sediments, while both were active in freshwaters. Aerobes were the most significant demethylators in estuarine sediments, but were unimportant in freshwater sediments. Products of anaerobic demethylation were mainly 14CO2 as well as lesser amounts of 14CH4. Acetogenic activity resulted in fixation of some 14CO2 produced from 14CH3HgI into acetate. Aerobic demethylation in estuarine sediments produced only 14CH4, while aerobic demethylation in freshwater sediments produced small amounts of both 14CH4 and 14CO2. Two species of Desulfovibrio produced only traces of 14CH4 from 14CH3HgI, while a culture of a methylotrophic methanogen formed traces of 14CO2 and 14CH4 when grown on trimethylamine in the presence of the 14CH3HgI. These results indicate that both aerobes and anaerobes demethylate mercury in sediments, but that either group may dominate in a particular sediment type. Aerobic demethylation in the estuarine sediments appeared to proceed by the previously characterized organomercurial-lyase pathway, because methane was the sole product. However, aerobic demethylation in freshwater sediments as well as anaerobic demethylation in all sediments studied produced primarily carbon dioxide. This indicates the presence of an oxidative pathway, possibly one in which methylmercury serves as an analog of one-carbon substrates.

  13. Oxidative demethylation of 2-picolines on vanadium oxide catalysts

    SciTech Connect

    Suvorov, B.V.; Glubokovskikh, L.K.; Demin, V.V.; Kan, I.I.

    1988-07-10

    One of the known methods for the preparation of pyridine is based on the dealkylation of alkylpyridines in the presence of vanadium-containing catalysts, molecular oxygen and steam. By using the oxidative demethylation of 2-picoline in the presence of steam on a fused vanadium(V) oxide, pyridine can be obtained in a yield of up to 88% of theory. To lower the consumption of vanadium(V) oxide and increase the thermostability of the catalyst, they studied the possible use of V/sub 2/O/sub 5/ catalysts on various carriers (diatomite, silica gel, porcelain balls), including the industrially produced brand SVD and SVS catalysts. The SVS brand catalyst has a satisfactory activity and selectivity in the oxidation demethylation of 2-picoline into pyridine. Under optimal conditions, pyridine is formed on this catalyst in a yield of 88% of the theoretical.

  14. DNA methylation and demethylation as targets for antipsychotic therapy.

    PubMed

    Guidotti, Alessandro; Grayson, Dennis R

    2014-09-01

    Schizophrenia (SZ) and bipolar disorder (BPD) patients show a downregulation of GAD67, reelin (RELN), brain-derived neurotrophic factor (BDNF), and other genes expressed in telencephalic GABAergic and glutamatergic neurons. This downregulation is associated with the enrichment of 5-methylcytosine and 5-hydroxymethylcytosine proximally at gene regulatory domains at the respective genes. A pharmacological strategy to reduce promoter hypermethylation and to induce a more permissive chromatin conformation is to administer drugs, such as the histone deacetylase (HDAC) inhibitor valproate (VPA), that facilitate chromatin remodeling. Studies in mouse models of SZ indicate that clozapine induces DNA demethylation at relevant promoters, and that this action is potentiated by VPA. By activating DNA demethylation, clozapine or its derivatives with VPA or other more potent and selective HDAC inhibitors may be a promising treatment strategy to correct the gene expression deficits detected in postmortem brain of SZ and BPD patients.

  15. Method for recovering and using lignin in adhesive resins by extracting demethylated lignin

    DOEpatents

    Schroeder, Herbert A.

    1991-01-01

    Lignin, or a lignin derived material, which has been significantly demethylated (e.g., the demethylated lignin found in the raffinate produced as a by-product of dimethyl sulfide production which can be carried out using the spent liquor from wood pulping operations) can be isolated by a process wherein an organic solvent is added to a lignin-containing aqueous solution. The organic solvent is typically a polar, and at least a partially water-immiscible substance such as, for example, ethyl acetate. The resulting lignin-containing aqueous solution/organic solvent mixture is acidified to produce a water layer which is discarded and an organic solvent layer which contains the demethylated lignin. Upon its recovery, the demethylated lignin is preferably dried and stored until it is used (along with an alkali, an aldehyde and an adhesive filler) in compounding an adhesive of the type generally used in the manufacture of plywood.

  16. Method for recovering and using lignin in adhesive resins by extracting demethylated lignin

    DOEpatents

    Schroeder, Herbert A.

    1991-01-01

    Lignin, or a lignin derived material, which has been significantly demethylated (e.g., the demethylated lignin found in the raffinate produced as a by-product of dimethyl sulfide production which can be carried out using the spent liquor from wood pulping operations) can be isolated by a process wherein an organic solvent is added to a lignin-containing aqueous solution. The organic solvent is typically a polar, and at least a partially water-immiscible substance such as, for example, ethyl acetate. The resulting lignin-containing aqueous solution/organic solvent mixture is acidified to produce a water layer which is discarded and an organic solvent layer which contains the demethylated lignin. Upon its recovery, the demethylated lignin is dissolved in an alkaline solution to which an aldehyde source is added to produce a resol-type resin. The aldehyde source may be formaldehyde in solution, paraformaldehyde, hexamethylenetetramine, or other aldehydes including acetaldehyde, furfural, and their derivatives.

  17. Demethylation initiated by ROS1 glycosylase involves random sliding along DNA

    PubMed Central

    Ponferrada-Marín, María Isabel; Roldán-Arjona, Teresa; Ariza, Rafael R.

    2012-01-01

    Active DNA demethylation processes play a critical role in shaping methylation patterns, yet our understanding of the mechanisms involved is still fragmented and incomplete. REPRESSOR OF SILENCING 1 (ROS1) is a prototype member of a family of plant 5-methylcytosine DNA glycosylases that initiate active DNA demethylation through a base excision repair pathway. As ROS1 binds DNA non-specifically, we have critically tested the hypothesis that facilitated diffusion along DNA may contribute to target location by the enzyme. We have found that dissociation of ROS1 from DNA is severely restricted when access to both ends is obstructed by tetraloops obstacles. Unblocking any end facilitates protein dissociation, suggesting that random surface sliding is the main route to a specific target site. We also found that removal of the basic N-terminal domain of ROS1 significantly impairs the sliding capacity of the protein. Finally, we show that sliding increases the catalytic efficiency of ROS1 on 5-meC:G pairs, but not on T:G mispairs, thus suggesting that the enzyme achieves recognition and excision of its two substrate bases by different means. A model is proposed to explain how ROS1 finds its potential targets on DNA. PMID:23034804

  18. Demethylation initiated by ROS1 glycosylase involves random sliding along DNA.

    PubMed

    Ponferrada-Marín, María Isabel; Roldán-Arjona, Teresa; Ariza, Rafael R

    2012-12-01

    Active DNA demethylation processes play a critical role in shaping methylation patterns, yet our understanding of the mechanisms involved is still fragmented and incomplete. REPRESSOR OF SILENCING 1 (ROS1) is a prototype member of a family of plant 5-methylcytosine DNA glycosylases that initiate active DNA demethylation through a base excision repair pathway. As ROS1 binds DNA non-specifically, we have critically tested the hypothesis that facilitated diffusion along DNA may contribute to target location by the enzyme. We have found that dissociation of ROS1 from DNA is severely restricted when access to both ends is obstructed by tetraloops obstacles. Unblocking any end facilitates protein dissociation, suggesting that random surface sliding is the main route to a specific target site. We also found that removal of the basic N-terminal domain of ROS1 significantly impairs the sliding capacity of the protein. Finally, we show that sliding increases the catalytic efficiency of ROS1 on 5-meC:G pairs, but not on T:G mispairs, thus suggesting that the enzyme achieves recognition and excision of its two substrate bases by different means. A model is proposed to explain how ROS1 finds its potential targets on DNA.

  19. Monkey liver cytochrome P450 2C9 is involved in caffeine 7-N-demethylation to form theophylline.

    PubMed

    Utoh, Masahiro; Murayama, Norie; Uno, Yasuhiro; Onose, Yui; Hosaka, Shinya; Fujino, Hideki; Shimizu, Makiko; Iwasaki, Kazuhide; Yamazaki, Hiroshi

    2013-12-01

    Caffeine (1,3,7-trimethylxanthine) is a phenotyping substrate for human cytochrome P450 1A2. 3-N-Demethylation of caffeine is the main human metabolic pathway, whereas monkeys extensively mediate the 7-N-demethylation of caffeine to form pharmacological active theophylline. Roles of monkey P450 enzymes in theophylline formation from caffeine were investigated using individual monkey liver microsomes and 14 recombinantly expressed monkey P450 enzymes, and the results were compared with those for human P450 enzymes. Caffeine 7-N-demethylation activity in microsomes from 20 monkey livers was not strongly inhibited by α-naphthoflavone, quinidine or ketoconazole, and was roughly correlated with diclofenac 4'-hydroxylation activities. Monkey P450 2C9 had the highest activity for caffeine 7-N-demethylation. Kinetic analysis revealed that monkey P450 2C9 had a high Vmax/Km value for caffeine 7-N-demethylation, comparable to low Km value for monkey liver microsomes. Caffeine could dock favorably with monkey P450 2C9 modeled for 7-N-demethylation and with human P450 1A2 for 3-N-demethylation. The primary metabolite theophylline was oxidized to 8-hydroxytheophylline in similar ways by liver microsomes and by recombinant P450s in both humans and monkeys. These results collectively suggest a high activity for monkey liver P450 2C9 toward caffeine 7-N-demethylation, whereas, in humans, P450 1A2-mediated caffeine 3-N-demethylation is dominant.

  20. DNA demethylation is initiated in the central cells of Arabidopsis and rice

    PubMed Central

    Park, Kyunghyuk; Kim, M. Yvonne; Vickers, Martin; Park, Jin-Sup; Hyun, Youbong; Okamoto, Takashi; Zilberman, Daniel; Fischer, Robert L.; Feng, Xiaoqi; Choi, Yeonhee; Scholten, Stefan

    2016-01-01

    Cytosine methylation is a DNA modification with important regulatory functions in eukaryotes. In flowering plants, sexual reproduction is accompanied by extensive DNA demethylation, which is required for proper gene expression in the endosperm, a nutritive extraembryonic seed tissue. Endosperm arises from a fusion of a sperm cell carried in the pollen and a female central cell. Endosperm DNA demethylation is observed specifically on the chromosomes inherited from the central cell in Arabidopsis thaliana, rice, and maize, and requires the DEMETER DNA demethylase in Arabidopsis. DEMETER is expressed in the central cell before fertilization, suggesting that endosperm demethylation patterns are inherited from the central cell. Down-regulation of the MET1 DNA methyltransferase has also been proposed to contribute to central cell demethylation. However, with the exception of three maize genes, central cell DNA methylation has not been directly measured, leaving the origin and mechanism of endosperm demethylation uncertain. Here, we report genome-wide analysis of DNA methylation in the central cells of Arabidopsis and rice—species that diverged 150 million years ago—as well as in rice egg cells. We find that DNA demethylation in both species is initiated in central cells, which requires DEMETER in Arabidopsis. However, we do not observe a global reduction of CG methylation that would be indicative of lowered MET1 activity; on the contrary, CG methylation efficiency is elevated in female gametes compared with nonsexual tissues. Our results demonstrate that locus-specific, active DNA demethylation in the central cell is the origin of maternal chromosome hypomethylation in the endosperm. PMID:27956642

  1. A replication-dependent passive mechanism modulates DNA demethylation in mouse primordial germ cells.

    PubMed

    Ohno, Rika; Nakayama, Megumi; Naruse, Chie; Okashita, Naoki; Takano, Osamu; Tachibana, Makoto; Asano, Masahide; Saitou, Mitinori; Seki, Yoshiyuki

    2013-07-01

    Germline cells reprogramme extensive epigenetic modifications to ensure the cellular totipotency of subsequent generations and to prevent the accumulation of epimutations. Notably, primordial germ cells (PGCs) erase genome-wide DNA methylation and H3K9 dimethylation marks in a stepwise manner during migration and gonadal periods. In this study, we profiled DNA and histone methylation on transposable elements during PGC development, and examined the role of DNA replication in DNA demethylation in gonadal PGCs. CpGs in short interspersed nuclear elements (SINEs) B1 and B2 were substantially demethylated in migrating PGCs, whereas CpGs in long interspersed nuclear elements (LINEs), such as LINE-1, were resistant to early demethylation. By contrast, CpGs in both LINE-1 and SINEs were rapidly demethylated in gonadal PGCs. Four major modifiers of DNA and histone methylation, Dnmt3a, Dnmt3b, Glp and Uhrf1, were actively repressed at distinct stages of PGC development. DNMT1 was localised at replication foci in nascent PGCs, whereas the efficiency of recruitment of DNMT1 into replication foci was severely impaired in gonadal PGCs. Hairpin bisulphite sequencing analysis showed that strand-specific hemi-methylated CpGs on LINE-1 were predominant in gonadal PGCs. Furthermore, DNA demethylation in SINEs and LINE-1 was impaired in Cbx3-deficient PGCs, indicating abnormalities in G1 to S phase progression. We propose that PGCs employ active and passive mechanisms for efficient and widespread erasure of genomic DNA methylation.

  2. DNA demethylation and invasive cancer: implications for therapeutics.

    PubMed

    Cheishvili, David; Boureau, Lisa; Szyf, Moshe

    2015-06-01

    One of the hallmarks of cancer is aberrant DNA methylation, which is associated with abnormal gene expression. Both hypermethylation and silencing of tumour suppressor genes as well as hypomethylation and activation of prometastatic genes are characteristic of cancer cells. As DNA methylation is reversible, DNA methylation inhibitors were tested as anticancer drugs with the idea that such agents would demethylate and reactivate tumour suppressor genes. Two cytosine analogues, 5-azacytidine (Vidaza) and 5-aza-2'-deoxycytidine, were approved by the Food and Drug Administration as antitumour agents in 2004 and 2006 respectively. However, these agents might cause activation of a panel of prometastatic genes in addition to activating tumour suppressor genes, which might lead to increased metastasis. This poses the challenge of how to target tumour suppressor genes and block cancer growth with DNA-demethylating drugs while avoiding the activation of prometastatic genes and precluding the morbidity of cancer metastasis. This paper reviews current progress in using DNA methylation inhibitors in cancer therapy and the potential promise and challenges ahead.

  3. DNA demethylation and histone H3K9 acetylation determine the active transcription of the NKG2D gene in human CD8+ T and NK cells

    PubMed Central

    Fernández-Sánchez, Alba; Baragaño Raneros, Aroa; Carvajal Palao, Reyes; Sanz, Ana B.; Ortiz, Alberto; Ortega, Francisco; Suárez-Álvarez, Beatriz; López-Larrea, Carlos

    2013-01-01

    The human activating receptor NKG2D is mainly expressed by NK, NKT, γδ T and CD8+ T cells and, under certain conditions, by CD4+ T cells. This receptor recognizes a diverse family of ligands (MICA, MICB and ULBPs 1–6) leading to the activation of effector cells and triggering the lysis of target cells. The NKG2D receptor-ligand system plays an important role in the immune response to infections, tumors, transplanted graft and autoantigens. Elucidation of the regulatory mechanisms of NKG2D is therefore essential for therapeutic purposes. In this study, we speculate whether epigenetic mechanisms, such as DNA methylation and histone acetylation, participate in NKG2D gene regulation in T lymphocytes and NK cells. DNA methylation in the NKG2D gene was observed in CD4+ T lymphocytes and T cell lines (Jurkat and HUT78), while this gene was unmethylated in NKG2D-positive cells (CD8+ T lymphocytes, NK cells and NKL cell line) and associated with high levels of histone H3 lysine 9 acetylation (H3K9Ac). Treatment with the histone acetyltransferase (HAT) inhibitor curcumin reduces H3K9Ac levels in the NKG2D gene, downregulates NKG2D transcription and leads to a marked reduction in the lytic capacity of NKG2D-mediated NKL cells. These findings suggest that differential NKG2D expression in the different cell subsets is regulated by epigenetic mechanisms and that its modulation by epigenetic treatments might provide a new strategy for treating several pathologies. PMID:23235109

  4. Neil DNA glycosylases promote substrate turnover by Tdg during DNA demethylation

    PubMed Central

    Arab, Khelifa; Kienhöfer, Sabine; von Seggern, Annika; Niehrs, Christof

    2016-01-01

    DNA 5-methylcytosine is a dynamic epigenetic mark which plays important roles in development and disease. In the Tet-Tdg demethylation pathway, methylated cytosine is iteratively oxidized by Tet dioxygenases and unmodified cytosine is restored via thymine DNA glycosylase (Tdg). Here we show that human NEIL1 and NEIL2 DNA glycosylases coordinate abasic site processing during TET–TDG DNA demethylation. NEIL1 and NEIL2 cooperate with TDG during base excision: TDG occupies the abasic site and is displaced by NEILs, which further process the baseless sugar, thereby stimulating TDG substrate turnover. In early Xenopus embryos Neil2 cooperates with Tdg to remove oxidized methylcytosines and to specify neural crest development together with Tet3. Thus, Neils function as AP lyases in the coordinated AP site hand-over during oxidative DNA demethylation. PMID:26751644

  5. Ligand binding affinities of arctigenin and its demethylated metabolites to estrogen receptor alpha.

    PubMed

    Jin, Jong-Sik; Lee, Jong-Hyun; Hattori, Masao

    2013-01-16

    Phytoestrogens are defined as plant-derived compounds with estrogen-like activities according to their chemical structures and activities. Plant lignans are generally categorized as phytoestrogens. It was reported that (-)-arctigenin, the aglycone of arctiin, was demethylated to (-)-dihydroxyenterolactone (DHENL) by Eubacterium (E.) sp. ARC-2. Through stepwise demethylation, E. sp. ARC-2 produced six intermediates, three mono-desmethylarctigenins and three di-desmethylarctigenins. In the present study, ligand binding affinities of (-)-arctigenin and its seven metabolites, including DHENL, were investigated for an estrogen receptor alpha, and found that demethylated metabolites had stronger binding affinities than (-)-arctigenin using a ligand binding screen assay method. The IC(50) value of (2R,3R)-2-(4-hydroxy-3-methoxybenzyl)-3-(3,4-dihydroxybenzyl)-butyrolactone was 7.9 × 10⁻⁴ M.

  6. Anaerobic Mercury Methylation and Demethylation by Geobacter bemidjiensis Bem

    SciTech Connect

    Lu, Xia; Liu, Yurong; Johs, Alexander; Zhao, Linduo; Wang, Tieshan; Yang, Ziming; Lin, Hui; Elias, Dwayne A.; Pierce, Eric M.; Liang, Liyuan; Barkay, Tamar; Gu, Baohua

    2016-03-28

    Two competing processes controlling the net production and bioaccumulation of neurotoxic methylmercury (MeHg) in natural ecosystems are microbial methylation and demethylation. Though mercury (Hg) methylation by anaerobic microorganisms and demethylation by aerobic Hg-resistant bacteria have both been extensively studied, little attention has been given to MeHg degradation by anaerobic bacteria, particularly the iron-reducing bacterium Geobacter bemidjensis Bem. Here we report, for the first time, that the strain G. bemidjensis Bem can methylate inorganic Hg and degrade MeHg concurrently under anoxic conditions. Our results suggest that G. bemidjensis cells utilize a reductive demethylation pathway to degrade MeHg, with elemental Hg(0) as the major reaction product, possibly due to the presence of homologs encoding both organo-mercurial lyase (MerB) and mercuric reductase (MerA) in this organism. In addition, the cells can mediate multiple reactions including Hg/MeHg sorption, Hg reduction and oxidation, resulting in both time and concentration dependent Hg species transformations. Moderate concentrations (10 500 M) of Hg-binding ligands such as cysteine enhance Hg(II) methylation but inhibit MeHg degradation. These findings indicate a cycle of methylation and demethylation among anaerobic bacteria and suggest that mer-mediated demethylation may play a role in the net balance of MeHg production in anoxic water and sediments.

  7. A Comparative Analysis of 5-Azacytidine- and Zebularine-Induced DNA Demethylation

    PubMed Central

    Griffin, Patrick T.; Niederhuth, Chad E.; Schmitz, Robert J.

    2016-01-01

    The nonmethylable cytosine analogs, 5-azacytidine and zebularine, are widely used to inhibit DNA methyltransferase activity and reduce genomic DNA methylation. In this study, whole-genome bisulfite sequencing is used to construct maps of DNA methylation with single base pair resolution in Arabidopsis thaliana seedlings treated with each demethylating agent. We find that both inhibitor treatments result in nearly indistinguishable patterns of genome-wide DNA methylation and that 5-azacytidine had a slightly greater demethylating effect at higher concentrations across the genome. Transcriptome analyses revealed a substantial number of upregulated genes, with an overrepresentation of transposable element genes, in particular CACTA-like elements. This demonstrates that chemical demethylating agents have a disproportionately large effect on loci that are otherwise silenced by DNA methylation. PMID:27402357

  8. Inhibitors of DNA Methylation, Histone Deacetylation, and Histone Demethylation: A Perfect Combination for Cancer Therapy.

    PubMed

    Zahnow, C A; Topper, M; Stone, M; Murray-Stewart, T; Li, H; Baylin, S B; Casero, R A

    2016-01-01

    Epigenetic silencing and inappropriate activation of gene expression are frequent events during the initiation and progression of cancer. These events involve a complex interplay between the hypermethylation of CpG dinucleotides within gene promoter and enhancer regions, the recruitment of transcriptional corepressors and the deacetylation and/or methylation of histone tails. These epigenetic regulators act in concert to block transcription or interfere with the maintenance of chromatin boundary regions. However, DNA/histone methylation and histone acetylation states are reversible, enzyme-mediated processes and as such, have emerged as promising targets for cancer therapy. This review will focus on the potential benefits and synergistic/additive effects of combining DNA-demethylating agents and histone deacetylase inhibitors or lysine-specific demethylase inhibitors together in epigenetic therapy for solid tumors and will highlight what is known regarding the mechanisms of action that contribute to the antitumor response.

  9. Epigenetic targets and drug discovery Part 2: Histone demethylation and DNA methylation.

    PubMed

    Liu, Ke; Liu, Yanli; Lau, Johnathan L; Min, Jinrong

    2015-07-01

    Chromatin structure is dynamically modulated by various chromatin modifications, such as histone/DNA methylation and demethylation. We have reviewed histone methyltransferases and methyllysine binders in terms of small molecule screening and drug discovery in the first part of this review series. In this part, we will summarize recent progress in chemical probe and drug discovery of histone demethylases and DNA methyltransferases. Histone demethylation and DNA methylation have attracted a lot of attention regarding their biology and disease implications. Correspondingly, many small molecule compounds have been designed to modulate the activity of histone demethylases and DNA methyltransferases, and some of them have been developed into therapeutic drugs or put into clinical trials.

  10. Methylmercury decomposition in sediments and bacterial cultures: Involvement of methanogens and sulfate reducers in oxidative demethylation

    SciTech Connect

    Oremland, R.S.; Culbertson, C.W. ); Winfrey, M.R. )

    1991-01-01

    The biogeochemical cycling of mercury has received considerable attention because of the toxicity of methylmercury, its bioaccumulation in biota, and its biomagnification in aquatic food chains. The formation of methylmercury is mediated primarily by microorganisms. Demethylation of monomethylmercury in freshwater and estuarine sediments and in bacterial cultures was investigated with {sup 14}CH{sub 3}HgI. Under anaerobiosis, results with inhibitors indicated partial involvement of both sulfate reducers and methanogens, the former dominated estuarine sediments, while both were active in freshwaters. Aerobes were the most significant demethylators in estuarine sediments, but were unimportant in freshwater sediments. Products of anaerobic demthylation were mainly {sup 14}CO{sub 2} as well as lesser amounts of {sup 14}CH{sub 4}. Acetogenic activity resulted in fixation of some {sup 14}CO{sub 2} produced from {sup 14}CH{sub 3}HgI into acetate. Aerobic demethylation in estuarine sediments produced only {sup 14}CH{sub 4}, while aerobic demethylation in freshwater sediments produced small amounts of both {sup 14}CH{sub 4} and {sup 14}CO{sub 2}. Two species of Desulfovibrio produced only traces of {sup 14}CH{sub 4} from {sup 14}CH{sub 3}HgI, while a culture of a methylotrophic methanogen formed traces of {sup 14}CO{sub 2} and {sup 14}CH{sub 4} when grown on trimethylamine in the presence of the {sup 14}CH{sub 3}HgI. These results indicate that both aerobes and anaerobes demethylate mercury in sediments, but that either group may dominate in a particular sediment type. Aerobic demethylation in the estuarine sediments appeared to proceed by the previously characterized organomercurial-lyase pathway, because methane was the sole product. This indicates the presence of an oxidative pathway, possibly one in which methylmercury serves as an analog of one-carbon substrates.

  11. Clinical development of demethylating agents in hematology

    PubMed Central

    Navada, Shyamala C.; Steinmann, Juliane; Lübbert, Michael; Silverman, Lewis R.

    2014-01-01

    The term epigenetics refers to the heritable changes in gene expression that are not associated with a change in the actual DNA sequence. Epigenetic dysregulation is linked to the pathogenesis of a number of malignancies and has been studied extensively in myelodysplastic syndromes and acute myeloid leukemia. DNA methylation is frequently altered in cancerous cells and likely results in transcriptional silencing of tumor suppressor genes. Re-expression of these genes by inhibition of the DNA methyltransferases has been successful in the treatment of benign and malignant disease. In this Review, we discuss the clinical development of demethylating agents in hematology, with a focus on azacitidine and decitabine. PMID:24382388

  12. Burning off DNA methylation: new evidence for oxygen-dependent DNA demethylation.

    PubMed

    Jurkowski, Tomasz P; Jeltsch, Albert

    2011-11-25

    Where do you stop? Three recent publications have described how the oxidation of 5-methylcytosine by Tet dioxygenases does not stop at the 5-hydroxymethylcytosine (5hmC) state, rather further oxidation of 5hmC is involved in DNA demethylation. The nature of the enzymes involved in this process shed light on the dynamics of epigenetic signaling and its evolutionary origin.

  13. No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos

    PubMed Central

    Kaneto, Reiya; Izawa, Toshiaki; Yokoi, Hayato; Hashimoto, Naohiro; Kikuchi, Yutaka

    2014-01-01

    The mechanisms responsible for active DNA demethylation remain elusive in Metazoa. A previous study that utilized zebrafish embryos provided a potent mechanism for active demethylation in which three proteins, AID, MBD4, and GADD45 are involved. We recently found age-dependent DNA hypomethylation in zebrafish, and it prompted us to examine if AID and MBD4 could be involved in the phenomenon. Unexpectedly, however, we found that most of the findings in the previous study were not reproducible. First, the injection of a methylated DNA fragment into zebrafish eggs did not affect either the methylation of genomic DNA, injected methylated DNA itself, or several loci tested or the expression level of aid, which has been shown to play a role in demethylation. Second, aberrant methylation was not observed at certain CpG islands following the injection of antisense morpholino oligonucleotides against aid and mbd4. Furthermore, we demonstrated that zebrafish MBD4 cDNA lacked a coding region for the methyl-CpG binding domain, which was assumed to be necessary for guidance to target regions. Taken together, we concluded that there is currently no evidence to support the proposed roles of AID and MBD4 in active demethylation in zebrafish embryos. PMID:25536520

  14. JMJD3 promotes SAHF formation in senescent WI38 cells by triggering an interplay between demethylation and phosphorylation of RB protein.

    PubMed

    Zhao, L; Zhang, Y; Gao, Y; Geng, P; Lu, Y; Liu, X; Yao, R; Hou, P; Liu, D; Lu, J; Huang, B

    2015-10-01

    Primary human fibroblasts undergoing oncogene-induced or replicative senescence are known to form senescence-associated heterochromatin foci (SAHF), which can stabilize the state of senescence. The retinoblastoma (RB) protein has an important role in SAHF; cells that lack active RB pathway fail to form SAHF. It has been known that the posttranslational modifications of RB, for example, phosphorylation, regulate its function. To date, whether methylation of RB impacts on the SAHF formation is unknown. Here we report that JMJD3, a histone demethylase catalyzing the tri-methylation of H3K27 (H3K27me3), can demethylate the non-histone protein RB at the lysine810 residue (K810), which is a target of the methyltransferase Set7/9. We detected a significant upregulation of JMJD3 during cellular senescence and SAHF formation in WI38 cells induced by H-RasV(12), and we found that ectopic expression of JMJD3 promoted cellular senescence and SAHF formation in WI38 cells. Furthermore, during the process of SAHF assembly, JMJD3 was transported to the cytoplasm and interacted with RB through its demethylase domain JmjC. Significantly, our data demonstrated that the JMJD3-mediated demethylation of RB at K810 impeded the interaction of RB with the protein kinase CDK4 and resulted in reduced level of phosphorylation of RB at Serine807/811 (S807/811), implicating an important role of the interplay between the demethylation and phosphorylation of RB in SAHF assembly. This study highlights the role of JMJD3 as a novel inducer of SAHF formation through demethylating RB and provides new insights into the mechanisms of cellular senescence and SAHF assembly.

  15. Active braze process

    SciTech Connect

    Levine, I.L.; Pike, R.A.

    1990-11-02

    Active metal bonding using Cusil (silver-copper) braze alloys is a well established method used at GE Neutron Devices (GEND) for bonding metal to metal, metal to ceramics, and ceramics to ceramics. However, there are many instances in which using a silver alloy for bonding is undesirable (e.g., in vacuum tube envelopes, or where sequential braze steps at different temperatures are required to complete an assembly). The Material and Processes Laboratory at GEND has discovered a new method of active brazing with non-silver alloys which has proved especially successful in ceramic-to-ceramic joints. This method has the added advantage of eliminating several steps which are required in conventional bonding techniques. 2 figs., 10 tabs.

  16. Curcuminoid Demethylation as an Alternative Metabolism by Human Intestinal Microbiota.

    PubMed

    Burapan, Supawadee; Kim, Mihyang; Han, Jaehong

    2017-04-14

    Curcumin and other curcuminoids from Curcuma longa are important bioactive compounds exhibiting various pharmacological activities. In addition to the known reductive metabolism of curcuminoids, an alternative biotransformation of curcuminoids by human gut microbiota is reported herein. A curcuminoid mixture, composed of curcumin (1), demethoxycurcumin (2), and bisdemethoxycurcumin (3), was metabolized by the human intestinal bacterium Blautia sp. MRG-PMF1. 1 and 2 were converted to new metabolites by the methyl aryl ether cleavage reaction. Two metabolites, demethylcurcumin (4) and bisdemethylcurcumin (5), were sequentially produced from 1, and demethyldemethoxycurcumin (6) was produced from 2. Until now, sequential reduction of the heptadienone backbone of curcuminoids was the only known metabolism to occur in the human intestine. In this study, a new intestinal metabolism of curcuminoids was discovered. Demethylation of curcuminoids produced three new colonic metabolites that were already known as promising synthetic curcumin analogues. The results could explain the observed beneficial effects of turmeric.

  17. Expression of Caveolin 1 is enhanced by DNA demethylation during adipocyte differentiation. status of insulin signaling.

    PubMed

    Palacios-Ortega, Sara; Varela-Guruceaga, Maider; Milagro, Fermín Ignacio; Martínez, José Alfredo; de Miguel, Carlos

    2014-01-01

    Caveolin 1 (Cav-1) is an essential constituent of adipocyte caveolae which binds the beta subunit of the insulin receptor (IR) and is implicated in the regulation of insulin signaling. We have found that, during adipocyte differentiation of 3T3-L1 cells the promoter, exon 1 and first intron of the Cav-1 gene undergo a demethylation process that is accompanied by a strong induction of Cav-1 expression, indicating that epigenetic mechanisms must have a pivotal role in this differentiation process. Furthermore, IR, PKB-Akt and Glut-4 expression are also increased during the differentiation process suggesting a coordinated regulation with Cav-1. Activation of Cav-1 protein by phosphorylation arises during the differentiation process, yet in fully mature adipocytes insulin is no longer able to significantly increase Cav-1 phosphorylation. However, these long-term differentiated cells are still able to respond adequately to insulin, increasing IR and PKB-Akt phosphorylation and glucose uptake. The activation of Cav-1 during the adipocyte differentiation process could facilitate the maintenance of insulin sensitivity by these fully mature adipocytes isolated from additional external stimuli. However, under the influence of physiological conditions associated to obesity, such as chronic inflammation and hypoxia, insulin sensitivity would finally be compromised.

  18. Expression of Caveolin 1 Is Enhanced by DNA Demethylation during Adipocyte Differentiation. Status of Insulin Signaling

    PubMed Central

    Palacios-Ortega, Sara; Varela-Guruceaga, Maider; Milagro, Fermín Ignacio; Martínez, José Alfredo; de Miguel, Carlos

    2014-01-01

    Caveolin 1 (Cav-1) is an essential constituent of adipocyte caveolae which binds the beta subunit of the insulin receptor (IR) and is implicated in the regulation of insulin signaling. We have found that, during adipocyte differentiation of 3T3-L1 cells the promoter, exon 1 and first intron of the Cav-1 gene undergo a demethylation process that is accompanied by a strong induction of Cav-1 expression, indicating that epigenetic mechanisms must have a pivotal role in this differentiation process. Furthermore, IR, PKB-Akt and Glut-4 expression are also increased during the differentiation process suggesting a coordinated regulation with Cav-1. Activation of Cav-1 protein by phosphorylation arises during the differentiation process, yet in fully mature adipocytes insulin is no longer able to significantly increase Cav-1 phosphorylation. However, these long-term differentiated cells are still able to respond adequately to insulin, increasing IR and PKB-Akt phosphorylation and glucose uptake. The activation of Cav-1 during the adipocyte differentiation process could facilitate the maintenance of insulin sensitivity by these fully mature adipocytes isolated from additional external stimuli. However, under the influence of physiological conditions associated to obesity, such as chronic inflammation and hypoxia, insulin sensitivity would finally be compromised. PMID:24751908

  19. Demethylation and cleavage of dimethylsulfoniopropionate in marine intertidal sediments

    USGS Publications Warehouse

    Visscher, P.T.; Kiene, R.P.; Taylor, B.F.

    1994-01-01

    Demethylation and cleavage of dimethylsulfoniopropionate (DMSP) was measured in three different types of,intertidal marine sediments: a cyanobacterial mat, a diatom-covered tidal flat and a carbonate sediment. Consumption rates of added DMSP were highest in cyanobacterial mat slurries (59 ?? mol DMSP l-1 slurry h-1) and lower in slurries from a diatom mat and a carbonate tidal sediment (24 and 9 ??mol DMSP l-1 h-1, respectively). Dimethyl sulfide (DMS) and 3-mercaptopropionate (MPA) were produced simultaneously during DMSP consumption, indicating that cleavage and demethylation occurred at the same time. Viable counts of DMSP-utilizing bacteria revealed a population of 2 x 107 cells cm-3 sediment (90% of these cleaved DMSP to DMS, 10% demethylated DMSP to MPA) in the cyanobacterial mat, 7 x 105 cells cm-3 in the diatom mat (23% cleavers, 77% demethylators), and 9 x 104 cells cm-3 (20% cleavers and 80% demethylators) in the carbonate sediment. In slurries of the diatom mat, the rate of MPA production from added 3-methiolpropionate (MMPA) was 50% of the rate of MPA formation from DMSP. The presence of a large population of demethylating bacteria and the production of MPA from DMSP suggest that the demethylation pathway, in addition to cleavage, contributes significantly to DMSP consumption in coastal sediments.

  20. Structural Mechanism of Demethylation and Inactivation of Protein Phosphatase 2A

    SciTech Connect

    Xing,Y.; Li, Z.; Chen, Y.; Stock, J.; Jeffrey, P.; Shi, Y.

    2008-01-01

    Protein phosphatase 2A (PP2A) is an important serine/threonine phosphatase that plays a role in many biological processes. Reversible carboxyl methylation of the PP2A catalytic subunit is an essential regulatory mechanism for its function. Demethylation and negative regulation of PP2A is mediated by a PP2A-specific methylesterase PME-1, which is conserved from yeast to humans. However, the underlying mechanism of PME-1 function remains enigmatic. Here we report the crystal structures of PME-1 by itself and in complex with a PP2A heterodimeric core enzyme. The structures reveal that PME-1 directly binds to the active site of PP2A and that this interaction results in the activation of PME-1 by rearranging the catalytic triad into an active conformation. Strikingly, these interactions also lead to inactivation of PP2A by evicting the manganese ions that are required for the phosphatase activity of PP2A. These observations identify a dual role of PME-1 that regulates PP2A activation, methylation, and holoenzyme assembly in cells.

  1. Promoter demethylation of Keap1 gene in human diabetic cataractous lenses

    SciTech Connect

    Palsamy, Periyasamy; Ayaki, Masahiko; Elanchezhian, Rajan; Shinohara, Toshimichi

    2012-07-06

    Highlights: Black-Right-Pointing-Pointer We found significant Keap1 promoter demethylation in diabetic cataractous lenses. Black-Right-Pointing-Pointer Demethylation of Keap1 gene upregulated the expression of Keap1 mRNA and protein. Black-Right-Pointing-Pointer Elevated levels of Keap1 are known to decrease the levels of Nrf2. Black-Right-Pointing-Pointer Thereby, the levels of antioxidant enzymes are suppressed by decreased Nrf2 level. -- Abstract: Age-related cataracts (ARCs) are the major cause of visual impairments worldwide, and diabetic adults tend to have an earlier onset of ARCs. Although age is the strongest risk factor for cataracts, little is known how age plays a role in the development of ARCs. It is known that oxidative stress in the lens increases with age and more so in the lenses of diabetics. One of the central adaptive responses against the oxidative stresses is the activation of the nuclear transcriptional factor, NF-E2-related factor 2 (Nrf2), which then activates more than 20 different antioxidative enzymes. Kelch-like ECH associated protein 1 (Keap1) targets and binds to Nrf2 for proteosomal degradation. We hypothesized that hyperglycemia will lead to a dysfunction of the Nrf2-dependent antioxidative protection in the lens of diabetics. We studied the methylation status of the CpG islands in 15 clear and 21 diabetic cataractous lenses. Our results showed significant levels of demethylated DNA in the Keap1 promoter in the cataractous lenses from diabetic patients. In contrast, highly methylated DNA was found in the clear lens and tumorized human lens epithelial cell (HLEC) lines (SRA01/04). HLECs treated with a demethylation agent, 5-aza-2 Prime deoxycytidine (5-Aza), had a 10-fold higher levels of Keap1 mRNA, 3-fold increased levels of Keap1 protein, produced higher levels of ROS, and increased cell death. Our results indicated that demethylation of the CpG islands in the Keap1 promoter will activate the expression of Keap1 protein, which

  2. Cumene hydroperoxide supported demethylation of N,N-dimethylaniline by cytochrome P-450 from adrenal cortex mitochondria.

    PubMed

    Akhrem, A A; Khatyleva SYu; Shkumatov, V M; Chashchin, V L; Kiselev, P A

    1982-01-01

    The interaction of highly purified cytochrome P-450 from bovine adrenal cortex mitochondria (cytochrome P-450scc) with N,N-dimethylaniline (DMA), aniline, N-dimethylcyclohexylamine and cumene hydroperoxide (CHP) has been investigated. The formation of complexes between cytochrome P-450scc and the above listed compounds could be demonstrated. The reaction of oxidative demethylation of DMA by cumene hydroperoxide involving cytochrome P-450scc has been carried out at 37 degrees C; the mechanism of this process is discussed. Incubation of cytochrome P-450scc with negatively charged phospholipids, phosphatidylglycerol (PG), and phosphatidylinosite (PI) exerts an inhibiting effect on the reaction of oxidative demethylation. The interaction of cytochrome P-450scc with CHP is accompanied by hemoprotein destruction in a complex biphasic way. The process of oxidative demethylation of DMA in the system of cytochrome P-450scc-CHP has been concluded to have a predominantly radical character.

  3. Methylation-independent DNA Binding Modulates Specificity of Repressor of Silencing 1 (ROS1) and Facilitates Demethylation in Long Substrates*

    PubMed Central

    Ponferrada-Marín, María Isabel; Martínez-Macías, María Isabel; Morales-Ruiz, Teresa; Roldán-Arjona, Teresa; Ariza, Rafael R.

    2010-01-01

    DNA cytosine methylation is an epigenetic mark that promotes gene silencing and performs critical roles during reproduction and development in both plants and animals. The genomic distribution of DNA methylation is the dynamic outcome of opposing methylation and demethylation processes. In plants, active demethylation occurs through a base excision repair pathway initiated by 5-methycytosine (5-meC) DNA glycosylases of the REPRESSOR OF SILENCING 1 (ROS1)/DEMETER (DME) family. To gain insight into the mechanism by which Arabidopsis ROS1 recognizes and excises 5-meC, we have identified those protein regions that are required for efficient DNA binding and catalysis. We have found that a short N-terminal lysine-rich domain conserved in members of the ROS1/DME family mediates strong methylation-independent binding of ROS1 to DNA and is required for efficient activity on 5-meC·G, but not for T·G processing. Removal of this domain does not significantly affect 5-meC excision from short molecules, but strongly decreases ROS1 activity on long DNA substrates. This region is not required for product binding and is not involved in the distributive behavior of the enzyme on substrates containing multiple 5-meC residues. Altogether, our results suggest that methylation-independent DNA binding allows ROS1 to perform a highly redundant search for efficient excision of a nondamaged, correctly paired base such as 5-meC in long stretches of DNA. These findings may have implications for understanding the evolution of structure and target specificity in DNA glycosylases. PMID:20489198

  4. Simulated vibrational spectra of aflatoxins and their demethylated products and the estimation of the energies of the demethylation reactions

    NASA Astrophysics Data System (ADS)

    Billes, Ferenc; Móricz, Ágnes M.; Tyihák, Ernő; Mikosch, Hans

    2006-06-01

    The structure of four natural mycotoxins, the aflatoxin B 1, B 2, G 1 and G 2 and their demethylated products were optimized with quantum chemical method. The energies and the thermodynamic functions of the molecules were calculated and applied to calculation of the reaction energies of the demethylations. Further results of the calculations are the vibrational force constants, the infrared spectra of the molecules and the assignments of the spectral bands.

  5. The Histone Demethylase UTX Promotes Brown Adipocyte Thermogenic Program Via Coordinated Regulation of H3K27 Demethylation and Acetylation*

    PubMed Central

    Zha, Lin; Li, Fenfen; Wu, Rui; Artinian, Liana; Rehder, Vincent; Yu, Liqing; Liang, Houjie; Xue, Bingzhong; Shi, Hang

    2015-01-01

    Brown adipocytes function to dissipate energy as heat through adaptive thermogenesis. Understanding the molecular mechanisms underlying the brown fat thermogenic program may provide insights for the development of therapeutic approaches in the treatment of obesity. Most studies investigating the mechanisms underlying brown fat development focus on genetic mechanisms; little is known about the epigenetic mechanisms in this process. We have discovered that ubiquitously transcribed tetratricopeptide repeat on chromosome X (UTX), a histone demethylase for di- or tri-methylated histone 3 lysine 27 (H3K27me2/3), plays a potential role in regulating brown adipocyte thermogenic program. We found that UTX is up-regulated during brown adipocyte differentiation and by cold exposure in both brown adipose tissue (BAT) and white adipose tissue (WAT) of mice, suggesting a potential role in thermogenesis. Inactivation of UTX down-regulates brown fat specific gene expression, while overexpression of UTX does the opposite. Notably, activation of β adrenergic signaling recruits UTX to the UCP1 and PGC1α promoters, leading to decreased H3K27me3, a histone transcriptional repressive mark. UTX demethylates H3K27me3 and subsequently interacts with the histone acetyltransferase (HAT) protein CBP, resulting in increased H3K27 acetylation (H3K27ac), a histone transcriptional active mark. UTX positively regulate brown adipocyte thermogenic program through coordinated control of demethylating H3K27me3 and acetylating H3K27, switching the transcriptional repressive state to the transcriptional active state at the promoters of UCP1 and PGC1α. We conclude that UTX may play a potential role in regulation of brown adipocyte gene expression and may mediate β adrenergic activation of brown fat function. PMID:26306033

  6. Simultaneous determination of the novel tyrosine kinase inhibitor meditinib and its active metabolite demethylation meditinib in monkey plasma by liquid chromatography-tandem mass spectrometry and its application to pharmacokinetic studies.

    PubMed

    Liang, Feng; Kong, Qi; Guo, Yongqi; Wang, Yu; Sun, Dejie; Liu, Shi; Cai, Jinling; Guan, Yongbiao; Ding, Rigao

    2015-08-01

    Meditinib (ME) is a novel tyrosine kinase inhibitor used as an antichronic myeloid leukemia drug. A simple, sensitive and specific LC/MS/MS method was developed and validated for the analysis of ME and its metabolite demethylation meditinib (PI) in monkey plasma using naltrexone as the internal standard. Sample preparation involved protein precipitation with methanol. The analysis was carried out on an Agilent C8 column (3.5 µm, 2.1 × 50 mm). Elution was achieved with a mobile phase gradient varying the proportion of a water solution containing 0.1% formic acid (solvent A) and a 0.1% formic acid in methanol solution (solvent B) at a flow rate of 300 μL/min. The method had a linear calibration curve over the concentration range of 2-1000 ng/mL for ME and 2-1000 ng/mL for PI. The lower limits of quantification of ME and PI were 2 and 2 ng/mL, respectively. The intra- and inter-day precision values were <15% and accuracy values were within ±10.0%. The mean recoveries of ME and PI from plasma were >85%. The assay has been successfully used for pharmacokinetic evaluation of ME and PI using the monkey as an animal model, and those data are reported for the first time.

  7. Further investigation of the N-demethylation of tertiary amine alkaloids using the non-classical Polonovski reaction.

    PubMed

    Thavaneswaran, Shanti; Scammells, Peter J

    2006-06-01

    The iron salt-mediated Polonovski reaction efficiently N-demethylates certain opiate alkaloids. In this process, the use of the hydrochloride salt of the tertiary N-methyl amine oxide was reported to give better yields of the desired N-demethylated product. Herein, we report further investigation into the use of N-oxide salts in the iron salt-mediated Polonovski reaction. An efficient approach for the removal of iron salts that greatly facilitates isolation and purification of the N-nor product is also described.

  8. Influence of HIV antiretrovirals on methadone N-demethylation and transport.

    PubMed

    Campbell, Scott D; Gadel, Sarah; Friedel, Christina; Crafford, Amanda; Regina, Karen J; Kharasch, Evan D

    2015-05-15

    Drug interactions involving methadone and/or HIV antiretrovirals can be problematic. Mechanisms whereby antiretrovirals induce clinical methadone clearance are poorly understood. Methadone is N-demethylated to 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) by CYP2B6 and CYP3A4 in vitro, but by CYP2B6 in vivo. This investigation evaluated human hepatocytes as a model for methadone induction, and tested the hypothesis that methadone and EDDP are substrates for human drug transporters. Human hepatocyte induction by several antiretrovirals of methadone N-demethylation, and CYP2B6 and CYP3A4 transcription, protein expression and catalytic activity, and pregnane X receptor (PXR) activation were evaluated. Methadone and EDDP uptake and efflux by overexpressed transporters were also determined. Methadone N-demethylation was generally not significantly increased by the antiretrovirals. CYP2B6 mRNA and activity (bupropion N-demethylation) were induced by several antiretrovirals, as were CYP3A4 mRNA and protein expression, but only indinavir increased CYP3A activity (alfentanil dealkylation). CYP upregulation appeared related to PXR activation. Methadone was not a substrate for uptake (OCT1, OCT2, OCT3, OATP1A2, OATP1B1, OATP1B3, OATP2B1) or efflux (P-gp, BCRP) transporters. EDDP was a good substrate for P-gp, BCRP, OCT1, OCT3, OATP1A2, and OATP1B1. OATP1A2- and OCT3-mediated EDDP uptake, and BCRP-mediated EDDP efflux transport, was inhibited by several antiretrovirals. Results show that hepatocyte methadone N-demethylation resembles expressed and liver microsomal metabolism more than clinical metabolism. Compared with clinical studies, hepatocytes underreport induction of methadone metabolism by HIV drugs. Hepatocytes are not a good predictive model for clinical antiretroviral induction of methadone metabolism and not a substitute for clinical studies. EDDP is a transporter substrate, and is susceptible to transporter-mediated interactions.

  9. Re-expression of Selected Epigenetically Silenced Candidate Tumor Suppressor Genes in Cervical Cancer by TET2-directed Demethylation.

    PubMed

    Huisman, Christian; van der Wijst, Monique G P; Schokker, Matthijs; Blancafort, Pilar; Terpstra, Martijn M; Kok, Klaas; van der Zee, Ate G J; Schuuring, Ed; Wisman, G Bea A; Rots, Marianne G

    2016-03-01

    DNA hypermethylation is extensively explored as therapeutic target for gene expression modulation in cancer. Here, we re-activated hypermethylated candidate tumor suppressor genes (TSGs) (C13ORF18, CCNA1, TFPI2, and Maspin) by TET2-induced demethylation in cervical cancer cell lines. To redirect TET2 to hypermethylated TSGs, we engineered zinc finger proteins (ZFPs), which were first fused to the transcriptional activator VP64 to validate effective gene re-expression and confirm TSG function. ChIP-Seq not only revealed enriched binding of ZFPs to their intended sequence, but also considerable off-target binding, especially at promoter regions. Nevertheless, results obtained by targeted re-expression using ZFP-VP64 constructs were in line with cDNA overexpression; both revealed strong growth inhibition for C13ORF18 and TFPI2, but not for CCNA1 and Maspin. To explore effectivity of locus-targeted demethylation, ZFP-TET2 fusions were constructed which efficiently demethylated genes with subsequent gene re-activation. Moreover, targeting TET2 to TFPI2 and C13ORF18, but not CCNA1, significantly decreased cell growth, viability, and colony formation in cervical cancer cells compared to a catalytically inactive mutant of TET2. These data underline that effective re-activation of hypermethylated genes can be achieved through targeted DNA demethylation by TET2, which can assist in realizing sustained re-expression of genes of interest.

  10. Sensory rhodopsins I and II modulate a methylation/demethylation system in Halobacterium halobium phototaxis

    SciTech Connect

    Spudich, E.N.; Takahashi, T.; Spudich, J.L. )

    1989-10-01

    This work demonstrates that phototaxis stimuli in the archaebacterium Halobacterium halobium control a methylation/demethylation system in vivo through photoactivation of sensory rhodopsin I (SR-I) in either its attractant or repellent signaling form as well as through the repellent receptor sensory rhodopsin II (SR-II, also called phoborhodopsin). The effects of positive stimuli that suppress swimming reversals (i.e., an increase in attractant or decrease in repellent light) and negative stimuli that induce swimming reversals (i.e., a decrease in attractant or increase in repellent light) through each photoreceptor were monitored by assaying release of volatile (3H)methyl groups. This assay has been used to measure (3H)methanol produced during the process of adaptation to chemotactic stimuli in eubacteria. In H. halobium positive photostimuli produce a transient increase in the rate of demethylation followed by a decrease below the unstimulated value, whereas negative photostimuli cause an increase followed by a rate similar to that of the unstimulated value. Photoactivation of the SR-I attractant and simultaneous photoactivation of the SR-II repellent receptors cancel in their effects on demethylation, demonstrating the methylation system is regulated by an integrated signal. Analysis of mutants indicates that the source for the volatile methyl groups is intrinsic membrane proteins distinct from the chromoproteins that share the membrane. A methyl-accepting protein (94 kDa) previously correlated in amount with the SR-I chromoprotein (25 kDa) is shown here to be missing in a recently isolated SR-I-SR-II+ mutant (Flx3b), thus confirming the association of this protein with SR-I. Photoactivated SR-II in mutant Flx3b controls demethylation, predicting the existence of a photomodulated methyl-accepting component distinct from the 94-kDa protein of SR-I.

  11. Genistein promotes DNA demethylation of the steroidogenic factor 1 (SF-1) promoter in endometrial stromal cells

    SciTech Connect

    Matsukura, Hiroshi; Aisaki, Ken-ichi; Igarashi, Katsuhide; Matsushima, Yuko; Kanno, Jun; Muramatsu, Masaaki; Sudo, Katsuko; Sato, Noriko

    2011-08-26

    Highlights: {yields} Genistein (GEN) is a phytoestrogen found in soy products. {yields} GEN demethylated/unsilenced the steroidogenic factor 1 gene in endometrial tissue. {yields} GEN thus altered mRNA expression in uteri of ovariectomized (OVX) mice. {yields} A high-resolution melting assay was used to screen for epigenetic change. {yields} We isolated an endometrial cell clone that was epigenetically modulated by GEN. -- Abstract: It has recently been demonstrated that genistein (GEN), a phytoestrogen in soy products, is an epigenetic modulator in various types of cells; but its effect on endometrium has not yet been determined. We investigated the effects of GEN on mouse uterine cells, in vivo and in vitro. Oral administration of GEN for 1 week induced mild proliferation of the endometrium in ovariectomized (OVX) mice, which was accompanied by the induction of steroidogenic factor 1 (SF-1) gene expression. GEN administration induced demethylation of multiple CpG sites in the SF-1 promoter; these sites are extensively methylated and thus silenced in normal endometrium. The GEN-mediated promoter demethylation occurred predominantly on the luminal side, as opposed to myometrium side, indicating that the epigenetic change was mainly shown in regenerated cells. Primary cultures of endometrial stromal cell colonies were screened for GEN-mediated alterations of DNA methylation by a high-resolution melting (HRM) method. One out of 20 colony-forming cell clones showed GEN-induced demethylation of SF-1. This clone exhibited a high proliferation capacity with continuous colony formation activity through multiple serial clonings. We propose that only a portion of endometrial cells are capable of receiving epigenetic modulation by GEN.

  12. Pluripotency Transcription Factor Oct4 Mediates Stepwise Nucleosome Demethylation and Depletion

    PubMed Central

    Shakya, Arvind; Callister, Catherine; Goren, Alon; Yosef, Nir; Garg, Neha; Khoddami, Vahid; Nix, David; Regev, Aviv

    2015-01-01

    The mechanisms whereby the crucial pluripotency transcription factor Oct4 regulates target gene expression are incompletely understood. Using an assay system based on partially differentiated embryonic stem cells, we show that Oct4 opposes the accumulation of local H3K9me2 and subsequent Dnmt3a-mediated DNA methylation. Upon binding DNA, Oct4 recruits the histone lysine demethylase Jmjd1c. Chromatin immunoprecipitation (ChIP) time course experiments identify a stepwise Oct4 mechanism involving Jmjd1c recruitment and H3K9me2 demethylation, transient FACT (facilitates chromatin transactions) complex recruitment, and nucleosome depletion. Genome-wide and targeted ChIP confirms binding of newly synthesized Oct4, together with Jmjd1c and FACT, to the Pou5f1 enhancer and a small number of other Oct4 targets, including the Nanog promoter. Histone demethylation is required for both FACT recruitment and H3 depletion. Jmjd1c is required to induce endogenous Oct4 expression and fully reprogram fibroblasts to pluripotency, indicating that the assay system identifies functional Oct4 cofactors. These findings indicate that Oct4 sequentially recruits activities that catalyze histone demethylation and depletion. PMID:25582194

  13. Metabolic demethylation of the insecticide dimethylvinphos in rats, in dogs, and in vitro.

    PubMed

    Crawford, M J; Hutson, D H; King, P A

    1976-12-01

    1. The alkenyl phosphate insecticide, dimethylvinphos, is rapidly metabolized and eliminated by rats and dogs. 2. Metabolism proceeds via demethylation followed by the hydrolysis of desmethyl dimethylvinphos to 2,4-dichlorophenacyl chloride which is further metabolized mainly to 2,4-dichloromandelic acid, 1-(2,4-dichlorophenyl)ethanol (glucuronide) and 2,4-dichlorphenylethanediol (glucuronide). 3. The dechlorination of 2,4-dichlorophenacyl chloride to 2,4-dichloroacetophenone proceeds via the spontaneous formation of S-(2,4-dichlorophenacyl) glutathione which is converted to the ketone by an enzyme-catalysed glutathione-dependent reaction. 4. Demethylation of dimethylvinphos occurs in liver fractions via the action of two enzymes: glutathione S-methyl transferase in the cytosol, and microsomal mono-oxygenase. The relatively high activities of both enzymes in dog liver (compared with rat liver) partly account for the observed differences in metabolism and toxicity of dimethylvinphos in the two species. 5. The glutathione transferase is enhanced twofold by pre-treatment of rats with 0-1% phenobarbital in their drinking water. This treatment also induces the microsomal demethylation 45-fold and results in a greater than 13-fold protective effect against the acute toxic effects of dimethylvinphos.

  14. Cytochrome P450 3A Enzymes Catalyze the O6-Demethylation of Thebaine, a Key Step in Endogenous Mammalian Morphine Biosynthesis.

    PubMed

    Kramlinger, Valerie M; Alvarado Rojas, Mónica; Kanamori, Tatsuyuki; Guengerich, F Peter

    2015-08-14

    Morphine, first characterized in opium from the poppy Papaver somniferum, is one of the strongest known analgesics. Endogenous morphine has been identified in several mammalian cells and tissues. The synthetic pathway of morphine in the opium poppy has been elucidated. The presence of common intermediates in plants and mammals suggests that biosynthesis occurs through similar pathways (beginning with the amino acid L-tyrosine), and the pathway has been completely delineated in plants. Some of the enzymes in the mammalian pathway have been identified and characterized. Two of the latter steps in the morphine biosynthesis pathway are demethylation of thebaine at the O(3)- and the O(6)-positions, the latter of which has been difficult to demonstrate. The plant enzymes responsible for both the O(3)-demethylation and the O(6)-demethylation are members of the Fe(II)/α-ketoglutarate-dependent dioxygenase family. Previous studies showed that human cytochrome P450 (P450) 2D6 can catalyze thebaine O(3)-demethylation. We report that demethylation of thebaine at the O(6)-position is selectively catalyzed by human P450s 3A4 and 3A5, with the latter being more efficient, and rat P450 3A2. Our results do not support O(6)-demethylation of thebaine by an Fe(II)/α-ketoglutarate-dependent dioxygenase. In rat brain microsomes, O(6)-demethylation was inhibited by ketoconazole, but not sulfaphenazole, suggesting that P450 3A enzymes are responsible for this activity in the brain. An alternate pathway to morphine, oripavine O(6)-demethylation, was not detected. The major enzymatic steps in mammalian morphine synthesis have now been identified.

  15. New methodology for the N-demethylation of opiate alkaloids.

    PubMed

    Dong, Zemin; Scammells, Peter J

    2007-12-21

    N-Demethylation is a key step in the preparation of a number of semisynthetic opiate pharmaceuticals. Herein we report a high-yielding, catalytic procedure for the N-demethylation of opiates which has a number of advantages over existing methods. For example, tetrasodium 5,10,15,20-tetra(4-sulfophenyl)porphyrinatoiron(II) (0.3 molar equiv) effected the transformation of codeine methyl ether to the corresponding N-nor analogue in 91% yield. The catalyst was readily removed and recycled.

  16. Decaffeination and measurement of caffeine content by addicted Escherichia coli with a refactored N-demethylation operon from Pseudomonas putida CBB5.

    PubMed

    Quandt, Erik M; Hammerling, Michael J; Summers, Ryan M; Otoupal, Peter B; Slater, Ben; Alnahhas, Razan N; Dasgupta, Aurko; Bachman, James L; Subramanian, Mani V; Barrick, Jeffrey E

    2013-06-21

    The widespread use of caffeine (1,3,7-trimethylxanthine) and other methylxanthines in beverages and pharmaceuticals has led to significant environmental pollution. We have developed a portable caffeine degradation operon by refactoring the alkylxanthine degradation (Alx) gene cluster from Pseudomonas putida CBB5 to function in Escherichia coli. In the process, we discovered that adding a glutathione S-transferase from Janthinobacterium sp. Marseille was necessary to achieve N 7 -demethylation activity. E. coli cells with the synthetic operon degrade caffeine to the guanine precursor, xanthine. Cells deficient in de novo guanine biosynthesis that contain the refactored operon are ″addicted″ to caffeine: their growth density is limited by the availability of caffeine or other xanthines. We show that the addicted strain can be used as a biosensor to measure the caffeine content of common beverages. The synthetic N-demethylation operon could be useful for reclaiming nutrient-rich byproducts of coffee bean processing and for the cost-effective bioproduction of methylxanthine drugs.

  17. Kinetic studies of Escherichia coli AlkB using a new fluorescence-based assay for DNA demethylation.

    PubMed

    Roy, Todd W; Bhagwat, A S

    2007-01-01

    The Escherichia coli AlkB protein catalyzes the direct reversal of alkylation damage to DNA; primarily 1-methyladenine (1mA) and 3-methylcytosine (3mC) lesions created by endogenous or environmental alkylating agents. AlkB is a member of the non-heme iron (II) alpha-ketoglutarate-dependent dioxygenase superfamily, which removes the alkyl group through oxidation eliminating a methyl group as formaldehyde. We have developed a fluorescence-based assay for the dealkylation activity of this family of enzymes. It uses formaldehyde dehydrogenase to convert formaldehyde to formic acid and monitors the creation of an NADH analog using fluorescence. This assay is a great improvement over the existing assays for DNA demethylation in that it is continuous, rapid and does not require radioactively labeled material. It may also be used to study other demethylation reactions including demethylation of histones. We used it to determine the kinetic constants for AlkB and found them to be somewhat different than previously reported values. The results show that AlkB demethylates 1mA and 3mC with comparable efficiencies and has only a modest preference for a single-stranded DNA substrate over its double-stranded DNA counterpart.

  18. Heterochromatin Protein 1a stimulates histone H3 lysine 36 demethylation by the Drosophila KDM4A demethylase

    PubMed Central

    Lin, Chia-Hui; Li, Bing; Swanson, Selene; Zhang, Ying; Florens, Laurence; Washburn, Michael P.; Abmayr, Susan M.; Workman, Jerry L.

    2008-01-01

    Summary Recent discoveries of histone demethylases demonstrate that histone methylation is reversible. However, mechanisms governing the targeting and regulation of histone demethylation remain elusive. Here we report that a Drosophila melanogaster JmjC domain-containing protein, dKDM4A, is a histone H3K36 demethylase. dKDM4A specifically demethylates H3K36me2 and me3 both in vitro and in vivo. Affinity purification and mass spectrometry analysis revealed that Heterochromatin Protein 1a (HP1a) associates with dKDMA4A. We found that the chromoshadow domain of HP1a and a HP1-interacting motif of dKDM4A are responsible for this interaction. HP1a stimulates the histone H3K36 demethylation activity of dKDM4A and this stimulation depends on the H3K9me binding motif of HP1a. Finally, we provide in vivo evidence suggesting that HP1a and dKDM4A interact with each other and loss of HP1a leads to increased level of histone H3K36me3. Collectively, these results suggest a function of HP1a in transcription facilitating H3K36 demethylation at transcribed and/or heterochromatin regions. PMID:19061644

  19. MES16, a member of the methylesterase protein family, specifically demethylates fluorescent chlorophyll catabolites during chlorophyll breakdown in Arabidopsis.

    PubMed

    Christ, Bastien; Schelbert, Silvia; Aubry, Sylvain; Süssenbacher, Iris; Müller, Thomas; Kräutler, Bernhard; Hörtensteiner, Stefan

    2012-02-01

    During leaf senescence, chlorophyll (Chl) is broken down to nonfluorescent chlorophyll catabolites (NCCs). These arise from intermediary fluorescent chlorophyll catabolites (FCCs) by an acid-catalyzed isomerization inside the vacuole. The chemical structures of NCCs from Arabidopsis (Arabidopsis thaliana) indicate the presence of an enzyme activity that demethylates the C13(2)-carboxymethyl group present at the isocyclic ring of Chl. Here, we identified this activity as methylesterase family member 16 (MES16; At4g16690). During senescence, mes16 leaves exhibited a strong ultraviolet-excitable fluorescence, which resulted from large amounts of different FCCs accumulating in the mutants. As confirmed by mass spectrometry, these FCCs had an intact carboxymethyl group, which slowed down their isomerization to respective NCCs. Like a homologous protein cloned from radish (Raphanus sativus) and named pheophorbidase, MES16 catalyzed the demethylation of pheophorbide, an early intermediate of Chl breakdown, in vitro, but MES16 also demethylated an FCC. To determine the in vivo substrate of MES16, we analyzed pheophorbide a oxygenase1 (pao1), which is deficient in pheophorbide catabolism and accumulates pheophorbide in the chloroplast, and a mes16pao1 double mutant. In the pao1 background, we additionally mistargeted MES16 to the chloroplast. Normally, MES16 localizes to the cytosol, as shown by analysis of a MES16-green fluorescent protein fusion. Analysis of the accumulating pigments in these lines revealed that pheophorbide is only accessible for demethylation when MES16 is targeted to the chloroplast. Together, these data demonstrate that MES16 is an integral component of Chl breakdown in Arabidopsis and specifically demethylates Chl catabolites at the level of FCCs in the cytosol.

  20. Importance of tetrahydrofolate and ATP in the anaerobic O-demethylation reaction for phenylmethylethers

    SciTech Connect

    Berman, M.H.; Frazer, A.C. )

    1992-03-01

    DL-Tetrahydrofolate (THF) and ATP were necessary for the anaerobic O-demethylation of phenylmethylethers in cell extracts of the type strain (ATCC 29683) of the homoacetogen Acetobacterium woodii. The reactants for this enzymatic activity have not been previously demonstrated in any system, nor has the mediating enzyme been studied. An assay using reaction mixtures containing 1 mM THF, 2 mM ATP, and 2 mM hydroferulate (i.e., 4-hydroxy,3-methoxyphenylpropionate) was developed and was performed under stringent anaerobic conditions. Pyridine nucleotides and several other possible cofactors were tested but had no effect on the activity. After centrifugation of disrupted cells at 27,000 x g, the activity was found primarily in the supernatant, which had a specific activity of 14.2 {plus minus} 0.5 nmol/min/mg of protein. At saturating levels of each of the other two substrates, apparent K{sub m} values for the variable substrate were 0.65 mM hydroferulate, 0.27 mM ATP, and 0.17 mM THF. Activity was significantly decreased when extract was preincubated at 60C and was completely lost after preincubation in air for 30 min. Thus, the soluble anaerobic O-demethylating enzyme system of A. woodii is oxygen sensitive. The THF- and ATP-dependent activity measurable in the soluble fraction of cell extracts constituted about 34% of the activity seen with intact cells.

  1. Methadone N-demethylation by the common CYP2B6 allelic variant CYP2B6.6.

    PubMed

    Gadel, Sarah; Crafford, Amanda; Regina, Karen; Kharasch, Evan D

    2013-04-01

    The long-acting opioid methadone displays considerable unexplained interindividual pharmacokinetic variability. Methadone metabolism clinically occurs primarily by N-demethylation to 2-ethyl-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), catalyzed predominantly by CYP2B6. Retrospective studies suggest that the common allele variant CYP2B6*6 may influence methadone plasma concentrations. The catalytic activity of CYP2B6.6, encoded by CYP2B6*6, is highly substrate-dependent. This investigation compared methadone N-demethylation by CYP2B6.6 with that by wild-type CYP2B6.1. Methadone enantiomer and racemate N-demethylation by recombinant-expressed CYP2B6.6 and CYP2B6.1 was determined. At substrate concentrations (0.25-2 µM) approximating plasma concentrations occurring clinically, rates of methadone enantiomer N-demethylation by CYP2B6.6, incubated individually or as the racemate, were one-third to one-fourth those by CYP2B6.1. For methadone individual enantiomers and metabolism by CYP2B6.6 compared with CYP2B6.1, Vmax was diminished, Ks was greater and the in vitro intrinsic clearance was diminished 5- to 6-fold. The intrinsic clearance for R- and S-EDDP formation from racemic methadone was diminished approximately 6-fold and 3-fold for R- and S-methadone, respectively. Both CYP2B6.6 and CYP2B6.1 showed similar stereoselectivity (S>R-methadone). Human liver microsomes with diminished CYP2B6 content due to a CYP2B6*6 allele had lower rates of methadone N-demethylation. Results show that methadone N-demethylation catalyzed by CYP2B6.6, the CYP2B6 variant encoded by the CYP2B6*6 polymorphism, is catalytically deficient compared with wild-type CYP2B6.1. Diminished methadone N-demethylation by CYP2B6.6 may provide a mechanistic explanation for clinical observations of altered methadone disposition in individuals carrying the CYP2B6*6 polymorphism.

  2. Hepatic microsomal N-oxidation and N-demethylation of N,N-dimethylaniline in red-winged blackbird compared with rat and other birds

    USGS Publications Warehouse

    Pan, H.P.; Fouts, J.R.; Devereux, T.R.

    1975-01-01

    Hepatic microsomes prepared from red-winged blackbirds and albino rats were incubated with N,N-dimethylaniline (DMA)_in complete incubation mixtures at pH 7.9 and 37?C for 10 min. Formaldehyde and N,N-dimethylaniline--oxide produced from DMA were measured. Redwings were found to have significantly lower N-demethylation activities than rats, and redwings had only marginal or no N-oxidation activities. Hepatic microsomes from redwings did not further metabolize the N-oxide. The N-oxidation and N-demethylation activities of brown-headed cowbirds, common grackles, and starlings were similar to those of redwings.

  3. Active Inference: A Process Theory.

    PubMed

    Friston, Karl; FitzGerald, Thomas; Rigoli, Francesco; Schwartenbeck, Philipp; Pezzulo, Giovanni

    2017-01-01

    This article describes a process theory based on active inference and belief propagation. Starting from the premise that all neuronal processing (and action selection) can be explained by maximizing Bayesian model evidence-or minimizing variational free energy-we ask whether neuronal responses can be described as a gradient descent on variational free energy. Using a standard (Markov decision process) generative model, we derive the neuronal dynamics implicit in this description and reproduce a remarkable range of well-characterized neuronal phenomena. These include repetition suppression, mismatch negativity, violation responses, place-cell activity, phase precession, theta sequences, theta-gamma coupling, evidence accumulation, race-to-bound dynamics, and transfer of dopamine responses. Furthermore, the (approximately Bayes' optimal) behavior prescribed by these dynamics has a degree of face validity, providing a formal explanation for reward seeking, context learning, and epistemic foraging. Technically, the fact that a gradient descent appears to be a valid description of neuronal activity means that variational free energy is a Lyapunov function for neuronal dynamics, which therefore conform to Hamilton's principle of least action.

  4. Demethylation of Polymethoxyflavones by Human Gut Bacterium, Blautia sp. MRG-PMF1.

    PubMed

    Burapan, Supawadee; Kim, Mihyang; Han, Jaehong

    2017-03-01

    Polymethoxyflavones (PMFs) were biotransformed to various demethylated metabolites in the human intestine by the PMF-metabolizing bacterium, Blautia sp. MRG-PMF1. Because the newly formed metabolites can have different biological activities, the pathways and regioselectivity of PMF bioconversion were investigated. Using an anaerobic in vitro study, 12 PMFs, 5,7-dimethoxyflavone (5,7-DMF), 5-hydroxy-7-methoxyflavone (5-OH-7-MF), 3,5,7-trimethoxyflavone (3,5,7-TMF), 5-hydroxy-3,7-dimethoxyflavone (5-OH-3,7-DMF), 5,7,4'-trimethoxyflavone (5,7,4'-TMF), 5-hydroxy-7,4'-dimethoxyflavone (5-OH-7,4'-DMF), 3,5,7,4'-tetramethoxyflavone (3,5,7,4'-TMF), 5-hydroxy-3,7,4'-trimethoxyflavone (5-OH-3,7,4'-TMF), 5,7,3',4'-tetramethoxyflavone (5,7,3',4'-TMF), 3,5,7,3',4'-pentamethoxyflavone (3,5,7,3',4'-PMF), 5-hydroxy-3,7,3',4'-tetramethoxyflavone (5-OH-3,7,3',4'-TMF), and 5,3'-dihydroxy-3,7,4'-trimethoxyflavone (5,3'-diOH-3,7,4'-TMF), were converted to chrysin, apigenin, galangin, kaempferol, luteolin, and quercetin after complete demethylation. The time-course monitoring of PMF biotransformations elucidated bioconversion pathways, including the identification of metabolic intermediates. As a robust flavonoid demethylase, regioselectivity of PMF demethylation generally followed the order C-7 > C-4' ≈ C-3' > C-5 > C-3. PMF demethylase in the MRG-PMF1 strain was suggested as a Co-corrinoid methyltransferase system, and this was supported by the experiments utilizing other methyl aryl ether substrates and inhibitors.

  5. Promoter demethylation of cystathionine-β-synthetase gene contributes to inflammatory pain in rats.

    PubMed

    Qi, Feihu; Zhou, Youlang; Xiao, Ying; Tao, Jin; Gu, Jianguo; Jiang, Xinghong; Xu, Guang-Yin

    2013-01-01

    Hydrogen sulfide (H(2)S), an endogenous gas molecule synthesized by cystathionine-β-synthetase (CBS), is involved in inflammation and nociceptive signaling. However, the molecular and epigenetic mechanisms of CBS-H(2)S signaling in peripheral nociceptive processing remain unknown. We demonstrated that peripheral inflammation induced by intraplantar injection of complete Freund adjuvant significantly up-regulated expression of CBS at both protein and mRNA levels in rat dorsal root ganglia (DRG). The CBS inhibitors hydroxylamine and aminooxyacetic acid attenuated mechanical hyperalgesia in a dose-dependent manner and reversed hyperexcitability of DRG neurons in inflamed rats. Intraplantar administration of NaHS (its addition mimics CBS production of H(2)S) or l-cysteine in healthy rats elicited mechanical hyperalgesia. Application of NaHS in vitro enhanced excitability and tetrodotoxin (TTX)-resistant sodium current of DRG neurons from healthy rats, which was attenuated by pretreatment of protein kinase A inhibitor H89. Methylation-specific PCR and bisulfite sequencing demonstrated that promoter region of cbs gene was less methylated in DRG samples from inflamed rats than that from controls. Peripheral inflammation did not alter expression of DNA methyltransferase 3a and 3b, the 2 major enzymes for DNA methylation, but led to a significant up-regulation of methyl-binding domain protein 4 and growth arrest and DNA damage inducible protein 45α, the enzymes involved in active DNA demethylation. Our findings suggest that epigenetic regulation of CBS expression may contribute to inflammatory hyperalgesia. H(2)S seems to increase TTX-resistant sodium channel current, which may be mediated by protein kinase A pathway, thus identifying a potential therapeutic target for the treatment of chronic pain.

  6. Emodin enhances the demethylation by 5-Aza-CdR of pancreatic cancer cell tumor-suppressor genes P16, RASSF1A and ppENK.

    PubMed

    Pan, Feng-Ping; Zhou, Hong-Kun; Bu, He-Qi; Chen, Zi-Qiang; Zhang, Hao; Xu, Lu-Ping; Tang, Jian; Yu, Qing-Jiang; Chu, Yong-Quan; Pan, Jie; Fei, Yong; Lin, Sheng-Zhang; Liu, Dian-Lei; Chen, Liang

    2016-04-01

    5-Aza-2'-deoxycytidine (5-Aza-CdR) is currently acknowledged as a demethylation drug, and causes a certain degree of demethylation in a variety of cancer cells, including pancreatic cancer cells. Emodin, a traditional Chinese medicine (TCM), is an effective monomer extracted from rhubarb and has been reported to exhibit antitumor activity in different manners in pancreatic cancer. In the present study, we examined whether emodin caused demethylation and increased the demethylation of three tumor-suppressor genes P16, RASSF1A and ppENK with a high degree of methylation in pancreatic cancer when combined with 5-Aza-CdR. Our research showed that emodin inhibited the growth of pancreatic cancer Panc-1 cells in a dose- and time-dependent manner. Dot-blot results showed that emodin combined with 5-Aza-CdR significantly suppressed the expression of genome 5mC in PANC-1 cells. In order to verify the effect of methylation, methylation-specific PCR (MSP) and bisulfite genomic sequencing PCR (BSP) combined with TA were selected for the cloning and sequencing. Results of MSP and BSP confirmed that emodin caused faint demethylation, and 5-Aza-CdR had a certain degree of demethylation. When emodin was combined with 5-Aza-CdR, the demethylation was more significant. At the same time, fluorescent quantitative PCR and western blot analysis results confirmed that when emodin was combined with 5-Aza-CdR, the expression levels of P16, RASSF1A and ppENK were increased more significantly compared to either treatment alone. In contrast, the expression levels of DNA methyltransferase 1 (DNMT1) and DNMT3a were more significantly reduced with the combination treatment than the control or either agent alone, further proving that emodin in combination with 5-Aza-CdR enhanced the demethylation effect of 5-Aza-CdR by reducing the expression of methyltransferases. In conclusion, the present study confirmed that emodin in combination with 5-Aza-CdR enhanced the demethylation by 5-Aza-CdR of tumor

  7. Emodin enhances the demethylation by 5-Aza-CdR of pancreatic cancer cell tumor-suppressor genes P16, RASSF1A and ppENK

    PubMed Central

    PAN, FENG-PING; ZHOU, HONG-KUN; BU, HE-QI; CHEN, ZI-QIANG; ZHANG, HAO; XU, LU-PING; TANG, JIAN; YU, QING-JIANG; CHU, YONG-QUAN; PAN, JIE; FEI, YONG; LIN, SHENG-ZHANG; LIU, DIAN-LEI; CHEN, LIANG

    2016-01-01

    5-Aza-2′-deoxycytidine (5-Aza-CdR) is currently acknowledged as a demethylation drug, and causes a certain degree of demethylation in a variety of cancer cells, including pancreatic cancer cells. Emodin, a traditional Chinese medicine (TCM), is an effective monomer extracted from rhubarb and has been reported to exhibit antitumor activity in different manners in pancreatic cancer. In the present study, we examined whether emodin caused demethylation and increased the demethylation of three tumor-suppressor genes P16, RASSF1A and ppENK with a high degree of methylation in pancreatic cancer when combined with 5-Aza-CdR. Our research showed that emodin inhibited the growth of pancreatic cancer Panc-1 cells in a dose- and time-dependent manner. Dot-blot results showed that emodin combined with 5-Aza-CdR significantly suppressed the expression of genome 5mC in PANC-1 cells. In order to verify the effect of methylation, methylation-specific PCR (MSP) and bisulfite genomic sequencing PCR (BSP) combined with TA were selected for the cloning and sequencing. Results of MSP and BSP confirmed that emodin caused faint demethylation, and 5-Aza-CdR had a certain degree of demethylation. When emodin was combined with 5-Aza-CdR, the demethylation was more significant. At the same time, fluorescent quantitative PCR and western blot analysis results confirmed that when emodin was combined with 5-Aza-CdR, the expression levels of P16, RASSF1A and ppENK were increased more significantly compared to either treatment alone. In contrast, the expression levels of DNA methyltransferase 1 (DNMT1) and DNMT3a were more significantly reduced with the combination treatment than the control or either agent alone, further proving that emodin in combination with 5-Aza-CdR enhanced the demethylation effect of 5-Aza-CdR by reducing the expression of meth-yltransferases. In conclusion, the present study confirmed that emodin in combination with 5-Aza-CdR enhanced the demethylation by 5-Aza-CdR of

  8. Dietary compound isoliquiritigenin prevents mammary carcinogenesis by inhibiting breast cancer stem cells through WIF1 demethylation

    PubMed Central

    Wang, Yu; Xie, Xiaoming; Shen, Jiangang; Peng, Cheng; You, Jieshu; Peng, Fu; Tang, Hailin; Guan, Xinyuan; Chen, Jianping

    2015-01-01

    Breast cancer stem cells (CSCs) are considered as the root of mammary tumorigenesis. Previous studies have demonstrated that ISL efficiently limited the activities of breast CSCs. However, the cancer prevention activities of ISL and its precise molecular mechanisms remain largely unknown. Here, we report a novel function of ISL as a natural demethylation agent targeting WIF1 to prevent breast cancer. ISL administration suppressed in vivo breast cancer initiation and progression, accompanied by reduced CSC-like populations. A global gene expression profile assay further identified WIF1 as the main response gene of ISL treatment, accompanied by the simultaneous downregulation of β-catenin signaling and G0/G1 phase arrest in breast CSCs. In addition, WIF1 inhibition significantly relieved the CSC-limiting effects of ISL and methylation analysis further revealed that ISL enhanced WIF1 gene expression via promoting the demethylation of its promoter, which was closely correlated with the inhibition of DNMT1 methyltransferase. Molecular docking analysis finally revealed that ISL could stably dock into the catalytic domain of DNMT1. Taken together, our findings not only provide preclinical evidence to demonstrate the use of ISL as a dietary supplement to inhibit mammary carcinogenesis but also shed novel light on WIF1 as an epigenetic target for breast cancer prevention. PMID:25918249

  9. Microbial Oxidation and Demethylation Processes in the Environmental Mercury Cycle

    SciTech Connect

    Summers, Anne O.

    2000-10-30

    This project demonstrated that bacterial catalase enzymes can convert unreactive Hg(0) to highly reactive Hg(II) ion. It also demonstrated the mechanism of the organomercural lyase, a bacterial enzyme which degrades methylmercury and other organomercurials. Lastly, it demonstrated the 3-dimensional structure of this enzyme by both solution NMR and by x-ray crystallography. These structures provide insights into the catalytic mechanism of the lyase that will allow engineering of variants with improved ability to degrade methylmercury.

  10. Glycogen synthase kinase-3β regulates leucine-309 demethylation of protein phosphatase-2A via PPMT1 and PME-1.

    PubMed

    Yao, Xiu-Qing; Li, Xia-Chun; Zhang, Xiao-Xue; Yin, Yang-Yang; Liu, Bin; Luo, Dan-Ju; Wang, Qun; Wang, Jian-Zhi; Liu, Gong-Ping

    2012-07-30

    Protein phosphatase-2A (PP2A) activity is significantly suppressed in Alzheimer's disease. We have reported that glycogen synthase kinase-3β (GSK-3β) inhibits PP2A via upregulating the phosphorylation of PP2A catalytic subunit (PP2A(C)). Here we studied the effects of GSK-3β on the inhibitory demethylation of PP2A at leucine-309 (dmL309-PP2A(C)). We found that GSK-3β regulates dmL309-PP2A(C) level by regulating PME-1 and PPMT1. Knockdown of PME-1 or PPMT1 eliminated the effects of GSK-3β on PP2A(C). GSK-3 could negatively regulate PP2A regulatory subunit protein level. We conclude that GSK-3β can inhibit PP2A by increasing the inhibitory L309-demethylation involving upregulation of PME-1 and inhibition of PPMT1.

  11. Dynamic and selective HERV RNA expression in neuroblastoma cells subjected to variation in oxygen tension and demethylation.

    PubMed

    Hu, Lijuan; Uzhameckis, Dmitrijs; Hedborg, Fredrik; Blomberg, Jonas

    2016-01-01

    We studied HERV expression in cell lines after hypoxia, mitogenic stimulation, and demethylation, to better understand if hypoxia may play a role in ERV activation also within the nervous system, as represented by neuroblastoma cell lines. The level of RNA of four human ERV groups (HERVs) (HERVE, I/T, H, and W), and three housekeeping genes, of different cell lines including A549, COS-1, Namalwa, RD-L and Vero-E6, as well as human neuroblastoma cell lines SH-SY5Y, SK-N-DZ, and SK-N-AS were studied using reverse transcription and real-time quantitative PCR (QPCR). During the course of recovery from hypoxia a pronounced and selective activation of RNA expression of HERVW-like sequences, but not of HERVE, I/T, H, and three housekeeping genes, was found in the neuroblastoma cell lines, most pronounced in SK-N-DZ. In the SK-N-DZ cell line, we also tested the expression of HERVs after chemical treatments. HERVW-like sequences were selectively upregulated by 5-azacytidine, a demethylating agent. Some HERVW loci seem especially responsive to hypoxia and demethylation. HERV expression in neuroblastoma cells is selectively and profoundly influenced by some physiological and chemical stimuli.

  12. Green tea polyphenol EGCG reverse cisplatin resistance of A549/DDP cell line through candidate genes demethylation.

    PubMed

    Zhang, Youwei; Wang, Xiang; Han, Liang; Zhou, Yizhou; Sun, Sanyuan

    2015-02-01

    Epigallocatechin-3-gallate (EGCG), the major polyphenol in green tea, has been extensively studied as a potential demethylating agent. Our hypothesis is that EGCG could resensitize non-small-cell lung cancer (NSCLC) cells to cisplatin (DDP) through candidate genes demethylation. The A549/DDP cell line was established by continuous exposure of A549 cells to increasing concentrations of DDP. MTT, colony formation assay, flow cytometric analysis, Hoechst staining, real time-PCR, quantitative methylation-specific PCR and in vivo experiments were performed in this study. EGCG+DDP treatment significantly caused proliferation inhibition, cell cycle arrest in G1 phase, increase of apoptosis in A549/DDP cells, along with inhibition of DNA methyltransferase (DNMT) activity and histone deacetylase (HDAC) activity, reversal of hypermethylated status and downregulated expression of GAS1, TIMP4, ICAM1 and WISP2 gene in A549/DDP cells. Furthermore, pre-treatment with EGCG followed by DDP caused significant tumor inhibition in vivo. Methylation levels of GAS1, TIMP4, ICAM1 and WISP2 were decreased and their expression levels were increased in EGCG-treatment groups, but only combinatorial treatment group caused growth inhibition. In conclusion, we identified EGCG pretreatment resensitized cells to DDP, along with the demethylation and restoration of expression of candidate genes.

  13. Parallel Activation in Bilingual Phonological Processing

    ERIC Educational Resources Information Center

    Lee, Su-Yeon

    2011-01-01

    In bilingual language processing, the parallel activation hypothesis suggests that bilinguals activate their two languages simultaneously during language processing. Support for the parallel activation mainly comes from studies of lexical (word-form) processing, with relatively less attention to phonological (sound) processing. According to…

  14. Evaluation of mercury methylation and methylmercury demethylation rates in vegetated and non-vegetated saltmarsh sediments from two Portuguese estuaries.

    PubMed

    Cesário, Rute; Hintelmann, Holger; Mendes, Ricardo; Eckey, Kevin; Dimock, Brian; Araújo, Beatriz; Mota, Ana Maria; Canário, João

    2017-04-05

    Neurotoxic methylmercury (MMHg) is formed from inorganic divalent mercury (Hg(2+)). However, it is poorly understood to what extent different mercury (Hg) pools contribute to existent MMHg levels. In this study, ambient concentrations of total Hg (THg) and MMHg as well as rates of methylation and demethylation were measured simultaneously in sediments with and without salt-marsh plant vegetation, which were collected in Guadiana and Tagus estuaries, Portugal. Concurrent processes of Hg methylation and MMHg demethylation were directly monitored and compared by spiking sediments cores with stable isotope tracers of (199)Hg(2+) and CH3(201)Hg(+) followed by gas chromatographic separation and isotope-specific detection using inductively coupled plasma mass spectrometry. Compared to the Guadiana estuary, where concentrations were comparatively low, THg and MMHg levels varied between vegetated and non-vegetated sediments collected at the Rosário site (ROS) of the Tagus estuary. Methylation (KM) and demethylation rates (KD) were also different between estuaries being dependent on the presence of vegetation. In addition, the type of macrophyte species influenced KM and KD values. In fact, the highest KM value was found in Sarcocornia fruticosa vegetated sediments at the Castro Marim site in Guadiana (CM, 0.160 day(-1)) and the lowest KM was observed in non-vegetated sediments at the Alcochete site in Tagus (ALC, 0.009 day(-1)). KD varied by a factor of three among sites with highest rates of demethylation observed in non-vegetated sediments in Guadiana (12 ± 1.3 day(-1), corresponding to a half-life of 1.4 ± 0.2 h). This study clearly shows that the presence of vegetation in sediments favors the formation of MMHg. Moreover, this effect might be site specific and further studies are needed to confirm the findings reported here.

  15. Simulation of human plasma concentration-time profiles of the partial glucokinase activator PF-04937319 and its disproportionate N-demethylated metabolite using humanized chimeric mice and semi-physiological pharmacokinetic modeling.

    PubMed

    Kamimura, Hidetaka; Ito, Satoshi; Chijiwa, Hiroyuki; Okuzono, Takeshi; Ishiguro, Tomohiro; Yamamoto, Yosuke; Nishinoaki, Sho; Ninomiya, Shin-Ichi; Mitsui, Marina; Kalgutkar, Amit S; Yamazaki, Hiroshi; Suemizu, Hiroshi

    2016-07-07

    1. The partial glucokinase activator N,N-dimethyl-5-((2-methyl-6-((5-methylpyrazin-2-yl)carbamoyl)benzofuran-4-yl)oxy)pyrimidine-2-carboxamide (PF-04937319) is biotransformed in humans to N-methyl-5-((2-methyl-6-((5-methylpyrazin-2-yl)carbamoyl)benzofuran-4-yl)oxy)pyrimidine-2-carboxamide (M1), accounting for ∼65% of total exposure at steady state. 2. As the disproportionately abundant nature of M1 could not be reliably predicted from in vitro metabolism studies, we evaluated a chimeric mouse model with humanized liver on TK-NOG background for its ability to retrospectively predict human disposition of PF-04937319. Since livers of chimeric mice were enlarged by hyperplasia and contained remnant mouse hepatocytes, hepatic intrinsic clearances normalized for liver weight, metabolite formation and liver to plasma concentration ratios were plotted against the replacement index by human hepatocytes and extrapolated to those in the virtual chimeric mouse with 100% humanized liver. 3. Semi-physiological pharmacokinetic analyses using the above parameters revealed that simulated concentration curves of PF-04937319 and M1 were approximately superimposed with the observed clinical data in humans. 4. Finally, qualitative profiling of circulating metabolites in humanized chimeric mice dosed with PF-04937319 or M1 also revealed the presence of a carbinolamide metabolite, identified in the clinical study as a human-specific metabolite. The case study demonstrates that humanized chimeric mice may be potentially useful in preclinical discovery towards studying disproportionate or human-specific metabolism of drug candidates.

  16. ROS1 5-methylcytosine DNA glycosylase is a slow-turnover catalyst that initiates DNA demethylation in a distributive fashion.

    PubMed

    Ponferrada-Marín, María Isabel; Roldán-Arjona, Teresa; Ariza, Rafael R

    2009-07-01

    Arabidopsis ROS1 belongs to a family of plant 5-methycytosine DNA glycosylases that initiate DNA demethylation through base excision. ROS1 displays the remarkable capacity to excise 5-meC, and to a lesser extent T, while retaining the ability to discriminate effectively against C and U. We found that replacement of the C5-methyl group by halogen substituents greatly decreased excision of the target base. Furthermore, 5-meC was excised more efficiently from mismatches, whereas excision of T only occurred when mispaired with G. These results suggest that ROS1 specificity arises by a combination of selective recognition at the active site and thermodynamic stability of the target base. We also found that ROS1 is a low-turnover catalyst because it binds tightly to the abasic site left after 5-meC removal. This binding leads to a highly distributive behaviour of the enzyme on DNA substrates containing multiple 5-meC residues, and may help to avoid generation of double-strand breaks during processing of bimethylated CG dinucleotides. We conclude that the biochemical properties of ROS1 are consistent with its proposed role in protecting the plant genome from excess methylation.

  17. ROS1 5-methylcytosine DNA glycosylase is a slow-turnover catalyst that initiates DNA demethylation in a distributive fashion

    PubMed Central

    Ponferrada-Marín, María Isabel; Roldán-Arjona, Teresa; Ariza, Rafael R.

    2009-01-01

    Arabidopsis ROS1 belongs to a family of plant 5-methycytosine DNA glycosylases that initiate DNA demethylation through base excision. ROS1 displays the remarkable capacity to excise 5-meC, and to a lesser extent T, while retaining the ability to discriminate effectively against C and U. We found that replacement of the C5-methyl group by halogen substituents greatly decreased excision of the target base. Furthermore, 5-meC was excised more efficiently from mismatches, whereas excision of T only occurred when mispaired with G. These results suggest that ROS1 specificity arises by a combination of selective recognition at the active site and thermodynamic stability of the target base. We also found that ROS1 is a low-turnover catalyst because it binds tightly to the abasic site left after 5-meC removal. This binding leads to a highly distributive behaviour of the enzyme on DNA substrates containing multiple 5-meC residues, and may help to avoid generation of double-strand breaks during processing of bimethylated CG dinucleotides. We conclude that the biochemical properties of ROS1 are consistent with its proposed role in protecting the plant genome from excess methylation. PMID:19443451

  18. Cytochrome P4502D6 catalyzes the O-demethylation of the psychoactive alkaloid ibogaine to 12-hydroxyibogamine.

    PubMed

    Obach, R S; Pablo, J; Mash, D C

    1998-08-01

    Ibogaine is a psychoactive alkaloid that possesses potential as an agent to treat opiate and cocaine addiction. The primary metabolite arises via O-demethylation at the 12-position to yield 12-hydroxyibogamine. In this report, evidence is presented that the O-demethylation of ibogaine observed in human hepatic microsomes is catalyzed primarily by the polymorphically expressed cytochrome P-4502D6 (CYP2D6). An enzyme kinetic examination of ibogaine O-demethylase activity in pooled human liver microsomes suggested that two (or more) enzymes are involved in this reaction: one with a low KMapp (1.1 microM) and the other with a high KMapp (>200 microM). The low KMapp activity comprised >95% of total intrinsic clearance. Human liver microsomes from three individual donors demonstrated similar enzyme kinetic parameters (mean KMapp = 0.55 +/- 0.09 microM and 310 +/- 10 microM for low and high KM activities, respectively). However, a fourth human microsome sample that appeared to be a phenotypic CYP2D6 poor metabolizer possessed only the high KMapp activity. In hepatic microsomes from a panel of human donors, the low KMapp ibogaine O-demethylase activity correlated with CYP2D6-catalyzed bufuralol 1'-hydroxylase activity but not with other P450 isoform-specific activities. Quinidine, a CYP2D6-specific inhibitor, inhibited ibogaine O-demethylase (IC50 = 0.2 microM), whereas other P450 isoform-specific inhibitors did not inhibit this activity. Also, of a battery of recombinant heterologously expressed human P450 isoforms, only rCYP2D6 possessed significant ibogaine O-demethylase activity. Thus, it is concluded that ibogaine O-demethylase is catalyzed by CYP2D6 and that this isoform is the predominant enzyme of ibogaine O-demethylation in humans. The potential pharmacological implications of these findings are discussed.

  19. Enhanced binding capability of nuclear factor-κB with demethylated P2X3 receptor gene contributes to cancer pain in rats.

    PubMed

    Zhou, You-Lang; Jiang, Guo-Qin; Wei, Jinrong; Zhang, Hong-Hong; Chen, Wei; Zhu, Hongyan; Hu, Shufen; Jiang, Xinghong; Xu, Guang-Yin

    2015-10-01

    Nuclear factor-kappa B (NF-κB) signaling is implicated in both cancer development and inflammation processes. However, the roles and mechanisms of NF-κB signaling in the development of cancer-induced pain (CIP) remain unknown. This study was designed to investigate the roles of the p65 subunit of NF-κB in regulation of the purinergic receptor (P2X3R) plasticity in dorsal root ganglion (DRG) of CIP rats. We showed here that tumor cell injection produced mechanical and thermal hyperalgesia, and an enhanced body weight-bearing difference, which was correlated with an upregulation of p65 and P2X3R expression in lumber DRGs and a potentiation of ATP-evoked responses of tibia-innervating DRG neurons. Inhibition of NF-κB signaling using p65 inhibitor pyrrolidine dithiocarbamate, BAY-11-7082, or lentiviral-p65 short-hairpin RNA significantly attenuated CIP and reversed the activities of P2X3R. Interestingly, tumor cell injection led to a significant demethylation of CpG island in p2x3r gene promoter and enhanced ability of p65 to bind the promoter of p2x3r gene. Our findings suggest that upregulation of P2X3R expression was mediated by the enhanced binding capability of p65 with demethylated promoter of p2x3r gene, thus contributing to CIP. NF-κBp65 might be a potential target for treating CIP, a neuropathic pain generated by tumor cell-induced injury to nerves that innervate the skin.

  20. Enhanced binding capability of nuclear factor-κB with demethylated P2X3 receptor gene contributes to cancer pain in rats

    PubMed Central

    Zhou, You-Lang; Jiang, Guo-Qin; Wei, Jinrong; Zhang, Hong-Hong; Chen, Wei; Zhu, Hongyan; Hu, Shufen; Jiang, Xinghong; Xu, Guang-Yin

    2015-01-01

    Abstract Nuclear factor-kappa B (NF-κB) signaling is implicated in both cancer development and inflammation processes. However, the roles and mechanisms of NF-κB signaling in the development of cancer-induced pain (CIP) remain unknown. This study was designed to investigate the roles of the p65 subunit of NF-κB in regulation of the purinergic receptor (P2X3R) plasticity in dorsal root ganglion (DRG) of CIP rats. We showed here that tumor cell injection produced mechanical and thermal hyperalgesia, and an enhanced body weight–bearing difference, which was correlated with an upregulation of p65 and P2X3R expression in lumber DRGs and a potentiation of ATP-evoked responses of tibia-innervating DRG neurons. Inhibition of NF-κB signaling using p65 inhibitor pyrrolidine dithiocarbamate, BAY-11-7082, or lentiviral-p65 short-hairpin RNA significantly attenuated CIP and reversed the activities of P2X3R. Interestingly, tumor cell injection led to a significant demethylation of CpG island in p2x3r gene promoter and enhanced ability of p65 to bind the promoter of p2x3r gene. Our findings suggest that upregulation of P2X3R expression was mediated by the enhanced binding capability of p65 with demethylated promoter of p2x3r gene, thus contributing to CIP. NF-κBp65 might be a potential target for treating CIP, a neuropathic pain generated by tumor cell–induced injury to nerves that innervate the skin. PMID:26049406

  1. DNA demethylation induced by 5-azacytidine does not affect fragile X expression.

    PubMed Central

    Glover, T W; Coyle-Morris, J; Pearce-Birge, L; Berger, C; Gemmill, R M

    1986-01-01

    Experiments were performed to determine the role of DNA demethylation in fragile X expression. Fragile X positive lymphoblastoid cells were treated with 5-azacytidine and harvested for analysis of fragile X expression both directly following treatment and after a recovery period in the absence of the drug. The effectiveness of 5-azacytidine treatment in inducing DNA demethylation was concurrently monitored by analysis of methylation changes at random autosomal loci in isolated DNA from treated cells. Under conditions where 5-azacytidine was found to inhibit fragile X expression, no DNA demethylation was observed. At the time when demethylation did occur, fragile X expression was not affected. These results strongly indicate that DNA demethylation is not involved in fragile X expression. Images Fig. 1 PMID:2420174

  2. Crystal Structure of Dicamba Monooxygenase: A Rieske Nonheme Oxygenase that Catalyzes Oxidative Demethylation

    SciTech Connect

    Dumitru, Razvan; Jiang, Wen Zhi; Weeks, Donald P.; Wilson, Mark A.

    2009-08-28

    Dicamba (3,6-dichloro-2-methoxybenzoic acid) is a widely used herbicide that is efficiently degraded by soil microbes. These microbes use a novel Rieske nonheme oxygenase, dicamba monooxygenase (DMO), to catalyze the oxidative demethylation of dicamba to 3,6-dichlorosalicylic acid (DCSA) and formaldehyde. We have determined the crystal structures of DMO in the free state, bound to its substrate dicamba, and bound to the product DCSA at 2.10-1.75 {angstrom} resolution. The structures show that the DMO active site uses a combination of extensive hydrogen bonding and steric interactions to correctly orient chlorinated, ortho-substituted benzoic-acid-like substrates for catalysis. Unlike other Rieske aromatic oxygenases, DMO oxygenates the exocyclic methyl group, rather than the aromatic ring, of its substrate. This first crystal structure of a Rieske demethylase shows that the Rieske oxygenase structural scaffold can be co-opted to perform varied types of reactions on xenobiotic substrates.

  3. Transcriptional regulation of 15-lipoxygenase expression by histone h3 lysine 4 methylation/demethylation.

    PubMed

    Liu, Cheng; Xu, Dawei; Han, Hongya; Fan, Yidong; Schain, Frida; Xu, Zhonghua; Claesson, Hans-Erik; Björkholm, Magnus; Sjöberg, Jan

    2012-01-01

    15-Lipoxygenase-1 (15-LOX-1) oxidizes polyunsaturated fatty acids to a rich spectrum of biologically active metabolites and is implicated in physiological membrane remodelling, inflammation and apoptosis. Its deregulation is involved in the pathogenesis of diverse cancer and immune diseases. Recent experimental evidence reveals that dynamic histone methylation/demethylation mediated by histone methyltransferases and demethylases plays a critical role in regulation of chromatin remodelling and gene expression. In the present study, we compared the histone 3 lysine 4 (H3-K4) methylation status of the 15-LOX-1 promoter region of the two Hodgkin lymphoma (HL) cell lines L1236 and L428 with abundant and undetectable 15-LOX-1 expression, respectively. We identified a potential role of H3-K4 methylation in positive regulation of 15-LOX-1 transcription. Furthermore, we found that histone methyltransferase SMYD3 inhibition reduced 15-LOX-1 expression by decreasing promoter activity in L1236 cells. SMYD3 knock down in these cells abolished di-/trimethylation of H3-K4, attenuated the occupancy by the transactivator STAT6, and led to diminished histone H3 acetylation at the 15-LOX-1 promoter. In contrast, inhibition of SMCX, a JmjC-domain-containing H3-K4 tri-demethylase, upregulated 15-LOX-1 expression through induction of H3-K4 trimethylation, histone acetylation and STAT6 recruitment at the 15-LOX-1 promoter in L428 cells. In addition, we observed strong SMYD3 expression in the prostate cancer cell line LNCaP and its inhibition led to decreased 15-LOX-1 expression. Taken together, our data suggest that regulation of histone methylation/demethylation at the 15-LOX-1 promoter is important in 15-LOX-1 expression.

  4. A re-investigation of the ribonuclease sensitivity of a DNA demethylation reaction in chicken embryo and G8 mouse myoblasts.

    PubMed

    Jost, J P; Siegmann, M; Thiry, S; Jost, Y C; Benjamin, D; Schwarz, S

    1999-04-23

    Recently published results (Nucleic Acids Res. 26, 5573-5580, 1998) suggest that the ribonuclease sensitivity of the DNA demethylation reaction may be an experimental artifact due to the possible tight binding of the nucleases to the methylated DNA substrate. Using an improved protocol we show for two different systems that demethylation of hemimethylated DNA is indeed sensitive to micrococcal nuclease, requires RNA and is not an experimental artifact. The purified 5-MeC-DNA glycosylase from chicken embryos and G8 mouse myoblasts was first incubated for 5 min at 37 degrees C with micrococcal nuclease in the presence of Ca2+ in the absence of the DNA substrate. Upon blocking the nuclease activity by the addition of 25 mM EGTA, the DNA demethylation reaction was initiated by adding the labeled hemimethylated DNA substrate to the reaction mixture. Under these conditions the DNA demethylation reaction was abolished. In parallel controls, where the purified 5-MeC-DNA glycosylase was pre-incubated at 37 degrees C with the nuclease, Ca2+ and EGTA or with the nuclease and EGTA, RNA was not degraded and no inhibition of the demethylation reaction was obtained. As has already been shown for chicken embryos, the loss of 5-MeC-DNA glycosylase activity from G8 myoblasts following nuclease treatment can also be restored by the addition of synthetic RNA complementary to the methylated strand of the substrate DNA. No reactivation of 5-MeC-DNA glycosylase is obtained by complementation with a random RNA sequence, the RNA sequence complementary to the non-methylated strand or DNA, thus ruling out a non-specific competition of the RNA for the binding of the nuclease to the labeled DNA substrate.

  5. O2 -independent demethylation of trimethylamine N-oxide by Tdm of Methylocella silvestris.

    PubMed

    Zhu, Yijun; Ksibe, Amira Z; Schäfer, Hendrik; Blindauer, Claudia A; Bugg, Timothy D H; Chen, Yin

    2016-11-01

    Bacterial trimethylamine N-oxide (TMAO) demethylase, Tdm, carries out an unusual oxygen-independent demethylation reaction, resulting in the formation of dimethylamine and formaldehyde. In this study, site-directed mutagenesis, homology modelling and metal analyses by inorganic mass spectrometry have been applied to gain insight into metal stoichiometry and underlying catalytic mechanism of Tdm of Methylocella silvestris BL2. Herein, we demonstrate that active Tdm has 1 molar equivalent of Zn(2+) and 1 molar equivalent of non-haem Fe(2+) . We further investigated Zn(2+) - and Fe(2+) -binding sites through homology modelling and site-directed mutagenesis and found that Zn(2+) is coordinated by a 3-sulfur-1-O motif. An aspartate residue (D198) likely bridges Fe(2+) and Zn(2+) centres, either directly or indirectly via H-bonding through a neighbouring H2 O molecule. H276 contributes to Fe(2+) binding, mutation of which results in an inactive enzyme, and the loss of iron, but not zinc. Site-directed mutagenesis of Tdm also led to the identification of three hydrophobic aromatic residues likely involved in substrate coordination (F259, Y305, W321), potentially through a cation-π interaction. Furthermore, a crossover experiment using a substrate analogue gave direct evidence that a trimethylamine-alike intermediate was produced during the Tdm catalytic cycle, suggesting TMAO has a dual role of being both a substrate and an oxygen donor for formaldehyde formation. Together, our results provide novel insight into the role of Zn(2+) and Fe(2+) in the catalysis of TMAO demethylation by this unique oxygen-independent enzyme.

  6. Two-step iron(0)-mediated N-demethylation of N-methyl alkaloids.

    PubMed

    Kok, Gaik B; Pye, Cory C; Singer, Robert D; Scammells, Peter J

    2010-07-16

    A mild and simple two-step Fe(0)-mediated N-demethylation of a number of tertiary N-methyl alkaloids is described. The tertiary N-methylamine is first oxidized to the corresponding N-oxide, which is isolated as the hydrochloride salt. Subsequent treatment of the N-oxide hydrochloride with iron powder readily provides the N-demethylated amine. Representative substrates include a number of opiate and tropane alkaloids. Key intermediates in the synthesis of semisynthetic 14-hydroxy pharmaceutical opiates such as oxycodone and oxymorphone are also readily N-demethylated using this method.

  7. Energetics of active transport processes.

    PubMed

    Essig, A; Caplan, S R

    1968-12-01

    Discussions of active transport usually assume stoichiometry between the rate of transport J(+) and the metabolic rate J(r). However, the observation of a linear relationship between J(+) and J(r) does not imply a stoichiometric relationship, i.e., complete coupling. Since coupling may possibly be incomplete, we examine systems of an arbitrary degree of coupling q, regarding stoichiometry as a limiting case. We consider a sodium pump, with J(+) and J(r) linear functions of the electrochemical potential difference, -X(+), and the chemical affinity of the metabolic driving reaction, A. The affinity is well defined even for various complex reaction pathways. Incorporation of a series barrier and a parallel leak does not affect the linearity of the composite observable system. The affinity of some region of the metabolic chain may be maintained constant, either by large pools of reactants or by regulation. If so, this affinity can be evaluated by two independent methods. Sodium transport is conveniently characterized by the open-circuit potential (Deltapsi)(I=0) and the natural limits, level flow (J(+))(X+=0), and static head X(0) (+) = (X(+))(J+=0). With high degrees of coupling -X(0) (+)/F approaches the electromotive force E(Na) (Ussing); -X(0) (+)/F cannot be identified with ((RT/F) ln f)(X+=0), where f is the flux ratio. The efficiency eta = -J(+)X(+)/J(r)A is of significance only when appreciable energy is being converted from one form to another. When either J(+) or -X(+) is small eta is low; the significant parameters are then the efficacies epsilon(J+) = J(+)/J(r)A and epsilon(X+) = -X(+)/J(r)A, respectively maximal at level flow and static head. Leak increases both J(+) and epsilon(J+) for isotonic saline reabsorption, but diminishes -X(0) (+) and epsilon(Xfemale symbol). Electrical resistance reflects both passive parameters and metabolism. Various fundamental relations are preserved despite coupling of passive ion and water flows.

  8. The CYP2B6*6 allele significantly alters the N-demethylation of ketamine enantiomers in vitro.

    PubMed

    Li, Yibai; Coller, Janet K; Hutchinson, Mark R; Klein, Kathrin; Zanger, Ulrich M; Stanley, Nathan J; Abell, Andrew D; Somogyi, Andrew A

    2013-06-01

    Ketamine is primarily metabolized to norketamine by hepatic CYP2B6 and CYP3A4-mediated N-demethylation. However, the relative contribution from each enzyme remains controversial. The CYP2B6*6 allele is associated with reduced enzyme expression and activity that may lead to interindividual variability in ketamine metabolism. We examined the N-demethylation of individual ketamine enantiomers using human liver microsomes (HLMs) genotyped for the CYP2B6*6 allele, insect cell-expressed recombinant CYP2B6 and CYP3A4 enzymes, and COS-1 cell-expressed recombinant CYP2B6.1 and CYP2B6.6 protein variant. Effects of CYP-selective inhibitors on norketamine formation were also determined in HLMs. The two-enzyme Michaelis-Menten model best fitted the HLM kinetic data. The Michaelis-Menten constants (K(m)) for the high-affinity enzyme and the low-affinity enzyme were similar to those for the expressed CYP2B6 and CYP3A4, respectively. The intrinsic clearance for both ketamine enantiomers by the high-affinity enzyme in HLMs with CYP2B6*1/*1 genotype were at least 2-fold and 6-fold higher, respectively, than those for CYP2B6*1/*6 genotype and CYP2B6*6/*6 genotype. The V(max) and K(m) values for CYP2B6.1 were approximately 160 and 70% of those for CYP2B6.6, respectively. N,N'N'-triethylenethiophosphoramide (thioTEPA) (CYP2B6 inhibitor, 25 μM) and the monoclonal antibody against CYP2B6 but not troleandomycin (CYP3A4 inhibitor, 25 μM) or the monoclonal antibody against CYP3A4 inhibited ketamine N-demethylation at clinically relevant concentrations. The degree of inhibition was significantly reduced in HLMs with the CYP2B6*6 allele (gene-dose P < 0.05). These results indicate a major role of CYP2B6 in ketamine N-demethylation in vitro and a significant impact of the CYP2B6*6 allele on enzyme-ketamine binding and catalytic activity.

  9. Co-expression of Cyanobacterial Genes for Arsenic Methylation and Demethylation in Escherichia coli Offers Insights into Arsenic Resistance

    PubMed Central

    Yan, Yu; Xue, Xi-Mei; Guo, Yu-Qing; Zhu, Yong-Guan; Ye, Jun

    2017-01-01

    Arsenite [As(III)] and methylarsenite [MAs(III)] are the most toxic inorganic and methylated arsenicals, respectively. As(III) and MAs(III) can be interconverted in the unicellular cyanobacterium Nostoc sp. PCC 7120 (Nostoc), which has both the arsM gene (NsarsM), which is responsible for arsenic methylation, and the arsI gene (NsarsI), which is responsible for MAs(III) demethylation. It is not clear how the cells prevent a futile cycle of methylation and demethylation. To investigate the relationship between arsenic methylation and demethylation, we constructed strains of Escherichia coli AW3110 (ΔarsRBC) expressing NsarsM or/and NsarsI. Expression of NsarsI conferred MAs(III) resistance through MAs(III) demethylation. Compared to NsArsI, NsArsM conferred higher resistance to As(III) and lower resistance to MAs(III) by methylating both As(III) and MAs(III). The major species found in solution was dimethylarsenate [DMAs(V)]. Co-expression of NsarsM and NsarsI conferred As(III) resistance at levels similar to that with NsarsM alone, although the main species found in solution after As(III) biotransformation was methylarsenate [MAs(V)] rather than DMAs(V). Co-expression of NsarsM and NsarsI conferred a higher level of resistance to MAs(III) than found with expression of NsarsM alone but lower than expression of only NsarsI. Cells co-expressing both genes converted MAs(III) to a mixture of As(III) and DMAs(V). In Nostoc NsarsM is constitutively expressed, while NsarsI is inducible by either As(III) or MAs(III). Thus, our results suggest that at low concentrations of arsenic, NsArsM activity predominates, while NsArsI activity predominates at high concentrations. We propose that coexistence of arsM and arsI genes in Nostoc could be advantageous for several reasons. First, it confers a broader spectrum of resistance to both As(III) and MAs(III). Second, at low concentrations of arsenic, the MAs(III) produced by NsArsM will possibly have antibiotic-like properties and

  10. Epigenetic choreography of stem cells: the DNA demethylation episode of development.

    PubMed

    Kar, Swayamsiddha; Parbin, Sabnam; Deb, Moonmoon; Shilpi, Arunima; Sengupta, Dipta; Rath, Sandip Kumar; Rakshit, Madhumita; Patra, Aditi; Patra, Samir Kumar

    2014-03-01

    Reversible DNA methylation is a fundamental epigenetic manipulator of the genomic information in eukaryotes. DNA demethylation plays a very significant role during embryonic development and stands out for its contribution in molecular reconfiguration during cellular differentiation for determining stem cell fate. DNA demethylation arbitrated extensive make-over of the genome via reprogramming in the early embryo results in stem cell plasticity followed by commitment to the principal cell lineages. This article attempts to highlight the sequential phases and hierarchical mode of DNA demethylation events during enactment of the molecular strategy for developmental transition. A comprehensive knowledge regarding the pattern of DNA demethylation during embryogenesis and organogenesis and study of the related lacunae will offer exciting avenues for future biomedical research and stem cell-based regenerative therapy.

  11. Cumene hydroperoxide-supported demethylation reactions catalyzed by cytochrome P450 2B4 lacking the NH2-terminal sequence.

    PubMed

    Zhang, Y; Pernecky, S J

    1999-04-29

    Catalytic activities of cytochrome P450 2B4 lacking NH2-terminal amino acids 2-27 (wt Delta2B4) and that of truncated 2B4 containing a Pro to Ser mutation at position 221 were examined in a system supported by cumene hydroperoxide. Demethylation activities of either truncated 2B4 with N-methylaniline, N,N-dimethylaniline, and d-benzphetamine were lower than those of liver microsomal 2B4, whereas the rate of 1-phenylethanol oxidation to acetophenone catalyzed by liver microsomal and truncated 2B4 enzymes was nearly the same. The Km and Vmax values for cumene hydroperoxide in the demethylation of N-methylaniline by wt Delta2B4 were 20% and 28%, respectively, of those obtained for 2B4. The reaction with wt Delta2B4 displayed a lesser dependence on phospholipid than did that with 2B4, and a complex relationship between activity and substrate concentration. The results suggest that the NH2-terminal region contributes to interaction of oxidant, substrate, and phospholipid in cumene hydroperoxide-supported reactions catalyzed by cytochrome P450 2B4.

  12. Efficient N-demethylation of opiate alkaloids using a modified nonclassical Polonovski reaction.

    PubMed

    McCamley, Kristy; Ripper, Justin A; Singer, Robert D; Scammells, Peter J

    2003-12-12

    A modified Polonovski reaction has been employed to N-demethylate several opiate alkaloids in moderate to high yield. This method provides an alternative to traditional N-demethylation procedures which utilize toxic reagents such as cyanogen bromide or expensive reagents such as vinyl chloroformate. The current synthesis involves N-oxide formation, isolation of the corresponding N-oxide hydrochloride, and an FeSO(4).7H(2)O mediated Polonovski reaction to afford the desired secondary amine.

  13. Functional characterization of CYP2B6 allelic variants in demethylation of antimalarial artemether.

    PubMed

    Honda, Masashi; Muroi, Yuka; Tamaki, Yuichiro; Saigusa, Daisuke; Suzuki, Naoto; Tomioka, Yoshihisa; Matsubara, Yoichi; Oda, Akifumi; Hirasawa, Noriyasu; Hiratsuka, Masahiro

    2011-10-01

    Artemether (AM) is one of the most effective antimalarial drugs. The elimination half-life of AM is very short, and it shows large interindividual variability in pharmacokinetic parameters. The aim of this study was to identify cytochrome P450 (P450) isozymes responsible for the demethylation of AM and to evaluate functional differences between 26 CYP2B6 allelic variants in vitro. Of 14 recombinant P450s examined in this study, CYP2B6 and CYP3A4 were primarily responsible for production of the desmethyl metabolite dihydroartemisinin. The intrinsic clearance (V(max)/K(m)) of CYP2B6 was 6-fold higher than that of CYP3A4. AM demethylation activity was correlated with CYP2B6 protein levels (P = 0.004); however, it was not correlated with CYP3A4 protein levels (P = 0.27) in human liver microsomes. Wild-type CYP2B6.1 and 25 CYP2B6 allelic variants (CYP2B6.2-CYP2B6.21 and CYP2B6.23-CYP2B6.27) were heterologously expressed in COS-7 cells. In vitro analysis revealed no enzymatic activity in 5 variants (CYP2B6.8, CYP2B6.12, CYP2B6.18, CYP2B6.21, and CYP2B6.24), lower activity in 7 variants (CYP2B6.10, CYP2B6.11, CYP2B6.14, CYP2B6.15, CYP2B6.16, CYP2B6.20, and CYP2B6.27), and higher activity in 4 variants (CYP2B6.2, CYP2B6.4, CYP2B6.6, and CYP2B6.19), compared with that of wild-type CYP2B6.1. In kinetic analysis, 3 variants (CYP2B6.2, CYP2B6.4, and CYP2B6.6) exhibited significantly higher V(max), and 3 variants (CYP2B6.14, CYP2B6.20 and CYP2B6.27) exhibited significantly lower V(max) compared with that of CYP2B6.1. This functional analysis of CYP2B6 variants could provide useful information for individualization of antimalarial drug therapy.

  14. Methylglyoxal induces endoplasmic reticulum stress and DNA demethylation in the Keap1 promoter of human lens epithelial cells and age-related cataracts.

    PubMed

    Palsamy, Periyasamy; Bidasee, Keshore R; Ayaki, Masahiko; Augusteyn, Robert C; Chan, Jefferson Y; Shinohara, Toshimichi

    2014-07-01

    Age-related cataracts are a leading cause of blindness. Previously, we have demonstrated the association of the unfolded protein response with various cataractogenic stressors. However, DNA methylation alterations leading to suppression of lenticular antioxidant protection remains unclear. Here, we report the methylglyoxal-mediated sequential events responsible for Keap1 promoter DNA demethylation in human lens epithelial cells, because Keap1 is a negative regulatory protein that regulates the Nrf2 antioxidant protein. Methylglyoxal induces endoplasmic reticulum stress and activates the unfolded protein response leading to overproduction of reactive oxygen species before human lens epithelial cell death. Methylglyoxal also suppresses Nrf2 and DNA methyltransferases but activates the DNA demethylation pathway enzyme TET1. Bisulfite genomic DNA sequencing confirms the methylglyoxal-mediated Keap1 promoter DNA demethylation leading to overexpression of Keap1 mRNA and protein. Similarly, bisulfite genomic DNA sequencing shows that human clear lenses (n = 15) slowly lose 5-methylcytosine in the Keap1 promoter throughout life, at a rate of 1% per year. By contrast, diabetic cataractous lenses (n = 21) lose an average of 90% of the 5-methylcytosine regardless of age. Overexpressed Keap1 protein is responsible for decreasing Nrf2 by proteasomal degradation, thereby suppressing Nrf2-dependent stress protection. This study demonstrates for the first time the associations of unfolded protein response activation, Nrf2-dependent antioxidant system failure, and loss of Keap1 promoter methylation because of altered active and passive DNA demethylation pathway enzymes in human lens epithelial cells by methylglyoxal. As an outcome, the cellular redox balance is altered toward lens oxidation and cataract formation.

  15. AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N(6)-methyladenosine.

    PubMed

    Wu, Weiche; Feng, Jie; Jiang, Denghu; Zhou, Xihong; Jiang, Qin; Cai, Min; Wang, Xinxia; Shan, Tizhong; Wang, Yizhen

    2017-02-08

    Skeletal muscle plays important roles in whole-body energy homeostasis. Excessive skeletal muscle lipid accumulation is associated with some metabolic diseases such as obesity and Type 2 Diabetes. The energy sensor AMPK (AMP-activated protein kinase) is a key regulator of skeletal muscle lipid metabolism, but the precise regulatory mechanism remains to be elucidated. Here, we provide a novel mechanism by which AMPK regulates skeletal muscle lipid accumulation through fat mass and obesity-associated protein (FTO)-dependent demethylation of N(6)-methyladenosine (m(6)A). We confirmed an inverse correlation between AMPK and skeletal muscle lipid content. Moreover, inhibition of AMPK enhanced lipid accumulation, while activation of AMPK reduced lipid accumulation in skeletal muscle cells. Notably, we found that mRNA m(6)A methylation levels were inversely correlated with lipid content in skeletal muscle. Furthermore, AMPK positively regulated the m(6)A methylation levels of mRNA, which could negatively regulate lipid accumulation in C2C12. At the molecular level, we demonstrated that AMPK regulated lipid accumulation in skeletal muscle cells by regulating FTO expression and FTO-dependent demethylation of m(6)A. Together, these results provide a novel regulatory mechanism of AMPK on lipid metabolism in skeletal muscle cells and suggest the possibility of controlling skeletal muscle lipid deposition by targeting AMPK or using m(6)A related drugs.

  16. AMPK regulates lipid accumulation in skeletal muscle cells through FTO-dependent demethylation of N6-methyladenosine

    PubMed Central

    Wu, Weiche; Feng, Jie; Jiang, Denghu; Zhou, Xihong; Jiang, Qin; Cai, Min; Wang, Xinxia; Shan, Tizhong; Wang, Yizhen

    2017-01-01

    Skeletal muscle plays important roles in whole-body energy homeostasis. Excessive skeletal muscle lipid accumulation is associated with some metabolic diseases such as obesity and Type 2 Diabetes. The energy sensor AMPK (AMP-activated protein kinase) is a key regulator of skeletal muscle lipid metabolism, but the precise regulatory mechanism remains to be elucidated. Here, we provide a novel mechanism by which AMPK regulates skeletal muscle lipid accumulation through fat mass and obesity-associated protein (FTO)-dependent demethylation of N6-methyladenosine (m6A). We confirmed an inverse correlation between AMPK and skeletal muscle lipid content. Moreover, inhibition of AMPK enhanced lipid accumulation, while activation of AMPK reduced lipid accumulation in skeletal muscle cells. Notably, we found that mRNA m6A methylation levels were inversely correlated with lipid content in skeletal muscle. Furthermore, AMPK positively regulated the m6A methylation levels of mRNA, which could negatively regulate lipid accumulation in C2C12. At the molecular level, we demonstrated that AMPK regulated lipid accumulation in skeletal muscle cells by regulating FTO expression and FTO-dependent demethylation of m6A. Together, these results provide a novel regulatory mechanism of AMPK on lipid metabolism in skeletal muscle cells and suggest the possibility of controlling skeletal muscle lipid deposition by targeting AMPK or using m6A related drugs. PMID:28176824

  17. Arginine demethylation is catalysed by a subset of JmjC histone lysine demethylases

    PubMed Central

    Walport, Louise J.; Hopkinson, Richard J.; Chowdhury, Rasheduzzaman; Schiller, Rachel; Ge, Wei; Kawamura, Akane; Schofield, Christopher J.

    2016-01-01

    While the oxygen-dependent reversal of lysine Nɛ-methylation is well established, the existence of bona fide Nω-methylarginine demethylases (RDMs) is controversial. Lysine demethylation, as catalysed by two families of lysine demethylases (the flavin-dependent KDM1 enzymes and the 2-oxoglutarate- and oxygen-dependent JmjC KDMs, respectively), proceeds via oxidation of the N-methyl group, resulting in the release of formaldehyde. Here we report detailed biochemical studies clearly demonstrating that, in purified form, a subset of JmjC KDMs can also act as RDMs, both on histone and non-histone fragments, resulting in formaldehyde release. RDM catalysis is studied using peptides of wild-type sequences known to be arginine-methylated and sequences in which the KDM's methylated target lysine is substituted for a methylated arginine. Notably, the preferred sequence requirements for KDM and RDM activity vary even with the same JmjC enzymes. The demonstration of RDM activity by isolated JmjC enzymes will stimulate efforts to detect biologically relevant RDM activity. PMID:27337104

  18. Dioxin induces Ahr-dependent robust DNA demethylation of the Cyp1a1 promoter via Tdg in the mouse liver

    NASA Astrophysics Data System (ADS)

    Amenya, Hesbon Z.; Tohyama, Chiharu; Ohsako, Seiichiroh

    2016-10-01

    The aryl hydrocarbon receptor (Ahr) is a highly conserved nuclear receptor that plays an important role in the manifestation of toxicity induced by polycyclic aromatic hydrocarbons. As a xenobiotic sensor, Ahr is involved in chemical biotransformation through activation of drug metabolizing enzymes. The activated Ahr cooperates with coactivator complexes to induce epigenetic modifications at target genes. Thus, it is conceivable that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a potent Ahr ligand, may elicit robust epigenetic changes in vivo at the Ahr target gene cytochrome P450 1a1 (Cyp1a1). A single dose of TCDD administered to adult mice induced Ahr-dependent CpG hypomethylation, changes in histone modifications, and thymine DNA glycosylase (Tdg) recruitment at the Cyp1a1 promoter in the liver within 24 hrs. These epigenetic changes persisted until 40 days post-TCDD treatment and there was Cyp1a1 mRNA hyperinduction upon repeat administration of TCDD at this time-point. Our demethylation assay using siRNA knockdown and an in vitro methylated plasmid showed that Ahr, Tdg, and the ten-eleven translocation methyldioxygenases Tet2 and Tet3 are required for the TCDD-induced DNA demethylation. These results provide novel evidence of Ahr-driven active DNA demethylation and epigenetic memory. The epigenetic alterations influence response to subsequent chemical exposure and imply an adaptive mechanism to xenobiotic stress.

  19. Dioxin induces Ahr-dependent robust DNA demethylation of the Cyp1a1 promoter via Tdg in the mouse liver

    PubMed Central

    Amenya, Hesbon Z.; Tohyama, Chiharu; Ohsako, Seiichiroh

    2016-01-01

    The aryl hydrocarbon receptor (Ahr) is a highly conserved nuclear receptor that plays an important role in the manifestation of toxicity induced by polycyclic aromatic hydrocarbons. As a xenobiotic sensor, Ahr is involved in chemical biotransformation through activation of drug metabolizing enzymes. The activated Ahr cooperates with coactivator complexes to induce epigenetic modifications at target genes. Thus, it is conceivable that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a potent Ahr ligand, may elicit robust epigenetic changes in vivo at the Ahr target gene cytochrome P450 1a1 (Cyp1a1). A single dose of TCDD administered to adult mice induced Ahr-dependent CpG hypomethylation, changes in histone modifications, and thymine DNA glycosylase (Tdg) recruitment at the Cyp1a1 promoter in the liver within 24 hrs. These epigenetic changes persisted until 40 days post-TCDD treatment and there was Cyp1a1 mRNA hyperinduction upon repeat administration of TCDD at this time-point. Our demethylation assay using siRNA knockdown and an in vitro methylated plasmid showed that Ahr, Tdg, and the ten-eleven translocation methyldioxygenases Tet2 and Tet3 are required for the TCDD-induced DNA demethylation. These results provide novel evidence of Ahr-driven active DNA demethylation and epigenetic memory. The epigenetic alterations influence response to subsequent chemical exposure and imply an adaptive mechanism to xenobiotic stress. PMID:27713569

  20. miR-29b induces SOCS-1 expression by promoter demethylation and negatively regulates migration of multiple myeloma and endothelial cells.

    PubMed

    Amodio, Nicola; Bellizzi, Dina; Leotta, Marzia; Raimondi, Lavinia; Biamonte, Lavinia; D'Aquila, Patrizia; Di Martino, Maria Teresa; Calimeri, Teresa; Rossi, Marco; Lionetti, Marta; Leone, Emanuela; Passarino, Giuseppe; Neri, Antonino; Giordano, Antonio; Tagliaferri, Pierosandro; Tassone, Pierfrancesco

    2013-12-01

    Epigenetic silencing of tumor suppressor genes frequently occurs and may account for their inactivation in cancer cells. We previously demonstrated that miR-29b is a tumor suppressor microRNA (miRNA) that targets de novo DNA methyltransferases and reduces the global DNA methylation of multiple myeloma (MM) cells. Here, we provide evidence that epigenetic activity of miR-29b leads to promoter demethylation of suppressor of cytokine signaling-1 (SOCS-1), a hypermethylated tumor suppressor gene. Enforced expression of synthetic miR-29b mimics in MM cell lines resulted in SOCS-1 gene promoter demethylation, as assessed by Sequenom MassARRAY EpiTYPER analysis, and SOCS-1 protein upregulation. miR-29b-induced SOCS-1 demethylation was associated with reduced STAT3 phosphorylation and impaired NFκB activity. Downregulation of VEGF-A and IL-8 mRNAs could be detected in MM cells transfected with miR-29b mimics as well as in endothelial (HUVEC) or stromal (HS-5) cells treated with conditioned medium from miR-29b-transfected MM cells. Notably, enforced expression of miR-29b mimics increased adhesion of MM cells to HS-5 and reduced migration of both MM and HUVEC cells. These findings suggest that miR-29b is a negative regulator of either MM or endothelial cell migration. Finally, the proteasome inhibitor bortezomib, which induces the expression of miR-29b, decreased global DNA methylation by a miR-29b-dependent mechanism and induced SOCS-1 promoter demethylation and protein upregulation. In conclusion, our data indicate that miR-29b is endowed with epigenetic activity and mediates previously unknown functions of bortezomib in MM cells.

  1. miR-29b induces SOCS-1 expression by promoter demethylation and negatively regulates migration of multiple myeloma and endothelial cells

    PubMed Central

    Amodio, Nicola; Bellizzi, Dina; Leotta, Marzia; Raimondi, Lavinia; Biamonte, Lavinia; D’Aquila, Patrizia; Di Martino, Maria Teresa; Calimeri, Teresa; Rossi, Marco; Lionetti, Marta; Leone, Emanuela; Passarino, Giuseppe; Neri, Antonino; Giordano, Antonio; Tagliaferri, Pierosandro; Tassone, Pierfrancesco

    2013-01-01

    Epigenetic silencing of tumor suppressor genes frequently occurs and may account for their inactivation in cancer cells. We previously demonstrated that miR-29b is a tumor suppressor microRNA (miRNA) that targets de novo DNA methyltransferases and reduces the global DNA methylation of multiple myeloma (MM) cells. Here, we provide evidence that epigenetic activity of miR-29b leads to promoter demethylation of suppressor of cytokine signaling-1 (SOCS-1), a hypermethylated tumor suppressor gene. Enforced expression of synthetic miR-29b mimics in MM cell lines resulted in SOCS-1 gene promoter demethylation, as assessed by Sequenom MassARRAY EpiTYPER analysis, and SOCS-1 protein upregulation. miR-29b-induced SOCS-1 demethylation was associated with reduced STAT3 phosphorylation and impaired NFκB activity. Downregulation of VEGF-A and IL-8 mRNAs could be detected in MM cells transfected with miR-29b mimics as well as in endothelial (HUVEC) or stromal (HS-5) cells treated with conditioned medium from miR-29b-transfected MM cells. Notably, enforced expression of miR-29b mimics increased adhesion of MM cells to HS-5 and reduced migration of both MM and HUVEC cells. These findings suggest that miR-29b is a negative regulator of either MM or endothelial cell migration. Finally, the proteasome inhibitor bortezomib, which induces the expression of miR-29b, decreased global DNA methylation by a miR-29b-dependent mechanism and induced SOCS-1 promoter demethylation and protein upregulation. In conclusion, our data indicate that miR-29b is endowed with epigenetic activity and mediates previously unknown functions of bortezomib in MM cells. PMID:24091729

  2. Hydrogen Sulfide Maintains Mitochondrial DNA Replication via Demethylation of TFAM

    PubMed Central

    Li, Shuangshuang

    2015-01-01

    Abstract Aims: Hydrogen sulfide (H2S) exerts a wide range of actions in the body, especially in the modulation of mitochondrial functions. The normal replication of mitochondrial DNA (mtDNA) is critical for cellular energy metabolism and mitochondrial biogenesis. The aim of this study was to investigate whether H2S affects mtDNA replication and the underlying mechanisms. We hypothesize that H2S maintains mtDNA copy number via inhibition of Dnmt3a transcription and TFAM promoter methylation. Results: Here, we demonstrated that deficiency of cystathionine gamma-lyase (CSE), a major H2S-producing enzyme, reduces mtDNA copy number and mitochondrial contents, and it inhibits the expressions of mitochondrial transcription factor A (TFAM) and mitochondrial marker genes in both smooth muscle cells and aorta tissues from mice. Supply of exogenous H2S stimulated mtDNA copy number and strengthened the expressions of TFAM and mitochondrial marker genes. TFAM knockdown diminished H2S-enhanced mtDNA copy number. In addition, CSE deficiency induced the expression of DNA methyltransferase 3a (Dnmt3a) and TFAM promoter DNA methylation, and H2S repressed Dnmt3a expression, resulting in TFAM promoter demethylation. We further found that H2S S-sulfhydrates transcription repressor interferon regulatory factor 1 (IRF-1) and enhances the binding of IRF-1 with Dnmt3a promoter after reduced Dnmt3a transcription. H2S had little effects on the expression of Dnmt1 and Dnmt3b as well as on ten-eleven translocation methylcytosine dioxygenase 1, 2, and 3. Innovation: A sufficient level of H2S is able to inhibit TFAM promoter methylation and maintain mtDNA copy number. Conclusion: CSE/H2S system contributes to mtDNA replication and cellular bioenergetics and provides a novel therapeutic avenue for cardiovascular diseases. Antioxid. Redox Signal. 23, 630–642. PMID:25758951

  3. Redistribution of demethylated RNA helicase A during foot-and-mouth disease virus infection: role of Jumonji C-domain containing protein 6 in RHA demethylation.

    PubMed

    Lawrence, Paul; Conderino, Joseph S; Rieder, Elizabeth

    2014-03-01

    Previously, RNA helicase A (RHA) re-localization from the nucleus to the cytoplasm in foot-and-mouth disease virus (FMDV) infected cells was shown to coincide with loss of RHA methylated arginine residues at its C-terminus. The potential interaction between RHA and Jumonji C-domain (JmjC) protein 6 (JMJD6) arginine demethylase in infected cells was investigated. Treatment with N-oxalylglycine (NOG) inhibitor of JmjC demethylases prevented FMDV-induced RHA demethylation and re-localization, and also decreased viral protein synthesis and virus titers. Physical interaction between JMJD6 and RHA was demonstrated via reciprocal co-immunoprecipitation, where RHA preferentially bound JMJD6 monomers. Nuclear efflux of demethylated RHA (DM-RHA) coincided with nuclear influx of JMJD6, which was not observed using another picornavirus. A modified biochemical assay demonstrated JMJD6 induced dose-dependent demethylation of RHA and two RHA-derived isoforms, which could be inhibited by NOG. We propose a role for JMJD6 in RHA demethylation stimulated by FMDV, that appears to facilitate virus replication.

  4. Redistribution of demethylated RNA helicase A during foot-and-mouth disease virus infection: role of jumonji C-domain containing protein 6 in RHA demethylation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We previously reported that RNA Helicase A (RHA) re-localized from the nucleus to the cytoplasm in foot-and-mouth disease virus (FMDV) infected cells, coincident with a reduction in methylation of arginine residues in the RHA C-terminus. To further define the mechanism of RHA demethylation in FMDV-...

  5. H3K4me3 demethylation by the histone demethylase KDM5C/JARID1C promotes DNA replication origin firing.

    PubMed

    Rondinelli, Beatrice; Schwerer, Hélène; Antonini, Elena; Gaviraghi, Marco; Lupi, Alessio; Frenquelli, Michela; Cittaro, Davide; Segalla, Simona; Lemaitre, Jean-Marc; Tonon, Giovanni

    2015-03-11

    DNA replication is a tightly regulated process that initiates from multiple replication origins and leads to the faithful transmission of the genetic material. For proper DNA replication, the chromatin surrounding origins needs to be remodeled. However, remarkably little is known on which epigenetic changes are required to allow the firing of replication origins. Here, we show that the histone demethylase KDM5C/JARID1C is required for proper DNA replication at early origins. JARID1C dictates the assembly of the pre-initiation complex, driving the binding to chromatin of the pre-initiation proteins CDC45 and PCNA, through the demethylation of the histone mark H3K4me3. Fork activation and histone H4 acetylation, additional early events involved in DNA replication, are not affected by JARID1C downregulation. All together, these data point to a prominent role for JARID1C in a specific phase of DNA replication in mammalian cells, through its demethylase activity on H3K4me3.

  6. DNA Demethylation of the Foxp3 Enhancer Is Maintained through Modulation of Ten-Eleven-Translocation and DNA Methyltransferases

    PubMed Central

    Nair, Varun Sasidharan; Song, Mi Hye; Ko, Myunggon; Oh, Kwon Ik

    2016-01-01

    Stable expression of Foxp3 is ensured by demethylation of CpG motifs in the Foxp3 intronic element, the conserved non-coding sequence 2 (CNS2), which persists throughout the lifespan of regulatory T cells (Tregs). However, little is known about the mechanisms on how CNS2 demethylation is sustained. In this study, we found that Ten-Eleven-Translocation (Tet) DNA dioxygenase protects the CpG motifs of CNS2 from re-methylation by DNA methyltransferases (Dnmts) and prevents Tregs from losing Foxp3 expression under inflammatory conditions. Upon stimulation of Tregs by interleukin-6 (IL6), Dnmt1 was recruited to CNS2 and induced methylation, which was inhibited by Tet2 recruited by IL2. Tet2 prevented CNS2 re-methylation by not only the occupancy of the CNS2 locus but also by its enzymatic activity. These results show that the CNS2 methylation status is dynamically regulated by a balance between Tets and Dnmts which influences the expression of Foxp3 in Tregs. PMID:27989104

  7. Mercury methylation and demethylation in highly contaminated sediments from the Deûle River in Northern France using species-specific enriched stable isotopes.

    PubMed

    Ouddane, Baghdad; Monperrus, Mathilde; Kadlecova, Milada; Daye, Mirna; Amouroux, David

    2015-01-01

    The methylation-demethylation processes in sediments of the Deûle River were determined using well-established isotope experiments. For this purpose, species-specific isotopically enriched tracers in the form of inorganic mercury IHg ((199)Hg) and methylmercury MeHg (Me(201)Hg) were used to determine Hg dynamics in the Deûle River. Sediment cores were collected at two sampling locations chosen in the most polluted zone of the Deûle River (Northern France) in proximity of a Zn, Pb, Cu, and Ni smelter called "Metaleurop" that had closed in 2003. Site I was chosen in the vicinity of the historic smelter site and site II upstream of the Deûle River. The incubation was realized directly in the sediment cores during the 24 hour experiment under environmental conditions close to the real natural systems (the same temperature, pH, humidity, light/dark conditions, oxygen levels…). The enriched isotopes were injected by needle into different sections of the core. After incubation, the core was sliced and the concentration of Hg species was determined in each section. The highest methylation potentials were found at sediment depths away from the sediment-water-interface. At site I, the methylation potential varied between 0.02-0.9% and at site II between 0.001-0.2%. The demethylation potentials fluctuated between 0.001-60% at site I and between 4-53% at site II. In both sites, negative net methylation potentials were obtained in several sediment depths, representing a net sink for MeHg. The average net methylation potential in site I demonstrated a negative value of 1919 ng g(-1) day(-1). It seems that in site I the demethylation process predominates methylation. Whereas, in site II, the average net methylation potential was a positive value of 138 ng g(-1) day(-1), demonstrating the dominance of methylation over demethylation.

  8. AlkB homolog 3-mediated tRNA demethylation promotes protein synthesis in cancer cells

    PubMed Central

    Ueda, Yuko; Ooshio, Ikumi; Fusamae, Yasuyuki; Kitae, Kaori; Kawaguchi, Megumi; Jingushi, Kentaro; Hase, Hiroaki; Harada, Kazuo; Hirata, Kazumasa; Tsujikawa, Kazutake

    2017-01-01

    The mammalian AlkB homolog (ALKBH) family of proteins possess a 2-oxoglutarate- and Fe(II)-dependent oxygenase domain. A similar domain in the Escherichia coli AlkB protein catalyzes the oxidative demethylation of 1-methyladenine (1-meA) and 3-methylcytosine (3-meC) in both DNA and RNA. AlkB homolog 3 (ALKBH3) was also shown to demethylate 1-meA and 3-meC (induced in single-stranded DNA and RNA by a methylating agent) to reverse the methylation damage and retain the integrity of the DNA/RNA. We previously reported the high expression of ALKBH3 in clinical tumor specimens and its involvement in tumor progression. In this study, we found that ALKBH3 effectively demethylated 1-meA and 3-meC within endogenously methylated RNA. Moreover, using highly purified recombinant ALKBH3, we identified N6-methyladenine (N6-meA) in mammalian transfer RNA (tRNA) as a novel ALKBH3 substrate. An in vitro translation assay showed that ALKBH3-demethylated tRNA significantly enhanced protein translation efficiency. In addition, ALKBH3 knockdown in human cancer cells impaired cellular proliferation and suppressed the nascent protein synthesis that is usually accompanied by accumulation of the methylated RNAs. Thus, our data highlight a novel role for ALKBH3 in tumor progression via RNA demethylation and subsequent protein synthesis promotion. PMID:28205560

  9. Time-course gene profiling and networks in demethylated retinoblastoma cell line.

    PubMed

    Malusa, Federico; Taranta, Monia; Zaki, Nazar; Cinti, Caterina; Capobianco, Enrico

    2015-09-15

    Retinoblastoma, a very aggressive cancer of the developing retina, initiatiates by the biallelic loss of RB1 gene, and progresses very quickly following RB1 inactivation. While its genome is stable, multiple pathways are deregulated, also epigenetically. After reviewing the main findings in relation with recently validated markers, we propose an integrative bioinformatics approach to include in the previous group new markers obtained from the analysis of a single cell line subject to epigenetic treatment. In particular, differentially expressed genes are identified from time course microarray experiments on the WERI-RB1 cell line treated with 5-Aza-2'-deoxycytidine (decitabine; DAC). By inducing demethylation of CpG island in promoter genes that are involved in biological processes, for instance apoptosis, we performed the following main integrative analysis steps: i) Gene expression profiling at 48h, 72h and 96h after DAC treatment; ii) Time differential gene co-expression networks and iii) Context-driven marker association (transcriptional factor regulated protein networks, master regulatory paths). The observed DAC-driven temporal profiles and regulatory connectivity patterns are obtained by the application of computational tools, with support from curated literature. It is worth emphasizing the capacity of networks to reconcile multi-type evidences, thus generating testable hypotheses made available by systems scale predictive inference power. Despite our small experimental setting, we propose through such integrations valuable impacts of epigenetic treatment in terms of gene expression measurements, and then validate evidenced apoptotic effects.

  10. Time-course gene profiling and networks in demethylated retinoblastoma cell line

    PubMed Central

    Malusa, Federico; Taranta, Monia; Zaki, Nazar; Cinti, Caterina; Capobianco, Enrico

    2015-01-01

    Retinoblastoma, a very aggressive cancer of the developing retina, initiatiates by the biallelic loss of RB1 gene, and progresses very quickly following RB1 inactivation. While its genome is stable, multiple pathways are deregulated, also epigenetically. After reviewing the main findings in relation with recently validated markers, we propose an integrative bioinformatics approach to include in the previous group new markers obtained from the analysis of a single cell line subject to epigenetic treatment. In particular, differentially expressed genes are identified from time course microarray experiments on the WERI-RB1 cell line treated with 5-Aza-2′-deoxycytidine (decitabine; DAC). By inducing demethylation of CpG island in promoter genes that are involved in biological processes, for instance apoptosis, we performed the following main integrative analysis steps: i) Gene expression profiling at 48h, 72h and 96h after DAC treatment; ii) Time differential gene co-expression networks and iii) Context-driven marker association (transcriptional factor regulated protein networks, master regulatory paths). The observed DAC-driven temporal profiles and regulatory connectivity patterns are obtained by the application of computational tools, with support from curated literature. It is worth emphasizing the capacity of networks to reconcile multi-type evidences, thus generating testable hypotheses made available by systems scale predictive inference power. Despite our small experimental setting, we propose through such integrations valuable impacts of epigenetic treatment in terms of gene expression measurements, and then validate evidenced apoptotic effects. PMID:26143641

  11. Curcumin inhibits growth of human breast cancer cells through demethylation of DLC1 promoter.

    PubMed

    Liu, Yufei; Zhou, Jun; Hu, Yuchang; Wang, Junjie; Yuan, Chengfu

    2017-01-01

    The heterogeneity of breast cancer makes it a challenging solid tumor to diagnose and treat. A tumor suppressor Deleted in Liver Cancer 1 (DLC1) has been reported to be down-regulated or even silenced in several kinds of cancer including breast cancer. Curcumin has been reported to modulate the growth of tumor cells through regulation of multiple cell signaling pathways and modulate epigenetic changes by CpG demethylation of many tumor suppressor genes. This study was designed to investigate the effect of curcumin on the expression of Deleted in Liver Cancer 1 (DLC1) in human breast cancer cell line MDA-MB-361 and the underlying mechanism in vitro and in vivo. Curcumin induced DLC1 expression in a dose-dependent manner. In curcumin-treated cells, methylation of DLC1 promoter was reduced and active forms of RhoA and Cdc42 were also decreased. DLC1 expression was closely related to tumor cell growth, demonstrated by Ki67 staining. Curcumin inhibited DNA methyltransferase 1 expression through down-regulation of transcription factor Sp1. Consistent with the in vitro data, in vivo administration of curcumin inhibited the growth of implanted MDA-MB-361 cells and induced DLC1 expression in tumor tissue. In MDA-MB-361 cells, curcumin down-regulates the expression of Sp1 to inhibit the expression of DNA methyltransferase 1, thus subsequently reducing hypermethylation of DLC1 promoter to induce DLC1 expression.

  12. Mitochondrial control through nutritionally regulated global histone H3 lysine-4 demethylation

    PubMed Central

    Soloveychik, Maria; Xu, Mengshu; Zaslaver, Olga; Lee, Kwanyin; Narula, Ashrut; Jiang, River; Rosebrock, Adam P.; Caudy, Amy A.; Meneghini, Marc D.

    2016-01-01

    Histone demethylation by Jumonji-family proteins is coupled with the decarboxylation of α-ketoglutarate (αKG) to yield succinate, prompting hypotheses that their activities are responsive to levels of these metabolites in the cell. Consistent with this paradigm we show here that the Saccharomyces cerevisiae Jumonji demethylase Jhd2 opposes the accumulation of H3K4me3 in fermenting cells only when they are nutritionally manipulated to contain an elevated αKG/succinate ratio. We also find that Jhd2 opposes H3K4me3 in respiratory cells that do not exhibit such an elevated αKG/succinate ratio. While jhd2∆ caused only limited gene expression defects in fermenting cells, transcript profiling and physiological measurements show that JHD2 restricts mitochondrial respiratory capacity in cells grown in non-fermentable carbon in an H3K4me-dependent manner. In association with these phenotypes, we find that JHD2 limits yeast proliferative capacity under physiologically challenging conditions as measured by both replicative lifespan and colony growth on non-fermentable carbon. JHD2’s impact on nutrient response may reflect an ancestral role of its gene family in mediating mitochondrial regulation. PMID:27897198

  13. Epigenetic DNA Demethylation Causes Inner Ear Stem Cell Differentiation into Hair Cell-Like Cells

    PubMed Central

    Zhou, Yang; Hu, Zhengqing

    2016-01-01

    The DNA methyltransferase (DNMT) inhibitor 5-azacytidine (5-aza) causes genomic demethylation to regulate gene expression. However, it remains unclear whether 5-aza affects gene expression and cell fate determination of stem cells. In this study, 5-aza was applied to mouse utricle sensory epithelia-derived progenitor cells (MUCs) to investigate whether 5-aza stimulated MUCs to become sensory hair cells. After treatment, MUCs increased expression of hair cell genes and proteins. The DNA methylation level (indicated by percentage of 5-methylcytosine) showed a 28.57% decrease after treatment, which causes significantly repressed DNMT1 protein expression and DNMT activity. Additionally, FM1-43 permeation assays indicated that the permeability of 5-aza-treated MUCs was similar to that of sensory hair cells, which may result from mechanotransduction channels. This study not only demonstrates a possible epigenetic approach to induce tissue specific stem/progenitor cells to become sensory hair cell-like cells, but also provides a cell model to epigenetically modulate stem cell fate determination. PMID:27536218

  14. Synergistic Cytotoxic Effect of L-Asparaginase Combined with Decitabine as a Demethylating Agent in Pediatric T-ALL, with Specific Epigenetic Signature

    PubMed Central

    Melchionda, Fraia; Pession, Andrea

    2016-01-01

    T-Acute Lymphoblastic Leukemia (T-ALL) remains a subgroup of pediatric ALL, with a lower response to standard chemotherapy. Some recent studies established the fundamental role of epigenetic aberrations such as DNA hypermethylation, to influence patients' outcome and response to chemotherapy. Moreover, L-asparaginase is an important chemotherapeutic agent for treatment of ALL and resistance to this drug has been linked to ASNS expression, which can be silenced through methylation. Therefore, we tested whether the sensitivity of T-ALL cell lines towards L-asparaginase is correlated to the epigenetic status of ASNS gene and whether the sensitivity can be modified by concurrent demethylating treatment. Hence we treated different T-ALL cell lines with L-asparaginase and correlated different responses to the treatment with ASNS expression. Then we demonstrated that the ASNS expression was dependent on the methylation status of the promoter. Finally we showed that, despite the demethylating effect on the ASNS gene expression, the combined treatment with the demethylating agent Decitabine could synergistically improve the L-asparaginase sensitivity in those T-ALL cell lines characterized by hypermethylation of the ASNS gene. In conclusion, this preclinical study identified an unexpected synergistic activity of L-asparaginase and Decitabine in the subgroup of T-ALL with low ASNS expression due to hypermethylation of the ASNS promoter, while it did not restore sensitivity in the resistant cell lines characterized by higher ASNS expression. PMID:28003999

  15. Black Raspberry-Derived Anthocyanins Demethylate Tumor Suppressor Genes Through the Inhibition of DNMT1 and DNMT3B in Colon Cancer Cells

    PubMed Central

    Wang, Li-Shu; Kuo, Chieh-Ti; Cho, Seung-Ju; Seguin, Claire; Siddiqui, Jibran; Stoner, Kristen; Weng, Yu-I; Huang, Tim H.-M.; Tichelaar, Jay; Yearsley, Martha; Stoner, Gary D.; Huang, Yi-Wen

    2013-01-01

    We previously reported that oral administration of black raspberry powder decreased promoter methylation of tumor suppressor genes in tumors from patients with colorectal cancer. The anthocyanins (ACs) in black raspberries are responsible, at least in part, for their cancer-inhibitory effects. In the present study, we asked if ACs are responsible for the demethylation effects observed in colorectal cancers. Three days of treatment of ACs at 0.5, 5, and 25 μg/ml suppressed activity and protein expression of DNMT1 and DNMT3B in HCT116, Caco2 and SW480 cells. Promoters of CDKN2A, and SFRP2, SFRP5, and WIF1, upstream of Wnt pathway, were demethylated by ACs. mRNA expression of some of these genes was increased. mRNA expression of β-catenin and c-Myc, downstream of Wnt pathway, and cell proliferation were decreased; apoptosis was increased. ACs were taken up into HCT116 cells and were differentially localized with DNMT1 and DNMT3B in the same cells visualized using confocal laser scanning microscopy. Although it was reported that DNMT3B is regulated by c-Myc in mouse lymphoma, DNMT3B did not bind with c-Myc in HCT116 cells. In conclusion, our results suggest that ACs are responsible, at least in part, for the demethylation effects of whole black raspberries in colorectal cancers. PMID:23368921

  16. Processing of Color Words Activates Color Representations

    ERIC Educational Resources Information Center

    Richter, Tobias; Zwaan, Rolf A.

    2009-01-01

    Two experiments were conducted to investigate whether color representations are routinely activated when color words are processed. Congruency effects of colors and color words were observed in both directions. Lexical decisions on color words were faster when preceding colors matched the color named by the word. Color-discrimination responses…

  17. Speech perception as an active cognitive process.

    PubMed

    Heald, Shannon L M; Nusbaum, Howard C

    2014-01-01

    One view of speech perception is that acoustic signals are transformed into representations for pattern matching to determine linguistic structure. This process can be taken as a statistical pattern-matching problem, assuming realtively stable linguistic categories are characterized by neural representations related to auditory properties of speech that can be compared to speech input. This kind of pattern matching can be termed a passive process which implies rigidity of processing with few demands on cognitive processing. An alternative view is that speech recognition, even in early stages, is an active process in which speech analysis is attentionally guided. Note that this does not mean consciously guided but that information-contingent changes in early auditory encoding can occur as a function of context and experience. Active processing assumes that attention, plasticity, and listening goals are important in considering how listeners cope with adverse circumstances that impair hearing by masking noise in the environment or hearing loss. Although theories of speech perception have begun to incorporate some active processing, they seldom treat early speech encoding as plastic and attentionally guided. Recent research has suggested that speech perception is the product of both feedforward and feedback interactions between a number of brain regions that include descending projections perhaps as far downstream as the cochlea. It is important to understand how the ambiguity of the speech signal and constraints of context dynamically determine cognitive resources recruited during perception including focused attention, learning, and working memory. Theories of speech perception need to go beyond the current corticocentric approach in order to account for the intrinsic dynamics of the auditory encoding of speech. In doing so, this may provide new insights into ways in which hearing disorders and loss may be treated either through augementation or therapy.

  18. Controlling contagion processes in activity driven networks.

    PubMed

    Liu, Suyu; Perra, Nicola; Karsai, Márton; Vespignani, Alessandro

    2014-03-21

    The vast majority of strategies aimed at controlling contagion processes on networks consider the connectivity pattern of the system either quenched or annealed. However, in the real world, many networks are highly dynamical and evolve, in time, concurrently with the contagion process. Here, we derive an analytical framework for the study of control strategies specifically devised for a class of time-varying networks, namely activity-driven networks. We develop a block variable mean-field approach that allows the derivation of the equations describing the coevolution of the contagion process and the network dynamic. We derive the critical immunization threshold and assess the effectiveness of three different control strategies. Finally, we validate the theoretical picture by simulating numerically the spreading process and control strategies in both synthetic networks and a large-scale, real-world, mobile telephone call data set.

  19. Further investigations into the N-demethylation of oripavine using iron and stainless steel.

    PubMed

    Kok, Gaik B; Scammells, Peter J

    2011-02-21

    Further investigations into the direct synthesis of N-nororipavine from oripavine using iron powder under non-classical Polonovski conditions have been conducted. The stoichiometry, solvents and iron oxidation rates were found to have a dramatic effect on the rate of N-demethylation as well as product yield. Herein, we also present high-yield access to the N-demethylated product simply by employing stainless steel rather than iron powder as redox catalyst. To our knowledge, this is the first time stainless steel has been used to moderate the redox chemistry of iron in organic synthesis.

  20. Antroquinonol D, isolated from Antrodia camphorata, with DNA demethylation and anticancer potential.

    PubMed

    Wang, Sheng-Chao; Lee, Tzong-Huei; Hsu, Chun-Hua; Chang, Yu-Jia; Chang, Man-Shan; Wang, Yi-Ching; Ho, Yuan-Soon; Wen, Wu-Che; Lin, Ruo-Kai

    2014-06-18

    DNA methyltransferase 1 (DNMT1) catalyzes DNA methylation and is overexpressed in various human diseases, including cancer. A rational approach to preventing tumorigenesis involves the use of pharmacologic inhibitors of DNA methylation; these inhibitors should reactivate tumor suppressor genes (TSGs) in tumor cells and restore tumor suppressor pathways. Antroquinonol D (3-demethoxyl antroquinonol), a new DNMT1 inhibitor, was isolated from Antrodia camphorata and identified using nuclear magnetic resonance. Antroquinonol D inhibited the growth of MCF7, T47D, and MDA-MB-231 breast cancer cells without harming normal MCF10A and IMR-90 cells. The SRB assay showed that the 50% growth inhibition (GI50) in MCF7, T47D, and MDA-MB-231 breast cancer cells following treatment with antroquinonol D was 8.01, 3.57, and 25.08 μM, respectively. d-Antroquinonol also inhibited the migratory ability of MDA-MB-231 breast cancer cells in wound healing and Transwell assays. In addition, antroquinonol D inhibited DNMT1 activity, as assessed by the DNMT1 methyltransferase activity assay. As the cofactor SAM level increased, the inhibitory effects of d-antroquinonol on DNMT1 gradually decreased. An enzyme activity assay and molecular modeling revealed that antroquinonol D is bound to the catalytic domain of DNMT1 and competes for the same binding pocket in the DNMT1 enzyme as the cofactor SAM, but does not compete for the binding pocket in the DNMT3B enzyme. An Illumina Methylation 450 K array-based assay and real-time PCR assay revealed that antroquinonol D decreased the methylation status and reactivated the expression of multiple TSGs in MDA-MB-231 breast cancer cells. In conclusion, we showed that antroquinonol D induces DNA demethylation and the recovery of multiple tumor suppressor genes, while inhibiting breast cancer growth and migration potential.

  1. Process for preparing active oxide powders

    DOEpatents

    Berard, Michael F.; Hunter, Jr., Orville; Shiers, Loren E.; Dole, Stephen L.; Scheidecker, Ralph W.

    1979-02-20

    An improved process for preparing active oxide powders in which cation hydroxide gels, prepared in the conventional manner are chemically dried by alternately washing the gels with a liquid organic compound having polar characteristics and a liquid organic compound having nonpolar characteristics until the mechanical water is removed from the gel. The water-free cation hydroxide is then contacted with a final liquid organic wash to remove the previous organic wash and speed drying. The dried hydroxide treated in the conventional manner will form a highly sinterable active oxide powder.

  2. Active voltammetric microsensors with neural signal processing.

    SciTech Connect

    Vogt, M. C.

    1998-12-11

    Many industrial and environmental processes, including bioremediation, would benefit from the feedback and control information provided by a local multi-analyte chemical sensor. For most processes, such a sensor would need to be rugged enough to be placed in situ for long-term remote monitoring, and inexpensive enough to be fielded in useful numbers. The multi-analyte capability is difficult to obtain from common passive sensors, but can be provided by an active device that produces a spectrum-type response. Such new active gas microsensor technology has been developed at Argonne National Laboratory. The technology couples an electrocatalytic ceramic-metallic (cermet) microsensor with a voltammetric measurement technique and advanced neural signal processing. It has been demonstrated to be flexible, rugged, and very economical to produce and deploy. Both narrow interest detectors and wide spectrum instruments have been developed around this technology. Much of this technology's strength lies in the active measurement technique employed. The technique involves applying voltammetry to a miniature electrocatalytic cell to produce unique chemical ''signatures'' from the analytes. These signatures are processed with neural pattern recognition algorithms to identify and quantify the components in the analyte. The neural signal processing allows for innovative sampling and analysis strategies to be employed with the microsensor. In most situations, the whole response signature from the voltammogram can be used to identify, classify, and quantify an analyte, without dissecting it into component parts. This allows an instrument to be calibrated once for a specific gas or mixture of gases by simple exposure to a multi-component standard rather than by a series of individual gases. The sampled unknown analytes can vary in composition or in concentration, the calibration, sensing, and processing methods of these active voltammetric microsensors can detect, recognize, and

  3. Proteolytic Processing Regulates Placental Growth Factor Activities*

    PubMed Central

    Hoffmann, Daniel C.; Willenborg, Sebastian; Koch, Manuel; Zwolanek, Daniela; Müller, Stefan; Becker, Ann-Kathrin A.; Metzger, Stephanie; Ehrbar, Martin; Kurschat, Peter; Hellmich, Martin; Hubbell, Jeffrey A.; Eming, Sabine A.

    2013-01-01

    Placental growth factor (PlGF) is a critical mediator of blood vessel formation, yet mechanisms of its action and regulation are incompletely understood. Here we demonstrate that proteolytic processing regulates the biological activity of PlGF. Specifically, we show that plasmin processing of PlGF-2 yields a protease-resistant core fragment comprising the vascular endothelial growth factor receptor-1 binding site but lacking the carboxyl-terminal domain encoding the heparin-binding domain and an 8-amino acid peptide encoded by exon 7. We have identified plasmin cleavage sites, generated a truncated PlGF118 isoform mimicking plasmin-processed PlGF, and explored its biological function in comparison with that of PlGF-1 and -2. The angiogenic responses induced by the diverse PlGF forms were distinct. Whereas PlGF-2 increased endothelial cell chemotaxis, vascular sprouting, and granulation tissue formation upon skin injury, these activities were abrogated following plasmin digestion. Investigation of PlGF/Neuropilin-1 binding and function suggests a critical role for heparin-binding domain/Neuropilin-1 interaction and its regulation by plasmin processing. Collectively, here we provide new mechanistic insights into the regulation of PlGF-2/Neuropilin-1-mediated tissue vascularization and growth. PMID:23645683

  4. Activation of consolidation processes of alumina ceramics

    NASA Astrophysics Data System (ADS)

    Matrenin, S. V.; Zenin, B. S.; Tayukin, R. V.

    2016-02-01

    The methods for activating sintering ceramics based on Al2O3 by mechanical activation in the planetary mill, by adding in the mixture of nanopowders (NP) Al, Al2O3, and submicron powder TiO2, and by applying the technology of spark plasma sintering (SPS) are developed. It has been shown that adding the nanopowder up to 20 wt. % Al2O3 in a coarse powder α-Al2O3 activates the sintering process resulting in increased density and hardness of the sintered alumina ceramics. Substantial effect of increasing density of alumina ceramics due to adding the submicron powder TiO2 in the compound of initial powder mixtures has been established.

  5. [Inhibitory action of divalent copper compounds on cumene hydroperoxide oxidative demethylation of N,N-dimethylaniline by cytochrome P-450].

    PubMed

    Kurchenko, V P; Usanov, S A; Metelitsa, D I

    1980-07-01

    The inhibitory action of divalent copper compounds on hydroperoxide-dependent oxidative demethylation of N,N-demethylaniline involving rabbit liver microsomes and highly purified cytochrome P-450 has been studied. CuCl2 is a non-competitive inhibitor, whereas copper tyrosine and lysine complexes are characterized by a mixed type inhibition. The inhibitory action of copper complexes is based on a decrease of cumene hydroperoxide concentration. The reaction results in formation of RO and RO2 radicals destroying cytochrome P-450 CuCl2 (0,001 M) also destroys cytochrome P-450 in the absence of cumene hydroperoxide; the destruction process is characterized by two phases with different rate constants. The nature of the inhibitory action of CuCl2 on N,N-demethylaniline oxidation by hydroperoxides is discussed.

  6. A Tetrahydrofolate-Dependent Methyltransferase Catalyzing the Demethylation of Dicamba in Sphingomonas sp. Strain Ndbn-20

    PubMed Central

    Yao, Li; Yu, Lin-Lu; Zhang, Jun-Jie; Xie, Xiang-Ting; Tao, Qing; Yan, Xin; Hong, Qing; Qiu, Ji-Guo

    2016-01-01

    ABSTRACT Sphingomonas sp. strain Ndbn-20 degrades and utilizes the herbicide dicamba as its sole carbon and energy source. In the present study, a tetrahydrofolate (THF)-dependent dicamba methyltransferase gene, dmt, was cloned from the strain, and three other genes, metF, dhc, and purU, which are involved in THF metabolism, were found to be located downstream of dmt. A transcriptional study revealed that the four genes constituted one transcriptional unit that was constitutively transcribed. Lysates of cells grown with glucose or dicamba exhibited almost the same activities, which further suggested that the dmt gene is constitutively expressed in the strain. Dmt shared 46% and 45% identities with the methyltransferases DesA and LigM from Sphingomonas paucimobilis SYK-6, respectively. The purified Dmt catalyzed the transfer of methyl from dicamba to THF to form the herbicidally inactive metabolite 3,6-dichlorosalicylic acid (DCSA) and 5-methyl-THF. The activity of Dmt was inhibited by 5-methyl-THF but not by DCSA. The introduction of a codon-optimized dmt gene into Arabidopsis thaliana enhanced resistance against dicamba. In conclusion, this study identified a THF-dependent dicamba methyltransferase, Dmt, with potential applications for the genetic engineering of dicamba-resistant crops. IMPORTANCE Dicamba is a very important herbicide that is widely used to control more than 200 types of broadleaf weeds and is a suitable target herbicide for the engineering of herbicide-resistant transgenic crops. A study of the mechanism of dicamba metabolism by soil microorganisms will benefit studies of its dissipation, transformation, and migration in the environment. This study identified a THF-dependent methyltransferase, Dmt, capable of catalyzing dicamba demethylation in Sphingomonas sp. Ndbn-20, and a preliminary study of its enzymatic characteristics was performed. Introduction of a codon-optimized dmt gene into Arabidopsis thaliana enhanced resistance against dicamba

  7. Synthesis and characterization of N-demethylated metabolites of malachite green and leucomalachite green.

    PubMed

    Cho, Bongsup P; Yang, Tianle; Blankenship, Lonnie R; Moody, Joanna D; Churchwell, Mona; Beland, Frederick A; Culp, Sandra J

    2003-03-01

    Malachite green (MG), a triphenylmethane dye used to treat fungal and protozoan infections in fish, undergoes sequential oxidation to produce various N-demethylated derivatives (monodes-, dides(sym)-, dides(unsym)-, trides-, and tetrades-) both before and after reduction to leucomalachite green (LMG). The close structure resemblance of the metabolites with aromatic amine carcinogens implicates a potential genotoxicity from exposure to MG. The availability of the synthetic standards is important for metabolic and DNA adduct studies of MG. This paper describes a simple and versatile method for the synthesis of MG, LMG, and their N-demethylated metabolites. The synthesis involves a coupling of 4-(dimethylamino)benzophenone or 4-nitrobenzophenone with the aryllithium reagents derived from appropriately substituted 4-bromoaniline derivatives, followed by treatment with HCl in methanol. The resulting cationic MG and their leuco analogues showed systematic UV/vis spectral and tandem mass fragmentation patterns consistent with sequential N-demethylation. The extensive (1)H and (13)C spectral assignments of the metabolites were aided by the availability of (13)C(7)-labeled MG and LMG. The results indicate the existence of a resonance structure with the cationic charge located in the central methane carbon (C(7)). The synthetic procedure is general in scope so that it can be extended to the preparation of N-demethylated metabolites of other structurally related N-methylated triphenylmethane dyes.

  8. Testosterone Depletion Induces Demethylation of Murine Reelin Promoter CpG Dinucleotides: A Preliminary Study

    PubMed Central

    da Silva, Victor Augusto Moraes; Dantas, Marília de Souza; Silva, Leonardo Agostinho de Castro; Carneiro, Juliana Garcia; Schamber-Reis, Bruno Luiz Fonseca

    2015-01-01

    Schizophrenia (SZ) is a debilitating mental disorder characterized by psychotic events, abnormal social behavior, false beliefs, and auditory hallucinations. Hypermethylation of the promoter region of reelin (RELN), a gene involved in regulation of neuronal positioning during telencephalic development, is strongly associated with low protein expression in several cortical structures and promoter hypermethylation in brain from postmortem SZ subjects. Recent experimental data suggests that testosterone is able to promote RELN demethylation, although no direct evidence of hormonal influence on reelin promoter methylation was obtained. We investigated if reduced levels of plasma testosterone in adult male mice lead to Reln promoter demethylation. Animals were administered with flutamide, an antiandrogenic compound, and reelin promoter methylation was assessed using methylationspecific PCR using bisulfite DNA from cerebellum. We found that flutamide was able to significantly lower plasma testosterone when compared to control mice, and treatment did not influence animal survival and body weight. We also show that low plasma testosterone was associated with demethylation of a cytosine residue located at −860 in reelin promoter region. These preliminary data suggest that androgenic hormones can influence cerebral reelin demethylation. To our knowledge, this is the first experimental approach directly linking testosterone depletion and RELN promoter methylation. PMID:26526966

  9. Profiling of methylation and demethylation pathways during brain development and ageing.

    PubMed

    Kraus, Theo F J; Kilinc, Selma; Steinmaurer, Martina; Stieglitz, Marc; Guibourt, Virginie; Kretzschmar, Hans A

    2016-03-01

    Numerous signal pathways are epigenetically controlled during brain development and ageing. Thereby, both 5-methylcytosine (5mC) and the newly described 5-hydroxymethylcytosine (5hmC) are highly exhibited in the brain. As there is an uneven distribution of 5hmC in the brain depending on age and region, there is the need to investigate the underlying mechanisms being responsible for 5hmC generation and decline. The aim of this study was to quantify expression levels of genes that are associated with DNA methylation/demethylation in different brain regions and at different ages. Therefore, we investigated frontal cortex and cerebellum of 40 mice (strain C57BL/6), each eight mice sacrificed at day 0, 7, 15, 30 and 120 after birth. We performed expression profiling of methylation/demethylation genes depending on age and brain region. Interestingly, we see significant expression differences of genes being responsible for methylation/demethylation with a significant reduction of expression levels during ageing. Validating selected expression data on protein level using immunohistochemistry verified the expression data. In conclusion, our findings demonstrate that the regulation of methylation/demethylation pathways is highly controlled depending on brain region and age. Thus our data will help to better understand the complexity and plasticity of the brain epigenome.

  10. Demethylation of Methylated Arsenic Species during Generation of Arsanes with Tetrahydridoborate(1-) in Acidic Media.

    PubMed

    Marschner, Karel; Musil, Stanislav; Dědina, Jiří

    2016-06-21

    Demethylation during generation of volatile hydrides (HG), i.e. formation of noncorresponding arsanes from monomethylarsonic acid (MAs(V)), dimethylarsinic acid (DMAs(V)), and trimethylarsine oxide (TMAs(V)O) by the reaction of sodium tetrahydridoborate(1-) (THB) with different acids under analytical conditions, was investigated and characterized. Pronounced demethylation of MAs(V), DMAs(V), and TMAs(V)O was found during the reaction of THB with HCl, H2SO4, and HClO4, while HG from CH3COOH or TRIS buffer after prereduction with l-cysteine resulted in the formation of only the corresponding hydrides. In the case of HNO3 formation of corresponding hydrides was preserved for MAs(V) and DMAs(V) but not for TMAs(V)O. The extent of demethylation strongly depends on concentration of the acid and THB. It can be strongly suppressed in HCl medium by partial hydrolysis of THB with optimal concentration of acid before it reacts with MAs(V), DMAs(V), or TMAs(V)O. It appears that the demethylation is due to the action of specific hydrolytic products of THB (most probably by the first and second one).

  11. Modeling of an Active Tablet Coating Process.

    PubMed

    Toschkoff, Gregor; Just, Sarah; Knop, Klaus; Kleinebudde, Peter; Funke, Adrian; Djuric, Dejan; Scharrer, Georg; Khinast, Johannes G

    2015-12-01

    Tablet coating is a common unit operation in the pharmaceutical industry, during which a coating layer is applied to tablet cores. The coating uniformity of tablets in a batch is especially critical for active coating, that is, coating that contains an active pharmaceutical ingredient. In recent years, discrete element method (DEM) simulations became increasingly common for investigating tablet coating. In this work, DEM was applied to model an active coating process as closely as possible, using measured model parameters and non-spherical particles. We studied how operational conditions (rotation speed, fill level, number of nozzles, and spray rate) influence the coating uniformity. To this end, simulation runs were planned and interpreted according to a statistical design of (simulation) experiments. Our general goal was to achieve a deeper understanding of the process in terms of residence times and dimensionless scaling laws. With that regard, the results were interpreted in light of analytical models. The results were presented at various detail levels, ranging from an overview of all variations to in-depth considerations. It was determined that the biggest uniformity improvement in a realistic setting was achieved by increasing the number of spray nozzles, followed by increasing the rotation speed and decreasing the fill level.

  12. Active PZT fibers: a commercial production process

    NASA Astrophysics Data System (ADS)

    Strock, Harold B.; Pascucci, Marina R.; Parish, Mark V.; Bent, Aaron A.; Shrout, Thomas R.

    1999-07-01

    Lead Zirconate Titanate (PZT) active fibers, from 80 to 250 micrometers in diameter, are produced for the AFOSR/DARPA funded Active Fiber Composites Consortium (AFCC) Program and commercial customers. CeraNova has developed a proprietary ceramics-based technology to produce PZT mono-filaments of the required purity, composition, straightness, and piezoelectric properties for use in active fiber composite structures. CeraNova's process begins with the extrusion of continuous lengths of mono-filament precursor fiber from a plasticized mix of PZT-5A powder. The care that must be taken to avoid mix contamination is described using illustrations form problems experiences with extruder wear and metallic contamination. Corrective actions are described and example microstructures are shown. The consequences of inadequate lead control are also shown. Sintered mono- filament mechanical strength and piezoelectric properties data approach bulk values but the validity of such a benchmark is questioned based on variable correlation with composite performance measures. Comb-like ceramic preform structures are shown that are being developed to minimize process and handling costs while maintaining the required mono-filament straightness necessary for composite fabrication. Lastly, actuation performance data are presented for composite structures fabricated and tested by Continuum Control Corporation. Free strain actuation in excess of 2000 microstrain are observed.

  13. Promoted Interaction of C/EBPα with Demethylated Cxcr3 Gene Promoter Contributes to Neuropathic Pain in Mice.

    PubMed

    Jiang, Bao-Chun; He, Li-Na; Wu, Xiao-Bo; Shi, Hui; Zhang, Wen-Wen; Zhang, Zhi-Jun; Cao, De-Li; Li, Chun-Hua; Gu, Jun; Gao, Yong-Jing

    2017-01-18

    DNA methylation has been implicated in the pathogenesis of chronic pain. However, the specific genes regulated by DNA methylation under neuropathic pain condition remain largely unknown. Here we investigated how chemokine receptor CXCR3 is regulated by DNA methylation and how it contributes to neuropathic pain induced by spinal nerve ligation (SNL) in mice. SNL increased Cxcr3 mRNA and protein expression in the neurons of the spinal cord. Meanwhile, the CpG (5'-cytosine-phosphate-guanine-3') island in the Cxcr3 gene promoter region was demethylated, and the expression of DNA methyltransferase 3b (DNMT3b) was decreased. SNL also increased the binding of CCAAT (cytidine-cytidine-adenosine-adenosine-thymidine)/enhancer binding protein α (C/EBPα) with Cxcr3 promoter and decreased the binding of DNMT3b with Cxcr3 promoter in the spinal cord. C/EBPα expression was increased in spinal neurons after SNL, and inhibition of C/EBPα by intrathecal small interfering RNA attenuated SNL-induced pain hypersensitivity and reduced Cxcr3 expression. Furthermore, SNL-induced mechanical allodynia and heat hyperalgesia were markedly reduced in Cxcr3(-/-) mice. Spinal inhibition of Cxcr3 by shRNA or CXCR3 antagonist also attenuated established neuropathic pain. Moreover, CXCL10, the ligand of CXCR3, was increased in spinal neurons and astrocytes after SNL. Superfusing spinal cord slices with CXCL10 enhanced spontaneous EPSCs and potentiated NMDA-induced and AMPA-induced currents of lamina II neurons. Finally, intrathecal injection of CXCL10 induced CXCR3-dependent pain hypersensitivity in naive mice. Collectively, our results demonstrated that CXCR3, increased by DNA demethylation and the enhanced interaction with C/EBPα, can be activated by CXCL10 to facilitate excitatory synaptic transmission and contribute to the maintenance of neuropathic pain.

  14. Promoted interaction of C/EBPα with demethylated Cxcr3 gene promoter contributes to neuropathic pain in mice.

    PubMed

    Jiang, Bao-Chun; He, Li-Na; Wu, Xiao-Bo; Shi, Hui; Zhang, Wen-Wen; Zhang, Zhi-Jun; Cao, De-Li; Li, Chun-Hua; Gu, Jun; Gao, Yong-Jing

    2016-12-09

    DNA methylation has been implicated in the pathogenesis of chronic pain. However, the specific genes that are regulated by DNA methylation under neuropathic pain condition remain largely unknown. Here we investigated how chemokine receptor CXCR3 is regulated by DNA methylation and its contribution to neuropathic pain induced by spinal nerve ligation (SNL) in mice. SNL increased Cxcr3 mRNA and protein expression in the neurons of spinal cord. Meanwhile, the CpG island in the Cxcr3 gene promoter region was demethylated, and the expression of DNA methyltransferase 3b (DNMT3b) was decreased. SNL also increased the binding of CCAAT/enhancer binding protein α (C/EBPα) with Cxcr3 promoter and decreased the binding of DNMT3b with Cxcr3 promoter in the spinal cord. C/EBPα expression was increased in spinal neurons after SNL, and inhibition of C/EBPα by intrathecal siRNA attenuated SNL-induced pain hypersensitivity and reduced Cxcr3 expression. Furthermore, SNL-induced mechanical allodynia and heat hyperalgesia were markedly reduced in Cxcr3(-/-) mice. Spinal inhibition of Cxcr3 by shRNA or CXCR3 antagonist also attenuated established neuropathic pain. Moreover, CXCL10, the ligand of CXCR3 was increased in spinal neurons and astrocytes after SNL. Superfusing spinal cord slices with CXCL10 enhanced spontaneous EPSCs and potentiated NMDA- and AMPA-induced currents of lamina II neurons. Finally, intrathecal injection of CXCL10 induced CXCR3-dependent pain hypersensitivity in naïve mice. Collectively, our results demonstrated that CXCR3, increased by DNA demethylation and the enhanced interaction with C/EBPα, can be activated by CXCL10 to facilitate excitatory synaptic transmission and contribute to the maintenance of neuropathic pain.

  15. 15 CFR 400.31 - Manufacturing and processing activity; criteria.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 15 Commerce and Foreign Trade 2 2010-01-01 2010-01-01 false Manufacturing and processing activity... ZONES BOARD Manufacturing and Processing Activity-Reviews § 400.31 Manufacturing and processing activity....” When evaluating zone and subzone manufacturing and processing activity, either as proposed in...

  16. 15 CFR 400.31 - Manufacturing and processing activity; criteria.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 15 Commerce and Foreign Trade 2 2011-01-01 2011-01-01 false Manufacturing and processing activity... ZONES BOARD Manufacturing and Processing Activity-Reviews § 400.31 Manufacturing and processing activity....” When evaluating zone and subzone manufacturing and processing activity, either as proposed in...

  17. 15 CFR 400.31 - Manufacturing and processing activity; criteria.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 15 Commerce and Foreign Trade 2 2012-01-01 2012-01-01 false Manufacturing and processing activity... ZONES BOARD Manufacturing and Processing Activity-Reviews § 400.31 Manufacturing and processing activity....” When evaluating zone and subzone manufacturing and processing activity, either as proposed in...

  18. Tet3 and DNA replication mediate demethylation of both the maternal and paternal genomes in mouse zygotes.

    PubMed

    Shen, Li; Inoue, Azusa; He, Jin; Liu, Yuting; Lu, Falong; Zhang, Yi

    2014-10-02

    With the exception of imprinted genes and certain repeats, DNA methylation is globally erased during preimplantation development. Recent studies have suggested that Tet3-mediated oxidation of 5-methylcytosine (5mC) and DNA replication-dependent dilution both contribute to global paternal DNA demethylation, but demethylation of the maternal genome occurs via replication. Here we present genome-scale DNA methylation maps for both the paternal and maternal genomes of Tet3-depleted and/or DNA replication-inhibited zygotes. In both genomes, we found that inhibition of DNA replication blocks DNA demethylation independently from Tet3 function and that Tet3 facilitates DNA demethylation largely by coupling with DNA replication. For both genomes, our data indicate that replication-dependent dilution is the major contributor to demethylation, but Tet3 plays an important role, particularly at certain loci. Our study thus defines the respective functions of Tet3 and DNA replication in paternal DNA demethylation and reveals an unexpected contribution of Tet3 to demethylation of the maternal genome.

  19. Correlated activity supports efficient cortical processing

    PubMed Central

    Hung, Chou P.; Cui, Ding; Chen, Yueh-peng; Lin, Chia-pei; Levine, Matthew R.

    2015-01-01

    Visual recognition is a computational challenge that is thought to occur via efficient coding. An important concept is sparseness, a measure of coding efficiency. The prevailing view is that sparseness supports efficiency by minimizing redundancy and correlations in spiking populations. Yet, we recently reported that “choristers”, neurons that behave more similarly (have correlated stimulus preferences and spontaneous coincident spiking), carry more generalizable object information than uncorrelated neurons (“soloists”) in macaque inferior temporal (IT) cortex. The rarity of choristers (as low as 6% of IT neurons) indicates that they were likely missed in previous studies. Here, we report that correlation strength is distinct from sparseness (choristers are not simply broadly tuned neurons), that choristers are located in non-granular output layers, and that correlated activity predicts human visual search efficiency. These counterintuitive results suggest that a redundant correlational structure supports efficient processing and behavior. PMID:25610392

  20. Diazirine photocrosslinking recruits activated FTO demethylase complexes for specific N(6)-methyladenosine recognition.

    PubMed

    Jeong, Hyun Seok; Hayashi, Gosuke; Okamoto, Akimitsu

    2015-06-19

    N(6)-methyladenosine (m(6)A) is a prevalent modification of RNAs. m(6)A exists in mRNA and plays an important role in RNA biological pathways and in RNA epigenetic regulation. We applied diazirine photocrosslinking to the event of m(6)A recognition mediated by the fat mass and obesity associated (FTO) demethylase. A highly photoreactive diazirine adjacent to m(6)A on the RNA successfully recruited activated FTO complexes with an m(6)A preference. The process of recognition of m(6)A via FTO using diazirine photocrosslinking was controlled by the α-ketoglutarate (α-KG) cosubstrate and the Fe(II) cofactor, which are involved in m(6)A oxidative demethylation. In addition, FTO bound to ssRNAs prior to the m(6)A recognition process. Diazirine photocrosslinking contributes to increasing the chances of capturing activated FTO complexes with specific m(6)A recognition and provides new insights into the dynamic FTO oxidative demethylation process.

  1. Methylation and demethylation of intermediates selenide and methylselenol in the metabolism of selenium.

    PubMed

    Ohta, Yuki; Suzuki, Kazuo T

    2008-01-15

    All nutritional selenium sources are transformed into the assumed common intermediate selenide for the syntheses of selenoproteins for utilization and/or of selenosugar for excretion. Methylselenol [monomethylselenide, MMSe] is the assumed intermediate leading to other methylated metabolites, dimethylselenide (DMSe) and trimethylselenonium (TMSe) for excretion, and also to the intermediate selenide from methylselenocysteine and methylseleninic acid (MSA). Here, related methylation and demethylation reactions were studied in vitro by providing chemically reactive starting substrates (76Se-selenide, 77Se-MMSe and 82Se-DMSe) which were prepared in situ by the reduction of the corresponding labeled proximate precursors (76Se-selenite, 77Se-MSA and 82Se-dimethylselenoxide (DMSeO), respectively) with glutathione, the three substrates being incubated simultaneously in rat organ supernatants and homogenates. The resulting chemically labile reaction products were detected simultaneously by speciation analysis with HPLC-ICP-MS after converting the products and un-reacted substrates to the corresponding oxidized derivatives (selenite, MSA and DMSeO). The time-related changes in selenium isotope profiles showed that demethylation of MMSe to selenide was efficient but that of DMSe to MMSe was negligible, whereas methylation of selenide to MMSe, and MMSe to DMSe were efficient, and that of DMSe to TMSe occurred less efficiently. The present methylation and demethylation reactions on equilibrium between selenide, MMSe and DMSe without producing selenosugar and selenoproteins indicated that DMSe rather than TMSe is produced as the end product, suggesting that DMSe is to be excreted more abundantly than TMSe. Organ-dependent differences in the methylation and demethylation reactions were characterized for the liver, kidney and lung.

  2. Methylation and demethylation of intermediates selenide and methylselenol in the metabolism of selenium

    SciTech Connect

    Ohta, Yuki; Suzuki, Kazuo T.

    2008-01-15

    All nutritional selenium sources are transformed into the assumed common intermediate selenide for the syntheses of selenoproteins for utilization and/or of selenosugar for excretion. Methylselenol [monomethylselenide, MMSe] is the assumed intermediate leading to other methylated metabolites, dimethylselenide (DMSe) and trimethylselenonium (TMSe) for excretion, and also to the intermediate selenide from methylselenocysteine and methylseleninic acid (MSA). Here, related methylation and demethylation reactions were studied in vitro by providing chemically reactive starting substrates ({sup 76}Se-selenide, {sup 77}Se-MMSe and {sup 82}Se-DMSe) which were prepared in situ by the reduction of the corresponding labeled proximate precursors ({sup 76}Se-selenite, {sup 77}Se-MSA and {sup 82}Se-dimethylselenoxide (DMSeO), respectively) with glutathione, the three substrates being incubated simultaneously in rat organ supernatants and homogenates. The resulting chemically labile reaction products were detected simultaneously by speciation analysis with HPLC-ICP-MS after converting the products and un-reacted substrates to the corresponding oxidized derivatives (selenite, MSA and DMSeO). The time-related changes in selenium isotope profiles showed that demethylation of MMSe to selenide was efficient but that of DMSe to MMSe was negligible, whereas methylation of selenide to MMSe, and MMSe to DMSe were efficient, and that of DMSe to TMSe occurred less efficiently. The present methylation and demethylation reactions on equilibrium between selenide, MMSe and DMSe without producing selenosugar and selenoproteins indicated that DMSe rather than TMSe is produced as the end product, suggesting that DMSe is to be excreted more abundantly than TMSe. Organ-dependent differences in the methylation and demethylation reactions were characterized for the liver, kidney and lung.

  3. Gene structure and transcription in mouse cells with extensively demethylated DNA.

    PubMed Central

    Michalowsky, L A; Jones, P A

    1989-01-01

    In previous work, three clonal cell lines with extremely low DNA methylation levels were derived by multiple consecutive treatments of C3H 10T1/2 C18 (10T1/2) cells with 5-aza-2'-deoxycytidine (5-aza-CdR). In this study we examined the methylation status of genes in these three methyl-deficient clones to assess the specificity of the induced hypomethylation. Complete demethylation of virtually all 5'-CCGG-3' sites was observed in four genes examined, but some sites common to all three clones were persistently methylated even after further exhaustive 5-aza-CdR treatment. Thus, there is a subset of methylation sites within these cells which can never be stably demethylated. The extensive demethylation was not always associated with changes in the level of RNA expression of the genes examined but was strongly correlated with an altered chromatin structure of the unexpressed alpha 1-globin gene and the muscle determination gene MyoD1. These results provide a direct correlation between hypomethylation and the induction of a transcriptionally competent chromatin state. Images PMID:2471061

  4. Antibotulinal activity of process cheese ingredients.

    PubMed

    Glass, Kathleen A; Johnson, Eric A

    2004-08-01

    Ingredients used in the manufacture of reduced-fat process cheese products were screened for their ability to inhibit growth of Clostridium botulinum serotypes A and B in media. Reinforced clostridial medium (RCM) supplemented with 0, 0.5, 1, 2, 3, 5, or 10% (wt/vol) of various ingredients, including a carbohydrate-based fat replacer, an enzyme-modified cheese (EMC) derived from a Blue cheese, sweet whey, modified whey protein, or whey protein concentrate, did not inhibit botulinal growth and toxin production when stored at 30 degrees C for 1 week. In contrast, RCM supplemented with 10% soy-based flavor enhancer, 10% Parmesan EMC, or 5 or 10% Cheddar EMC inhibited botulinal toxin production in media for at least 6 weeks of storage at 30 degrees C. Subsequent trials revealed that the antibotulinal effect varied significantly among 13 lots of EMC and that the antimicrobial effect was not correlated with the pH or water activity of the EMC.

  5. Degassing Processes at Persistently Active Explosive Volcanoes

    NASA Astrophysics Data System (ADS)

    Smekens, Jean-Francois

    Among volcanic gases, sulfur dioxide (SO2) is by far the most commonly measured. More than a monitoring proxy for volcanic degassing, SO 2 has the potential to alter climate patterns. Persistently active explosive volcanoes are characterized by short explosive bursts, which often occur at periodic intervals numerous times per day, spanning years to decades. SO 2 emissions at those volcanoes are poorly constrained, in large part because the current satellite monitoring techniques are unable to detect or quantify plumes of low concentration in the troposphere. Eruption plumes also often show high concentrations of ash and/or aerosols, which further inhibit the detection methods. In this work I focus on quantifying volcanic gas emissions at persistently active explosive volcanoes and their variations over short timescales (minutes to hours), in order to document their contribution to natural SO2 flux as well as investigate the physical processes that control their behavior. In order to make these measurements, I first develop and assemble a UV ground-based instrument, and validate it against an independently measured source of SO2 at a coal-burning power plant in Arizona. I establish a measurement protocol and demonstrate that the instrument measures SO 2 fluxes with < 20 % error. Using the same protocol, I establish a record of the degassing patterns at Semeru volcano (Indonesia), a volcano that has been producing cycles of repeated explosions with periods of minutes to hours for the past several decades. Semeru produces an average of 21-71 tons of SO2 per day, amounting to a yearly output of 8-26 Mt. Using the Semeru data, along with a 1-D transient numerical model of magma ascent, I test the validity of a model in which a viscous plug at the top of the conduit produces cycles of eruption and gas release. I find that it can be a valid hypothesis to explain the observed patterns of degassing at Semeru. Periodic behavior in such a system occurs for a very narrow range

  6. Global identification of genes regulated by estrogen signaling and demethylation in MCF-7 breast cancer cells

    SciTech Connect

    Putnik, Milica; Zhao, Chunyan; Gustafsson, Jan-Ake; Dahlman-Wright, Karin

    2012-09-14

    Highlights: Black-Right-Pointing-Pointer Estrogen signaling and demethylation can both control gene expression in breast cancers. Black-Right-Pointing-Pointer Cross-talk between these mechanisms is investigated in human MCF-7 breast cancer cells. Black-Right-Pointing-Pointer 137 genes are influenced by both 17{beta}-estradiol and demethylating agent 5-aza-2 Prime -deoxycytidine. Black-Right-Pointing-Pointer A set of genes is identified as targets of both estrogen signaling and demethylation. Black-Right-Pointing-Pointer There is no direct molecular interplay of mediators of estrogen and epigenetic signaling. -- Abstract: Estrogen signaling and epigenetic modifications, in particular DNA methylation, are involved in regulation of gene expression in breast cancers. Here we investigated a potential regulatory cross-talk between these two pathways by identifying their common target genes and exploring underlying molecular mechanisms in human MCF-7 breast cancer cells. Gene expression profiling revealed that the expression of approximately 140 genes was influenced by both 17{beta}-estradiol (E2) and a demethylating agent 5-aza-2 Prime -deoxycytidine (DAC). Gene ontology (GO) analysis suggests that these genes are involved in intracellular signaling cascades, regulation of cell proliferation and apoptosis. Based on previously reported association with breast cancer, estrogen signaling and/or DNA methylation, CpG island prediction and GO analysis, we selected six genes (BTG3, FHL2, PMAIP1, BTG2, CDKN1A and TGFB2) for further analysis. Tamoxifen reverses the effect of E2 on the expression of all selected genes, suggesting that they are direct targets of estrogen receptor. Furthermore, DAC treatment reactivates the expression of all selected genes in a dose-dependent manner. Promoter CpG island methylation status analysis revealed that only the promoters of BTG3 and FHL2 genes are methylated, with DAC inducing demethylation, suggesting DNA methylation directs repression of

  7. UTX demethylase activity is required for satellite cell–mediated muscle regeneration

    PubMed Central

    Wang, Chaochen; Nakka, Kiran; Benyoucef, Aissa; Sebastian, Soji; Zhuang, Lenan; Chu, Alphonse; Palii, Carmen G.; Camellato, Brendan; Brand, Marjorie

    2016-01-01

    The X chromosome–encoded histone demethylase UTX (also known as KDM6A) mediates removal of repressive trimethylation of histone H3 lysine 27 (H3K27me3) to establish transcriptionally permissive chromatin. Loss of UTX in female mice is embryonic lethal. Unexpectedly, male UTX-null mice escape embryonic lethality due to expression of UTY, a paralog that lacks H3K27 demethylase activity, suggesting an enzyme-independent role for UTX in development and thereby challenging the need for active H3K27 demethylation in vivo. However, the requirement for active H3K27 demethylation in stem cell–mediated tissue regeneration remains untested. Here, we employed an inducible mouse KO that specifically ablates Utx in satellite cells (SCs) and demonstrated that active H3K27 demethylation is necessary for muscle regeneration. Loss of UTX in SCs blocked myofiber regeneration in both male and female mice. Furthermore, we demonstrated that UTX mediates muscle regeneration through its H3K27 demethylase activity, as loss of demethylase activity either by chemical inhibition or knock-in of demethylase-dead UTX resulted in defective muscle repair. Mechanistically, dissection of the muscle regenerative process revealed that the demethylase activity of UTX is required for expression of the transcription factor myogenin, which in turn drives differentiation of muscle progenitors. Thus, we have identified a critical role for the enzymatic activity of UTX in activating muscle-specific gene expression during myofiber regeneration and have revealed a physiological role for active H3K27 demethylation in vivo. PMID:26999603

  8. Analysis of the machinery and intermediates of the 5hmC-mediated DNA demethylation pathway in aging on samples from the MARK-AGE Study

    PubMed Central

    Valentini, Elisabetta; Zampieri, Michele; Malavolta, Marco; Bacalini, Maria Giulia; Calabrese, Roberta; Guastafierro, Tiziana; Reale, Anna; Franceschi, Claudio; Hervonen, Antti; Koller, Bernhard; Bernhardt, Jürgen; Slagboom, P. Eline; Toussaint, Olivier; Sikora, Ewa; Gonos, Efstathios S.; Breusing, Nicolle; Grune, Tilman; Jansen, Eugène; Dollé, Martijn E.T.; Moreno-Villanueva, María; Sindlinger, Thilo; Bürkle, Alexander; Ciccarone, Fabio; Caiafa, Paola

    2016-01-01

    Gradual changes in the DNA methylation landscape occur throughout aging virtually in all human tissues. A widespread reduction of 5-methylcytosine (5mC), associated with highly reproducible site-specific hypermethylation, characterizes the genome in aging. Therefore, an equilibrium seems to exist between general and directional deregulating events concerning DNA methylation controllers, which may underpin the age-related epigenetic changes. In this context, 5mC-hydroxylases (TET enzymes) are new potential players. In fact, TETs catalyze the stepwise oxidation of 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), driving the DNA demethylation process based on thymine DNA glycosylase (TDG)-mediated DNA repair pathway. The present paper reports the expression of DNA hydroxymethylation components, the levels of 5hmC and of its derivatives in peripheral blood mononuclear cells of age-stratified donors recruited in several European countries in the context of the EU Project ‘MARK-AGE’. The results provide evidence for an age-related decline of TET1, TET3 and TDG gene expression along with a decrease of 5hmC and an accumulation of 5caC. These associations were independent of confounding variables, including recruitment center, gender and leukocyte composition. The observed impairment of 5hmC-mediated DNA demethylation pathway in blood cells may lead to aberrant transcriptional programs in the elderly. PMID:27587280

  9. Analysis of the machinery and intermediates of the 5hmC-mediated DNA demethylation pathway in aging on samples from the MARK-AGE Study.

    PubMed

    Valentini, Elisabetta; Zampieri, Michele; Malavolta, Marco; Bacalini, Maria Giulia; Calabrese, Roberta; Guastafierro, Tiziana; Reale, Anna; Franceschi, Claudio; Hervonen, Antti; Koller, Bernhard; Bernhardt, Jürgen; Slagboom, P Eline; Toussaint, Olivier; Sikora, Ewa; Gonos, Efstathios S; Breusing, Nicolle; Grune, Tilman; Jansen, Eugène; Dollé, Martijn E T; Moreno-Villanueva, María; Sindlinger, Thilo; Bürkle, Alexander; Ciccarone, Fabio; Caiafa, Paola

    2016-08-29

    Gradual changes in the DNA methylation landscape occur throughout aging virtually in all human tissues. A widespread reduction of 5-methylcytosine (5mC), associated with highly reproducible site-specific hypermethylation, characterizes the genome in aging. Therefore, an equilibrium seems to exist between general and directional deregulating events concerning DNA methylation controllers, which may underpin the age-related epigenetic changes. In this context, 5mC-hydroxylases (TET enzymes) are new potential players. In fact, TETs catalyze the stepwise oxidation of 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC), driving the DNA demethylation process based on thymine DNA glycosylase (TDG)-mediated DNA repair pathway. The present paper reports the expression of DNA hydroxymethylation components, the levels of 5hmC and of its derivatives in peripheral blood mononuclear cells of age-stratified donors recruited in several European countries in the context of the EU Project 'MARK-AGE'. The results provide evidence for an age-related decline of TET1, TET3 and TDG gene expression along with a decrease of 5hmC and an accumulation of 5caC. These associations were independent of confounding variables, including recruitment center, gender and leukocyte composition. The observed impairment of 5hmC-mediated DNA demethylation pathway in blood cells may lead to aberrant transcriptional programs in the elderly.

  10. Tissue Inhibitor of Metalloproteinase 1 Expression Associated with Gene Demethylation Confers Anoikis Resistance in Early Phases of Melanocyte Malignant Transformation1

    PubMed Central

    Ricca, Tatiana I; Liang, Gangning; Suenaga, Ana Paula M; Han, Sang W; Jones, Peter A; Jasiulionis, Miriam G

    2009-01-01

    Although anoikis resistance has been considered a hallmark of malignant phenotype, the causal relation between neoplastic transformation and anchorage-independent growth remains undefined. We developed an experimental model of murine melanocyte malignant transformation, where a melanocyte lineage (melan-a) was submitted to sequential cycles of anchorage blockade, resulting in progressive morphologic alterations, and malignant transformation. Throughout this process, cells corresponding to premalignant melanocytes and melanoma cell lines were established and show progressive anoikis resistance and increased expression of Timp1. In melan-a melanocytes, Timp1 expression is suppressed by DNA methylation as indicated by its reexpression after 5-aza-2′-deoxycytidine treatment. Methylation-sensitive single-nucleotide primer extension analysis showed increased demethylation in Timp1 in parallel with its expression along malignant transformation. Interestingly, TIMP1 expression has already been related with negative prognosis in some human cancers. Although described as a MMP inhibitor, this protein has been associated with apoptosis resistance in different cell types. Melan-a cells overexpressing Timp1 showed increased survival in suspension but were unable to form tumors in vivo, whereas Timp1-overexpressing melanoma cells showed reduced latency time for tumor appearance and increased metastatic potential. Here, we demonstrated for the first time an increment in Timp1 expression since the early phases of melanocyte malignant transformation, associated to a progressive gene demethylation, which confers anoikis resistance. In this way, Timp1 might be considered as a valued marker for melanocyte malignant transformation. PMID:19956395

  11. Automated Quantification of DNA Demethylation Effects in Cells via 3D Mapping of Nuclear Signatures and Population Homogeneity Assessment1

    PubMed Central

    Gertych, Arkadiusz; Wawrowsky, Kolja A.; Lindsley, Erik; Vishnevsky, Eugene; Farkas, Daniel L.; Tajbakhsh, Jian

    2009-01-01

    Background Today’s advanced microscopic imaging applies to the preclinical stages of drug discovery that employ high-throughput and high-content three-dimensional (3D) analysis of cells to more efficiently screen candidate compounds. Drug efficacy can be assessed by measuring response homogeneity to treatment within a cell population. In this study topologically quantified nuclear patterns of methylated cytosine and global nuclear DNA are utilized as signatures of cellular response to the treatment of cultured cells with the demethylating anti-cancer agents: 5-azacytidine (5-AZA) and octreotide (OCT). Methods Mouse pituitary folliculostellate TtT-GF cells treated with 5-AZA and OCT for 48 hours, and untreated populations, were studied by immunofluorescence with a specific antibody against 5-methylcytosine (MeC), and 4,6-diamidino-2-phenylindole (DAPI) for delineation of methylated sites and global DNA in nuclei (n=163). Cell images were processed utilizing an automated 3D analysis software that we developed by combining seeded watershed segmentation to extract nuclear shells with measurements of Kullback-Leibler’s (K-L) divergence to analyze cell population homogeneity in the relative nuclear distribution patterns of MeC versus DAPI stained sites. Each cell was assigned to one of the four classes: similar, likely similar, unlikely similar and dissimilar. Results Evaluation of the different cell groups revealed a significantly higher number of cells with similar or likely similar MeC/DAPI patterns among untreated cells (~100%), 5-AZA-treated cells (90%), and a lower degree of same type of cells (64%) in the OCT-treated population. The latter group contained (28%) of unlikely similar or dissimilar (7%) cells. Conclusion Our approach was successful in the assessment of cellular behavior relevant to the biological impact of the applied drugs, i.e. the reorganization of MeC/DAPI distribution by demethylation. In a comparison with other metrics, K-L divergence has

  12. N-demethylation and N-oxidation of imipramine in rat thoracic aortic endothelial cells.

    PubMed

    Ueda, Yukari; Yaginuma, Toshihiko; Sakurai, Eiko; Sakurai, Eiichi

    2014-06-01

    The aim of this study was to examine whether cultured rat thoracic aortic endothelial cells (TAECs) have the ability to metabolize the tertiary amine, imipramine. In rat TAECs, imipramine was biotransformed into N-demethylate and N-oxide by cytochrome P450 (CYP) and flavin-containing monooxygenase (FMO), respectively. The intrinsic clearance (V max/K m) for the N-oxide formation was approximately five times as high as that for the N-demethylate formation, indicating that oxidation by CYP was much higher than that by FMO. Moreover, we suggest that CYP2C11 and CYP3A2 are key players in the metabolism to N-demethylate in rat TAECs using the respective anti-rat CYP antibodies (anti-CYP2C11 and anti-CYP3A2). The presence of CYP2C11 and CYP3A2 proteins was also confirmed in cultured rat TAECs using a polyclonal anti-CYP antibody and immunofluorescence microscopy. In contrast, the formation rate of N-oxide at pH 8.4 was higher than that at pH 7.4. Inhibition of N-oxide formation by methimazole was found to be the best model of competitive inhibition yielding an apparent K i value of 0.80 μmol/L, demonstrating that N-oxidation was catalyzed by FMO in rat TAECs. These results suggest that rat TAEC enzymes can convert substrates of exogenous origin such as imipramine, indicating that TAECs have an important function for metabolic products, besides hepatic cells.

  13. IDH mutation impairs histone demethylation and results in a block to cell differentiation.

    PubMed

    Lu, Chao; Ward, Patrick S; Kapoor, Gurpreet S; Rohle, Dan; Turcan, Sevin; Abdel-Wahab, Omar; Edwards, Christopher R; Khanin, Raya; Figueroa, Maria E; Melnick, Ari; Wellen, Kathryn E; O'Rourke, Donald M; Berger, Shelley L; Chan, Timothy A; Levine, Ross L; Mellinghoff, Ingo K; Thompson, Craig B

    2012-02-15

    Recurrent mutations in isocitrate dehydrogenase 1 (IDH1) and IDH2 have been identified in gliomas, acute myeloid leukaemias (AML) and chondrosarcomas, and share a novel enzymatic property of producing 2-hydroxyglutarate (2HG) from α-ketoglutarate. Here we report that 2HG-producing IDH mutants can prevent the histone demethylation that is required for lineage-specific progenitor cells to differentiate into terminally differentiated cells. In tumour samples from glioma patients, IDH mutations were associated with a distinct gene expression profile enriched for genes expressed in neural progenitor cells, and this was associated with increased histone methylation. To test whether the ability of IDH mutants to promote histone methylation contributes to a block in cell differentiation in non-transformed cells, we tested the effect of neomorphic IDH mutants on adipocyte differentiation in vitro. Introduction of either mutant IDH or cell-permeable 2HG was associated with repression of the inducible expression of lineage-specific differentiation genes and a block to differentiation. This correlated with a significant increase in repressive histone methylation marks without observable changes in promoter DNA methylation. Gliomas were found to have elevated levels of similar histone repressive marks. Stable transfection of a 2HG-producing mutant IDH into immortalized astrocytes resulted in progressive accumulation of histone methylation. Of the marks examined, increased H3K9 methylation reproducibly preceded a rise in DNA methylation as cells were passaged in culture. Furthermore, we found that the 2HG-inhibitable H3K9 demethylase KDM4C was induced during adipocyte differentiation, and that RNA-interference suppression of KDM4C was sufficient to block differentiation. Together these data demonstrate that 2HG can inhibit histone demethylation and that inhibition of histone demethylation can be sufficient to block the differentiation of non-transformed cells.

  14. Cell-Wide DNA De-Methylation and Re-Methylation of Purkinje Neurons in the Developing Cerebellum

    PubMed Central

    Zhou, Feng C.; Resendiz, Marisol; Lo, Chiao-Ling; Chen, Yuanyuan

    2016-01-01

    Global DNA de-methylation is thought to occur only during pre-implantation and gametogenesis in mammals. Scalable, cell-wide de-methylation has not been demonstrated beyond totipotent stages. Here, we observed a large scale de-methylation and subsequent re-methylation (CDR) (including 5-methylcytosine (5mC) and 5-hydroxylmethylcytosine (5hmC)) in post-mitotic cerebellar Purkinje cells (PC) through the course of normal development. Through single cell immuno-identification and cell-specific quantitative methylation assays, we demonstrate that the CDR event is an intrinsically scheduled program, occurring in nearly every PC. Meanwhile, cerebellar granule cells and basket interneurons adopt their own DNA methylation program, independent of PCs. DNA de-methylation was further demonstrated at the gene level, on genes pertinent to PC development. The PC, being one of the largest neurons in the brain, may showcase an amplified epigenetic cycle which may mediate stage transformation including cell cycle arrest, vast axonal-dendritic growth, and synaptogenesis at the onset of neuronal specificity. This discovery is a key step toward better understanding the breadth and role of DNA methylation and de-methylation during neural ontology. PMID:27583369

  15. Laboratory Activities for Developing Process Skills.

    ERIC Educational Resources Information Center

    Institute for Services to Education, Inc., Washington, DC.

    This workbook contains laboratory exercises designed for use in a college introductory biology course. Each exercise helps the student develop a basic science skill. The exercises are arranged in a hierarchical sequence suggesting the scientific method. Each skill facilitates the development of succeeding ones. Activities include Use of the…

  16. Dioxygenases catalyze O-demethylation and O,O-demethylenation with widespread roles in benzylisoquinoline alkaloid metabolism in opium poppy.

    PubMed

    Farrow, Scott C; Facchini, Peter J

    2013-10-04

    In opium poppy, the antepenultimate and final steps in morphine biosynthesis are catalyzed by the 2-oxoglutarate/Fe(II)-dependent dioxygenases, thebaine 6-O-demethylase (T6ODM) and codeine O-demethylase (CODM). Further investigation into the biochemical functions of CODM and T6ODM revealed extensive and unexpected roles for such enzymes in the metabolism of protopine, benzo[c]phenanthridine, and rhoeadine alkaloids. When assayed with a wide range of benzylisoquinoline alkaloids, CODM, T6ODM, and the functionally unassigned paralog DIOX2, renamed protopine O-dealkylase, showed novel and efficient dealkylation activities, including regio- and substrate-specific O-demethylation and O,O-demethylenation. Enzymes catalyzing O,O-demethylenation, which cleave a methylenedioxy bridge leaving two hydroxyl groups, have previously not been reported in plants. Similar cleavage of methylenedioxy bridges on substituted amphetamines is catalyzed by heme-dependent cytochromes P450 in mammals. Preferred substrates for O,O-demethylenation by CODM and protopine O-dealkylase were protopine alkaloids that serve as intermediates in the biosynthesis of benzo[c]phenanthridine and rhoeadine derivatives. Virus-induced gene silencing used to suppress the abundance of CODM and/or T6ODM transcripts indicated a direct physiological role for these enzymes in the metabolism of protopine alkaloids, and they revealed their indirect involvement in the formation of the antimicrobial benzo[c]phenanthridine sanguinarine and certain rhoeadine alkaloids in opium poppy.

  17. Modelling the Active Hearing Process in Mosquitoes

    NASA Astrophysics Data System (ADS)

    Avitabile, Daniele; Homer, Martin; Jackson, Joe; Robert, Daniel; Champneys, Alan

    2011-11-01

    A simple microscopic mechanistic model is described of the active amplification within the Johnston's organ of the mosquito species Toxorhynchites brevipalpis. The model is based on the description of the antenna as a forced-damped oscillator coupled to a set of active threads (ensembles of scolopidia) that provide an impulsive force when they twitch. This twitching is in turn controlled by channels that are opened and closed if the antennal oscillation reaches a critical amplitude. The model matches both qualitatively and quantitatively with recent experiments. New results are presented using mathematical homogenization techniques to derive a mesoscopic model as a simple oscillator with nonlinear force and damping characteristics. It is shown how the results from this new model closely resemble those from the microscopic model as the number of threads approach physiologically correct values.

  18. Capturing Cognitive Processing Time for Active Authentication

    DTIC Science & Technology

    2014-02-01

    biometrics, extracted from keystroke dynamics , as “something a user is” for active authentication. This scheme performs continual verification in the...fingerprint for continuous authentication. Its effectiveness has been verified through a large-scale dataset. 2.0 INTRODUCTION Keystroke dynamics —the...measure the similarity. A recent survey on biometric authentication using keystroke dynamics classified research papers on the basis of their

  19. Uav Data Processing for Rapid Mapping Activities

    NASA Astrophysics Data System (ADS)

    Tampubolon, W.; Reinhardt, W.

    2015-08-01

    During disaster and emergency situations, geospatial data plays an important role to serve as a framework for decision support system. As one component of basic geospatial data, large scale topographical maps are mandatory in order to enable geospatial analysis within quite a number of societal challenges. The increasing role of geo-information in disaster management nowadays consequently needs to include geospatial aspects on its analysis. Therefore different geospatial datasets can be combined in order to produce reliable geospatial analysis especially in the context of disaster preparedness and emergency response. A very well-known issue in this context is the fast delivery of geospatial relevant data which is expressed by the term "Rapid Mapping". Unmanned Aerial Vehicle (UAV) is the rising geospatial data platform nowadays that can be attractive for modelling and monitoring the disaster area with a low cost and timely acquisition in such critical period of time. Disaster-related object extraction is of special interest for many applications. In this paper, UAV-borne data has been used for supporting rapid mapping activities in combination with high resolution airborne Interferometric Synthetic Aperture Radar (IFSAR) data. A real disaster instance from 2013 in conjunction with Mount Sinabung eruption, Northern Sumatra, Indonesia, is used as the benchmark test for the rapid mapping activities presented in this paper. On this context, the reliable IFSAR dataset from airborne data acquisition in 2011 has been used as a comparable dataset for accuracy investigation and assessment purpose in 3 D reconstructions. After all, this paper presents a proper geo-referencing and feature extraction method of UAV data to support rapid mapping activities.

  20. Processing abstract language modulates motor system activity.

    PubMed

    Glenberg, Arthur M; Sato, Marc; Cattaneo, Luigi; Riggio, Lucia; Palumbo, Daniele; Buccino, Giovanni

    2008-06-01

    Embodiment theory proposes that neural systems for perception and action are also engaged during language comprehension. Previous neuroimaging and neurophysiological studies have only been able to demonstrate modulation of action systems during comprehension of concrete language. We provide neurophysiological evidence for modulation of motor system activity during the comprehension of both concrete and abstract language. In Experiment 1, when the described direction of object transfer or information transfer (e.g., away from the reader to another) matched the literal direction of a hand movement used to make a response, speed of responding was faster than when the two directions mismatched (an action-sentence compatibility effect). In Experiment 2, we used single-pulse transcranial magnetic stimulation to study changes in the corticospinal motor pathways to hand muscles while reading the same sentences. Relative to sentences that do not describe transfer, there is greater modulation of activity in the hand muscles when reading sentences describing transfer of both concrete objects and abstract information. These findings are discussed in relation to the human mirror neuron system.

  1. Genome-wide CpG island methylation and intergenic demethylation propensities vary among different tumor sites

    PubMed Central

    Lee, Seung-Tae; Wiemels, Joseph L.

    2016-01-01

    The epigenetic landscape of cancer includes both focal hypermethylation and broader hypomethylation in a genome-wide manner. By means of a comprehensive genomic analysis on 6637 tissues of 21 tumor types, we here show that the degrees of overall methylation in CpG island (CGI) and demethylation in intergenic regions, defined as ‘backbone’, largely vary among different tumors. Depending on tumor type, both CGI methylation and backbone demethylation are often associated with clinical, epidemiological and biological features such as age, sex, smoking history, anatomic location, histological type and grade, stage, molecular subtype and biological pathways. We found connections between CGI methylation and hypermutability, microsatellite instability, IDH1 mutation, 19p gain and polycomb features, and backbone demethylation with chromosomal instability, NSD1 and TP53 mutations, 5q and 19p loss and long repressive domains. These broad epigenetic patterns add a new dimension to our understanding of tumor biology and its clinical implications. PMID:26464434

  2. Continuous Zebularine Treatment Effectively Sustains Demethylation in Human Bladder Cancer Cells

    PubMed Central

    Cheng, Jonathan C.; Weisenberger, Daniel J.; Gonzales, Felicidad A.; Liang, Gangning; Xu, Guo-Liang; Hu, Ye-Guang; Marquez, Victor E.; Jones, Peter A.

    2004-01-01

    During tumorigenesis, tumor suppressor and cancer-related genes are commonly silenced by aberrant DNA methylation in their promoter regions. Recently, we reported that zebularine [1-(β-d-ribofuranosyl)-1,2-dihydropyrimidin-2-one] acts as an inhibitor of DNA methylation and exhibits chemical stability and minimal cytotoxicity both in vitro and in vivo. Here we show that continuous application of zebularine to T24 cells induces and maintains p16 gene expression and sustains demethylation of the 5′ region for over 40 days, preventing remethylation. In addition, continuous zebularine treatment effectively and globally demethylated various hypermethylated regions, especially CpG-poor regions. The drug caused a complete depletion of extractable DNA methyltransferase 1 (DNMT1) and partial depletion of DNMT3a and DNMT3b3. Last, sequential treatment with 5-aza-2′-deoxycytidine followed by zebularine hindered the remethylation of the p16 5′ region and gene resilencing, suggesting the possible combination use of both drugs as a potential anticancer regimen. PMID:14729971

  3. DNMT3B gene amplification predicts resistance to DNA demethylating drugs.

    PubMed

    Simó-Riudalbas, Laia; Melo, Sónia A; Esteller, Manel

    2011-07-01

    Disruption of the DNA methylation landscape is one of the most common features of human tumors. However, genetic alterations of DNA methyltransferases (DNMTs) have not been described in carcinogenesis. Herein, we show that pancreatic and breast cancer cells undergo gene amplification of the DNA methyltransferase 3B (DNMT3B). The presence of extra copies of the DNMT3B gene is linked to higher levels of the corresponding mRNA and protein. Most importantly, the elevated gene dosage of DNMT3B is associated with increased resistance to the growth-inhibitory effect mediated by DNA demethylating agents. In particular, cancer cells harboring DNMT3B gene amplification are less sensitive to the decrease in cell viability caused by 5-azacytidine (Vidaza), 5-aza-2-deoxycytidine (Decitabine), and SGI-1027. Overall, the data confirm DNMT3B as a bona fide oncogene in human cancer and support the incorporation of the DNMT3B copy number assay into current clinical trials assessing the efficacy of DNA demethylating drugs in solid tumors.

  4. Herbivore-Induced DNA Demethylation Changes Floral Signalling and Attractiveness to Pollinators in Brassica rapa

    PubMed Central

    Schlüter, Philipp M.; Schiestl, Florian P.

    2016-01-01

    Plants have to fine-tune their signals to optimise the trade-off between herbivore deterrence and pollinator attraction. An important mechanism in mediating plant-insect interactions is the regulation of gene expression via DNA methylation. However, the effect of herbivore-induced DNA methylation changes on pollinator-relevant plant signalling has not been systematically investigated. Here, we assessed the impact of foliar herbivory on DNA methylation and floral traits in the model crop plant Brassica rapa. Methylation-sensitive amplified fragment length polymorphism (MSAP) analysis showed that leaf damage by the caterpillar Pieris brassicae was associated with genome-wide methylation changes in both leaves and flowers of B. rapa as well as a downturn in flower number, morphology and scent. A comparison to plants with jasmonic acid-induced defence showed similar demethylation patterns in leaves, but both the floral methylome and phenotype differed significantly from P. brassicae infested plants. Standardised genome-wide demethylation with 5-azacytidine in five different B. rapa full-sib groups further resulted in a genotype-specific downturn of floral morphology and scent, which significantly reduced the attractiveness of the plants to the pollinator bee Bombus terrestris. These results suggest that DNA methylation plays an important role in adjusting plant signalling in response to changing insect communities. PMID:27870873

  5. Allele-specific FKBP5 DNA demethylation mediates gene–childhood trauma interactions

    PubMed Central

    Klengel, Torsten; Mehta, Divya; Anacker, Christoph; Rex-Haffner, Monika; Pruessner, Jens C; Pariante, Carmine M; Pace, Thaddeus W W; Mercer, Kristina B; Mayberg, Helen S; Bradley, Bekh; Nemeroff, Charles B; Holsboer, Florian; Heim, Christine M; Ressler, Kerry J; Rein, Theo; Binder, Elisabeth B

    2014-01-01

    Although the fact that genetic predisposition and environmental exposures interact to shape development and function of the human brain and, ultimately, the risk of psychiatric disorders has drawn wide interest, the corresponding molecular mechanisms have not yet been elucidated. We found that a functional polymorphism altering chromatin interaction between the transcription start site and long-range enhancers in the FK506 binding protein 5 (FKBP5) gene, an important regulator of the stress hormone system, increased the risk of developing stress-related psychiatric disorders in adulthood by allele-specific, childhood trauma–dependent DNA demethylation in functional glucocorticoid response elements of FKBP5. This demethylation was linked to increased stress-dependent gene transcription followed by a long-term dysregulation of the stress hormone system and a global effect on the function of immune cells and brain areas associated with stress regulation. This identification of molecular mechanisms of genotype-directed long-term environmental reactivity will be useful for designing more effective treatment strategies for stress-related disorders. PMID:23201972

  6. DNA Demethylation Upregulated Nrf2 Expression in Alzheimer’s Disease Cellular Model

    PubMed Central

    Cao, Huimin; Wang, Li; Chen, Beibei; Zheng, Peng; He, Yi; Ding, Yubin; Deng, Yushuang; Lu, Xi; Guo, Xiuming; Zhang, Yuping; Li, Yu; Yu, Gang

    2016-01-01

    Nuclear factor erythroid 2-related factor 2 (Nrf2) is an important transcription factor in the defense against oxidative stress. Cumulative evidence has shown that oxidative stress plays a key role in the pathogenesis of Alzheimer’s disease (AD). Previous animal and clinical studies had observed decreased expression of Nrf2 in AD. However, the underlying regulation mechanisms of Nrf2 in AD remain unclear. Here, we used the DNA methyltransferases (Dnmts) inhibitor 5-aza-2′-deoxycytidine (5-Aza) to test whether Nrf2 expression was regulated by methylation in N2a cells characterizing by expressing human Swedish mutant amyloid precursor protein (N2a/APPswe). We found 5-Aza treatment increased Nrf2 at both messenger RNA and protein levels via downregulating the expression of Dnmts and DNA demethylation. In addition, 5-Aza-mediated upregulation of Nrf2 expression was concomitant with increased nuclear translocation of Nrf2 and higher expression of Nrf2 downstream target gene NAD(P)H:quinone oxidoreductas (NQO1). Our study showed that DNA demethylation promoted the Nrf2 cell signaling pathway, which may enhance the antioxidant system against AD development. PMID:26779013

  7. Clomipramine demethylation rate is important on the outcome of obsessive-compulsive disorder treatment.

    PubMed

    Marcourakis, Tania; Bernik, Márcio A; Lotufo Neto, Francisco; Gedanke Shavitt, Roseli; Gorenstein, Clarice

    2015-01-01

    The aim of this study was to investigate the influence of demethylation rate on the outcome of obsessive-compulsive disorder patients treated with clomipramine. Eighteen patients meeting the DSM-IV criteria for obsessive-compulsive disorder received 150-300 mg of clomipramine daily in a single-blind design for 12 weeks. The patients were evaluated with the Clinical Global Impression scale and the Yale-Brown Obsessive-Compulsive Scale (YBOCS). Clinical assessment and serum measurements of clomipramine and desmethylclomipramine were carried out at baseline and after 3, 6, 8, 10, and 12 weeks. A greater improvement in Clinical Global Impression scale rating was associated with a lower desmethylclomipramine/daily dose and the total clomipramine and desmethylclomipramine/daily dose. Moreover, an improved response on the YBOCS-obsession score was associated with higher serum levels of clomipramine and the total clomipramine and desmethylclomipramine/daily dose. Patients with a greater reduction in baseline YBOCS rating had a lower desmethylclomipramine/clomipramine ratio. These data suggest that a lower demethylation rate correlates with better clinical outcome.

  8. Process for reducing beta activity in uranium

    DOEpatents

    Briggs, Gifford G.; Kato, Takeo R.; Schonegg, Edward

    1986-10-07

    This invention is a method for lowering the beta radiation hazards associated with the casting of uranium. The method reduces the beta radiation emitted from the as-cast surfaces of uranium ingots. The method also reduces the amount of beta radiation emitters retained on the interiors of the crucibles that have been used to melt the uranium charges and which have undergone cleaning in a remote handling facility. The lowering of the radioactivity is done by scavenging the beta emitters from the molten uranium with a molten mixture containing the fluorides of magnesium and calcium. The method provides a means of collection and disposal of the beta emitters in a manner that reduces radiation exposure to operating personnel in the work area where the ingots are cast and processed.

  9. Process for reducing beta activity in uranium

    DOEpatents

    Briggs, Gifford G.; Kato, Takeo R.; Schonegg, Edward

    1986-01-01

    This invention is a method for lowering the beta radiation hazards associated with the casting of uranium. The method reduces the beta radiation emitted from the as-cast surfaces of uranium ingots. The method also reduces the amount of beta radiation emitters retained on the interiors of the crucibles that have been used to melt the uranium charges and which have undergone cleaning in a remote handling facility. The lowering of the radioactivity is done by scavenging the beta emitters from the molten uranium with a molten mixture containing the fluorides of magnesium and calcium. The method provides a means of collection and disposal of the beta emitters in a manner that reduces radiation exposure to operating personnel in the work area where the ingots are cast and processed.

  10. Process for reducing beta activity in uranium

    DOEpatents

    Briggs, G.G.; Kato, T.R.; Schonegg, E.

    1985-04-11

    This invention is a method for lowering the beta radiation hazards associated with the casting of uranium. The method reduces the beta radiation emitted from the as-cast surfaces of uranium ingots. The method also reduces the amount of beta radiation emitters retained on the interiors of the crucibles that have been used to melt the uranium charges and which undergone cleaning in a remote handling facility. The lowering of the radioactivity is done by scavenging the beta emitters from the molten uranium with a molten mixture containing the fluorides of magnesium and calcium. The method provides a means of collection and disposal of the beta emitters in a manner that reduces radiation exposure to operating personnel in the work area where the ingots are cast and processed. 5 tabs.

  11. 43 CFR 3420.3 - Activity planning: The leasing process.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 2 2013-10-01 2013-10-01 false Activity planning: The leasing process... LAND MANAGEMENT, DEPARTMENT OF THE INTERIOR MINERALS MANAGEMENT (3000) COMPETITIVE LEASING Competitive Leasing § 3420.3 Activity planning: The leasing process....

  12. 43 CFR 3420.3 - Activity planning: The leasing process.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 2 2012-10-01 2012-10-01 false Activity planning: The leasing process... LAND MANAGEMENT, DEPARTMENT OF THE INTERIOR MINERALS MANAGEMENT (3000) COMPETITIVE LEASING Competitive Leasing § 3420.3 Activity planning: The leasing process....

  13. 43 CFR 3420.3 - Activity planning: The leasing process.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 43 Public Lands: Interior 2 2014-10-01 2014-10-01 false Activity planning: The leasing process... LAND MANAGEMENT, DEPARTMENT OF THE INTERIOR MINERALS MANAGEMENT (3000) COMPETITIVE LEASING Competitive Leasing § 3420.3 Activity planning: The leasing process....

  14. 12 CFR 211.604 - Data processing activities.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 2 2013-01-01 2013-01-01 false Data processing activities. 211.604 Section 211... processing activities under Regulation K (12 CFR part 211). This question has arisen as a result of the fact... authority in Regulation K was somewhat broader than that permissible in the United States under Regulation...

  15. 12 CFR 211.604 - Data processing activities.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 2 2011-01-01 2011-01-01 false Data processing activities. 211.604 Section 211... processing activities under Regulation K (12 CFR part 211). This question has arisen as a result of the fact... authority in Regulation K was somewhat broader than that permissible in the United States under Regulation...

  16. 12 CFR 211.604 - Data processing activities.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 2 2014-01-01 2014-01-01 false Data processing activities. 211.604 Section 211... processing activities under Regulation K (12 CFR part 211). This question has arisen as a result of the fact... authority in Regulation K was somewhat broader than that permissible in the United States under Regulation...

  17. 12 CFR 211.604 - Data processing activities.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 2 2012-01-01 2012-01-01 false Data processing activities. 211.604 Section 211... processing activities under Regulation K (12 CFR part 211). This question has arisen as a result of the fact... authority in Regulation K was somewhat broader than that permissible in the United States under Regulation...

  18. 12 CFR 211.604 - Data processing activities.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 2 2010-01-01 2010-01-01 false Data processing activities. 211.604 Section 211... processing activities under Regulation K (12 CFR part 211). This question has arisen as a result of the fact... authority in Regulation K was somewhat broader than that permissible in the United States under Regulation...

  19. 43 CFR 3420.3 - Activity planning: The leasing process.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Activity planning: The leasing process... LAND MANAGEMENT, DEPARTMENT OF THE INTERIOR MINERALS MANAGEMENT (3000) COMPETITIVE LEASING Competitive Leasing § 3420.3 Activity planning: The leasing process....

  20. Epigenetic alteration by DNA-demethylating treatment restores apoptotic response to glucocorticoids in dexamethasone-resistant human malignant lymphoid cells

    PubMed Central

    2014-01-01

    Background Glucocorticoids (GCs) are often included in the therapy of lymphoid malignancies because they kill several types of malignant lymphoid cells. GCs activate the glucocorticoid receptor (GR), to regulate a complex genetic network, culminating in apoptosis. Normal lymphoblasts and many lymphoid malignancies are sensitive to GC-driven apoptosis. Resistance to GCs can be a significant clinical problem, however, and correlates with resistance to several other major chemotherapeutic agents. Methods We analyzed the effect of treatment with the cytosine analogue 5 aza-2’ deoxycytidine (AZA) on GC resistance in two acute lymphoblastic leukemia (T or pre-T ALL) cell lines- CEM and Molt-4- and a (B-cell) myeloma cell line, RPMI 8226. Methods employed included tissue culture, flow cytometry, and assays for clonogenicity, cytosine extension, immunochemical identification of proteins, and gene transactivation. High throughput DNA sequencing was used to confirm DNA methylation status. Conclusions Treatment of these cells with AZA resulted in altered DNA methylation and restored GC-evoked apoptosis in all 3 cell lines. In CEM cells the altered epigenetic state resulted in site-specific phosphorylation of the GR, increased GR potency, and GC-driven induction of the GR from promoters that lie in CpG islands. In RPMI 8226 cells, expression of relevant coregulators of GR function was altered. Activation of p38 mitogen-activated protein kinase (MAPK), which is central to a feed-forward mechanism of site-specific GR phosphorylation and ultimately, apoptosis, occurred in all 3 cell lines. These data show that in certain malignant hematologic B- and T-cell types, epigenetically controlled GC resistance can be reversed by cell exposure to a compound that causes DNA demethylation. The results encourage studies of application to in vivo systems, looking towards eventual clinical applications. PMID:24795534

  1. Measurement of action spectra of light-activated processes

    NASA Astrophysics Data System (ADS)

    Ross, Justin; Zvyagin, Andrei V.; Heckenberg, Norman R.; Upcroft, Jacqui; Upcroft, Peter; Rubinsztein-Dunlop, Halina H.

    2006-01-01

    We report on a new experimental technique suitable for measurement of light-activated processes, such as fluorophore transport. The usefulness of this technique is derived from its capacity to decouple the imaging and activation processes, allowing fluorescent imaging of fluorophore transport at a convenient activation wavelength. We demonstrate the efficiency of this new technique in determination of the action spectrum of the light mediated transport of rhodamine 123 into the parasitic protozoan Giardia duodenalis.

  2. Speech Perception as a Cognitive Process: The Interactive Activation Model.

    DTIC Science & Technology

    1983-04-01

    one another in COHORT, the nodes for sell, your, light, and cellulite , wil all bc in active competition with one another. The system will have no way...7 AD-AI28 787 SPEECH PERCEPTION AS A COGNITIVE PROCESS: THE INTERACTIVE ACTIVATION MODE..(U) CALIFORNIA UNIV SAN D IEGO LA dOLLA INST FOR COGNITIVE...TYPE OF REPORT & PERIOD COVERED Speech Perception as a Cognitive Process: Technical Report The Interactive Activation Model S. PERFORMING ORG. REPORT

  3. Increased expression of prion protein gene is accompanied by demethylation of CpG sites in a mouse embryonal carcinoma cell line, P19C6

    PubMed Central

    DALAI, Wuyun; MATSUO, Eiko; TAKEYAMA, Natsumi; KAWANO, Junichi; SAEKI, Keiichi

    2017-01-01

    Elucidation of the processes regulating the prion protein gene (Prnp) is an important key to understanding the development of prion disorders. In this study, we explored the involvement of DNA methylation in Prnp transcriptional regulation during neuronal differentiation of embryonic carcinoma P19C6 cells. When P19C6 cells were differentiated into neuronal cells, the expression of Prnp was markedly increased, while CpG methylation was significantly demethylated at the nucleotide region between −599 and −238 from the transcription start site. In addition, when P19C6 cells were applied in a DNA methyltransferase inhibitor, RG108, Prnp transcripts were also significantly increased in relation to the decreased methylation statuses. These findings helped to elucidate the DNA methylation-mediated regulation of Prnp expression during neuronal differentiation. PMID:28132962

  4. Process of activation of a palladium catalyst system

    SciTech Connect

    Sobolevskiy, Anatoly; Rossin, Joseph A.; Knapke, Michael J.

    2011-08-02

    Improved processes for activating a catalyst system used for the reduction of nitrogen oxides are provided. In one embodiment, the catalyst system is activated by passing an activation gas stream having an amount of each of oxygen, water vapor, nitrogen oxides, and hydrogen over the catalyst system and increasing a temperature of the catalyst system to a temperature of at least 180.degree. C. at a heating rate of from 1-20.degree./min. Use of activation processes described herein leads to a catalyst system with superior NOx reduction capabilities.

  5. Human concentrative nucleoside transporter 1-mediated uptake of 5-azacytidine enhances DNA demethylation.

    PubMed

    Rius, Maria; Stresemann, Carlo; Keller, Daniela; Brom, Manuela; Schirrmacher, Esther; Keppler, Dietrich; Lyko, Frank

    2009-01-01

    The DNA methyltransferase inhibitors 5-azacytidine (5-azaCyd) and 5-aza-2'-deoxycytidine have found increasing use for the treatment of myeloid leukemias and solid tumors. Both nucleoside analogues must be transported into cells and phosphorylated before they can be incorporated into DNA and inactivate DNA methyltransferases. The members of the human equilibrative and concentrative nucleoside transporter families mediate transport of natural nucleosides and some nucleoside analogues into cells. However, the molecular identity of the transport proteins responsible for mediating the uptake of 5-azanucleosides has remained unknown. To this end, we have generated a stably transfected Madin-Darby canine kidney strain II cell line expressing recombinant hCNT1. An antiserum directed against hCNT1 specifically detected the protein in the apical membrane of hCNT1-expressing Madin-Darby canine kidney cells. Using [14C]5-azaCyd, we show here that hCNT1 mediated the Na+-dependent uptake of this drug with a Km value of 63 micromol/L. Na+-dependent transport of radiolabeled cytidine, uridine, and 5-fluoro-5'-deoxyuridine further showed the functionality of the transporter. hCNT1-expressing cells were significantly more sensitive to 5-azaCyd, and drug-dependent covalent trapping of DNA methyltransferase 1 was substantially more pronounced. Importantly, these results correlated with a significant sensitization of hCNT1-expressing cells toward the demethylating effects of 5-azaCyd and 5-aza-2'-deoxycytidine. In conclusion, our study identifies 5-azaCyd as a novel substrate for hCNT1 and provides direct evidence that hCNT1 is involved in the DNA-demethylating effects of this drug.

  6. Preliminary studies of a canine 13C-aminopyrine demethylation blood test.

    PubMed Central

    Moeller, E M; Steiner, J M; Williams, D A; Klein, P D

    2001-01-01

    The objectives of this study were to determine whether a 13C-aminopyrine demethylation blood test is technically feasible in clinically healthy dogs, whether oral administration of 13C-aminopyrine causes a detectable increase in percent dose/min (PCD) of 13C administered as 13C-aminopyrine and recovered in gas extracted from blood, and whether gas extraction efficiency has an impact on PCD. A dose of 2 mg/kg body weight of 13C-aminopyrine dissolved in deionized water was administered orally to 6 clinically healthy dogs. Blood samples were taken from each dog 0, 30, 60, and 120 min after administration of the 13C-aminopyrine. Carbon dioxide was extracted from blood samples by addition of acid and analyzed by fractional mass spectrometry. None of the 6 dogs showed any side effects after 13C-aminopyrine administration. All 6 dogs showed a measurable increase of the PCD in gas samples extracted from blood samples at 30 min, 60 min, and 120 min after 13C-aminopyrine administration. Coefficients of variation between the triplicate samples were statistically significantly higher for the %CO2, a measure of extraction efficiency, than for PCD values (P < 0.0001). The 13C-aminopyrine demethylation blood test described here is technically feasible. Oral administration of 13C-aminopyrine did not lead to gross side effects in the 6 dogs. Clinically healthy dogs show a measurable increase of PCD in gas extracted from blood samples after oral administration of 13C-aminopyrine. Efficiency of CO2 extraction from blood samples does not have an impact on PCD determined from these blood samples. This test may prove useful to evaluate hepatic function in dogs. PMID:11227194

  7. Hold My Calls: An Activity for Introducing the Statistical Process

    ERIC Educational Resources Information Center

    Abel, Todd; Poling, Lisa

    2015-01-01

    Working with practicing teachers, this article demonstrates, through the facilitation of a statistical activity, how to introduce and investigate the unique qualities of the statistical process including: formulate a question, collect data, analyze data, and interpret data.

  8. Evaluation of Control Parameters for the Activated Sludge Process

    ERIC Educational Resources Information Center

    Stall, T. Ray; Sherrard, Josephy H.

    1978-01-01

    An evaluation of the use of the parameters currently being used to design and operate the activated sludge process is presented. The advantages and disadvantages for the use of each parameter are discussed. (MR)

  9. Insights into the epigenetic mechanisms involving histone lysine methylation and demethylation in ischemia induced damage and repair has therapeutic implication.

    PubMed

    Chakravarty, Sumana; Jhelum, Priya; Bhat, Unis Ahmad; Rajan, Wenson D; Maitra, Swati; Pathak, Salil S; Patel, Anant B; Kumar, Arvind

    2017-01-01

    Cerebral ischemic stroke is one of the leading causes of death and disability worldwide. Therapeutic interventions to minimize ischemia-induced neural damage are limited due to poor understanding of molecular mechanisms mediating complex pathophysiology in stroke. Recently, epigenetic mechanisms mostly histone lysine (K) acetylation and deacetylation have been implicated in ischemic brain damage and have expanded the dimensions of potential therapeutic intervention to the systemic/local administration of histone deacetylase inhibitors. However, the role of other epigenetic mechanisms such as histone lysine methylation and demethylation in stroke-induced damage and subsequent recovery process is elusive. Here, we established an Internal Carotid Artery Occlusion (ICAO) model in CD1 mouse that resulted in mild to moderate level of ischemic damage to the striatum, as suggested by magnetic resonance imaging (MRI), TUNEL and histopathological staining along with an evaluation of neurological deficit score (NDS), grip strength and rotarod performance. The molecular investigations show dysregulation of a number of histone lysine methylases (KMTs) and few of histone lysine demethylases (KDMs) post-ICAO with significant global attenuation in the transcriptionally repressive epigenetic mark H3K9me2 in the striatum. Administration of Dimethyloxalylglycine (DMOG), an inhibitor of KDM4 or JMJD2 class of histone lysine demethylases, significantly ameliorated stroke-induced NDS by restoring perturbed H3K9me2 levels in the ischemia-affected striatum. Overall, these results highlight the novel role of epigenetic regulatory mechanisms controlling the epigenetic mark H3K9me2 in mediating the stroke-induced striatal damage and subsequent repair following mild to moderate cerebral ischemia.

  10. Implication of DNA demethylation and bivalent histone modification for selective gene regulation in mouse primordial germ cells.

    PubMed

    Mochizuki, Kentaro; Tachibana, Makoto; Saitou, Mitinori; Tokitake, Yuko; Matsui, Yasuhisa

    2012-01-01

    Primordial germ cells (PGCs) sequentially induce specific genes required for their development. We focused on epigenetic changes that regulate PGC-specific gene expression. mil-1, Blimp1, and Stella are preferentially expressed in PGCs, and their expression is upregulated during PGC differentiation. Here, we first determined DNA methylation status of mil-1, Blimp1, and Stella regulatory regions in epiblast and in PGCs, and found that they were hypomethylated in differentiating PGCs after E9.0, in which those genes were highly expressed. We used siRNA to inhibit a maintenance DNA methyltransferase, Dnmt1, in embryonic stem (ES) cells and found that the flanking regions of all three genes became hypomethylated and that expression of each gene increased 1.5- to 3-fold. In addition, we also found 1.5- to 5-fold increase of the PGC genes in the PGCLCs (PGC-like cells) induced form ES cells by knockdown of Dnmt1. We also obtained evidence showing that methylation of the regulatory region of mil-1 resulted in 2.5-fold decrease in expression in a reporter assay. Together, these results suggested that DNA demethylation does not play a major role on initial activation of the PGC genes in the nascent PGCs but contributed to enhancement of their expression in PGCs after E9.0. However, we also found that repression of representative somatic genes, Hoxa1 and Hoxb1, and a tissue-specific gene, Gfap, in PGCs was not dependent on DNA methylation; their flanking regions were hypomethylated, but their expression was not observed in PGCs at E13.5. Their promoter regions showed the bivalent histone modification in PGCs, that may be involved in repression of their expression. Our results indicated that epigenetic status of PGC genes and of somatic genes in PGCs were distinct, and suggested contribution of epigenetic mechanisms in regulation of the expression of a specific gene set in PGCs.

  11. Combination of PDT and a DNA demethylating agent produces anti-tumor immune response in a mouse tumor model

    NASA Astrophysics Data System (ADS)

    Mroz, Pawel; Hamblin, Michael R.

    2009-06-01

    Epigenetic mechanisms, which involve DNA methylation and histone modifications, result in the heritable silencing of genes without a change in their coding sequence. However, these changes must be actively maintained after each cell division rendering them a promising target for pharmacologic inhibition. DNA methyltransferase inhibitors like 5-aza-deoxycytidine (5-aza-dC) induce and/or up-regulate the expression of MAGE-type antigens in human and mice cancer cells. Photodynamic therapy (PDT) has been shown to be an effective locally ablative anti-cancer treatment that has the additional advantage of stimulating tumor-directed immune response. We studied the effects of a new therapy that combined the demethylating agent 5-aza-dC with PDT in the breast cancer model 4T1 syngenic to immunocompetent BALB/c mice. PDT was used as a locally ablating tumor treatment that is capable of eliciting strong and tumor directed immune response while 5-aza-dC pretreatment was used promote de novo induction of the expression of P1A.protein. This is the mouse homolog of human MAGE family antigens and is reported to function as a tumor rejection antigen in certain mouse tumors. This strategy led to an increase in PDT-mediated immune response and better treatment outcome. These results strongly suggest that the MAGE family antigens are important target for PDT mediated immune response but that their expression can be silenced by epigenetic mechanisms. Therefore the possibility that PDT can be combined with epigenetic strategies to elicit anti-tumor immunity in MAGE-positive tumor models is highly clinically significant and should be studied in detail.

  12. Cascading Activation across Levels of Representation in Children's Lexical Processing

    ERIC Educational Resources Information Center

    Huang, Yi Ting; Snedeker, Jesse

    2011-01-01

    Recent work in adult psycholinguistics has demonstrated that activation of semantic representations begins long before phonological processing is complete. This incremental propagation of information across multiple levels of analysis is a hallmark of adult language processing but how does this ability develop? In two experiments, we elicit…

  13. Instructional Transaction Theory: Knowledge Relationships among Processes, Entities, and Activities.

    ERIC Educational Resources Information Center

    Merrill, M. David; And Others

    1993-01-01

    Discussion of instructional transaction theory focuses on knowledge representation in an automated instructional design expert system. A knowledge structure called PEA-Net (processes, entities, and activities) is explained; the refrigeration process is used as an example; text resources and graphic resources are described; and simulations are…

  14. Students' Learning Activities While Studying Biological Process Diagrams

    ERIC Educational Resources Information Center

    Kragten, Marco; Admiraal, Wilfried; Rijlaarsdam, Gert

    2015-01-01

    Process diagrams describe how a system functions (e.g. photosynthesis) and are an important type of representation in Biology education. In the present study, we examined students' learning activities while studying process diagrams, related to their resulting comprehension of these diagrams. Each student completed three learning tasks. Verbal…

  15. Enhanced Multistatic Active Sonar via Innovative Signal Processing

    DTIC Science & Technology

    2014-09-30

    DATES COVERED (From - To) Oct. 01. 2013-Sept. 30, 2014 4. TITLE AND SUBTITLE Enhanced Multistatic Active Sonar via Innovative Signal Processing 5a...DISTRIBUTION AVAILABILITY STATEMENT Approved for Public Release; Distribution is Unlimited. 13. SUPPLEMENTARY NOTES 14. ABSTRACT Pulsed active sonar ...PAS) and continuous active sonar (CAS) in the presence of strong direct blast are studied for the Doppler-tolerant linear frequency modulation

  16. Dissecting gamma frequency activity during human memory processing.

    PubMed

    Kucewicz, Michal T; Berry, Brent M; Kremen, Vaclav; Brinkmann, Benjamin H; Sperling, Michael R; Jobst, Barbara C; Gross, Robert E; Lega, Bradley; Sheth, Sameer A; Stein, Joel M; Das, Sandthitsu R; Gorniak, Richard; Stead, S Matthew; Rizzuto, Daniel S; Kahana, Michael J; Worrell, Gregory A

    2017-03-13

    Gamma frequency activity (30-150 Hz) is induced in cognitive tasks and is thought to reflect underlying neural processes. Gamma frequency activity can be recorded directly from the human brain using intracranial electrodes implanted in patients undergoing treatment for drug-resistant epilepsy. Previous studies have independently explored narrowband oscillations in the local field potential and broadband power increases. It is not clear, however, which processes contribute to human brain gamma frequency activity, or their dynamics and roles during memory processing. Here a large dataset of intracranial recordings obtained during encoding of words from 101 patients was used to detect, characterize and compare induced gamma frequency activity events. Individual bursts of gamma frequency activity were isolated in the time-frequency domain to determine their spectral features, including peak frequency, amplitude, frequency span, and duration. We found two distinct types of gamma frequency activity events that showed either narrowband or broadband frequency spans revealing characteristic spectral properties. Narrowband events, the predominant type, were induced by word presentations following an initial induction of broadband events, which were temporally separated and selectively correlated with evoked response potentials, suggesting that they reflect different neural activities and play different roles during memory encoding. The two gamma frequency activity types were differentially modulated during encoding of subsequently recalled and forgotten words. In conclusion, we found evidence for two distinct activity types induced in the gamma frequency range during cognitive processing. Separating these two gamma frequency activity components contributes to the current understanding of electrophysiological biomarkers, and may prove useful for emerging neurotechnologies targeting, mapping and modulating distinct neurophysiological processes in normal and epileptogenic brain.

  17. Investigation of oxidation process of mechanically activated ultrafine iron powders

    NASA Astrophysics Data System (ADS)

    Lysenko, E. N.; Nikolaev, E. V.; Vlasov, V. A.; Zhuravkov, S. P.

    2016-02-01

    The oxidation of mechanically activated ultrafine iron powders was studied using X- ray powder diffraction and thermogravimetric analyzes. The powders with average particles size of 100 nm were made by the electric explosion of wire, and were subjected to mechanical activation in planetary ball mill for 15 and 40 minutes. It was shown that a certain amount of FeO phase is formed during mechanical activation of ultrafine iron powders. According to thermogravimetric analysis, the oxidation process of non-milled ultrafine iron powders is a complex process and occurs in three stages. The preliminary mechanical activation of powders considerably changes the nature of the iron powders oxidation, leads to increasing in the temperature of oxidation onset and shifts the reaction to higher temperatures. For the milled powders, the oxidation is more simple process and occurs in a single step.

  18. Fingerprinting DNA oxidation processes: IR characterization of the 5-methyl-2'-deoxycytidine radical cation.

    PubMed

    Bucher, Dominik B; Pilles, Bert M; Pfaffeneder, Toni; Carell, Thomas; Zinth, Wolfgang

    2014-02-24

    Methylated cytidine plays an important role as an epigenetic signal in gene regulation. Its oxidation products are assumed to be involved in active demethylation processes but also in damaging DNA. Here, we report the photochemical production of the 5-methyl-2'-deoxycytidine radical cation via a two-photon ionization process. The radical cation is detected by time-resolved IR spectroscopy and identified by band assignment using density functional theory calculations. Two final oxidation products are characterized with liquid chromatography coupled to mass spectrometry.

  19. Digital image processing and analysis for activated sludge wastewater treatment.

    PubMed

    Khan, Muhammad Burhan; Lee, Xue Yong; Nisar, Humaira; Ng, Choon Aun; Yeap, Kim Ho; Malik, Aamir Saeed

    2015-01-01

    Activated sludge system is generally used in wastewater treatment plants for processing domestic influent. Conventionally the activated sludge wastewater treatment is monitored by measuring physico-chemical parameters like total suspended solids (TSSol), sludge volume index (SVI) and chemical oxygen demand (COD) etc. For the measurement, tests are conducted in the laboratory, which take many hours to give the final measurement. Digital image processing and analysis offers a better alternative not only to monitor and characterize the current state of activated sludge but also to predict the future state. The characterization by image processing and analysis is done by correlating the time evolution of parameters extracted by image analysis of floc and filaments with the physico-chemical parameters. This chapter briefly reviews the activated sludge wastewater treatment; and, procedures of image acquisition, preprocessing, segmentation and analysis in the specific context of activated sludge wastewater treatment. In the latter part additional procedures like z-stacking, image stitching are introduced for wastewater image preprocessing, which are not previously used in the context of activated sludge. Different preprocessing and segmentation techniques are proposed, along with the survey of imaging procedures reported in the literature. Finally the image analysis based morphological parameters and correlation of the parameters with regard to monitoring and prediction of activated sludge are discussed. Hence it is observed that image analysis can play a very useful role in the monitoring of activated sludge wastewater treatment plants.

  20. Canada's physical activity guides: background, process, and development.

    PubMed

    Sharratt, Michael T; Hearst, William E

    2007-01-01

    This historical background paper chronicles the major events leading to the development of Canada's physical activity guides (for children, youth, adults, and older adults). The paper outlines the process and the steps used, including information (where applicable) regarding national partners, project administration, Health Canada communications, product development, endorsement, distribution and implementation, collateral activities, media relations and evaluation framework. Brief summaries of the science that led to the recommended guidelines are included. The paper also summarizes the various physical activity guide assessment and evaluation projects and their findings, particularly as they relate to research carried out on Canada's physical activity guides for children and youth (and the associated support resources).

  1. Nonoxidative removal of organics in the activated sludge process

    PubMed Central

    Modin, Oskar; Persson, Frank; Wilén, Britt-Marie; Hermansson, Malte

    2016-01-01

    ABSTRACT The activated sludge process is commonly used to treat wastewater by aerobic oxidation of organic pollutants into carbon dioxide and water. However, several nonoxidative mechanisms can also contribute to removal of organics. Sorption onto activated sludge can remove a large fraction of the colloidal and particulate wastewater organics. Intracellular storage of, e.g., polyhydroxyalkanoates (PHA), triacylglycerides (TAG), or wax esters can convert wastewater organics into precursors for high-value products. Recently, several environmental, economic, and technological drivers have stimulated research on nonoxidative removal of organics for wastewater treatment. In this paper, we review these nonoxidative removal mechanisms as well as the existing and emerging process configurations that make use of them for wastewater treatment. Better utilization of nonoxidative processes in activated sludge could reduce the wasteful aerobic oxidation of organic compounds and lead to more resource-efficient wastewater treatment plants. PMID:27453679

  2. Impact of activation process on fog life cycle

    NASA Astrophysics Data System (ADS)

    Mazoyer, Marie; Burnet, Frédéric; Lac, Christine; Roberts, Greg; Dupont, Jean-Charles; Haeffelin, Martial; Elias, Thierry

    2015-04-01

    Fogs are complex meteorological system dealing with fine scale processes. Subtle interaction between radiative, dynamic, turbulent and microphysic processes can lead to different fog life cycle, which make prediction difficult. The droplets that composed fogs are formed trough the activation of aerosol particles called CCN (cloud condensation nuclei) described by the Köhler theory (Köhler, 1936). The number and distribution of the droplets activated during fog formation is determined by the aerosols particles properties and number and the ambient vapor supersaturation of the atmosphere. In the frame of the PreViBOSS project, an in-situ measurement platform of fog properties at ground level was deployed at SIRTA (Instrumented Site for Atmospheric Remote Sensing Research) during winter 2010 to 2013. Microphysics data supply a detailed characterization of number size spectrum from dry to wet aerosols particles and inform on the abilities of the aerosols particles to act as a CCN. 48 fog events have been studied. Supersaturation critical values and concentrations of CCN have been determined and linked to aerosols properties. The main impact of aerosols size distribution on activation have been pointed out. The study of droplets spectra evolution reveals the major physical processes into fogs and suggests that even if thermodynamic dominates the fog life cycle, activation process seems to have a significant effect. Large eddy simulation of fog run with Meso-NH model allow to explore precisely the interaction between fog physical processes and to quantify activation impact. Supersaturation modelling is a key point, a new pseudo-prognostic scheme (Thouron et al., 2012) is used. Confrontation between a detailed experimental study and three-dimensional fine scale simulation in LES provides an accurate investigation of the impact of activation process on fog life cycle.

  3. Active Cellular Mechanics and Information Processing in the Living Cell

    NASA Astrophysics Data System (ADS)

    Rao, M.

    2014-07-01

    I will present our recent work on the organization of signaling molecules on the surface of living cells. Using novel experimental and theoretical approaches we have found that many cell surface receptors are organized as dynamic clusters driven by active currents and stresses generated by the cortical cytoskeleton adjoining the cell surface. We have shown that this organization is optimal for both information processing and computation. In connecting active mechanics in the cell with information processing and computation, we bring together two of the seminal works of Alan Turing.

  4. Active non-volatile memory post-processing

    DOEpatents

    Kannan, Sudarsun; Milojicic, Dejan S.; Talwar, Vanish

    2017-04-11

    A computing node includes an active Non-Volatile Random Access Memory (NVRAM) component which includes memory and a sub-processor component. The memory is to store data chunks received from a processor core, the data chunks comprising metadata indicating a type of post-processing to be performed on data within the data chunks. The sub-processor component is to perform post-processing of said data chunks based on said metadata.

  5. An effective O-demethylation of some C19-diterpenoid alkaloids with HBr-glacial acetic acid.

    PubMed

    Zou, Chun-Lan; Ji, Hong; Xie, Guang-Bo; Chen, Dong-Lin; Wang, Feng-Peng

    2008-01-01

    The aconitine-type alkaloids talatisamine (1), 8,14-diacetyltalatisamine (11), and compound 3, the lycoctonine-type alkaloid deltaline (5), and the 7,17-seco C(19)-diterpenoid alkaloids 7 and 9 were treated with HBr-glacial acetic acid to give useful O-demethylated derivatives 2, 2, 4, 6, 8, and 10 respectively in good to high yields (49-90%).

  6. O-Demethylation and Successive Oxidative Dechlorination of Methoxychlor by Bradyrhizobium sp. Strain 17-4, Isolated from River Sediment

    PubMed Central

    Masuda, Minoru; Sato, Kiyoshi

    2012-01-01

    O-Demethylation of insecticide methoxychlor is well known as a phase I metabolic reaction in various eukaryotic organisms. Regarding prokaryotic organisms, however, no individual species involved in such reaction have been specified and characterized so far. Here we successfully isolated a bacterium that mediates oxidative transformation of methoxychlor, including O-demethylation and dechlorination, from river sediment. The isolate was found to be closely related to Bradyrhizobium elkanii at the 16S rRNA gene sequence level (100% identical). However, based on some differences in the physiological properties of this bacterium, we determined that it was actually a different species, Bradyrhizobium sp. strain 17-4. The isolate mediated O-demethylation of methoxychlor to yield a monophenolic derivative [Mono-OH; 1,1,1-trichloro-2-(4-hydroxyphenyl)-2-(4-methoxyphenyl)ethane] as the primary degradation product. The chiral high-performance liquid chromatography (HPLC) analysis revealed that the isolate possesses high enantioselectivity favoring the formation of (S)-Mono-OH (nearly 100%). Accompanied by the sequential O-demethylation to form the bis-phenolic derivative Bis-OH [1,1,1-trichloro-2,2-bis(4-hydroxyphenyl)ethane], oxidative dechlorination of the side chain proceeded, and monophenolic carboxylic acid accumulated, followed by the formation of multiple unidentified polar degradation products. The breakdown proceeded more rapidly when reductively dechlorinated (dichloro-form) methoxychlor was applied as the initial substrate. The resultant carboxylic acids and polar degradation products are likely further biodegraded by ubiquitous bacteria. The isolate possibly plays an important role for complete degradation (mineralization) of methoxychlor by providing the readily biodegradable substrates. PMID:22635993

  7. Meclofenamic acid selectively inhibits FTO demethylation of m6A over ALKBH5

    PubMed Central

    Huang, Yue; Yan, Jingli; Li, Qi; Li, Jiafei; Gong, Shouzhe; Zhou, Hu; Gan, Jianhua; Jiang, Hualiang; Jia, Gui-Fang; Luo, Cheng; Yang, Cai-Guang

    2015-01-01

    Two human demethylases, the fat mass and obesity-associated (FTO) enzyme and ALKBH5, oxidatively demethylate abundant N6-methyladenosine (m6A) residues in mRNA. Achieving a method for selective inhibition of FTO over ALKBH5 remains a challenge, however. Here, we have identified meclofenamic acid (MA) as a highly selective inhibitor of FTO. MA is a non-steroidal, anti-inflammatory drug that mechanistic studies indicate competes with FTO binding for the m6A-containing nucleic acid. The structure of FTO/MA has revealed much about the inhibitory function of FTO. Our newfound understanding, revealed herein, of the part of the nucleotide recognition lid (NRL) in FTO, for example, has helped elucidate the principles behind the selectivity of FTO over ALKBH5. Treatment of HeLa cells with the ethyl ester form of MA (MA2) has led to elevated levels of m6A modification in mRNA. Our collective results highlight the development of functional probes of the FTO enzyme that will (i) enable future biological studies and (ii) pave the way for the rational design of potent and specific inhibitors of FTO for use in medicine. PMID:25452335

  8. Registered report: IDH mutation impairs histone demethylation and results in a block to cell differentiation.

    PubMed

    Richarson, Adam D; Scott, David A; Zagnitko, Olga; Aza-Blanc, Pedro; Chang, Chih-Cheng; Russler-Germain, David A

    2016-03-11

    The Reproducibility Project: Cancer Biology seeks to address growing concerns about reproducibility in scientific research by conducting replications of selected experiments from a number of high-profile papers in the field of cancer biology. The papers, which were published between 2010 and 2012, were selected on the basis of citations and Altmetric scores (Errington et al., 2014). This Registered Report describes the proposed replication plan of key experiments from "IDH mutation impairs histone demethylation and results in a block to cell differentiation" by Lu and colleagues, published in Nature in 2012 (Lu et al., 2012). The experiments that will be replicated are those reported in Figures 1B, 2A, 2B, 2D and 4D. Lu and colleagues demonstrated that expression of mutant forms of IDH1 or IDH2 caused global increases in histone methylation and increased levels of 2 hydroxyglutarate (Figure 1B). This was correlated with a block in differentiation (Figures 2A, B and D). This effect appeared to be mediated by the histone demethylase KDM4C (Figure 4D). The Reproducibility Project: Cancer Biology is a collaboration between the Center for Open Scienceand Science Exchange, and the results of the replications will be published by eLife.

  9. Immunomediated Pan-cancer Regulation Networks are Dominant Fingerprints After Treatment of Cell Lines with Demethylation

    PubMed Central

    El Baroudi, Mariama; Cinti, Caterina; Capobianco, Enrico

    2016-01-01

    Pan-cancer studies are particularly relevant not only for addressing the complexity of the inherently observed heterogeneity but also for identifying clinically relevant features that may be common to the cancer types. Immune system regulations usually reveal synergistic modulation with other cancer mechanisms and in combination provide insights on possible advances in cancer immunotherapies. Network inference is a powerful approach to decipher pan-cancer systems dynamics. The methodology proposed in this study elucidates the impacts of epigenetic treatment on the drivers of complex pan-cancer regulation circuits involving cell lines of five cancer types. These patterns were observed from differential gene expression measurements following demethylation with 5-azacytidine. Networks were built to establish associations of phenotypes at molecular level with cancer hallmarks through both transcriptional and post-transcriptional regulation mechanisms. The most prominent feature that emerges from our integrative network maps, linking pathway landscapes to disease and drug-target associations, refers primarily to a mosaic of immune-system crosslinked influences. Therefore, characteristics initially evidenced in single cancer maps become motifs well summarized by network cores and fingerprints. PMID:27147816

  10. Induced DNA demethylation can reshape chromatin topology at the IGF2-H19 locus

    PubMed Central

    Ito, Yoko; Nativio, Raffaella; Murrell, Adele

    2013-01-01

    Choriocarcinomas are embryonal tumours with loss of imprinting and hypermethylation at the insulin-like growth factor 2 (IGF2)-H19 locus. The DNA methyltransferase inhibitor, 5-Aza-2′deoxycytidine (5-AzaCdR) is an approved epigenetic cancer therapy. However, it is not known to what extent 5-AzaCdR influences other epigenetic marks. In this study, we set out to determine whether 5-AzaCdR treatment can reprogram the epigenomic organization of the IGF2-H19 locus in a choriocarcinoma cancer cell line (JEG3). We found that localized DNA demethylation at the H19 imprinting control region (ICR) induced by 5-AzaCdR, reduced IGF2, increased H19 expression, increased CTCF and cohesin recruitment and changed histone modifications. Furthermore chromatin accessibility was increased locus-wide and chromatin looping topography was altered such that a CTCF site downstream of the H19 enhancers switched its association with the CTCF site upstream of the IGF2 promoters to associate with the ICR. We identified a stable chromatin looping domain, which forms independently of DNA methylation. This domain contains the IGF2 gene and is marked by a histone H3 lysine 27 trimethylation block between CTCF site upstream of the IGF2 promoters and the Centrally Conserved Domain upstream of the ICR. Together, these data provide new insights into the responsiveness of chromatin topography to DNA methylation changes. PMID:23585276

  11. Active Mining from Process Time Series by Learning Classifier System

    NASA Astrophysics Data System (ADS)

    Kurahashi, Setsuya; Terano, Takao

    Continuation processes in chemical and/or biotechnical plants always generate a large amount of time series data. However, since conventional process models are described as a set of control models, it is difficult to explain the complicated and active plant behaviors. Based on the background, this research proposes a novel method to develop a process response model from continuous time-series data. The method consists of the following phases: 1) Collect continuous process data at each tag point in a target plant; 2) Normalize the data in the interval between zero and one; 3) Get the delay time, which maximizes the correlation between given two time series data; 4) Select tags with the higher correlation; 5) Develop a process response model to describe the relations among the process data using the delay time and the correlation values; 6) Develop a process prediction model via several tag points data using a neural network; 1) Discover control rules from the process prediction model using Learning Classifier system. The main contribution of the research is to establish a method to mine a set of meaningful control rules from Learning Classifier System using the Minimal Description Length criteria. The proposed method has been applied to an actual process of a biochemical plant and has shown the validity and the effectiveness.

  12. Enhanced Passive and Active Processing of Syllables in Musician Children

    ERIC Educational Resources Information Center

    Chobert, Julie; Marie, Celine; Francois, Clement; Schon, Daniele; Besson, Mireille

    2011-01-01

    The aim of this study was to examine the influence of musical expertise in 9-year-old children on passive (as reflected by MMN) and active (as reflected by discrimination accuracy) processing of speech sounds. Musician and nonmusician children were presented with a sequence of syllables that included standards and deviants in vowel frequency,…

  13. Ambient and focal visual processing of naturalistic activity.

    PubMed

    Eisenberg, Michelle L; Zacks, Jeffrey M

    2016-01-01

    When people inspect a picture, they progress through two distinct phases of visual processing: an ambient, or exploratory, phase that emphasizes input from peripheral vision and rapid acquisition of low-frequency information, followed by a focal phase that emphasizes central vision, salient objects, and high-frequency information. Does this qualitative shift occur during dynamic scene viewing? If so, when? One possibility is that shifts to exploratory processing are triggered at subjective event boundaries. This shift would be adaptive, because event boundaries typically occur when activity features change and when activity becomes unpredictable. Here, we used a perceptual event segmentation task, in which people identified boundaries between meaningful units of activity, to test this hypothesis. In two studies, an eye tracker recorded eye movements and pupil size while participants first watched movies of actors engaged in everyday activities and then segmented them into meaningful events. Saccade amplitudes and fixation durations during the initial viewings suggest that event boundaries function much like the onset of a new picture during static picture presentation: Viewers initiate an ambient processing phase and then progress to focal viewing as the event progresses. These studies suggest that this shift in processing mode could play a role in the formation of mental representations of the current environment.

  14. The Land Processes Distributed Active Archive Center (LP DAAC)

    USGS Publications Warehouse

    Golon, Danielle K.

    2016-10-03

    The Land Processes Distributed Active Archive Center (LP DAAC) operates as a partnership with the U.S. Geological Survey and is 1 of 12 DAACs within the National Aeronautics and Space Administration (NASA) Earth Observing System Data and Information System (EOSDIS). The LP DAAC ingests, archives, processes, and distributes NASA Earth science remote sensing data. These data are provided to the public at no charge. Data distributed by the LP DAAC provide information about Earth’s surface from daily to yearly intervals and at 15 to 5,600 meter spatial resolution. Data provided by the LP DAAC can be used to study changes in agriculture, vegetation, ecosystems, elevation, and much more. The LP DAAC provides several ways to access, process, and interact with these data. In addition, the LP DAAC is actively archiving new datasets to provide users with a variety of data to study the Earth.

  15. Differential Processing for Actively Ignored Pictures and Words

    PubMed Central

    Ciraolo, Margeaux

    2017-01-01

    Previous work suggests that, when attended, pictures may be processed more readily than words. The current study extends this research to assess potential differences in processing between these stimulus types when they are actively ignored. In a dual-task paradigm, facilitated recognition for previously ignored words was found provided that they appeared frequently with an attended target. When adapting the same paradigm here, previously unattended pictures were recognized at high rates regardless of how they were paired with items during the primary task, whereas unattended words were later recognized at higher rates only if they had previously been aligned with primary task targets. Implicit learning effects obtained by aligning unattended items with attended task-targets may apply only to conceptually abstract stimulus types, such as words. Pictures, on the other hand, may maintain direct access to semantic information, and are therefore processed more readily than words, even when being actively ignored. PMID:28122022

  16. N-Glycosylation Regulates ADAM8 Processing and Activation*

    PubMed Central

    Srinivasan, Srimathi; Romagnoli, Mathilde; Bohm, Andrew; Sonenshein, Gail E.

    2014-01-01

    The transmembrane ADAM8 (A Disintegrin And Metalloproteinase 8) protein is abundantly expressed in human breast tumors and derived metastases compared with normal breast tissue, and plays critical roles in aggressive Triple-Negative breast cancers (TNBCs). During ADAM8 maturation, the inactive proform dimerizes or multimerizes and autocatalytically removes the prodomain leading to the formation of the active, processed form. ADAM8 is a glycoprotein; however, little was known about the structure or functional role of these sugar moieties. Here, we report that in estrogen receptor (ER)α-negative, but not -positive, breast cancer cells ADAM8 contains N-glycosylation, which is required for its correct processing and activation. Consistently ADAM8 dimers were detected on the surface of ERα-negative breast cancer cells but not on ERα-positive ones. Site-directed mutagenesis confirmed four N-glycosylazhytion sites (Asn-67, Asn-91, Asn-436, and Asn-612) in human ADAM8. The Asn-67 and Asn-91 prodomain sites contained high mannose, whereas complex type N-glycosylation was observed on Asn-436 and Asn-612 in the active and remnant forms. The Asn-91 and Asn-612 sites were essential for its correct processing and cell surface localization, in particular its exit from the Golgi and endoplasmic reticulum, respectively. The N436Q mutation led to decreased ADAM8 stability due to enhanced lysosomal degradation. In contrast, mutation of the Asn-67 site had only modest effects on enzyme stability and processing. Thus, N-glycosylation is essential for processing, localization, stability, and activity of ADAM8. PMID:25336660

  17. Dynamic Stimuli And Active Processing In Human Visual Perception

    NASA Astrophysics Data System (ADS)

    Haber, Ralph N.

    1990-03-01

    Theories of visual perception traditionally have considered a static retinal image to be the starting point for processing; and has considered processing both to be passive and a literal translation of that frozen, two dimensional, pictorial image. This paper considers five problem areas in the analysis of human visually guided locomotion, in which the traditional approach is contrasted to newer ones that utilize dynamic definitions of stimulation, and an active perceiver: (1) differentiation between object motion and self motion, and among the various kinds of self motion (e.g., eyes only, head only, whole body, and their combinations); (2) the sources and contents of visual information that guide movement; (3) the acquisition and performance of perceptual motor skills; (4) the nature of spatial representations, percepts, and the perceived layout of space; and (5) and why the retinal image is a poor starting point for perceptual processing. These newer approaches argue that stimuli must be considered as dynamic: humans process the systematic changes in patterned light when objects move and when they themselves move. Furthermore, the processing of visual stimuli must be active and interactive, so that perceivers can construct panoramic and stable percepts from an interaction of stimulus information and expectancies of what is contained in the visual environment. These developments all suggest a very different approach to the computational analyses of object location and identification, and of the visual guidance of locomotion.

  18. Gaussian Process for Activity Modeling and Anomaly Detection

    NASA Astrophysics Data System (ADS)

    Liao, W.; Rosenhahn, B.; Yang, M. Ying

    2015-08-01

    Complex activity modeling and identification of anomaly is one of the most interesting and desired capabilities for automated video behavior analysis. A number of different approaches have been proposed in the past to tackle this problem. There are two main challenges for activity modeling and anomaly detection: 1) most existing approaches require sufficient data and supervision for learning; 2) the most interesting abnormal activities arise rarely and are ambiguous among typical activities, i.e. hard to be precisely defined. In this paper, we propose a novel approach to model complex activities and detect anomalies by using non-parametric Gaussian Process (GP) models in a crowded and complicated traffic scene. In comparison with parametric models such as HMM, GP models are nonparametric and have their advantages. Our GP models exploit implicit spatial-temporal dependence among local activity patterns. The learned GP regression models give a probabilistic prediction of regional activities at next time interval based on observations at present. An anomaly will be detected by comparing the actual observations with the prediction at real time. We verify the effectiveness and robustness of the proposed model on the QMUL Junction Dataset. Furthermore, we provide a publicly available manually labeled ground truth of this data set.

  19. Processing activities for STS-91 continue in OPF Bay 2

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Processing activities for STS-91 continue in KSC's Orbiter Processing Facility Bay 2. Two Get Away Special (GAS) canisters are shown after their installation into Discovery's payload bay. At left is G-648, an Canadian Space Agency-sponsored study of manufactured organic thin film by the physical vapor transport method, and the can on the right contains commemorative flags to be flown during the mission. STS-91 is scheduled to launch aboard the Space Shuttle Discovery for the ninth and final docking with the Russian Space Station Mir from KSC's Launch Pad 39A on June 2 with a launch window opening around 6:04 p.m. EDT.

  20. Processing activities for STS-91 continue in OPF Bay 2

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Processing activities for STS-91 continue in KSC's Orbiter Processing Facility Bay 2. Two Get Away Special (GAS) canisters are shown after their installation into Discovery's payload bay. At left is G-090, containing three educational experiments sponsored by Utah State University, and at right is G-743, an experiment sponsored by Broward Community College in Florida to test DNA exposed to cosmic radiation in a microgravity environment. STS-91 is scheduled to launch aboard the Space Shuttle Discovery for the ninth and final docking with the Russian Space Station Mir from KSC's Launch Pad 39A on June 2 with a launch window opening around 6:04 p.m. EDT.

  1. Hanford's Simulated Low Activity Waste Cast Stone Processing

    SciTech Connect

    Kim, Young

    2013-08-20

    Cast Stone is undergoing evaluation as the supplemental treatment technology for Hanford’s (Washington) high activity waste (HAW) and low activity waste (LAW). This report will only cover the LAW Cast Stone. The programs used for this simulated Cast Stone were gradient density change, compressive strength, and salt waste form phase identification. Gradient density changes show a favorable outcome by showing uniformity even though it was hypothesized differently. Compressive strength exceeded the minimum strength required by Hanford and greater compressive strength increase seen between the uses of different salt solution The salt waste form phase is still an ongoing process as this time and could not be concluded.

  2. Suppression of Wnt signaling by the miR-29 family is mediated by demethylation of WIF-1 in non-small-cell lung cancer

    SciTech Connect

    Tan, Min; Wu, Junjie; Cai, Yong

    2013-09-06

    Highlights: •Dnmt3A and Dnmt3B are involved in the down-regulation of WIF-1 expression in non-small-cell lung cancer. •MiR-29 family members could restore WIF-1 expression through demethylation. •MiR-29s suppress Wnt/β-catenin signaling pathway and inhibit tumor growth. •The expression of miR-29a and miR-29b could be regulated partially in a positive feedback loop. -- Abstract: Wnt inhibitory factor-1 (WIF-1) silencing induced by promoter hypermethylation is a common mechanism of aberrant activation of the Wnt signaling pathway in non-small-cell lung cancer (NSCLC). However, the activity of regulators associated with the methylation of the WIF-1 gene remains unclear. Here, we investigated the role of three DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) in the expression of WIF-1. The three DNMTs were up-regulated in NSCLC tumor tissues and suppression of DNMT3A and DNMT3B restored the expression of WIF-1 in NSCLC cells. The miR-29 family (miR-29a, -29b, and -29c), which negatively regulates DNMT3A and DNMT3B, was examined in association with the Wnt/β-catenin signaling pathway. A positive correlation between the expression of WIF-1 and that of MiR-29s was observed in NSCLC tissues. Methylation-specific PCR and Western blotting indicated that miR-29s positively regulate WIF-1 expression by inhibiting the methylation of its promoter. Furthermore, miR-29 overexpression downregulated β-catenin expression, inhibited cell proliferation and induced apoptosis. The expression of miR-29a and miR-29b was partially regulated by DNMT3A and DNMT3B in a positive feedback loop. Taken together, our findings show that miR-29s suppress the Wnt signaling pathway through demethylation of WIF-1 in NSCLC.

  3. Enhancement of activated sludge disintegration and dewaterability by Fenton process

    NASA Astrophysics Data System (ADS)

    Heng, G. C.; Isa, M. H.

    2016-06-01

    Municipal and industrial wastewater treatment plants produce large amounts of sludge. This excess sludge is an inevitable drawback inherent to the activated sludge process. In this study, the waste activated sludge was obtained from the campus wastewater treatment plant at Universiti Teknologi PETRONAS (UTP), Malaysia. Fenton pretreatment was optimized by using the response surface methodology (RSM) to study the effects of three operating conditions including the dosage of H2O2 (g H2O2/kg TS), the molar ratio of H2O2/Fe2+ and reaction time. The optimum operating variables to achieve MLVSS removal 65%, CST reduction 28%, sCOD 11000 mg/L and EPS 500 mg/L were: 1000 g H2O2/kg TS, H2O2/Fe2+ molar ratio 70 and reaction time 45 min. Fenton process was proved to be able to enhance the sludge disintegration and dewaterability.

  4. Cross-fostering study of methyl mercury retention, demethylation and excretion in the neonatal hamster.

    PubMed

    Nordenhäll, K; Dock, L; Vahter, M

    1998-03-01

    The cross-fostering technique was used in order to compare methyl mercury (MeHg) metabolism in hamsters following prenatal (in utero) and neonatal (lactational) exposure. Pregnant Syrian golden hamsters were administered radiolabeled MeHg on day 12 of gestation. The offspring was nursed by foster mothers unexposed to MeHg, while the pups from the unexposed animals were nursed by the MeHg-administered animals. Under these conditions, each pup in the litter received a dose of MeHg in utero corresponding to 0.9% of the maternal dose. The average amount of mercury found in the pups exposed via milk corresponded to 4.5% of the total body burden of the foster dam at the onset of lactation. This was about half the amount received by the pups exposed in utero. The total body burden of mercury, and the amount of mercury in the liver, brain and kidney of the pups exposed in utero began to decrease at seven days of age. The rate of decrease differed among the tissues and was lowest in the kidney. The amount of mercury in pups exposed via milk reached a peak level when the pups were 10-15 days old. The total body burden of mercury showed a slow decrease while the liver, brain and kidney levels decreased rapidly. In both groups of animals, up to 80% of the total body burden of mercury was found in the pelt. These data show that milk may be a significant exposure route for mercury and that neonatal hamsters are unable to demethylate MeHg and excrete mercury in urine and faeces.

  5. Unraveling the processing and activation of snake venom metalloproteinases.

    PubMed

    Portes-Junior, José A; Yamanouye, Norma; Carneiro, Sylvia M; Knittel, Paloma S; Sant'Anna, Sávio S; Nogueira, Fabio C S; Junqueira, Magno; Magalhães, Geraldo S; Domont, Gilberto B; Moura-da-Silva, Ana M

    2014-07-03

    Snake venom metalloproteinases (SVMPs) are zinc-dependent enzymes responsible for most symptoms of human envenoming. Like matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinase (ADAM) proteins, SVMPs are synthesized as zymogens, and enzyme activation is regulated by hydrolysis of their prodomain, but the processing of SVMPs is still unclear. In this study, we attempted to identify the presence of prodomain in different compartments of snake venom glands as zymogens or in the free form to elucidate some mechanism involved in SVMP activation. Using antibodies obtained by immunization with a recombinant prodomain, bands of zymogen molecular mass and prodomain peptides were detected mostly in gland extracts all along the venom production cycle and in the venom collected from the lumen at the peak of venom production. Prodomain was detected in secretory cells mostly in the secretory vesicles near the Golgi. We hypothesize that the processing of SVMPs starts within secretory vesicles and continues in the lumen of the venom gland just after enzyme secretion and involves different steps compared to ADAMs and MMPs but can be used as a model for studying the relevance of peptides resulting from prodomain processing and degradation for controlling the activity of metalloproteinases.

  6. Land processes distributed active archive center product lifecycle plan

    USGS Publications Warehouse

    Daucsavage, John C.; Bennett, Stacie D.

    2014-01-01

    The U.S. Geological Survey (USGS) Earth Resources Observation and Science (EROS) Center and the National Aeronautics and Space Administration (NASA) Earth Science Data System Program worked together to establish, develop, and operate the Land Processes (LP) Distributed Active Archive Center (DAAC) to provide stewardship for NASA’s land processes science data. These data are critical science assets that serve the land processes science community with potential value beyond any immediate research use, and therefore need to be accounted for and properly managed throughout their lifecycle. A fundamental LP DAAC objective is to enable permanent preservation of these data and information products. The LP DAAC accomplishes this by bridging data producers and permanent archival resources while providing intermediate archive services for data and information products.

  7. Materials and Process Activities for NASA's Composite Crew Module

    NASA Technical Reports Server (NTRS)

    Polis, Daniel L.

    2012-01-01

    In January 2007, the NASA Administrator and Associate Administrator for the Exploration Systems Mission Directorate chartered the NASA Engineering and Safety Center (NESC) to design, build, and test a full-scale Composite Crew Module (CCM). The overall goal of the CCM project was to develop a team from the NASA family with hands-on experience in composite design, manufacturing, and testing in anticipation of future space exploration systems being made of composite materials. The CCM project was planned to run concurrently with the Orion project s baseline metallic design within the Constellation Program so that features could be compared and discussed without inducing risk to the overall Program. The materials and process activities were prioritized based on a rapid prototype approach. This approach focused developmental activities on design details with greater risk and uncertainty, such as out-of-autoclave joining, over some of the more traditional lamina and laminate building block levels. While process development and associated building block testing were performed, several anomalies were still observed at the full-scale level due to interactions between process robustness and manufacturing scale-up. This paper describes the process anomalies that were encountered during the CCM development and the subsequent root cause investigations that led to the final design solutions. These investigations highlight the importance of full-scale developmental work early in the schedule of a complex composite design/build project.

  8. Spectral dynamics of electroencephalographic activity during auditory information processing.

    PubMed

    Cacace, Anthony T; McFarland, Dennis J

    2003-02-01

    Dynamics of electroencephalographic (EEG) activity during auditory information processing were evaluated in response to changes in stimulus complexity, stimulus discriminability and attention using the oddball paradigm. In comparison to pre-stimulus baseline conditions, auditory stimulation synchronized EEG activity in delta, theta and alpha frequency bands. Event-related synchronization (ERS) effects were greatest at approximately 3 Hz (theta frequency band), and their magnitude depended on stimulus and task demands. Event-related desynchronization (ERD) of EEG activity was observed in the beta frequency band. This effect was greatest at approximately 21 Hz but occurred only for easily discriminable stimuli in attention-related target conditions. Because active discrimination tasks also required a button-press response with the right hand, ERDs involved more complex responses that may be related to a combination of perceptual, motor and cognitive processes. These results demonstrate that oddball and attention-related EEG responses to auditory stimulation could be characterized in the frequency domain. The specific design and analysis features described herein may prove useful since they provide a simple index of the brain's response to stimulation while at the same time provide powerful information not contained in typical time domain analysis.

  9. Demethylation of oligogalacturonides by FaPE1 in the fruits of the wild strawberry Fragaria vesca triggers metabolic and transcriptional changes associated with defence and development of the fruit.

    PubMed

    Osorio, Sonia; Bombarely, Aureliano; Giavalisco, Patrick; Usadel, Björn; Stephens, Camilla; Aragüez, Irene; Medina-Escobar, Nieves; Botella, Miguel A; Fernie, Alisdair R; Valpuesta, Victoriano

    2011-05-01

    Ectopic expression of the strawberry (Fragaria×ananassa) gene FaPE1 encoding pectin methyl esterase produced in the wild species Fragaria vesca partially demethylated oligogalacturonides (OGAs), which conferred partial resistance of ripe fruits to the fungus Botrytis cinerea. Analyses of metabolic and transcriptional changes in the receptacle of the transgenic fruits revealed channelling of metabolites to aspartate and aromatic amino acids as well as phenolics, flavanones, and sesquiterpenoids, which was in parallel with the increased expression of some genes related to plant defence. The results illustrate the changes associated with resistance to B. cinerea in the transgenic F. vesca. These changes were accompanied by a significant decrease in the auxin content of the receptacle of the ripe fruits of transgenic F. vesca, and enhanced expression of some auxin-repressed genes. The role of these OGAs in fruit development was revealed by the larger size of the ripe fruits in transgenic F. vesca. When taken together these results show that in cultivated F. ananassa FaPE1 participates in the de-esterification of pectins and the generation of partially demethylated OGAs, which might reinforce the plant defence system and play an active role in fruit development.

  10. Active vision system integrating fast and slow processes

    NASA Astrophysics Data System (ADS)

    Castrillon-Santana, Modesto; Guerra-Artal, C.; Hernandez-Sosa, J.; Dominguez-Brito, A.; Isern-Gonzalez, J.; Cabrera-Gamez, Jorge; Hernandez-Tejera, F. M.

    1998-10-01

    This paper describes an Active Vision System whose design assumes a distinction between fast or reactive and slow or background processes. Fast processes need to operate in cycles with critical timeouts that may affect system stability. While slow processes, though necessary, do not compromise system stability if its execution is delayed. Based on this simple taxonomy, a control architecture has been proposed and a prototype implemented that is able to track people in real-time with a robotic head while trying to identify the target. In this system, the tracking mobile is considered as the reactive part of the system while person identification is considered a background task. This demonstrator has been developed using a new generation DSP (TMS320C80) as a specialized coprocessor to deal with fast processes, and a commercial robotic head with a dedicated DSP-based motor controller. These subsystems are hosted by a standard Pentium-Pro PC running Windows NT where slow processes are executed. The flexibility achieved in the design phase and the preliminary results obtained so far seem to validate the approach followed to integrate time- critical and slow tasks on a heterogeneous hardware platform.

  11. Role of sympathetic nerve activity in the process of fainting

    PubMed Central

    Iwase, Satoshi; Nishimura, Naoki; Mano, Tadaaki

    2014-01-01

    Syncope is defined as a transient loss of consciousness and postural tone, characterized by rapid onset, short duration, and spontaneous recovery, and the process of syncope progression is here described with two types of sympathetic change. Simultaneous recordings of microneurographically-recorded muscle sympathetic nerve activity (MSNA) and continuous and noninvasive blood pressure measurement has disclosed what is going on during the course of syncope progression. For vasovagal or neurally mediated syncope, three stages are identified in the course of syncope onset, oscillation, imbalance, and catastrophe phases. Vasovagal syncope is characterized by sympathoexcitation, followed by vagal overcoming via the Bezold-Jarisch reflex. Orthostatic syncope is caused by response failure or a lack of sympathetic nerve activity to the orthostatic challenge, followed by fluid shift and subsequent low cerebral perfusion. Four causes are considered for the compensatory failure that triggers orthostatic syncope: hypovolemia, increased pooling in the lower body, failure to activate sympathetic activity, and failure of vasoconstriction against sympathetic vasoconstrictive stimulation. Many pathophysiological conditions have been described from the perspectives of (1) exaggerated sympathoexcitation and (2) failure to activate the sympathetic nerve. We conclude that the sympathetic nervous system can control cardiovascular function, and its failure results in syncope; however, responses of the system obtained by microneurographically-recorded MSNA would determine the pathophysiology of the onset and progression of syncope, explaining the treatment effect that could be achieved by the analysis of this mechanism. PMID:25309444

  12. Drilling to investigate processes in active tectonics and magmatism

    NASA Astrophysics Data System (ADS)

    Shervais, J.; Evans, J.; Toy, V.; Kirkpatrick, J.; Clarke, A.; Eichelberger, J.

    2014-12-01

    Coordinated drilling efforts are an important method to investigate active tectonics and magmatic processes related to faults and volcanoes. The US National Science Foundation (NSF) recently sponsored a series of workshops to define the nature of future continental drilling efforts. As part of this series, we convened a workshop to explore how continental scientific drilling can be used to better understand active tectonic and magmatic processes. The workshop, held in Park City, Utah, in May 2013, was attended by 41 investigators from seven countries. Participants were asked to define compelling scientific justifications for examining problems that can be addressed by coordinated programs of continental scientific drilling and related site investigations. They were also asked to evaluate a wide range of proposed drilling projects, based on white papers submitted prior to the workshop. Participants working on faults and fault zone processes highlighted two overarching topics with exciting potential for future scientific drilling research: (1) the seismic cycle and (2) the mechanics and architecture of fault zones. Recommended projects target fundamental mechanical processes and controls on faulting, and range from induced earthquakes and earthquake initiation to investigations of detachment fault mechanics and fluid flow in fault zones. Participants working on active volcanism identified five themes: the volcano eruption cycle; eruption sustainability, near-field stresses, and system recovery; eruption hazards; verification of geophysical models; and interactions with other Earth systems. Recommended projects address problems that are transferrable to other volcanic systems, such as improved methods for identifying eruption history and constraining the rheological structure of shallow caldera regions. Participants working on chemical geodynamics identified four major themes: large igneous provinces (LIPs), ocean islands, continental hotspot tracks and rifts, and

  13. The role of highly purified forms of rat liver cytochrome P-450 in the dimethylation of dimethylnitrosamine and its activation to mutagens.

    PubMed

    Masson, H A; Ioannides, C; Gibson, G G

    1983-06-01

    Highly purified NADPH-cytochrome P-450 reductase and the major phenobarbital (PB) and beta-naphthoflavone (beta NF) forms of cytochrome P-450 were used in reconstituted systems to study the demethylation and subsequent activation of dimethylnitrosamine (DMN) to mutagenic intermediates. Both forms of cytochrome P-450 were active in the demethylation of DMN, cytochrome P-450 from PB-treated animals being more efficient, generating nearly twice as much formaldehyde per nmol of haemoprotein. Neither form of the cytochrome could activate DMN to mutagens in the Ames test. These findings indicate that DMN demethylation does not lead to its activation to mutagenic products.

  14. Recognition of Histone H3K4 Trimethylation by the Plant Homeodomain of PHF2 Modulates Histone Demethylation

    SciTech Connect

    Wen, Hong; Li, Jingzhi; Song, Tanjing; Lu, Ming; Kan, Pu-Yeh; Lee, Min Gyu; Sha, Bingdong; Shi, Xiaobing

    2010-10-28

    Distinct lysine methylation marks on histones create dynamic signatures deciphered by the 'effector' modules, although the underlying mechanisms remain unclear. We identified the plant homeodomain- and Jumonji C domain-containing protein PHF2 as a novel histone H3K9 demethylase. We show in biochemical and crystallographic analyses that PHF2 recognizes histone H3K4 trimethylation through its plant homeodomain finger and that this interaction is essential for PHF2 occupancy and H3K9 demethylation at rDNA promoters. Our study provides molecular insights into the mechanism by which distinct effector domains within a protein cooperatively modulate the 'cross-talk' of histone modifications.

  15. Processing activities for STS-91 continue in OPF Bay 2

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Processing activities for STS-91 continue in KSC's Orbiter Processing Facility Bay 2. The payload bay of Space Shuttle Discovery is relatively empty as installation of the Get Away Special (GAS) canisters begins. Two GAS canisters can be seen in the center of the photograph. On the left is G-648, a Canadian Space Agency-sponsored study on manufactured organic thin film by the physical vapor transport method, and on the right is a can with hundreds of commemorative flags to be flown on the mission. STS-91 is scheduled to launch aboard the Space Shuttle Discovery for the ninth and final docking with the Russian Space Station Mir from KSC's Launch Pad 39A on June 2 with a launch window opening around 6:04 p.m. EDT.

  16. Processing activities for STS-91 continue in OPF Bay 2

    NASA Technical Reports Server (NTRS)

    1998-01-01

    Processing activities for STS-91 continue in Orbiter Processing Facility Bay 2. Two Get Away Special (GAS) canisters are shown after their installation into Discovery's payload bay. The GAS payload G-765, in the canister on the left, is sponsored by the Canadian Space Agency and managed by C-CORE/Memorial University of Newfoundland. It is a study to understand the transport of fluids in porous media as it pertains to improving methods for enhanced oil recovery. The GAS canister on the right houses the Space Experiment Module (SEM-05), part of an educational initiative of NASA's Shuttle Small Payloads Project. STS-91 is scheduled to launch aboard the Space Shuttle Discovery for the ninth and final docking with the Russian Space Station Mir from KSC's Launch Pad 39A on June 2 with a launch window opening around 6:04 p.m. EDT.

  17. Rapid rotation and mixing in active OB stars - Physical processes

    NASA Astrophysics Data System (ADS)

    Zahn, Jean-Paul

    2011-07-01

    In the standard description of stellar interiors, O and B stars possess a thoroughly mixed convective core surrounded by a stable radiative envelope in which no mixing occurs. But as is well known, this model disagrees strongly with the spectroscopic diagnostic of these stars, which reveals the presence at their surface of chemical elements that have been synthesized in the core. Hence the radiation zone must be the seat of some mild mixing mechanisms. The most likely to operate there are linked with the rotation: these are the shear instabilites triggered by the differential rotation, and the meridional circulation caused by the changes in the rotation profile accompanying the non-homologous evolution of the star. In addition to these hydrodynamical processes, magnetic stresses may play an important role in active stars, which host a magnetic field. These physical processes will be critically examined, together with some others that have been suggested.

  18. A process activity monitor for AOS/VS

    NASA Technical Reports Server (NTRS)

    Mckosky, R. A.; Lindley, S. W.; Chapman, J. S.

    1986-01-01

    With the ever increasing concern for computer security, users of computer systems are becoming more sensitive to unauthorized access. One of the initial security concerns for the Shuttle Management Information System was the problem of users leaving their workstations unattended while still connected to the system. This common habit was a concern for two reasons: it ties up resources unnecessarily and it opens the way for unauthorized access to the system. The Data General MV/10000 does not come equipped with an automatic time-out option on interactive peripherals. The purpose of this memorandum is to describe a system which monitors process activity on the system and disconnects those users who show no activity for some time quantum.

  19. Periodic changes of the activity of processes in Jupiter's atmosphere

    NASA Astrophysics Data System (ADS)

    Vidmachenko, A. P.

    2016-10-01

    Variations of the Earth jovimagnetic latitude on Jupiter are preferred in solar-driven changes of reflective properties of clouds and haze on Jupiter. Because of the orbit eccentricity (e=0,048450) the northern hemisphere receives 21% greater solar energy flow to the atmosphere, because Jupiter is in the perihelia near the time of the summer solstice. Results of our studies showed that the ratio of the brightness of the northern and southern tropical and temperate regions is evident factor of the photometric activity of the Jupiter's atmospheric processes. The obtained from the analysis of observational data for the period from 1962 to 2015 existence of variations of activity factor of the planet hemispheres with a period of 11.86 years has allowed us to talk about an existence of the seasonal reconstruction of the physical parameters of Jupiter's atmosphere.

  20. The consent process: Enabling or disabling patients' active participation?

    PubMed

    Doherty, Carole; Stavropoulou, Charitini; Saunders, Mark Nk; Brown, Tracey

    2015-10-20

    Standards expected by doctors' regulatory bodies in respect of the process of consent to treatment have arguably sought to restructure the nature of the doctor-patient relationship from one of the paternalism to that of shared decision-making. Yet, few studies have explored empirically, from patients' perspectives, the extent to which the process of consent to treatment enables or disables patients' participation in medical decision-making. Our article examines patients' attitudes towards the consent process, exploring how and why these attitudes influence patients' active participation in decision-making and considering possible consequent medico-legal issues. Data were collected longitudinally using semi-structured interviews and field observations involving 35 patients and 19 of their caregivers, in an English hospital between February and November 2014. These indicate that generally patients defer to the doctor in respect of treatment decision-making. Although most patients and their caregivers wanted detailed information and discussion, they did not necessarily expect that this would be provided. Furthermore, patients perceived that signing the consent form was an obligatory routine principally to protect doctors from legal action should something go wrong. Our study suggests that patients' predominantly paternalistic perceptions of the consent process can not only undermine attempts by doctors to involve them in decision-making but, as patients are now considered in law as informed actors, their perceptions of the consent form as not being in their interests could be a self-fulfilling prophecy if signing is undertaken without due consideration to the content.

  1. Vitamin C down-regulate apo(a) expression via Tet2-dependent DNA demethylation in HepG2 cells.

    PubMed

    Qu, Kai; Ma, Xiao-Feng; Li, Guo-Hua; Zhang, Hai; Liu, Ya-Mi; Zhang, Kai; Zeng, Jun-Fa; Lei, Jian-Jun; Wei, Dang-Heng; Wang, Zuo

    2017-05-01

    Lipoprotein(a)[Lp(a)] is a risk factor for coronary heart diseases. However, the metabolism of this protein remains poorly understood. Efficient and specific drugs that can decrease high plasma levels of Lp(a) have not been developed yet. Vitamin C is responsible for maintaining the catalytic activity of a group of iron and 2-oxoglutarate (2OG)-dependent dioxygenases and induces the generation of 5-hydroxymethylcytosine (5hmC) via Ten-eleven translocation (Tet) dioxygenases. In addition, It has been reported vitamin C deficiency induces atherosclerosis and increases Lp(a) and apo(a) plasma levels in Lp(a)+ mice. However, the mechanism is still unclear. In this study, we investigated the effects of vitamin C on apo(a) expression and the possible molecular mechanism of vitamin C that influences apolipoprotein(a) [apo(a)] biosynthesis in HepG2 cells. Results showed that vitamin C significantly inhibited the expression and secretion levels of apo(a). Vitamin C can also increase ELK1 expression and hydroxymethylation of ELK1 promoter and the globle DNA in HepG2 cells. In addition, the effects of vitamin C inhibiting the apo(a) expression were attenuated by ELK1siRNA and Tet2siRNA. These results suggested vitamin C down-regulate apo(a) expression via Tet2-dependent DNA demethylation in HepG2 cells.

  2. The red sport of 'Zaosu' pear and its red-striped pigmentation pattern are associated with demethylation of the PyMYB10 promoter.

    PubMed

    Qian, Minjie; Sun, Yongwang; Allan, Andrew C; Teng, Yuanwen; Zhang, Dong

    2014-11-01

    'Zaosu' pear, a hybrid of Pyrus pyrifolia and Pyrus communis, is a popular cultivar developed in China. 'Zaosu Red' is a bud sport of 'Zaosu' with red shoots, young leaves, and fruit. After grafting of 'Zaosu Red', reverse mutations in some branches lead to a loss of colour in leaves and stems. Also, the mature fruit of 'Zaosu Red' exhibits two phenotypes; fully red and striped. The aim of this study was to establish the mechanism of the red colour mutation in 'Zaosu' and the striped pigmentation pattern in fruit of 'Zaosu Red'. The accumulation of anthocyanins and transcript levels of the genes PpUFGT2 and PyMYB10 were highly correlated. The open reading frames (ORF) and promoter regions of these two key genes were cloned and compared between 'Zaosu' and its bud sports, but no sequence differences were found. The R2R3 MYB, PyMYB10, can activate expression of genes encoding enzymes of the anthocyanin biosynthetic pathway. A yeast one-hybrid assay showed that PyMYB10 was associated with the -658 to -172bp fragment of the PpUFGT2 promoter, probably via a MYB binding site (MBS) located at -466bp. The PyMYB10 promoter had lower methylation levels in anthocyanin-rich tissues, indicating that the red bud sport of 'Zaosu' pear and the striped pigmentation pattern of 'Zaosu Red' pear are associated with demethylation of the PyMYB10 promoter.

  3. DOE Solar Process Heat Program: FY1991 Solar Process Heat Prefeasibility Studies activity

    SciTech Connect

    Hewett, R.

    1992-11-01

    During fiscal year (FY) 1991, the US Department of Energy (DOE) Solar Process Heat Program implemented a Solar Process Heat Prefeasibility Studies activity. For Program purposes, a prefeasibility study is an engineering assessment that investigates the technical and economic feasibility of a solar system for a specific application for a specific end-user. The study includes an assessment of institutional issues (e.g., financing, availability of insurance, etc.) that impact the feasibility of the proposed solar project. Solar process heat technology covers solar thermal energy systems (utilizing flat plate or concentrating solar Collectors) for water heating, water preheating, cooling/refrigeration, steam generation, ventilation air heating/preheating, etc. for applications in industry, commerce, and government. The studies are selected for funding through a competitive solicitation. For FY 1991, six projects were selected for funding. As of August 31, 1992, three teams had completed their studies. This paper describes the prefeasibility studies activity, presents the results from the study performed by United Solar Technologies, and summarizes the conclusions from the studies that have been completed to date and their implications for the Solar Process Heat Program.

  4. Department of Energy solar process heat program: FY 1991 solar process heat prefeasibility studies activity

    NASA Astrophysics Data System (ADS)

    Hewett, R.

    1992-11-01

    During fiscal year (FY) 1991, the US Department of Energy (DOE) Solar Process Heat Program implemented a Solar Process Heat Prefeasibility Studies activity. For Program purposes, a prefeasibility study is an engineering assessment that investigates the technical and economic feasibility of a solar system for a specific application for a specific end-user. The study includes an assessment of institutional issues (e.g., financing, availability of insurance, etc.) that impact the feasibility of the proposed solar project. Solar process heat technology covers solar thermal energy systems (utilizing flat plate or concentrating solar collectors) for water heating, water preheating, cooling/refrigeration, steam generation, ventilation air heating/preheating, etc., for applications in industry, commerce, and government. The studies are selected for funding through a competitive solicitation. For FY-91, six projects were selected for funding. As of 31 Aug. 1992, three teams had completed their studies. This paper describes the prefeasibility studies activity, presents the results from the study performed by United Solar Technologies, and summarizes the conclusions from the studies that have been completed to date and their implications for the Solar Process Heat Program.

  5. Behavioral activation system modulation on brain activation during appetitive and aversive stimulus processing

    PubMed Central

    Ventura-Campos, Noelia; Sanjuán-Tomás, Ana; Belloch, Vicente; Parcet, Maria-Antònia; Ávila, César

    2010-01-01

    The reinforcement sensitivity theory (RST) proposed the behavioral activation system (BAS) as a neurobehavioral system that is dependent on dopamine-irrigated structures and that mediates the individual differences in sensitivity and reactivity to appetitive stimuli associated with BAS-related personality traits. Theoretical developments propose that high BAS sensitivity is associated with both enhanced appetitive stimuli processing and the diminished processing of aversive stimuli. The objective of this study was to analyze how individual differences in BAS functioning were associated with brain activation during erotic and aversive picture processing while subjects were involved in a simple goal-directed task. Forty-five male participants took part in this study. The task activation results confirm the activation of the reward and punishment brain-related structures while viewing erotic and aversive pictures, respectively. The SR scores show a positive correlation with activation of the left lateral prefrontal cortex, the mesial prefrontal cortex and the right occipital cortex while viewing erotic pictures, and a negative correlation with the right lateral prefrontal cortex and the left occipital cortex while viewing aversive pictures. In summary, the SR scores modulate the activity of the cortical areas in the prefrontal and the occipital cortices that are proposed to modulate the BAS and the BIS-FFFS. PMID:20147458

  6. Fractionating soluble microbial products in the activated sludge process.

    PubMed

    Ni, Bing-Jie; Zeng, Raymond J; Fang, Fang; Xie, Wen-Ming; Sheng, Guo-Ping; Yu, Han-Qing

    2010-04-01

    Soluble microbial products (SMP) are the pool of organic compounds originating from microbial growth and decay, and are usually the major component of the soluble organic matters in effluents from biological treatment processes. In this work, SMP in activated sludge were characterized, fractionized, and quantified using integrated chemical analysis and mathematical approach. The utilization-associated products (UAP) in SMP, produced in the substrate-utilization process, were found to be carbonaceous compounds with a molecular weight (MW) lower than 290 kDa which were quantified separately from biomass-associated products (BAP). The BAP were mainly cellular macromolecules with an MW in a range of 290-5000 kDa, and for the first time were further classified into the growth-associated BAP (GBAP) with an MW of 1000 kDa, which were produced in the microbial growth phase, and the endogeny-associated BAP (EBAP) with an MW of 4500 kDa, which were generated in the endogenous phase. Experimental and modeling results reveal that the UAP could be utilized by the activated sludge and that the BAP would accumulate in the system. The GBAP and EBAP had different formation rates from the hydrolysis of extracellular polymeric substances and distinct biodegradation kinetics. This study provides better understanding of SMP formation mechanisms and becomes useful for subsequent effluent treatment.

  7. Methane recovery from water hyacinth through anaerobic activated sludge process

    SciTech Connect

    Savaswat, N.; Khana, P.

    1986-02-01

    The concepts of phase separation, anaerobic activated sludge process, and alkali pretreatment have been incorporated in this investigation with the objective of developing rational and cost-effective designs of diphasic anaerobic activated sludge systems, with and without alkali treatment, for methane recovery from water hyacinth (WH). Evaluation of process kinetics and optimization analyses of laboratory data reveal that a diphasic system with alkali treatment could be designed with an alkali pretreatment step (3.6% Na/sub 2/CO/sub 3/ + 2.5% Ca(OH)/sub 2/ (w/w) of WH, 24 h duration) followed by an open acid phase (2.1 days HRT) and closed methane reactor with sludge recycle (5.7 days HRT, 7.7 days MCRT) for gas yield of 50 L/kg WH/d at 35-37/sup 0/C. Likewise, a diphasic system without alkali treatment could be designed with an open acid phase (2 days HRT) followed by closed methane reactor with sludge recycle (3.2 days HRT, 6 days MCRT) for gas yield of 32.5 L/kg WH/d at 35-37/sup 0/C. Detailed economic analyses bring forth greater cost-efficacy of the diphasic system without alkali treatment and reveal that the advantage accrued in terms of higher gas yield is overshadowed by the cost of chemicals in the diphasic system with alkali treatment.

  8. Methane recovery from water hyacinth through anaerobic activated sludge process

    SciTech Connect

    Saraswat, N.; Khanna, P.

    1986-02-01

    The concepts of phase separation, anaerobic activated sludge process, and alkali pretreatment have been incorporated in this investigation with the objective of developing rational and cost-effective designs of diphasic anaerobic activated sludge systems, with and without alkali treatment, for methane recovery from water hyacinth (WH). Evaluation of process kinetics and optimization analyses of laboratory data reveal that a diphasic system with alkali treatment could be designed with an alkali pretreatment step (3.6% Na/sub 2/CO/sub 3/ + 2.5% Ca(OH)/sub 2/ (w/w) of WH, 24 h duration) followed by an open acid phase (2.1 days HRT) and closed methane reactor with sludge recycle (5.7 days HRT, 7.7 days MCRT) for gas yield of 50 l/kg WH/d at 35-37/sup 0/C. Likewise, a diphasic system without alkali treatment could be designed with an open acid phase (2 days HRT) followed by close methane reactor with sludge recycle (3.2 days HRT, 6 days MCRT) for gas yield of 32.5 l.kg WH/d at 35-37/sup 0/C. Detailed economic analyses bring forth greater cost-efficacy of the diphasic system without alkali treatment and reveal that the advantage accrued in terms of higher gas yield is overshadowed by the cost of chemicals in the diphasic system with alkali treatment.

  9. Cloud condensation nuclei activity and droplet activation kinetics of wet processed regional dust samples and minerals

    NASA Astrophysics Data System (ADS)

    Kumar, P.; Sokolik, I. N.; Nenes, A.

    2011-08-01

    This study reports laboratory measurements of particle size distributions, cloud condensation nuclei (CCN) activity, and droplet activation kinetics of wet generated aerosols from clays, calcite, quartz, and desert soil samples from Northern Africa, East Asia/China, and Northern America. The dependence of critical supersaturation, sc, on particle dry diameter, Ddry, is used to characterize particle-water interactions and assess the ability of Frenkel-Halsey-Hill adsorption activation theory (FHH-AT) and Köhler theory (KT) to describe the CCN activity of the considered samples. Wet generated regional dust samples produce unimodal size distributions with particle sizes as small as 40 nm, CCN activation consistent with KT, and exhibit hygroscopicity similar to inorganic salts. Wet generated clays and minerals produce a bimodal size distribution; the CCN activity of the smaller mode is consistent with KT, while the larger mode is less hydrophilic, follows activation by FHH-AT, and displays almost identical CCN activity to dry generated dust. Ion Chromatography (IC) analysis performed on regional dust samples indicates a soluble fraction that cannot explain the CCN activity of dry or wet generated dust. A mass balance and hygroscopicity closure suggests that the small amount of ions (from low solubility compounds like calcite) present in the dry dust dissolve in the aqueous suspension during the wet generation process and give rise to the observed small hygroscopic mode. Overall these results identify an artifact that may question the atmospheric relevance of dust CCN activity studies using the wet generation method. Based on the method of threshold droplet growth analysis, wet generated mineral aerosols display similar activation kinetics compared to ammonium sulfate calibration aerosol. Finally, a unified CCN activity framework that accounts for concurrent effects of solute and adsorption is developed to describe the CCN activity of aged or hygroscopic dusts.

  10. Supporting the process of removing humic substances on activated carbon.

    PubMed

    Olesiak, Paulina; Stępniak, Longina

    2014-01-01

    This study is focused on biosorption process used in water treatment. The process has a number of advantages and a lot of research has been done into its intensification by means of ultrasonic modification of solutions. The study carried out by the authors leads to the conclusion that sonication of organic solutions allows for extension of the time of operation of carbon beds. For the analysis of the results obtained during the sorption of humic substances (HS) from the solution dependencies UV/UV₀ or DOC/DOC₀ were used. In comparative studies the effectiveness of sorption and sonosorption (UV/UV₀) shows that the share of ultrasounds (US) is beneficial for extension of time deposit, both at a flow rate HS solution equal to 1 m/h and 5 m/h. Analysis of the US impact sorption on HS sorption in a biological fluidized bed, both prepared from biopreparat and the activated sludge confirms the higher efficiency compared to sonobiosorption than biosorption. These results confirm the degree of reduction UV₂₅₄/UV₀ and DOC/DOC₀ for the same processes. EMS index also confirms the improvement of HSbiodegradation by sludge microorganisms.

  11. RUNX3 gene promoter demethylation by 5-Aza-CdR induces apoptosis in breast cancer MCF-7 cell line.

    PubMed

    Kang, Hua-Feng; Dai, Zhi-Jun; Bai, He-Ping; Lu, Wang-Feng; Ma, Xiao-Bin; Bao, Xing; Lin, Shuai; Wang, Xi-Jing

    2013-01-01

    Runt-related transcription factor 3 (RUNX3) is a tumor suppressor gene, its inactivation due to hypermethylation related to carcinogenesis. The aim of this study was to investigate the effects of 5-aza-2'-deoxycytidine (5-Aza-CdR) on cell proliferation and apoptosis by demethylation of the promoter region and restoring the expression of RUNX3 in the breast cancer MCF-7 cell line. MCF-7 cells were cultured with different concentrations (0.4-102.4 μmol/L) of 5-Aza-CdR in vitro. MTT assay was used to determine the proliferation of MCF-7 cells. Flow cytometry and Hoechst staining were used for analyzing cell apoptosis. The methylation status and expression of RUNX3 in mRNA and protein levels were measured by methylation-specific polymerase chain reaction (PCR [MSP]), reverse transcription (RT)-PCR, and Western blot. It was shown that the RUNX3 gene downregulated and hypermethylated in MCF-7 cells. 5-Aza-CdR induced demethylation, upregulated the expression of RUNX3 on both mRNA and protein levels in cancer cells, and induced growth suppression and apoptosis in vitro in a dose- and time-dependent manner. The results demonstrate that RUNX3 downregulation in breast cancer is frequently due to hypermethylation, and that 5-Aza-CdR can inhibit cell proliferation and induce apoptosis by eliminating the methylation status of RUNX3 promoter and restoring its expression.

  12. The continuous kinetic determination of p-nitroanisole O-demethylation in hemoglobin-free perfused rat liver.

    PubMed

    Thurman, R G; Marazzo, D P; Jones, L S; Kauffman, F C

    1977-05-01

    P-Nitroanisole O-demethylation by perfused rat liver based on the spectral properties of the product, p-nitrophenolate, was determined continuously. Rates of p-nitrophenol production in this system were sensitive to inhibition by CO. p-Nitrophenolate production by livers of normal animals was linear for up to 30 minutes; however, rates were only linear for 1 to 2 minutes followed by a steady decline in induced (6-fold) livers from phenobarbital-treated rats. Only a small portion (24%) of this steady decline could be accounted for by the formation of conjugation products. Additionally, infusion of p-nitrophenol (14 micronM) was not associated with a decline in rate. The decline in rate in induced livers was reversed by glucose, suggesting that an intimate relationship may exist between drug and carbohydrate metabolism in the intact liver. Alteration in rates of p-nitroanisole metabolism with various inducing agents of the mixed-function oxidation system (phenobarbital; ethanol) produced parallel changes in rates of hepatic lactate production, most likely reflecting the aciton of p-nitrophenol to uncouple oxidative phosphorylation. The data support the hypothesis that the decline in rate in p-nitroanisole O-demethylation in livers from phenobarbital-treated rats is due to reduced availability of NADPH for mixed-function oxidation.

  13. Induced Pib Expression and Resistance to Magnaporthe grisea are Compromised by Cytosine Demethylation at Critical Promoter Regions in Rice.

    PubMed

    Li, Yuan; Xia, Qiong; Kou, Hongping; Wang, Dan; Lin, Xiuyun; Wu, Ying; Xu, Chunming; Xing, Shaochen; Liu, Bao

    2011-10-01

    Pib is a well-characterized rice blast-resistance gene belonging to the nucleotide binding site (NBS) and leucine-rich repeat (LRR) superfamily. Expression of Pib was low under non-challenged conditions, but strongly induced by the blast-causing fungal pathogen Magnaporthe grisea, thereby conferring resistance to the pathogen. It is generally established that cytosine methylation of the promoter-region often plays a repressive role in modulating expression of the gene in question. We report here that two critical regions of the Pib promoter were heavily CG cytosine-methylated in both cultivars studied. Surprisingly, induced expression of Pib by M. grisea infection did not entail its promoter demethylation, and partial demethylation by 5-azacytidine-treatment actually reduced Pib expression relative to wild-type plants. Accordingly, the blast disease-resistance was compromised in the 5'-azaC-treated plants relative to wild-type. In contrast, the disease susceptibility was not affected by the 5'-azaC treatment in another two rice cultivars that did not contain the Pib gene, ruling out effects of other R genes and non-specific genotoxic effects by the drug-treatment as a cause for the compromised Pib-conditioned blast-resistance. Taken together, our results suggest that promoter DNA methylation plays a novel enhancing role in conditioning high-level of induced expression of the Pib gene in times of M. grisea infection, and its conferred resistance to the pathogen.

  14. The consent process: Enabling or disabling patients’ active participation?

    PubMed Central

    Doherty, Carole; Stavropoulou, Charitini; Saunders, Mark NK; Brown, Tracey

    2015-01-01

    Standards expected by doctors’ regulatory bodies in respect of the process of consent to treatment have arguably sought to restructure the nature of the doctor–patient relationship from one of the paternalism to that of shared decision-making. Yet, few studies have explored empirically, from patients’ perspectives, the extent to which the process of consent to treatment enables or disables patients’ participation in medical decision-making. Our article examines patients’ attitudes towards the consent process, exploring how and why these attitudes influence patients’ active participation in decision-making and considering possible consequent medico-legal issues. Data were collected longitudinally using semi-structured interviews and field observations involving 35 patients and 19 of their caregivers, in an English hospital between February and November 2014. These indicate that generally patients defer to the doctor in respect of treatment decision-making. Although most patients and their caregivers wanted detailed information and discussion, they did not necessarily expect that this would be provided. Furthermore, patients perceived that signing the consent form was an obligatory routine principally to protect doctors from legal action should something go wrong. Our study suggests that patients’ predominantly paternalistic perceptions of the consent process can not only undermine attempts by doctors to involve them in decision-making but, as patients are now considered in law as informed actors, their perceptions of the consent form as not being in their interests could be a self-fulfilling prophecy if signing is undertaken without due consideration to the content. PMID:26487687

  15. A novel diterpene skeleton: identification of a highly aromatic, cytotoxic and antioxidant 5-methyl-10-demethyl-abietane-type diterpene from Premna serratifolia.

    PubMed

    Habtemariam, Solomon; Varghese, George K

    2015-01-01

    Premna serratifolia Linn. (syn: . P. corymbosa (Burm. f.) Merr., P. integrifolia L. and P. obtusifolia R. Br.) is a member of the Verbenaceae family that is extensively used in the Ayurvedic system of medicine in India. As part of our continuous pharmacological and phytochemical studies on medicinal plants, we have screened the methanolic extracts of leaves, root bark (RB) and root wood of P. serratifolia for cytotoxic activity against two cancer cell lines: SHSY-5Y neuroblastoma and B16 melanoma cells. The RB extract that showed promising activity was fractionated using solvents of increasing polarity followed by a combination of Sephadex LH-20 column and Combiflash chromatography as well as HPLC to afford the active principle. Comprehensive spectroscopic analysis including 1D and 2D NMR (COSY, HMQC, HMBC, NOESY) and MS analysis revealed the identity of the isolated compound as 11,12,16-trihydroxy-2-oxo-5-methyl-10-demethyl-abieta-1[10],6,8,11,13-pentene that appears to be a novel compound based on a new diterpene skeleton. The cytotoxic activity of the isolated compound was 21 and 23 times higher than the crude extract against the SHSY-5Y and B16 cells, respectively. The novel compound also possesses in vitro antioxidant effects as evidenced by the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging effect where an IC50 value of 20.4 ± 1.3 μM was obtained. In comparison, the positive control, caffeic acid, showed an IC50 value of 14.4 ± 1.6 μM.

  16. Imaging Active Surface Processes in Barnacle Adhesive Interfaces.

    PubMed

    Golden, Joel P; Burden, Daniel K; Fears, Kenan P; Barlow, Daniel E; So, Christopher R; Burns, Justin; Miltenberg, Benjamin; Orihuela, Beatriz; Rittshof, Daniel; Spillmann, Christopher M; Wahl, Kathryn J; Tender, Leonard M

    2016-01-19

    Surface plasmon resonance imaging (SPRI) and voltammetry were used simultaneously to monitor Amphibalanus (=Balanus) amphitrite barnacles reattached and grown on gold-coated glass slides in artificial seawater. Upon reattachment, SPRI revealed rapid surface adsorption of material with a higher refractive index than seawater at the barnacle/gold interface. Over longer time periods, SPRI also revealed secretory activity around the perimeter of the barnacle along the seawater/gold interface extending many millimeters beyond the barnacle and varying in shape and region with time. Ex situ experiments using attenuated total reflectance infrared (ATR-IR) spectroscopy confirmed that reattachment of barnacles was accompanied by adsorption of protein to surfaces on similar time scales as those in the SPRI experiments. Barnacles were grown through multiple molting cycles. While the initial reattachment region remained largely unchanged, SPRI revealed the formation of sets of paired concentric rings having alternately darker/lighter appearance (corresponding to lower and higher refractive indices, respectively) at the barnacle/gold interface beneath the region of new growth. Ex situ experiments coupling the SPRI imaging with optical and FTIR microscopy revealed that the paired rings coincide with molt cycles, with the brighter rings associated with regions enriched in amide moieties. The brighter rings were located just beyond orifices of cement ducts, consistent with delivery of amide-rich chemistry from the ducts. The darker rings were associated with newly expanded cuticle. In situ voltammetry using the SPRI gold substrate as the working electrode revealed presence of redox active compounds (oxidation potential approx 0.2 V vs Ag/AgCl) after barnacles were reattached on surfaces. Redox activity persisted during the reattachment period. The results reveal surface adsorption processes coupled to the complex secretory and chemical activity under barnacles as they construct

  17. Observations on Multi-Slug Activity - Implications for Volcanic Processes

    NASA Astrophysics Data System (ADS)

    Pering, T. D.; McGonigle, A. J. S.; James, M. R.; Lane, S. J.; Capponi, A.; Tamburello, G.; Aiuppa, A.

    2014-12-01

    The study of single gas slugs in volcanic conduits has received a large amount of focus within the literature. However, the more complex behaviour associated with the rise and burst of multiple slugs has yet to be considered in detail in a volcanic context. Here we combine observations and analyses of such activity using a three-pronged approach consisting of existing gas mass data collected during rapid slug driven activity at Mt. Etna, scaled laboratory analogue experiments, and computer simulations using the Ansys Fluent® fluid dynamics software. Particular focus was applied to the process of coalescence and wake capture during slug expansion and rise. The results indicate a variety of potential features and relationships, including: promotion of coalescence at distances further than predicted wake lengths, approximate maximum gas volume fraction and minimum magma viscosity values for the occurrence of stable multi-slug activity, and in the laboratory regimes a series of linear trends are associated with overall gas volume fraction and burst volume. A previously observed phenomenon at Mt. Etna, whereby larger slug bursting events are subject to a longer repose period prior to the following event, than smaller events, is also evident in the lab setting. By combining all acquired and modelled data, we derive an approximate relation, using existing formulae for slug base rise speed (Viana et al. 2003) and wake length (Campos and Guedes de Carvalho, 1988), to describe a minimum repose period which is likely to follow the burst of a slug at the surface. The outlined work has significant fluid dynamic implications for possible magma and conduit properties which can allow multi-slug activity at volcanic targets.

  18. AVIRIS and TIMS data processing and distribution at the land processes distributed active archive center

    NASA Technical Reports Server (NTRS)

    Mah, G. R.; Myers, J.

    1993-01-01

    The U.S. Government has initiated the Global Change Research program, a systematic study of the Earth as a complete system. NASA's contribution of the Global Change Research Program is the Earth Observing System (EOS), a series of orbital sensor platforms and an associated data processing and distribution system. The EOS Data and Information System (EOSDIS) is the archiving, production, and distribution system for data collected by the EOS space segment and uses a multilayer architecture for processing, archiving, and distributing EOS data. The first layer consists of the spacecraft ground stations and processing facilities that receive the raw data from the orbiting platforms and then separate the data by individual sensors. The second layer consists of Distributed Active Archive Centers (DAAC) that process, distribute, and archive the sensor data. The third layer consists of a user science processing network. The EOSDIS is being developed in a phased implementation. The initial phase, Version 0, is a prototype of the operational system. Version 0 activities are based upon existing systems and are designed to provide an EOSDIS-like capability for information management and distribution. An important science support task is the creation of simulated data sets for EOS instruments from precursor aircraft or satellite data. The Land Processes DAAC, at the EROS Data Center (EDC), is responsible for archiving and processing EOS precursor data from airborne instruments such as the Thermal Infrared Multispectral Scanner (TIMS), the Thematic Mapper Simulator (TMS), and Airborne Visible and Infrared Imaging Spectrometer (AVIRIS). AVIRIS, TIMS, and TMS are flown by the NASA-Ames Research Center ARC) on an ER-2. The ER-2 flies at 65000 feet and can carry up to three sensors simultaneously. Most jointly collected data sets are somewhat boresighted and roughly registered. The instrument data are being used to construct data sets that simulate the spectral and spatial

  19. Criminal profiling as a plotting activity based on abductive processes.

    PubMed

    Verde, Alfredo; Nurra, Antonio

    2010-10-01

    In this article the authors analyze the nature and aims of criminal profiling from a theoretical point of view. The need to become increasingly "scientific" has given rise to the modern approaches of profiling, which have been particularly successful in cases of serial homicides and sex crimes, given that compulsive (perverse) acts, because of their ritual nature, have been described as being more easily foreseeable and presumably linkable to the psychological and even personal characteristics of a given criminal. On this basis, the authors analyze profiling from an epistemological point of view and show how, in the concrete activity of profiling, profilers depart from the "certainty" of the scientific models (those that are based on deductive-inductive processes); the epistemological basis of reasoning changes as there is no longer an induction-deduction model but rather an abductive model (as conceived and explained by Peirce) in which the importance of plotting (the weaving of a narrative) becomes greater.

  20. Linking Plagioclase Zoning Patterns to Active Magma Processes

    NASA Astrophysics Data System (ADS)

    Izbekov, P. E.; Nicolaysen, K. P.; Neill, O. K.; Shcherbakov, V.; Plechov, P.; Eichelberger, J. C.

    2015-12-01

    Plagioclase, one of the most common and abundant mineral phases in volcanic products, will vary in composition in response to changes in temperature, pressure, composition of the ambient silicate melt, and melt H2O concentration. Changes in these parameters may cause dissolution or growth of plagioclase crystals, forming characteristic textural and compositional variations (zoning patterns), the complete core-to-rim sequence of which describes events experienced by an individual crystal from its nucleation to the last moments of its growth. Plagioclase crystals in a typical volcanic rock may look drastically dissimilar despite their spatial proximity and the fact that they have erupted together. Although they shared last moments of their growth during magma ascent and eruption, their prior experiences could be very different, as plagioclase crystals often come from different domains of the same magma system. Distinguishing similar zoning patterns, correlating them across the entire population of plagioclase crystals, and linking these patterns to specific perturbations in the magmatic system may provide additional perspective on the variety, extent, and timing of magma processes at active volcanic systems. Examples of magma processes, which may be distinguished based on plagioclase zoning patterns, include (1) cooling due to heat loss, (2) heating and/or pressure build up due to an input of new magmatic material, (3) pressure drop in response to magma system depressurization, and (4) crystal transfer between different magma domains/bodies. This review will include contrasting examples of zoning patters from recent eruptions of Karymsky, Bezymianny, and Tolbachik Volcanoes in Kamchatka, Augustine and Cleveland Volcanoes in Alaska, as well as from the drilling into an active magma body at Krafla, Iceland.

  1. Activation processes in a medical linear accelerator and spatial distribution of activation products.

    PubMed

    Fischer, Helmut W; Tabot, Ben E; Poppe, Björn

    2006-12-21

    Activation products have been identified by in situ gamma spectroscopy at the isocentre of a medical linear accelerator shortly after termination of a high energy photon beam irradiation with 15 x 15 cm field size. Spectra have been recorded either with an open or with a closed collimator. Whilst some activation products disappear from the spectrum with closed collimator or exhibit reduced count rates, others remain with identical intensity. The former isotopes are neutron-deficient and mostly decay by positron emission or electron capture; the latter have neutron excess and decay by beta(-) emission. This new finding is consistent with the assumption that photons in the primary beam produce activation products by (gamma, n) reactions in the treatment head and subsequently the neutrons created in these processes undergo (n, gamma) reactions creating activation products in a much larger area. These findings are expected to be generally applicable to all medical high energy linear accelerators.

  2. Coordinated activities of human dicer domains in regulatory RNA processing.

    PubMed

    Ma, Enbo; Zhou, Kaihong; Kidwell, Mary Anne; Doudna, Jennifer A

    2012-09-28

    The conserved ribonuclease Dicer generates microRNAs and short-interfering RNAs that guide gene silencing in eukaryotes. The specific contributions of human Dicer's structural domains to RNA product length and substrate preference are incompletely understood, due in part to the difficulties of Dicer purification. Here, we show that active forms of human Dicer can be assembled from recombinant polypeptides expressed in bacteria. Using this system, we find that three distinct modes of RNA recognition give rise to Dicer's fidelity and product length specificity. The first involves anchoring one end of a double-stranded RNA helix within the PAZ domain, which can assemble in trans with Dicer's catalytic domains to reconstitute an accurate but non-substrate-selective dicing activity. The second entails nonspecific RNA binding by the double-stranded RNA binding domain, an interaction that is essential for substrate recruitment in the absence of the PAZ domain. The third mode of recognition involves hairpin RNA loop recognition by the helicase domain, which ensures efficient processing of specific substrates. These results reveal distinct interactions of each Dicer domain with different RNA structural features and provide a facile system for investigating the molecular mechanisms of human microRNA biogenesis.

  3. Inhibition of H3K27me3 Histone Demethylase Activity Prevents the Proliferative Regeneration of Zebrafish Lateral Line Neuromasts

    PubMed Central

    Bao, Beier; He, Yingzi; Tang, Dongmei; Li, Wenyan; Li, Huawei

    2017-01-01

    The H3K27 demethylases are involved in a variety of biological processes, including cell differentiation, proliferation, and cell death by regulating transcriptional activity. However, the function of H3K27 demethylation in the field of hearing research is poorly understood. Here, we investigated the role of H3K27me3 histone demethylase activity in hair cell regeneration using an in vivo animal model. Our data showed that pharmacologic inhibition of H3K27 demethylase activity with the specific small-molecule inhibitor GSK-J4 decreased the number of regenerated hair cells in response to neomycin damage. Furthermore, inhibition of H3K27me3 histone demethylase activity dramatically suppressed cell proliferation and activated caspase-3 levels in the regenerating neuromasts of the zebrafish lateral line. GSK-J4 administration also increased the expression of p21 and p27 in neuromast cells and inhibited the ERK signaling pathway. Collectively, our findings indicate that H3K27me3 demethylation is a key epigenetic regulator in the process of hair cell regeneration in zebrafish and suggest that H3K27me3 histone demethylase activity might be a novel therapeutic target for the treatment of hearing loss. PMID:28348517

  4. A novel glycylcycline, 9-(N,N-dimethylglycylamido)-6-demethyl-6-deoxytetracycline, is neither transported nor recognized by the transposon Tn10-encoded metal-tetracycline/H+ antiporter.

    PubMed Central

    Someya, Y; Yamaguchi, A; Sawai, T

    1995-01-01

    A novel tetracycline derivative, DMG-DMDOT [9-(N,N-dimethylglycylamido)-6-demethyl-6-deoxytetracycline] , is one of the glycylcyclines which have a broad antibacterial spectrum, including many tetracyclineresistant bacteria (R.T. Testa, P.J. Petersen, N.V. Jacobus, P.-E. Sum, V.J. Lee, and F.P. Tally, Antimicrob. Agents Chemother. 37:2270-2277, 1993). The mechanism by which DMG-DMDOT overcomes efflux-based tetracycline resistance was investigated. Tetracycline-resistant Escherichia coli cells carrying an R plasmid encoding the tet(B) gene, which encodes the typical tetracycline efflux pump [TetA(B)] of gram-negative bacteria, were as susceptible to DMG-DMDOT as was the tetracycline-susceptible host. When mid-log-phase cells carrying the tet(B) gene were incubated with a subbactericidal concentration of DMG-DMDOT (0.5 micrograms/ml) for 2 h, a significant amount of the TetA(B) protein was detected in the cell membrane by Western blotting (immunoblotting) with an anti-carboxyl-terminal antibody, similar to the case in which tetracycline was used as the inducer, indicating that the tet repressor, TetR, can recognize DMG-DMDOT as an efficient inducer. Everted membrane vesicles prepared from cells producing the TetA(B) protein showed absolutely no transport activity for DMG-DMDOT. Furthermore, the presence of excess DMG-DMDOT had no effect on the tetracycline transport activity of the everted vesicles, indicating that DMG-DMDOT is not recognized as a substrate by the TetA(B) protein. PMID:7695316

  5. Genistein downregulates onco-miR-1260b and upregulates sFRP1 and Smad4 via demethylation and histone modification in prostate cancer cells

    PubMed Central

    Hirata, H; Hinoda, Y; Shahryari, V; Deng, G; Tanaka, Y; Tabatabai, Z L; Dahiya, R

    2014-01-01

    Background: Recently several microRNAs (miRNAs) have been found to be regulated by genistein in cancer cells. In this study, we focused on the gene regulatory effect of genistein on microRNA and its target genes in prostate cancer (PC). Methods: Initially, we investigated the effect of genistein on prostate cancer cells and identified that the expression of miRNA-1260b was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA-1260b expression and prostate cancer patient outcomes. Two target genes (sFRP1 and Smad4) of miR-1260b were identified based on computer algorithm and 3′UTR luciferase assay was carried out to determine direct miRNA regulation of the genes. Results: Genistein promoted apoptosis while inhibiting prostate cancer cell proliferation, invasion and TCF reporter activity in PC cells. MiR-1260b was highly expressed in prostate cancer tissues and significantly downregulated by genistein in PC cells. After knocking down miR-1260b, cell proliferation, invasion, migration and TCF reporter activity were decreased in PC cells. Western analysis and 3′UTR luciferase assay showed that the two target genes (sFRP1 and Smad4) were directly regulated by miR-1260b. The expression of sFRP1 and Smad4 was significantly decreased in prostate cancer tissues. Genistein also increased expression of these two genes via DNA demethylation and histone modifications. Conclusions: Our data suggest that genistein exerts its anti-tumour effect via downregulation of miR-1260b that targeted sRRP1 and Smad4 genes in prostate cancer cells. The expression of sFRP1 and Smad4 was also modulated by genistein via DNA methylation or histone modifications in PC cell lines. PMID:24504368

  6. Edge effect modeling and experiments on active lap processing.

    PubMed

    Liu, Haitao; Wu, Fan; Zeng, Zhige; Fan, Bin; Wan, Yongjian

    2014-05-05

    Edge effect is regarded as one of the most difficult technical issues for fabricating large primary mirrors, especially for large polishing tools. Computer controlled active lap (CCAL) uses a large size pad (e.g., 1/3 to 1/5 workpiece diameters) to grind and polish the primary mirror. Edge effect also exists in the CCAL process in our previous fabrication. In this paper the material removal rules when edge effects happen (i.e. edge tool influence functions (TIFs)) are obtained through experiments, which are carried out on a Φ1090-mm circular flat mirror with a 375-mm-diameter lap. Two methods are proposed to model the edge TIFs for CCAL. One is adopting the pressure distribution which is calculated based on the finite element analysis method. The other is building up a parametric equivalent pressure model to fit the removed material curve directly. Experimental results show that these two methods both effectively model the edge TIF of CCAL.

  7. Active microchannel fluid processing unit and method of making

    DOEpatents

    Bennett, Wendy D [Kennewick, WA; Martin, Peter M [Kennewick, WA; Matson, Dean W [Kennewick, WA; Roberts, Gary L [West Richland, WA; Stewart, Donald C [Richland, WA; Tonkovich, Annalee Y [Pasco, WA; Zilka, Jennifer L [Pasco, WA; Schmitt, Stephen C [Dublin, OH; Werner, Timothy M [Columbus, OH

    2002-12-10

    The present invention is an active microchannel fluid processing unit and method of making, both relying on having (a) at least one inner thin sheet; (b) at least one outer thin sheet; (c) defining at least one first sub-assembly for performing at least one first unit operation by stacking a first of the at least one inner thin sheet in alternating contact with a first of the at least one outer thin sheet into a first stack and placing an end block on the at least one inner thin sheet, the at least one first sub-assembly having at least a first inlet and a first outlet; and (d) defining at least one second sub-assembly for performing at least one second unit operation either as a second flow path within the first stack or by stacking a second of the at least one inner thin sheet in alternating contact with second of the at least one outer thin sheet as a second stack, the at least one second sub-assembly having at least a second inlet and a second outlet.

  8. Active microchannel fluid processing unit and method of making

    DOEpatents

    Bennett, Wendy D [Kennewick, WA; Martin, Peter M [Kennewick, WA; Matson, Dean W [Kennewick, WA; Roberts, Gary L [West Richland, WA; Stewart, Donald C [Richland, WA; Tonkovich, Annalee Y [Pasco, WA; Zilka, Jennifer L [Pasco, WA; Schmitt, Stephen C [Dublin, OH; Werner, Timothy M [Columbus, OH

    2001-01-01

    The present invention is an active microchannel fluid processing unit and method of making, both relying on having (a) at least one inner thin sheet; (b) at least one outer thin sheet; (c) defining at least one first sub-assembly for performing at least one first unit operation by stacking a first of the at least one inner thin sheet in alternating contact with a first of the at least one outer thin sheet into a first stack and placing an end block on the at least one inner thin sheet, the at least one first sub-assembly having at least a first inlet and a first outlet; and (d) defining at least one second sub-assembly for performing at least one second unit operation either as a second flow path within the first stack or by stacking a second of the at least one inner thin sheet in alternating contact with second of the at least one outer thin sheet as a second stack, the at least one second sub-assembly having at least a second inlet and a second outlet.

  9. Activation of Peroxymonosulfate by Benzoquinone: A Novel Nonradical Oxidation Process.

    PubMed

    Zhou, Yang; Jiang, Jin; Gao, Yuan; Ma, Jun; Pang, Su-Yan; Li, Juan; Lu, Xue-Ting; Yuan, Li-Peng

    2015-11-03

    The reactions between peroxymonosulfate (PMS) and quinones were investigated for the first time in this work, where benzoquinone (BQ) was selected as a model quinone. It was demonstrated that BQ could efficiently activate PMS for the degradation of sulfamethoxazole (SMX; a frequently detected antibiotic in the environments), and the degradation rate increased with solution pH from 7 to 10. Interestingly, quenching studies suggested that neither hydroxyl radical (•OH) nor sulfate radical (SO4•-) was produced therein. Instead, the generation of singlet oxygen (1O2) was proved by using two chemical probes (i.e., 2,2,6,6-tetramethyl-4-piperidinol and 9,10-diphenylanthracene) with the appearance of 1O2 indicative products detected by electron paramagnetic resonance spectrometry and liquid chromatography mass spectrometry, respectively. A catalytic mechanism was proposed involving the formation of a dioxirane intermediate between PMS and BQ and the subsequent decomposition of this intermediate into 1O2. Accordingly, a kinetic model was developed, and it well described the experimental observation that the pH-dependent decomposition rate of PMS was first-order with respect to BQ. These findings have important implications for the development of novel nonradical oxidation processes based on PMS, because 1O2 as a moderately reactive electrophile may suffer less interference from background organic matters compared with nonselective •OH and SO4•-.

  10. Ether Cleavage Re-Investigated: Elucidating the Mechanism of BBr3-Facilitated Demethylation of Aryl Methyl Ethers

    PubMed Central

    Kosak, Talon M; Conrad, Heidi A; Korich, Andrew L; Lord, Richard L

    2015-01-01

    One of the most well-known, highly utilized reagents for ether cleavage is boron tribromide (BBr3), and this reagent is frequently employed in a 1:1 stoichiometric ratio with ethers. Density functional theory calculations predict a new mechanistic pathway involving charged intermediates for ether cleavage in aryl methyl ethers. Moreover, these calculations predict that one equivalent of BBr3 can cleave up to three equivalents of anisole, producing triphenoxyborane [B(OPh)3] prior to hydrolysis. These predictions were validated by gas chromatography analysis of reactions where the BBr3:anisole ratio was varied. Not only do we confirm that sub-stoichiometric equivalents may be used for ether demethylation, but the findings also support our newly proposed three cycle mechanism for cleavage of aryl methyl ethers. PMID:26693209

  11. Methods for Individualized Determination of Methylmercury Elimination Rate and De-Methylation Status in Humans Following Fish Consumption

    PubMed Central

    Rand, Mathew D.; Vorojeikina, Daria; van Wijngaarden, Edwin; Jackson, Brian P.; Scrimale, Thomas; Zareba, Grazyna; Love, Tanzy M.; Myers, Gary J.; Watson, Gene E.

    2016-01-01

    Methylmercury (MeHg) exposure via fish in the diet remains a priority public health concern. Individual variation in response to a given MeHg exposure and the biotransformation of MeHg that follows complicate our understanding of this issue. MeHg elimination from the human body occurs slowly (elimination rate (kel) approximately 0.01 day−1 or approximately 70 days half-life [t1/2]) and is a major determinant of the Hg body burden resulting from fish consumption. The underlying mechanisms that control MeHg elimination from the human body remain poorly understood. We describe here improved methods to obtain a MeHg elimination rate via longitudinal Hg analysis in hair using laser ablation-inductively coupled plasma-mass spectrometry. We measured MeHg elimination rates in eight individuals following the consumption of 3 fish meals in two 75-day trials separated by a 4-month washout period. In addition, since MeHg biotransformation to inorganic Hg (I-Hg) is associated with Hg excretion, we speciated Hg in feces samples to estimate individual MeHg de-methylation status. We observed a wide range of MeHg elimination rates between individuals and within individuals over time (kel = 0.0163–0.0054 day−1; estimated t1/2 = 42.5–128.3 days). The ratio of MeHg and I-Hg in feces also varied widely among individuals. While the %I-Hg in feces was likely influenced by dental amalgams, findings with subjects who lacked amalgams suggest that faster MeHg elimination is associated with a higher %I-Hg in feces indicating more complete de-methylation. We anticipate these methods will contribute to future investigations of genetic and dietary factors that influence MeHg disposition in people. PMID:26572661

  12. Combination of lenalidomide with vitamin D3 induces apoptosis in mantle cell lymphoma via demethylation of BIK

    PubMed Central

    Brosseau, C; Dousset, C; Touzeau, C; Maïga, S; Moreau, P; Amiot, M; Le Gouill, S; Pellat-Deceunynck, C

    2014-01-01

    Mantle cell lymphoma (MCL) is a currently incurable B-cell malignancy. Lenalidomide (Len) has been demonstrated to be one of the most efficient new treatment options. Because Len and 1α,25-dihydroxyvitamin (VD3) synergize to kill breast cancer cells, we investigated whether VD3 could increase the ability of Len to induce MCL cell death. While MCL cells were weakly sensitive to Len (1 μM), the addition of VD3 at physiological dose (100 nM) strongly increased cell death, accompanied by slowdown in cell cycle progression in MCL cell lines (n=4 out of 6) and primary samples (n=5 out of 7). The Len/VD3 treatment markedly increased the expression of the BH3-only BCL2-interacting killer (Bik) without affecting the expression of other Bcl-2 molecules. Immunoprecipitation assays demonstrated that Bik was free from anti-apoptotic partners, Bcl-2 and Bcl-xL, in treated cells. Moreover, silencing of BIK prevented apoptosis induced by Len/VD3, confirming the direct involvement of Bik in cell death. Bik accumulation induced by Len/VD3 was related to an increase in BIK mRNA levels, which resulted from a demethylation of BIK CpG islands. The sensitivity of MCL cells to Len/VD3 was similar to the response to 5-azacytidine, which also induced demethylation of BIK CpG islands. These preclinical data provide the rationale to investigate the role of VD3 in vivo in the response to Len. PMID:25165875

  13. [Phenylhexyl isothiocyanate induces gene p15 re-expression by regulating histone methylation and DNA demethylation in Molt-4 cells].

    PubMed

    Ma, Xu-Dong; Huang, Yi-Qun; Jiang, Shao-Hong; Zheng, Rui-Ji

    2010-06-01

    This study was aimed to investigate the regulatory effect of phenylhexyl isothiocyanate (PHI) on methylation of histone H3K4, H3K9 and demethylation of p15 gene in acute leukemia cell line Molt-4, and to explore the possible mechanism inducing re-expression of silent gene. The methylation status of histone H3K4, H3K9 and the expression of P15 protein in the Molt-4 cells treated with PHI were detected by Western blot; the methylation status of p15 gene in the Molt-4 cells before and after treatment with PHI was determined by methylation specific polymerase chain reaction (MSP); the expression level of p15 gene mRNA in Molt-4 cells treated with PHI was assayed by semiquantitative reverse transcription-PCR. The results indicated that the PHI could increase methylation of histone H3K4 and decrease methylation of histone H3K9 in concentration-and time-dependent manners. After treatment of Molt-4 cells with PHI for 5 days, the methylation of p15 gene was reduced, the significant hypermethylation of p15 gene was reversed, the silenced p15 gene re-expressed; the expressions of p15 mRNA and P15 protein were enhanced in concentration-dependent manner. It is concluded that probably through specifically regulating the methylation level of histone H3K4 and H3K9, the PHI causes the changes of chromosome space structure and results in the demethylation of CPG island in p15 gene, thereby induces the re-expression of p15 gene which was silenced.

  14. Physicochemical and porosity characteristics of thermally regenerated activated carbon polluted with biological activated carbon process.

    PubMed

    Dong, Lihua; Liu, Wenjun; Jiang, Renfu; Wang, Zhansheng

    2014-11-01

    The characteristics of thermally regenerated activated carbon (AC) polluted with biological activated carbon (BAC) process were investigated. The results showed that the true micropore and sub-micropore volume, pH value, bulk density, and hardness of regenerated AC decreased compared to the virgin AC, but the total pore volume increased. XPS analysis displayed that the ash contents of Al, Si, and Ca in the regenerated AC respectively increased by 3.83%, 2.62% and 1.8%. FTIR spectrum showed that the surface functional groups of virgin and regenerated AC did not change significantly. Pore size distributions indicated that the AC regeneration process resulted in the decrease of micropore and macropore (D>10 μm) volume and the increase of mesopore and macropore (0.1 μmprocess.

  15. Multiplicative point process as a model of trading activity

    NASA Astrophysics Data System (ADS)

    Gontis, V.; Kaulakys, B.

    2004-11-01

    Signals consisting of a sequence of pulses show that inherent origin of the 1/ f noise is a Brownian fluctuation of the average interevent time between subsequent pulses of the pulse sequence. In this paper, we generalize the model of interevent time to reproduce a variety of self-affine time series exhibiting power spectral density S( f) scaling as a power of the frequency f. Furthermore, we analyze the relation between the power-law correlations and the origin of the power-law probability distribution of the signal intensity. We introduce a stochastic multiplicative model for the time intervals between point events and analyze the statistical properties of the signal analytically and numerically. Such model system exhibits power-law spectral density S( f)∼1/ fβ for various values of β, including β= {1}/{2}, 1 and {3}/{2}. Explicit expressions for the power spectra in the low-frequency limit and for the distribution density of the interevent time are obtained. The counting statistics of the events is analyzed analytically and numerically, as well. The specific interest of our analysis is related with the financial markets, where long-range correlations of price fluctuations largely depend on the number of transactions. We analyze the spectral density and counting statistics of the number of transactions. The model reproduces spectral properties of the real markets and explains the mechanism of power-law distribution of trading activity. The study provides evidence that the statistical properties of the financial markets are enclosed in the statistics of the time interval between trades. A multiplicative point process serves as a consistent model generating this statistics.

  16. Activated Sludge. Student Manual. Biological Treatment Process Control.

    ERIC Educational Resources Information Center

    Boe, Owen K.; Klopping, Paul H.

    This student manual contains the textual material for a seven-lesson unit on activated sludge. Topic areas addressed in the lessons include: (1) activated sludge concepts and components (including aeration tanks, aeration systems, clarifiers, and sludge pumping systems); (2) activated sludge variations and modes; (3) biological nature of activated…

  17. 15 CFR 400.33 - Restrictions on manufacturing and processing activity.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 15 Commerce and Foreign Trade 2 2012-01-01 2012-01-01 false Restrictions on manufacturing and...-TRADE ZONES BOARD Manufacturing and Processing Activity-Reviews § 400.33 Restrictions on manufacturing and processing activity. (a) In general. In approving manufacturing or processing activity for a...

  18. Demethylation drug 5-Aza-2′-deoxycytidine-induced upregulation of miR-200c inhibits the migration, invasion and epithelial-mesenchymal transition of clear cell renal cell carcinoma in vitro

    PubMed Central

    JIANG, JUAN; YI, BO; DING, SIQING; SUN, JIAN; CAO, WEI; LIU, MENGZI

    2016-01-01

    The microRNA (miR)-200 family has been found to be involved in the process of mesenchymal-epithelial transition during renal development. Deregulation of miR-200c has been suggested to be involved in clear cell renal cell carcinoma (ccRCC). However, the precise role of miR-200c in the regulation of ccRCC metastasis has not been previously reported. In the present study, it was observed that miR-200c was frequently downregulated in ccRCC tissue compared with matched adjacent normal tissue. The expression of miR-200c was additionally reduced in ccRCC cell lines when compared with levels in normal renal cells. The DNA demethylation drug 5-Aza-2′-deoxycytidine (Aza) was used to treat several ccRCC cell lines, and it was observed that the expression of miR-200c was significantly increased following Aza treatment. Furthermore, treatment with Aza markedly inhibited ccRCC cell invasion and migration, while treatment with miR-200c inhibitor significantly enhanced invasion and migration of ccRCC cells. In addition, Aza treatment significantly promoted expression of E-cadherin and inhibited the expression of N-cadherin, while the inhibition of miR-200c downregulated E-cadherin and upregulated the expression of N-cadherin, suggesting that miR-200c has a suppressive role in epithelial-mesenchymal transition (EMT) of ccRCC cells. In conclusion, it was suggested that demethylation drug Aza-induced upregulation of miR-200c may inhibit migration, invasion and EMT in ccRCC cells. PMID:27123083

  19. Hypomethylation at the Regulatory T Cell–Specific Demethylated Region in CD25hi T Cells Is Decoupled from FOXP3 Expression at the Inflamed Site in Childhood Arthritis

    PubMed Central

    Pesenacker, Anne M.; Ursu, Simona; Wu, Qiong; Lom, Hannah; Thirugnanabalan, Balathas; Wedderburn, Lucy R.

    2014-01-01

    The maintenance of FOXP3 expression in CD25hi regulatory T cells (Tregs) is crucial to the control of inflammation and essential for successful Treg transfer therapies. Coexpression of CD25 and FOXP3 in combination with a hypomethylated region within the FOXP3 gene, called the Treg-specific demethylated region (TSDR), is considered the hallmark of stable Tregs. The TSDR is an epigenetic motif that is important for stable FOXP3 expression and is used as a biomarker to measure Treg lineage commitment. In this study, we report that, unlike in peripheral blood, CD4+ T cell expression of CD25 and FOXP3 is frequently dissociated at the inflamed site in patients with juvenile idiopathic arthritis, which led us to question the stability of human Tregs in chronic inflammatory environments. We describe a novel CD4+CD127loCD25hi human T cell population that exhibits extensive TSDR and promoter demethylation in the absence of stable FOXP3 expression. This population expresses high levels of CTLA-4 and can suppress T conventional cell proliferation in vitro. These data collectively suggest that this population may represent a chronically activated FOXP3lo Treg population. We show that these cells have defects in IL-2 signaling and reduced expression of a deubiquitinase important for FOXP3 stability. Clinically, the proportions of these cells within the CD25hi T cell subset are increased in patients with the more severe courses of disease. Our study demonstrates, therefore, that hypomethylation at the TSDR can be decoupled from FOXP3 expression in human T cells and that environment-specific breakdown in FOXP3 stability may compromise the resolution of inflammation in juvenile idiopathic arthritis. PMID:25092890

  20. Enhanced Multistatic Active Sonar via Innovative Signal Processing

    DTIC Science & Technology

    2015-09-30

    Grove, CA, November, 2014. [in press, refereed]. C . Gianelli, L. Xu, and J. Li, " Active Sonar Systems in the Presence of Strong Direct Blast", Oceans...3. DATES COVERED (From - To) Oct. 01, 2014-Sept. 30, 2015 4. TITLE AND SUBTITLE Enhanced Multistatic Active Sonar via Innovative Signal... active sonar (CAS) in the presence of strong direct blast is studied for the Doppler-tolerant linear frequency modulation waveform. A receiver design

  1. Transformation of diclofenac in hybrid biofilm-activated sludge processes.

    PubMed

    Jewell, Kevin S; Falås, Per; Wick, Arne; Joss, Adriano; Ternes, Thomas A

    2016-11-15

    The biotransformation of diclofenac during wastewater treatment was investigated. Attached growth biomass from a carrier-filled compartment of a hybrid-MBBR at the wastewater treatment plant (WWTP) in Bad Ragaz, Switzerland was used to test the biotransformation. Laboratory-scale incubation experiments were performed with diclofenac and carriers and high-resolution LC-QTof-MS was implemented to monitor the biotransformation. Up to 20 diclofenac transformation products (TPs) were detected. Tentative structures were proposed for 16 of the TPs after characterization by MS(2) fragmentation and/or inferring the structure from the transformation pathway and the molecular formula given by the high resolution ionic mass. The remaining four TPs were unambiguously identified via analytical reference standards. The postulated reactions forming the TPs were: hydroxylation, decarboxylation, oxidation, amide formation, ring-opening and reductive dechlorination. Incubation experiments of individual TPs, those which were available as reference standards, provided a deeper look into the transformation pathways. It was found that the transformation consists of four main pathways but no pathway accounted for a clear majority of the transformation. A 10-day monitoring campaign of the full-scale plant confirmed an 88% removal of diclofenac (from approximately 1.6 μg/L in WWTP influent) and the formation of TPs as found in the laboratory was observed. One of the TPs, N-(2,6-dichlorophenyl)-2-indolinone detected at concentrations of around 0.25 μg/L in WWTP effluent, accounting for 16% of the influent diclofenac concentration. The biotransformation of carriers was compared to a second WWTP not utilising carriers. It was found that in contact with activated sludge, similar hydroxylation and decarboxylation reactions occurred but at much slower rates, whereas some reactions, e.g. reductive dechlorination, were not detected at all. Finally, incubation experiments were performed with

  2. Active-Learning Processes Used in US Pharmacy Education

    PubMed Central

    Brown, Stacy D.; Clavier, Cheri W.; Wyatt, Jarrett

    2011-01-01

    Objective To document the type and extent of active-learning techniques used in US colleges and schools of pharmacy as well as factors associated with use of these techniques. Methods A survey instrument was developed to assess whether and to what extent active learning was used by faculty members of US colleges and schools of pharmacy. This survey instrument was distributed via the American Association of Colleges of Pharmacy (AACP) mailing list. Results Ninety-five percent (114) of all US colleges and schools of pharmacy were represented with at least 1 survey among the 1179 responses received. Eighty-seven percent of respondents used active-learning techniques in their classroom activities. The heavier the teaching workload the more active-learning strategies were used. Other factors correlated with higher use of active-learning strategies included younger faculty member age (inverse relationship), lower faculty member rank (inverse relationship), and departments that focused on practice, clinical and social, behavioral, and/or administrative sciences. Conclusions Active learning has been embraced by pharmacy educators and is used to some extent by the majority of US colleges and schools of pharmacy. Future research should focus on how active-learning methods can be used most effectively within pharmacy education, how it can gain even broader acceptance throughout the academy, and how the effect of active learning on programmatic outcomes can be better documented. PMID:21769144

  3. Sensory processing and world modeling for an active ranging device

    NASA Technical Reports Server (NTRS)

    Hong, Tsai-Hong; Wu, Angela Y.

    1991-01-01

    In this project, we studied world modeling and sensory processing for laser range data. World Model data representation and operation were defined. Sensory processing algorithms for point processing and linear feature detection were designed and implemented. The interface between world modeling and sensory processing in the Servo and Primitive levels was investigated and implemented. In the primitive level, linear features detectors for edges were also implemented, analyzed and compared. The existing world model representations is surveyed. Also presented is the design and implementation of the Y-frame model, a hierarchical world model. The interfaces between the world model module and the sensory processing module are discussed as well as the linear feature detectors that were designed and implemented.

  4. A Multiscale Survival Process for Modeling Human Activity Patterns

    PubMed Central

    Zhang, Tianyang; Cui, Peng; Song, Chaoming; Zhu, Wenwu; Yang, Shiqiang

    2016-01-01

    Human activity plays a central role in understanding large-scale social dynamics. It is well documented that individual activity pattern follows bursty dynamics characterized by heavy-tailed interevent time distributions. Here we study a large-scale online chatting dataset consisting of 5,549,570 users, finding that individual activity pattern varies with timescales whereas existing models only approximate empirical observations within a limited timescale. We propose a novel approach that models the intensity rate of an individual triggering an activity. We demonstrate that the model precisely captures corresponding human dynamics across multiple timescales over five orders of magnitudes. Our model also allows extracting the population heterogeneity of activity patterns, characterized by a set of individual-specific ingredients. Integrating our approach with social interactions leads to a wide range of implications. PMID:27023682

  5. Valuation of OSA process and folic acid addition as excess sludge minimization alternatives applied in the activated sludge process.

    PubMed

    Martins, C L; Velho, V F; Ramos, S R A; Pires, A S C D; Duarte, E C N F A; Costa, R H R

    2016-01-01

    The aim of this study was to investigate the ability of the oxic-settling-anaerobic (OSA)-process and the folic acid addition applied in the activated sludge process to reduce the excess sludge production. The study was monitored during two distinct periods: activated sludge system with OSA-process, and activated sludge system with folic acid addition. The observed sludge yields (Yobs) were 0.30 and 0.08 kgTSS kg(-1) chemical oxygen demand (COD), control phase and OSA-process (period 1); 0.33 and 0.18 kgTSS kg(-1) COD, control phase and folic acid addition (period 2). The Yobs decreased by 73 and 45% in phases with the OSA-process and folic acid addition, respectively, compared with the control phases. The sludge minimization alternatives result in a decrease in excess sludge production, without negatively affecting the performance of the effluent treatment.

  6. Impact of Non-Poissonian Activity Patterns on Spreading Processes

    NASA Astrophysics Data System (ADS)

    Vazquez, Alexei; Rácz, Balázs; Lukács, András; Barabási, Albert-László

    2007-04-01

    Halting a computer or biological virus outbreak requires a detailed understanding of the timing of the interactions between susceptible and infected individuals. While current spreading models assume that users interact uniformly in time, following a Poisson process, a series of recent measurements indicates that the intercontact time distribution is heavy tailed, corresponding to a temporally inhomogeneous bursty contact process. Here we show that the non-Poisson nature of the contact dynamics results in prevalence decay times significantly larger than predicted by the standard Poisson process based models. Our predictions are in agreement with the detailed time resolved prevalence data of computer viruses, which, according to virus bulletins, show a decay time close to a year, in contrast with the 1 day decay predicted by the standard Poisson process based models.

  7. Saharan Dust, Transport Processes, and Possible Impacts on Hurricane Activities

    NASA Technical Reports Server (NTRS)

    Lau, William K. M.; Kim, K. M.

    2010-01-01

    In this paper, we present observational evidence of significant relationships between Saharan dust outbreak, and African Easterly wave activities and hurricane activities. We found two dominant paths of transport of Saharan dust: a northern path, centered at 25degN associated with eastward propagating 6-19 days waves over northern Africa, and a southern path centered at 15degN, associated with the AEW, and the Atlantic ITCZ. Seasons with stronger dust outbreak from the southern path are associated with a drier atmosphere over the Maximum Development Region (MDR) and reduction in tropical cyclone and hurricane activities in the MDR. Seasons with stronger outbreak from the northern path are associated with a cooler N. Atlantic, and suppressed hurricane in the western Atlantic basin.

  8. Detoxification of pesticide waste via activated carbon adsorption process.

    PubMed

    Foo, K Y; Hameed, B H

    2010-03-15

    Concern about environmental protection has increased over the years from a global viewpoint. To date, the percolation of pesticide waste into the groundwater tables and aquifer systems remains an aesthetic issue towards the public health and food chain interference. With the renaissance of activated carbon, there has been a consistent growing interest in this research field. Confirming the assertion, this paper presents a state of art review of pesticide agrochemical practice, its fundamental characteristics, background studies and environmental implications. Moreover, the key advance of activated carbon adsorption, its major challenges together with the future expectation are summarized and discussed. Conclusively, the expanding of activated carbon adsorption represents a plausible and powerful circumstance, leading to the superior improvement of environmental preservation.

  9. Basic Science Process Skills. An Inservice Workshop Kit: Outlines and Activities.

    ERIC Educational Resources Information Center

    Rowland, Paul; And Others

    A science process skill project was developed to help elementary teachers meet competency standards in New Mexico for teaching the process approach in their science classes. An outline of the process skills along with recommended activities are presented in this document. Performance objectives are identified and a sample activity form is…

  10. The Roles of Structured Input Activities in Processing Instruction and the Kinds of Knowledge They Promote

    ERIC Educational Resources Information Center

    Marsden, Emma; Chen, Hsin-Ying

    2011-01-01

    This study aimed to isolate the effects of the two input activities in Processing Instruction: referential activities, which force learners to focus on a form and its meaning, and affective activities, which contain exemplars of the target form and require learners to process sentence meaning. One hundred and twenty 12-year-old Taiwanese learners…

  11. Adding structure to the transition process to advanced mathematical activity

    NASA Astrophysics Data System (ADS)

    Engelbrecht, Johann

    2010-03-01

    The transition process to advanced mathematical thinking is experienced as traumatic by many students. Experiences that students had of school mathematics differ greatly to what is expected from them at university. Success in school mathematics meant application of different methods to get an answer. Students are not familiar with logical deductive reasoning, required in advanced mathematics. It is necessary to assist students in this transition process, in moving from general to mathematical thinking. In this article some structure is suggested for this transition period. This essay is an argumentative exposition supported by personal experience and international literature. This makes this study theoretical rather than empirical.

  12. A Model for Speech Processing in Second Language Listening Activities

    ERIC Educational Resources Information Center

    Zoghbor, Wafa Shahada

    2016-01-01

    Teachers' understanding of the process of speech perception could inform practice in listening classrooms. Catford (1950) developed a model for speech perception taking into account the influence of the acoustic features of the linguistic forms used by the speaker, whereby the listener "identifies" and "interprets" these…

  13. Informal Learning in the Workplace: Key Activities and Processes

    ERIC Educational Resources Information Center

    Cunningham, John; Hillier, Emilie

    2013-01-01

    Purpose: The purpose of this study is to define characteristics and processes that enhance informal learning in a public sector workplace. Design/methodology/approach: Based on interviews and questionnaires, the authors solicited examples of informal learning practices that 40 supervisors experienced during their careers. The examples were content…

  14. Adding Structure to the Transition Process to Advanced Mathematical Activity

    ERIC Educational Resources Information Center

    Engelbrecht, Johann

    2010-01-01

    The transition process to advanced mathematical thinking is experienced as traumatic by many students. Experiences that students had of school mathematics differ greatly to what is expected from them at university. Success in school mathematics meant application of different methods to get an answer. Students are not familiar with logical…

  15. Application of statistical process control and process capability analysis procedures in orbiter processing activities at the Kennedy Space Center

    NASA Technical Reports Server (NTRS)

    Safford, Robert R.; Jackson, Andrew E.; Swart, William W.; Barth, Timothy S.

    1994-01-01

    Successful ground processing at KSC requires that flight hardware and ground support equipment conform to specifications at tens of thousands of checkpoints. Knowledge of conformance is an essential requirement for launch. That knowledge of conformance at every requisite point does not, however, enable identification of past problems with equipment, or potential problem areas. This paper describes how the introduction of Statistical Process Control and Process Capability Analysis identification procedures into existing shuttle processing procedures can enable identification of potential problem areas and candidates for improvements to increase processing performance measures. Results of a case study describing application of the analysis procedures to Thermal Protection System processing are used to illustrate the benefits of the approaches described in the paper.

  16. Activated Sludge. Instructor's Guide. Biological Treatment Process Control.

    ERIC Educational Resources Information Center

    Boe, Owen K.

    This instructor's guide contains the materials needed to teach a seven-lesson unit on activated sludge. These materials include an overview of the unit, lesson plans, lecture outlines (keyed to slides designed for use with the lessons), student worksheets for each of the seven lessons (with answers), and two copies of a final quiz (with and…

  17. Age-Related Variability in Cortical Activity during Language Processing

    ERIC Educational Resources Information Center

    Fridriksson, Julius; Morrow, K. Leigh; Moser, Dana; Baylis, Gordon C.

    2006-01-01

    Purpose: The present study investigated the extent of cortical activity during overt picture naming using functional magnetic resonance imaging (fMRI). Method: Participants comprised 20 healthy, adult participants with ages ranging from 20 to 82 years. While undergoing fMRI, participants completed a picture-naming task consisting of 60…

  18. An Activity to Teach Students about Schematic Processing

    ERIC Educational Resources Information Center

    Isbell, Linda M.; Tyler, James M.; Burns, Kathleen C.

    2007-01-01

    We designed a classroom activity to foster students' understanding of what schemas are and how they function. We used a video of the instructor as an infant to illustrate how schemas influence gender stereotyping. Before the video, we told students that the baby was either a boy or a girl. After the video, students rated whether the baby would…

  19. English for Everyday Activities: A Picture Process Dictionary.

    ERIC Educational Resources Information Center

    Zwier, Lawrence J.

    These books are designed to help English-as-a-Second-Language (ESL) students learn the skills they need to communicate the step-by-step aspects of daily activities. Unlike most picture dictionaries, this is a verb-based multi-skills program that uses a student text with a clear and colorful pictorial detail as a starting point and focuses on the…

  20. Digital active material processing platform effort (DAMPER), SBIR phase 2

    NASA Technical Reports Server (NTRS)

    Blackburn, John; Smith, Dennis

    1992-01-01

    Applied Technology Associates, Inc., (ATA) has demonstrated that inertial actuation can be employed effectively in digital, active vibration isolation systems. Inertial actuation involves the use of momentum exchange to produce corrective forces which act directly on the payload being actively isolated. In a typical active vibration isolation system, accelerometers are used to measure the inertial motion of the payload. The signals from the accelerometers are then used to calculate the corrective forces required to counteract, or 'cancel out' the payload motion. Active vibration isolation is common technology, but the use of inertial actuation in such systems is novel, and is the focus of the DAMPER project. A May 1991 report was completed which documented the successful demonstration of inertial actuation, employed in the control of vibration in a single axis. In the 1 degree-of-freedom (1DOF) experiment a set of air bearing rails was used to suspend the payload, simulating a microgravity environment in a single horizontal axis. Digital Signal Processor (DSP) technology was used to calculate in real time, the control law between the accelerometer signals and the inertial actuators. The data obtained from this experiment verified that as much as 20 dB of rejection could be realized by this type of system. A discussion is included of recent tests performed in which vibrations were actively controlled in three axes simultaneously. In the three degree-of-freedom (3DOF) system, the air bearings were designed in such a way that the payload is free to rotate about the azimuth axis, as well as translate in the two horizontal directions. The actuator developed for the DAMPER project has applications beyond payload isolation, including structural damping and source vibration isolation. This report includes a brief discussion of these applications, as well as a commercialization plan for the actuator.

  1. Meltlets(®) of soy isoflavones: process optimization and the effect of extrusion spheronization process parameters on antioxidant activity.

    PubMed

    Deshmukh, Ketkee; Amin, Purnima

    2013-07-01

    In the current research work an attempt was made to develop "Melt in mouth pellets" (Meltlets(®)) containing 40% herbal extract of soy isoflavones that served to provide antioxidants activity in menopausal women. The process of extrusion-spheronization was optimized for extruder speed, extruder screen size, spheronization speed, and time. While doing so the herbal extract incorporated in the pellet matrix was subjected to various processing conditions such as the effect of the presence of other excipients, mixing or kneading to prepare wet mass, heat generated during the process of extrusion, spheronization, and drying. Thus, the work further investigates the effect of these processing parameters on the antioxidant activity of the soy isoflavone herbal extract incorporated in the formula. Thereby, the antioxidant activity of the soya bean herbal extract, Meltlets(®) and of the placebo pellets was evaluated using DPPH free radical scavenging assay and total reduction capacity.

  2. Disturbance detection and isolation in the activated sludge process.

    PubMed

    Yoo, C K; Choi, S W; Lee, I

    2002-01-01

    This paper proposes a new fault detection and isolation (FDI) method. This method monitors the distribution of process data and detects changes in this distribution, which reflect changes in the corresponding operating condition. A modified dissimilarity index and a FDI technique are defined to quantitatively evaluate the difference between data sets. This technique considers the importance of each transformed variable in the multivariate system. The FDI technique is applied to a benchmark simulation and to data from a real wastewater treatment plant. Simulation results show that it immediately detects disturbances and automatically distinguishes between serious and minor anomalies for various types of fault. The method not only detects the disturbances, but also isolates the scale of the disturbance, facilitating the interpretation of the disturbance source. The proposed monitoring technique is found to be appropriate for analyzing the biological wastewater treatment process, which is characterized by a variety of fault and disturbance sources and non-stationary characteristics.

  3. Active DNA unwinding dynamics during processive DNA replication.

    PubMed

    Morin, José A; Cao, Francisco J; Lázaro, José M; Arias-Gonzalez, J Ricardo; Valpuesta, José M; Carrascosa, José L; Salas, Margarita; Ibarra, Borja

    2012-05-22

    Duplication of double-stranded DNA (dsDNA) requires a fine-tuned coordination between the DNA replication and unwinding reactions. Using optical tweezers, we probed the coupling dynamics between these two activities when they are simultaneously carried out by individual Phi29 DNA polymerase molecules replicating a dsDNA hairpin. We used the wild-type and an unwinding deficient polymerase variant and found that mechanical tension applied on the DNA and the DNA sequence modulate in different ways the replication, unwinding rates, and pause kinetics of each polymerase. However, incorporation of pause kinetics in a model to quantify the unwinding reaction reveals that both polymerases destabilize the fork with the same active mechanism and offers insights into the topological strategies that could be used by the Phi29 DNA polymerase and other DNA replication systems to couple unwinding and replication reactions.

  4. Participation of CYP2C8 and CYP3A4 in the N-demethylation of imatinib in human hepatic microsomes

    PubMed Central

    Nebot, Noelia; Crettol, Severine; d'Esposito, Fabrizio; Tattam, Bruce; Hibbs, David E; Murray, Michael

    2010-01-01

    BACKGROUND AND PURPOSE Imatinib is a clinically important inhibitor of tyrosine kinases that are dysregulated in chronic myelogenous leukaemia and gastrointestinal stromal tumours. Inter-individual variation in imatinib pharmacokinetics is extensive, and influences drug safety and efficacy. Hepatic cytochrome P450 (CYP) 3A4 has been implicated in imatinib N-demethylation, but the clearance of imatinib decreases during prolonged therapy. CYP3A phenotype correlates with imatinib clearance at the commencement of therapy, but not at steady state. The present study evaluated the possibility that multiple CYPs may contribute to imatinib oxidation in liver. EXPERIMENTAL APPROACH Imatinib biotransformation in human liver microsomes (n = 20) and by cDNA-expressed CYPs was determined by LC–MS. Relationships between imatinib N-demethylation and other drug metabolizing CYPs were assessed. KEY RESULTS N-desmethylimatinib formation was correlated with microsomal oxidation of the CYP3A4 substrates testosterone (ρ= 0.60; P < 0.01) and midazolam (ρ= 0.46; P < 0.05), and the CYP2C8 substrate paclitaxel (ρ= 0.58; P < 0.01). cDNA-derived CYPs 2C8, 3A4, 3A5 and 3A7 supported imatinib N-demethylation, but 10 other CYPs were inactive; in kinetic studies, CYP2C8 was a high-affinity enzyme with a catalytic efficiency ∼15-fold greater than those of CYPs 3A4 and 3A5. The CYP3A inhibitors ketoconazole and troleandomycin, and the CYP2C8 inhibitors quercetin and paclitaxel decreased imatinib oxidation. From molecular modelling, the imatinib structure could be superimposed on a pharmacophore for CYP2C8 substrates. CONCLUSIONS AND IMPLICATIONS CYP2C8 and CYPs 3A contribute to imatinib N-demethylation in human liver. The involvement of CYP2C8 may account in part for the wide inter-patient variation in imatinib pharmacokinetics observed in clinical practice. PMID:20977456

  5. Characterization of air pollutants from an activated sludge process

    SciTech Connect

    Scheff, P.A.; Holden, J.A.; Wadden, R.A.

    1981-02-01

    An eight-month monitoring study was conducted to characterize air pollutants near a large activated sludge plant in a Chicago suburb. Air pollutants detected include aerobic bacteria-containing particles, total suspended particulates, nitrogen dioxide, sulfur dioxide, chloride, hydrogen sulfides, and trace elements. The wastewater treatment plant is concluded to be a significant source of total coliforms and atmospheric bacteria-containing particles. (6 maps, 23 references, 6 tables)

  6. Quantification of citalopram or escitalopram and their demethylated metabolites in neonatal hair samples by liquid chromatography-tandem mass spectrometry.

    PubMed

    Frison, Giampietro; Favretto, Donata; Vogliardi, Susanna; Terranova, Claudio; Ferrara, Santo Davide

    2008-08-01

    Citalopram and escitalopram are highly selective serotonin reuptake inhibitors widely used in the treatment of depression. They exhibit adverse drug reactions and side effects, however, and the development of specific methods for their determination is of great interest in clinical and forensic toxicology. A liquid chromatography-tandem mass spectrometry method has been developed and validated for the assay of citalopram, escitalopram, and their demethylated metabolites in 10-mg hair samples. The analytes were extracted by incubation in methanol and liquid/liquid extraction with diethyl ether/dichloromethane. Gradient elution on a narrow bore C18 column was realized using clomipramine-d3 as an internal standard. Positive ion electrospray ionization and tandem mass spectrometry determination by collision-induced dissociation were performed in an ion trap mass spectrometer. The method exhibited a linear range of 25 to 2000 pg/mg, a quantification limit of 25 pg/mg for all analytes, relative standard deviations in the range of 12.10 to 9.80 (intraassay), and 13.80 to 11.78 (interassay), and accuracies (as percent recovery of the spiked standards) in the range of 90% to 110%; it was applied to the determination of citalopram and escitalopram and their metabolites in hair samples of two newborns to document their in utero exposure to the drugs. The method proved suitable for neonatal hair analysis of citalopram or escitalopram and was applied to two real cases of gestational exposure.

  7. Intramolecular Oxidative O-Demethylation of an Oxoferryl Porphyrin Complexed with a Per-O-methylated β-Cyclodextrin Dimer.

    PubMed

    Kitagishi, Hiroaki; Kurosawa, Shun; Kano, Koji

    2016-11-22

    The intramolecular oxidation of ROCH3 to ROCH2 OH, where the latter compound spontaneously decomposed to ROH and HCHO, was observed during the reaction of the supramolecular complex (met-hemoCD3) with cumene hydroperoxide in aqueous solution. Met-hemoCD3 is composed of meso-tetrakis(4-sulfonatophenyl)porphinatoiron(III) (Fe(III) TPPS) and a per-O-methylated β-cyclodextrin dimer having an -OCH2 PyCH2 O- linker (Py=pyridine-3,5-diyl). The O=Fe(IV) TPPS complex was formed by the reaction of met-hemoCD3 with cumene hydroperoxide, and isolated by gel-filtration chromatography. Although the isolated O=Fe(IV) TPPS complex in the cyclodextrin cage was stable in aqueous solution at 25 °C, it was gradually converted to Fe(II) TPPS (t1/2 =7.6 h). This conversion was accompanied by oxidative O-demethylation of an OCH3 group in the cyclodextrin dimer. The results indicated that hydrogen abstraction by O=Fe(IV) TPPS from ROCH3 yields HO-Fe(III) TPPS and ROCH2(.) . This was followed by radical coupling to afford Fe(II) TPPS and ROCH2 OH. The hemiacetal (ROCH2 OH) immediately decomposed to ROH and HCHO. This study revealed the ability of oxoferryl porphyrin to induce two-electron oxidation.

  8. Process for producing an activated carbon adsorbent with integral heat transfer apparatus

    NASA Technical Reports Server (NTRS)

    Jones, Jack A. (Inventor); Yavrouian, Andre H. (Inventor)

    1996-01-01

    A process for producing an integral adsorbent-heat exchanger apparatus useful in ammonia refrigerant heat pump systems. In one embodiment, the process wets an activated carbon particles-solvent mixture with a binder-solvent mixture, presses the binder wetted activated carbon mixture on a metal tube surface and thereafter pyrolyzes the mixture to form a bonded activated carbon matrix adjoined to the tube surface. The integral apparatus can be easily and inexpensively produced by the process in large quantities.

  9. Impact of agricultural activities on anaerobic processes in stream sediments

    NASA Astrophysics Data System (ADS)

    Schade, J. D.; Ludwig, S.; Nelson, L. C.; Porterfield, J.; Sather, K. L.; Songpitak, M.; Spawn, S.; Weigel, B.

    2013-12-01

    Streams draining agriculture watersheds are subject to significant anthropogenic impacts, including sedimentation from soil erosion and high nitrate input from heavy fertilizer application. Sedimentation degrades habitat and can reduce hydrologic exchange between surface and subsurface waters. Disconnecting surface and subsurface flow reduces oxygen input to hyporheic water, increasing the extent of anoxic zones in stream sediments and creating hotspots for anaerobic processes like denitrification and methanogenesis that can be important sources of nitrous oxide and methane, both powerful greenhouse gases. Increased nitrate input may influence greenhouse gas fluxes from stream sediments by stimulating rates of denitrification and potentially reducing rates of methanogenesis, either through direct inhibition or by increasing competition for organic substrates from denitrifying bacteria. We hypothesized that accumulation of fine sediments in stream channels would result in high rates of methanogenesis in stream sediments, and that increased nitrate input from agricultural runoff would stimulate denitrification and reduce rates of methane production. Our work focused on streams in northern and central Minnesota, in particular on Rice Creek, a small stream draining an agricultural watershed. We used a variety of approaches to test our hypotheses, including surveys of methane concentrations in surface waters of streams ranging in sediment type and nitrate concentration, bottle incubations of sediment from several sites in Rice Creek, and the use of functional gene probes and RNA analyses to determine if genes for these processes are present and being expressed in stream sediments. We found higher methane concentrations in surface water from streams with large deposits of fine sediments, but significantly less methane in these streams when nitrate concentrations were high. We also found high potential for both methanogenesis and denitrification in sediment incubations

  10. Damage-inducible intragenic demethylation of the human TP53 tumor suppressor gene is associated with transcription from an alternative intronic promoter.

    PubMed

    Blackburn, James; Roden, Daniel L; Ng, Robert; Wu, Jianmin; Bosman, Alexis; Epstein, Richard J

    2016-12-01

    Wild-type TP53 exons 5-8 contain CpG dinucleotides that are prone to methylation-dependent mutation during carcinogenesis, but the regulatory effects of methylation affecting these CpG sites are unclear. To clarify this, we first assessed site-specific TP53 CpG methylation in normal and transformed cells. Both DNA damage and cell ageing were associated with site-specific CpG demethylation in exon 5 accompanied by induction of a truncated TP53 isoform regulated by an adjacent intronic promoter (P2). We then synthesized novel synonymous TP53 alleles with divergent CpG content but stable encodement of the wild-type polypeptide. Expression of CpG-enriched TP53 constructs selectively reduced production of the full-length transcript (P1), consistent with a causal relationship between intragenic demethylation and transcription. 450K methylation comparison of normal (TP53-wildtype) and cancerous (TP53-mutant) human cells and tissues revealed focal cancer-associated declines in CpG methylation near the P1 transcription start site, accompanied by rises near the alternate exon 5 start site. These data confirm that site-specific changes of intragenic TP53 CpG methylation are extrinsically inducible, and suggest that human cancer progression is mediated in part by dysregulation of damage-inducible intragenic CpG demethylation that alters TP53 P1/P2 isoform expression. © 2015 The Authors. Molecular Carcinogenesis Published by Wiley Periodicals, Inc.

  11. A new process for enriching nitrifiers in activated sludge through separate heterotrophic wasting from biofilm carriers.

    PubMed

    Parker, Denny S; Rusten, Bjørn; Wien, Asgeir; Siljudalen, Jon G

    2002-01-01

    A new process, the biofilm-activated sludge innovative nitrification (BASIN) process, consisting of a moving-bed biofilm reactor (MBBR) with separate heterotrophic wasting, followed by an activated-sludge process, has been proposed to reduce the volumetric requirements of the activated-sludge process for nitrification. The basic principle is to remove chemical oxygen demand on the biofilm carriers by heterotrophic organisms and then to waste a portion of the heterotrophic biomass before it can be released into the activated-sludge reactor. By this means, the amount of heterotrophic organisms grown in the activated-sludge reactor is reduced, thereby reducing the volume of that tank needed for nitrification. For nitrification applications, the simplest method for stripping biomass was to use an in-tank technique using high shearing rates with aeration. Bench-scale testing showed sludge yields in the BASIN process were one-half of that in a control activated-sludge process and twice that of a process line with intermediate settling between the MBBR and activated-sludge stage. Critical washout solids retention times for nitrifiers were the same for all three lines, so activated-sludge volumes for the BASIN process could be reduced by 50% compared with the control. Originally conceived process concepts for the BASIN process were confirmed by the experimental work.

  12. Deoxyribonucleic acid methyl transferases 3a and 3b associate with the nuclear orphan receptor COUP-TFI during gene activation.

    PubMed

    Gallais, Rozenn; Demay, Florence; Barath, Peter; Finot, Laurence; Jurkowska, Renata; Le Guével, Rémy; Gay, Frédérique; Jeltsch, Albert; Métivier, Raphaël; Salbert, Gilles

    2007-09-01

    Transcriptional activation of silent genes can require the erasure of epigenetic marks such as DNA methylation at CpGs (cytosine-guanine dinucleotide). Active demethylation events have been observed, and associated processes are repeatedly suspected to involve DNA glycosylases such as mCpG binding domain protein 4, thymine DNA glycosylase (TDG), Demeter, and repressor of silencing 1. A complete characterization of the molecular mechanisms occurring in metazoan is nonetheless awaited. Here, we report that activation of the endogenous vitronectin gene in P19 cells by the nuclear receptor chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI) is observed in parallel with the recruitment of TDG and p68 RNA helicase, two components of a putative demethylation complex. Interestingly, when activated, the vitronectin gene was loaded with DNA methyltransferases 3a and 3b (Dnmt3a/b), and a strand-biased decrease in CpG methylation was detected. Dnmt3a was further found to associate with COUP-TFI and TDG in vivo, and cotransfection experiments demonstrated that Dnmt3a/b can enhance COUP-TFI-mediated activation of a methylated reporter gene. These results suggest that Dnmt3a/b could cooperate with the orphan receptor COUP-TFI to regulate transcription of the vitronectin gene.

  13. DNA methylation/demethylation programming during peach flower bud dormancy release, development and blooming

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peach flower bud development undergoes a long, complex and temperature-dependent regulation process with cessation of growth in response to cool temperatures in late fall, a slow but gradual development during the chilling period in winter, and eventually blooming in early spring. It has been demon...

  14. Carbon activation process for increased surface accessibility in electrochemical capacitors

    DOEpatents

    Doughty, Daniel H.; Eisenmann, Erhard T.

    2001-01-01

    A process for making carbon film or powder suitable for double capacitor electrodes having a capacitance of up to about 300 F/cm.sup.3 is disclosed. This is accomplished by treating in aqueous nitric acid for a period of about 5 to 15 minutes thin carbon films obtained by carbonizing carbon-containing polymeric material having a high degree of molecular directionality, such as polyimide film, then heating the treated carbon film in a non-oxidizing atmosphere at a non-graphitizing temperature of at least 350.degree. C. for about 20 minutes, and repeating alternately the nitric acid step and the heating step from 7 to 10 times. Capacitors made with this carbon may find uses ranging from electronic devices to electric vehicle applications.

  15. GRID based Thermal Images Processing for volcanic activity monitoring

    NASA Astrophysics Data System (ADS)

    Mangiagli, S.; Coco, S.; Drago, L.; Laudani, A.,; Lodato, L.; Pollicino, G.; Torrisi, O.

    2009-04-01

    Since 2001, the Catania Section of the National Institute of Geophysics and Volcanology (INGV) has been running the video stations recording the volcanic activity of Mount Etna, Stromboli and the Fossa Crater of Vulcano island. The video signals of 11 video cameras (seven operating in the visible band and four in infrared) are sent in real time to INGV Control Centre where they are visualized on monitors and archived on a dedicated NAS storage. The video surveillance of the Sicilian volcanoes, situated near to densely populated areas, helps the volcanologists providing the Civil Protection authorities with updates in real time on the on-going volcanic activity. In particular, five video cameras are operating on Mt. Etna and they record the volcano from the south and east sides 24 hours a day. During emergencies, mobile video stations may also be used to better film the most important phases of the activity. Single shots are published on the Catania Section intranet and internet websites. On June 2006 a A 40 thermal camera was installed in Vulcano La Fossa Crater. The location was in the internal and opposite crater flank (S1), 400 m distant from the fumarole field. The first two-year of data on temperature distribution frequency were recorded with this new methodology of acquisition, and automatically elaborated by software at INGV Catania Section. In fact a dedicated software developed in IDL, denominated Volcano Thermo Analysis (VTA), was appositely developed in order to extract a set of important features, able to characterize with a good approssimation the volcanic activity. In particular the program first load and opportunely convert the thermal images, then according to the Region Of Interest (ROI) and the temperature ranges defined by the user provide to automatic spatial and statistic analysis. In addition the VTA is able to analysis all the temporal series of images available in order to achieve the time-event analysis and the dynamic of the volcanic

  16. The activation of modality-specific representations during discourse processing.

    PubMed

    Kurby, Christopher A; Zacks, Jeffrey M

    2013-09-01

    Previous research has shown that readers generate mental images of events. Most studies have investigated imagery during the reading of short texts, which also included explicit judgment tasks. In two fMRI studies, we assessed whether modality-specific imagery occurs during naturalistic, discourse comprehension. We identified clauses in the texts that elicited auditory, motor, or visual imagery. In both studies, reading motor imagery clauses was associated with increases in activity in left postcentral and precentral sulci, and reading auditory imagery clauses was associated with increases in left superior temporal gyrus and perisylvian language-related regions. Study 2 compared presentation of connected discourse to a condition in which unconnected sentences were presented, preventing the establishment of global coherence. Sensorimotor imagery was strongest when readers were able to generate a globally coherent discourse representation. Overall, these results suggest that modality-specific imagery occurs during discourse comprehension and it is dependent on the development of discourse-level representations.

  17. Epidemic process on activity-driven modular networks

    NASA Astrophysics Data System (ADS)

    Han, Dun; Sun, Mei; Li, Dandan

    2015-08-01

    In this paper, we propose two novel models of epidemic spreading by considering the activity-driven and the network modular. Firstly, we consider the susceptible-infected-susceptible (SIS) contagion model and derive analytically the epidemic threshold. The results indicate that the epidemic threshold only involves with the value of the spread rate and the recovery rate. In addition, the asymptotic refractory density of infected nodes in the different communities exhibits different trends with the change of the modularity-factor. Then, the infected-driven vaccination model is presented. Simulation results illustrate that the final density of vaccination will increase with the increase of the response strength of vaccination. Moreover, the final infected density in the original-infected-community shows different trends with the change of the response strength of vaccination and the spreading rate. The infected-driven vaccination is a good way to control the epidemic spreading.

  18. The Activation of Modality-Specific Representations During Discourse Processing

    PubMed Central

    Kurby, Christopher A.; Zacks, Jeffrey M.

    2013-01-01

    Previous research has shown that readers generate mental images of events. Most studies have investigated imagery during the reading of short texts, which also included explicit judgment tasks. In two fMRI studies, we assessed whether modality-specific imagery occurs during naturalistic, discourse comprehension. We identified clauses in the texts that elicited auditory, motor, or visual imagery. In both studies, reading motor imagery clauses was associated with increases in activity in left postcentral and precentral sulci, and reading auditory imagery clauses was associated with increases in left superior temporal gyrus and perisylvian language-related regions. Study 2 compared presentation of connected discourse to a condition in which unconnected sentences were presented, preventing the establishment of global coherence. Sensorimotor imagery was strongest when readers were able to generate a globally coherent discourse representation. Overall, these results suggest that modality-specific imagery occurs during discourse comprehension and it is dependent on the development of discourse-level representations. PMID:23933473

  19. Influence of process parameters on the surface and chemical properties of activated carbon obtained from biochar by chemical activation.

    PubMed

    Angın, Dilek; Altintig, Esra; Köse, Tijen Ennil

    2013-11-01

    Activated carbons were produced from biochar obtained through pyrolysis of safflower seed press cake by chemical activation with zinc chloride. The influences of process variables such as the activation temperature and the impregnation ratio on textural and chemical-surface properties of the activated carbons were investigated. Also, the adsorptive properties of activated carbons were tested using methylene blue dye as the targeted adsorbate. The experimental data indicated that the adsorption isotherms are well described by the Langmuir equilibrium isotherm equation. The optimum conditions resulted in activated carbon with a monolayer adsorption capacity of 128.21 mg g(-1) and carbon content 76.29%, while the BET surface area and total pore volume corresponded to 801.5m(2)g(-1) and 0.393 cm(3)g(-1), respectively. This study demonstrated that high surface area activated carbons can be prepared from the chemical activation of biochar with zinc chloride as activating agents.

  20. Life cycle analysis within pharmaceutical process optimization and intensification: case study of active pharmaceutical ingredient production.

    PubMed

    Ott, Denise; Kralisch, Dana; Denčić, Ivana; Hessel, Volker; Laribi, Yosra; Perrichon, Philippe D; Berguerand, Charline; Kiwi-Minsker, Lioubov; Loeb, Patrick

    2014-12-01

    As the demand for new drugs is rising, the pharmaceutical industry faces the quest of shortening development time, and thus, reducing the time to market. Environmental aspects typically still play a minor role within the early phase of process development. Nevertheless, it is highly promising to rethink, redesign, and optimize process strategies as early as possible in active pharmaceutical ingredient (API) process development, rather than later at the stage of already established processes. The study presented herein deals with a holistic life-cycle-based process optimization and intensification of a pharmaceutical production process targeting a low-volume, high-value API. Striving for process intensification by transfer from batch to continuous processing, as well as an alternative catalytic system, different process options are evaluated with regard to their environmental impact to identify bottlenecks and improvement potentials for further process development activities.

  1. Anticancer Activity of Metal Complexes: Involvement of Redox Processes

    PubMed Central

    Jungwirth, Ute; Kowol, Christian R.; Keppler, Bernhard K.; Hartinger, Christian G.; Berger, Walter; Heffeter, Petra

    2012-01-01

    Cells require tight regulation of the intracellular redox balance and consequently of reactive oxygen species for proper redox signaling and maintenance of metal (e.g., of iron and copper) homeostasis. In several diseases, including cancer, this balance is disturbed. Therefore, anticancer drugs targeting the redox systems, for example, glutathione and thioredoxin, have entered focus of interest. Anticancer metal complexes (platinum, gold, arsenic, ruthenium, rhodium, copper, vanadium, cobalt, manganese, gadolinium, and molybdenum) have been shown to strongly interact with or even disturb cellular redox homeostasis. In this context, especially the hypothesis of “activation by reduction” as well as the “hard and soft acids and bases” theory with respect to coordination of metal ions to cellular ligands represent important concepts to understand the molecular modes of action of anticancer metal drugs. The aim of this review is to highlight specific interactions of metal-based anticancer drugs with the cellular redox homeostasis and to explain this behavior by considering chemical properties of the respective anticancer metal complexes currently either in (pre)clinical development or in daily clinical routine in oncology. PMID:21275772

  2. Momentum Driving: Which Physical Processes Dominate Active Galactic Nucleus Feedback?

    NASA Astrophysics Data System (ADS)

    Ostriker, Jeremiah P.; Choi, Ena; Ciotti, Luca; Novak, Gregory S.; Proga, Daniel

    2010-10-01

    The deposition of mechanical feedback from a supermassive black hole (SMBH) in an active galactic nucleus into the surrounding galaxy occurs via broad-line winds which must carry mass and radial momentum as well as energy. The effect can be summarized by the dimensionless parameter η ={\\dot{M}_outf}/{\\dot{M}_acc}= {2 ɛ_wc^2}/{v_w^2} where epsilonw (≡ \\dot{E}_w/(\\dot{M}_accc^2)) is the efficiency with which accreted matter is turned into wind energy in the disk surrounding the central SMBH. The outflowing mass and momentum are proportional to η, and many prior treatments have essentially assumed that η = 0. We perform one- and two-dimensional simulations and find that the growth of the central SMBH is very sensitive to the inclusion of the mass and momentum driving but is insensitive to the assumed mechanical efficiency. For example in representative calculations, the omission of momentum and mass feedback leads to a hundred-fold increase in the mass of the SMBH to over 1010 M sun. When allowance is made for momentum driving, the final SMBH mass is much lower and the wind efficiencies that lead to the most observationally acceptable results are relatively low with epsilonw <~ 10-4.

  3. Predictive active disturbance rejection control for processes with time delay.

    PubMed

    Zheng, Qinling; Gao, Zhiqiang

    2014-07-01

    Active disturbance rejection control (ADRC) has been shown to be an effective tool in dealing with real world problems of dynamic uncertainties, disturbances, nonlinearities, etc. This paper addresses its existing limitations with plants that have a large transport delay. In particular, to overcome the delay, the extended state observer (ESO) in ADRC is modified to form a predictive ADRC, leading to significant improvements in the transient response and stability characteristics, as shown in extensive simulation studies and hardware-in-the-loop tests, as well as in the frequency response analysis. In this research, it is assumed that the amount of delay is approximately known, as is the approximated model of the plant. Even with such uncharacteristic assumptions for ADRC, the proposed method still exhibits significant improvements in both performance and robustness over the existing methods such as the dead-time compensator based on disturbance observer and the Filtered Smith Predictor, in the context of some well-known problems of chemical reactor and boiler control problems.

  4. 76 FR 30696 - Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-26

    ... Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites AGENCY: Department of... eligible active uranium and thorium processing site licensees for reimbursement under Title X of the Energy... requires DOE to reimburse eligible uranium and thorium licensees for certain costs of...

  5. 76 FR 24871 - Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-03

    ... Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites AGENCY: Department of... from eligible active uranium and thorium processing site licensees for reimbursement under Title X of...). Title X requires DOE to reimburse eligible uranium and thorium licensees for certain costs...

  6. Drosophila Kdm4 demethylases in histone H3 lysine 9 demethylation and ecdysteroid signaling

    PubMed Central

    Tsurumi, Amy; Dutta, Pranabanada; Yan, Shian-Jang; Sheng, Robin; Li, Willis X.

    2013-01-01

    The dynamic regulation of chromatin structure by histone post-translational modification is an essential regulatory mechanism that controls global gene transcription. The Kdm4 family of H3K9me2,3 and H3K36me2,3 dual specific histone demethylases has been implicated in development and tumorigenesis. Here we show that Drosophila Kdm4A and Kdm4B are together essential for mediating ecdysteroid hormone signaling during larval development. Loss of Kdm4 genes leads to globally elevated levels of the heterochromatin marker H3K9me2,3 and impedes transcriptional activation of ecdysone response genes, resulting in developmental arrest. We further show that Kdm4A interacts with the Ecdysone Receptor (EcR) and colocalizes with EcR at its target gene promoter. Our studies suggest that Kdm4A may function as a transcriptional co-activator by removing the repressive histone mark H3K9me2,3 from cognate promoters. PMID:24100631

  7. Metacaspase 2 of Trypanosoma brucei is a calcium-dependent cysteine peptidase active without processing.

    PubMed

    Moss, Catherine X; Westrop, Gareth D; Juliano, Luiz; Coombs, Graham H; Mottram, Jeremy C

    2007-12-11

    Metacaspases are cysteine peptidases that are distantly related to the caspases, for which proteolytic processing is central to their activation. Here, we show that recombinant metacaspase 2 (MCA2) from Trypanosoma brucei has arginine/lysine-specific, Ca(2+)-dependent proteolytic activity. Autocatalytic processing of MCA2 occurred after Lys55 and Lys268; however, this was shown not to be required for the enzyme to be proteolytically active. The necessity of Ca(2+), but not processing, for MCA2 enzymatic activity clearly distinguishes MCA2 from the caspases and would be consistent with different physiological roles.

  8. Dynamics of H3K27me3 methylation and demethylation in plant development

    PubMed Central

    Gan, Eng-Seng; Xu, Yifeng; Ito, Toshiro

    2015-01-01

    Epigenetic regulation controls multiple aspects of the plant development. The N-terminal tail of histone can be differently modified to regulate various chromatin activities. One of them, the trimethylation of histone H3 lysine 27 (H3K27me3) confers a repressive chromatin state with gene silencing. H3K27me3 is dynamically deposited and removed throughout development. While components of the H3K27me3 writer, Polycomb repressive complex 2 (PRC2), have been reported for almost 2 decades, it is only recently that JUMONJI (JMJ) proteins are reported as H3K27me3 demethylases, affirming the dynamic nature of histone modifications. This review highlights recent progress in plant epigenetic research, focusing on the H3K27me3 demethylases. PMID:26313233

  9. Timing of Transcriptional Quiescence during Gametogenesis Is Controlled by Global Histone H3K4 Demethylation

    PubMed Central

    Xu, Mengshu; Soloveychik, Maria; Ranger, Mathieu; Schertzberg, Michael; Shah, Zarna; Raisner, Ryan; Venkatasubrahmanyan, Shivkumar; Tsui, Kyle; Gebbia, Marinella; Hughes, Tim; van Bakel, Harm; Nislow, Corey; Madhani, Hiten D.; Meneghini, Marc D.

    2013-01-01

    SUMMARY Gametes are among the most highly specialized cells produced during development. Although gametogenesis culminates in transcriptional quiescence in plants and animals, regulatory mechanisms controlling this are unknown. Here, we confirm that gamete differentiation in the single-celled yeast Saccharomyces cerevisiae is accompanied by global transcriptional shutoff following the completion of meiosis. We show that Jhd2, a highly conserved JARID1-family histone H3K4 demethylase, activates protein-coding gene transcription in opposition to this programmed transcriptional shutoff, sustaining the period of productive transcription during spore differentiation. Moreover, using genome-wide nucleosome, H3K4me, and transcript mapping experiments, we demonstrate that JHD2 globally represses intergenic noncoding transcription during this period. The widespread transcriptional defects of JHD2 mutants are associated with precocious differentiation and the production of stress-sensitive spores, demonstrating that Jhd2 regulation of the global postmeiotic transcriptional program is critical for the production of healthy meiotic progeny. PMID:23123093

  10. Timing of transcriptional quiescence during gametogenesis is controlled by global histone H3K4 demethylation.

    PubMed

    Xu, Mengshu; Soloveychik, Maria; Ranger, Mathieu; Schertzberg, Michael; Shah, Zarna; Raisner, Ryan; Venkatasubrahmanyan, Shivkumar; Tsui, Kyle; Gebbia, Marinella; Hughes, Tim; van Bakel, Harm; Nislow, Corey; Madhani, Hiten D; Meneghini, Marc D

    2012-11-13

    Gametes are among the most highly specialized cells produced during development. Although gametogenesis culminates in transcriptional quiescence in plants and animals, regulatory mechanisms controlling this are unknown. Here, we confirm that gamete differentiation in the single-celled yeast Saccharomyces cerevisiae is accompanied by global transcriptional shutoff following the completion of meiosis. We show that Jhd2, a highly conserved JARID1-family histone H3K4 demethylase, activates protein-coding gene transcription in opposition to this programmed transcriptional shutoff, sustaining the period of productive transcription during spore differentiation. Moreover, using genome-wide nucleosome, H3K4me, and transcript mapping experiments, we demonstrate that JHD2 globally represses intergenic noncoding transcription during this period. The widespread transcriptional defects of JHD2 mutants are associated with precocious differentiation and the production of stress-sensitive spores, demonstrating that Jhd2 regulation of the global postmeiotic transcriptional program is critical for the production of healthy meiotic progeny.

  11. Nucleosome repositioning links DNA (de)methylation and differential CTCF binding during stem cell development

    PubMed Central

    Teif, Vladimir B.; Beshnova, Daria A.; Vainshtein, Yevhen; Marth, Caroline; Mallm, Jan-Philipp; Höfer, Thomas; Rippe, Karsten

    2014-01-01

    During differentiation of embryonic stem cells, chromatin reorganizes to establish cell type-specific expression programs. Here, we have dissected the linkages between DNA methylation (5mC), hydroxymethylation (5hmC), nucleosome repositioning, and binding of the transcription factor CTCF during this process. By integrating MNase-seq and ChIP-seq experiments in mouse embryonic stem cells (ESC) and their differentiated counterparts with biophysical modeling, we found that the interplay between these factors depends on their genomic context. The mostly unmethylated CpG islands have reduced nucleosome occupancy and are enriched in cell type-independent binding sites for CTCF. The few remaining methylated CpG dinucleotides are preferentially associated with nucleosomes. In contrast, outside of CpG islands most CpGs are methylated, and the average methylation density oscillates so that it is highest in the linker region between nucleosomes. Outside CpG islands, binding of TET1, an enzyme that converts 5mC to 5hmC, is associated with labile, MNase-sensitive nucleosomes. Such nucleosomes are poised for eviction in ESCs and become stably bound in differentiated cells where the TET1 and 5hmC levels go down. This process regulates a class of CTCF binding sites outside CpG islands that are occupied by CTCF in ESCs but lose the protein during differentiation. We rationalize this cell type-dependent targeting of CTCF with a quantitative biophysical model of competitive binding with the histone octamer, depending on the TET1, 5hmC, and 5mC state. PMID:24812327

  12. The contribution of activated processes to Q. [stress corrosion cracking in seismic wave attenuation

    NASA Technical Reports Server (NTRS)

    Spetzler, H. A.; Getting, I. C.; Swanson, P. L.

    1980-01-01

    The possible role of activated processes in seismic attenuation is investigated. In this study, a solid is modeled by a parallel and series configuration of dashpots and springs. The contribution of stress and temperature activated processes to the long term dissipative behavior of this system is analyzed. Data from brittle rock deformation experiments suggest that one such process, stress corrosion cracking, may make a significant contribution to the attenuation factor, Q, especially for long period oscillations under significant tectonic stress.

  13. An evaluation of process parameters to improve coating efficiency of an active tablet film-coating process.

    PubMed

    Wang, Jennifer; Hemenway, Jeffrey; Chen, Wei; Desai, Divyakant; Early, William; Paruchuri, Srinivasa; Chang, Shih-Ying; Stamato, Howard; Varia, Sailesh

    2012-05-10

    Effects of material and manufacturing process parameters on the efficiency of an aqueous active tablet film-coating process in a perforated pan coater were evaluated. Twenty-four batches representing various core tablet weights, sizes, and shapes were coated at the 350-500 kg scale. The coating process efficiency, defined as the ratio of the amount of active deposited on tablet cores to the amount of active sprayed, ranged from 86 to 99%. Droplet size and velocity of the coating spray were important for an efficient coating process. Factors governing them such as high ratios of the suspension spray rate to atomization air flow rate, suspension spray rate to pattern air flow rate, or atomization air flow rate to pattern air flow rate improved the coating efficiency. Computational fluid dynamics modeling of the droplets showed that reducing the fraction of the smaller droplets, especially those smaller than 10 μm, resulted in a marked improvement in the coating efficiency. Other material and process variables such as coating suspension solids concentration, pan speed, tablet velocity, exhaust air temperature, and the length of coating time did not affect the coating efficiency profoundly over the ranges examined here.

  14. DNA demethylation of the TIM-3 promoter is critical for its stable expression on T cells.

    PubMed

    Chou, F-C; Kuo, C-C; Chen, H-Y; Chen, H-H; Sytwu, H-K

    2016-04-01

    The T-cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) is selectively expressed on terminally differentiated T helper 1 (Th1) cells and acts as a negative regulator that terminates Th1 responses. The dysregulation of TIM-3 expression on T cells is associated with several autoimmune phenotypes and with chronic viral infections; however, the mechanism of this regulation is unclear. In this study, we investigated the effect of DNA methylation on the expression of TIM-3. By analyzing the sequences of TIM-3 promoter regions in human and mouse, we identified a CpG island within the TIM-3 promoter and demonstrated that the promoter activity was controlled by DNA methylation. Furthermore, treatment with 5-aza-2'-deoxycytidine enhanced TIM-3 expression on mouse primary CD4(+) T cells under Th0-, Th1- or Th2-polarizing conditions. Finally, pyrosequencing analysis revealed that the methylation level of the TIM-3 promoter gradually decreased after each round of T-cell polarization, and this decrease was inversely correlated with TIM-3 expression. These data suggest that the DNA methylation of the TIM-3 promoter cooperates with lineage-specific transcription factors in the control of Th-cell development. In conclusion, DNA methylation-based regulation of TIM-3 may provide novel insights into understanding the dysregulation of TIM-3 expression under pathogenic conditions.

  15. [Preparation and optimum process of walnut peel activated carbon by zinc chloride as activating agent].

    PubMed

    Liu, Xiao-hong; Wang, Xing-wei; Zhao, Bo; Lü, Jun-fang; Kang, Ni-na; Zhang, Yao-jun

    2014-12-01

    Walnut peel as raw material, zinc chloride was used as activating agent for preparation walnut peel activated carbon in the muffle furnace in this experiment, using orthogonal design. Yield, the specific surface area and iodine number of walnut peel activated carbon were determined at all designed experimental conditions and the optimum technological condition of preparation was obtained. By analysis of aperture, infrared spectra and the content of acidic group in surface with Boehm, walnut peel activated carbon of prepared at the optimum condition was characterized. The results showed the optimum technological parameters of preparation: activation temperature (600 °C), activation time (1 h), the concentration of zinc chloride (50%), the particle size (60 mesh). The specific surface area of walnut peel activated carbon obtained at optimum condition was mounting to 1258.05 m2 · g(-1), the ratio of medium porous 32.18%. Therefore, walnut peel can be used in the preparation of the high-quality activated carbon of large surface area. Agricultural wastes, as walnut peel, not only were implemented recycle, but also didn't make any pollution. Meanwhile, a cheap adsorbent was provided and it was of great significance to open a new source of activated carbon.

  16. Chemical and Biochemical Approaches in the Study of Histone Methylation and Demethylation

    PubMed Central

    Li, Keqin Kathy; Luo, Cheng; Wang, Dongxia; Jiang, Hualiang; Zheng, Y. George

    2014-01-01

    Histone methylation represents one of the most critical epigenetic events in DNA function regulation in eukaryotic organisms. Classic molecular biology and genetics tools provide significant knowledge about mechanisms and physiological roles of histone methyltransferases and demethylases in various cellular processes. In addition to this stream line, development and application of chemistry and chemistry-related techniques are increasingly involved in biological study, and provide information otherwise difficulty to obtain by standard molecular biology methods. Herein, we review recent achievements and progress in developing and applying chemical and biochemical approaches in the study of histone methylation, including chromatin immunoprecipitation (ChIP), chemical ligation, mass spectrometry (MS), biochemical assays, and inhibitor development. These technological advances allow histone methylation to be studied from genome-wide level to molecular and atomic levels. With ChIP technology, information can be obtained about precise mapping of histone methylation patterns at specific promoters, genes or other genomic regions. MS is particularly useful in detecting and analyzing methylation marks in histone and nonhistone protein substrates. Chemical approaches that permit site-specific incorporation of methyl groups into histone proteins greatly facilitate the investigation of the biological impacts of methylation at individual modification sites. Discovery and design of selective organic inhibitors of histone methyltransferases and demethylases provide chemical probes to interrogate methylation-mediated cellular pathways. Overall, these chemistry-related technological advances have greatly improved our understanding of the biological functions of histone methylation in normal physiology and diseased states, and also are of great potential to translate basic epigenetics research into diagnostic and therapeutic application in the clinic. PMID:22777714

  17. [INT-ETS activity change of activated sludge during nitrification, denirification and organism removal in SBR process].

    PubMed

    Yin, Jun; Wang, Jian-hui; Xie, Yan-cui; Huo, Yu-feng; Wang, Xue-feng

    2007-10-01

    Through nitrogen shock loading and organic shock loading experiments, we studied the changing rule and activity status of biological activity of activated sludge in organic matter biodegradation, nitrification and denitrification course in SBR process by the determination of INT-ETS activity. The experimental results show that INT-ETS activity could provide a good indication of the ongoing biological reactions of SBR process. Biological activity of organic matter biodegradation, nitrification and denitrification course in SBR process reduced orderly. INT-ETS activity reduced from 232.59 mg/(gxh) to 190.65 mg/(gxh) and resultly to 113.88 mg/(gxh) when the effluent concentration of COD and NH4(+)-N were 300 mg/L and 40 mg/L. Nitrogen shock loading (14.5 mg/L and 42.0 mg/L) and organic shock loading (COD are 293 mg/L and 685 mg/L) experiments cure verify that the operational conditions could not influence the changing rule of INT-ETS activity, but the time of the appearance of break points marking different reaction course in profile was influenced.

  18. Early neural activation during facial affect processing in adolescents with Autism Spectrum Disorder☆

    PubMed Central

    Leung, Rachel C.; Pang, Elizabeth W.; Cassel, Daniel; Brian, Jessica A.; Smith, Mary Lou; Taylor, Margot J.

    2014-01-01

    Impaired social interaction is one of the hallmarks of Autism Spectrum Disorder (ASD). Emotional faces are arguably the most critical visual social stimuli and the ability to perceive, recognize, and interpret emotions is central to social interaction and communication, and subsequently healthy social development. However, our understanding of the neural and cognitive mechanisms underlying emotional face processing in adolescents with ASD is limited. We recruited 48 adolescents, 24 with high functioning ASD and 24 typically developing controls. Participants completed an implicit emotional face processing task in the MEG. We examined spatiotemporal differences in neural activation between the groups during implicit angry and happy face processing. While there were no differences in response latencies between groups across emotions, adolescents with ASD had lower accuracy on the implicit emotional face processing task when the trials included angry faces. MEG data showed atypical neural activity in adolescents with ASD during angry and happy face processing, which included atypical activity in the insula, anterior and posterior cingulate and temporal and orbitofrontal regions. Our findings demonstrate differences in neural activity during happy and angry face processing between adolescents with and without ASD. These differences in activation in social cognitive regions may index the difficulties in face processing and in comprehension of social reward and punishment in the ASD group. Thus, our results suggest that atypical neural activation contributes to impaired affect processing, and thus social cognition, in adolescents with ASD. PMID:25610782

  19. Early neural activation during facial affect processing in adolescents with Autism Spectrum Disorder.

    PubMed

    Leung, Rachel C; Pang, Elizabeth W; Cassel, Daniel; Brian, Jessica A; Smith, Mary Lou; Taylor, Margot J

    2015-01-01

    Impaired social interaction is one of the hallmarks of Autism Spectrum Disorder (ASD). Emotional faces are arguably the most critical visual social stimuli and the ability to perceive, recognize, and interpret emotions is central to social interaction and communication, and subsequently healthy social development. However, our understanding of the neural and cognitive mechanisms underlying emotional face processing in adolescents with ASD is limited. We recruited 48 adolescents, 24 with high functioning ASD and 24 typically developing controls. Participants completed an implicit emotional face processing task in the MEG. We examined spatiotemporal differences in neural activation between the groups during implicit angry and happy face processing. While there were no differences in response latencies between groups across emotions, adolescents with ASD had lower accuracy on the implicit emotional face processing task when the trials included angry faces. MEG data showed atypical neural activity in adolescents with ASD during angry and happy face processing, which included atypical activity in the insula, anterior and posterior cingulate and temporal and orbitofrontal regions. Our findings demonstrate differences in neural activity during happy and angry face processing between adolescents with and without ASD. These differences in activation in social cognitive regions may index the difficulties in face processing and in comprehension of social reward and punishment in the ASD group. Thus, our results suggest that atypical neural activation contributes to impaired affect processing, and thus social cognition, in adolescents with ASD.

  20. Active pharmaceutical ingredient (API) production involving continuous processes--a process system engineering (PSE)-assisted design framework.

    PubMed

    Cervera-Padrell, Albert E; Skovby, Tommy; Kiil, Søren; Gani, Rafiqul; Gernaey, Krist V

    2012-10-01

    A systematic framework is proposed for the design of continuous pharmaceutical manufacturing processes. Specifically, the design framework focuses on organic chemistry based, active pharmaceutical ingredient (API) synthetic processes, but could potentially be extended to biocatalytic and fermentation-based products. The method exploits the synergic combination of continuous flow technologies (e.g., microfluidic techniques) and process systems engineering (PSE) methods and tools for faster process design and increased process understanding throughout the whole drug product and process development cycle. The design framework structures the many different and challenging design problems (e.g., solvent selection, reactor design, and design of separation and purification operations), driving the user from the initial drug discovery steps--where process knowledge is very limited--toward the detailed design and analysis. Examples from the literature of PSE methods and tools applied to pharmaceutical process design and novel pharmaceutical production technologies are provided along the text, assisting in the accumulation and interpretation of process knowledge. Different criteria are suggested for the selection of batch and continuous processes so that the whole design results in low capital and operational costs as well as low environmental footprint. The design framework has been applied to the retrofit of an existing batch-wise process used by H. Lundbeck A/S to produce an API: zuclopenthixol. Some of its batch operations were successfully converted into continuous mode, obtaining higher yields that allowed a significant simplification of the whole process. The material and environmental footprint of the process--evaluated through the process mass intensity index, that is, kg of material used per kg of product--was reduced to half of its initial value, with potential for further reduction. The case-study includes reaction steps typically used by the pharmaceutical

  1. Interactions of xanthines with activated carbon. I. Kinetics of the adsorption process

    NASA Astrophysics Data System (ADS)

    Navarrete Casas, R.; García Rodriguez, A.; Rey Bueno, F.; Espínola Lara, A.; Valenzuela Calahorro, C.; Navarrete Guijosa, A.

    2006-06-01

    Because of their pharmaceutical and industrial applications, we have studied the adsorption of xanthine derivates (caffeine and theophylline) by activated carbon. To this end, we examined kinetic, equilibrium and thermodynamic aspects of the process. This paper reports the kinetics results. The experimental results indicate that the process was first order in C and the overall process was assumed to involve a single, reversible adsorption-desorption process obeying a kinetic law postulated by us.

  2. Cognitive activity and physiological arousal: processes that mediate mood-congruent memory.

    PubMed

    Varner, L J; Ellis, H C

    1998-09-01

    This research proposes that the cognitive activity associated with the experience of an emotional state mediates the occurrence of mood-congruent processing. Two experiments examined the role of cognitive activity in selective processing of words in a mood congruence paradigm. Four induction procedures were used: a depressed-mood induction, a schema induction organized around the theme of writing a paper, an arousal induction, and a control neutral-mood induction. The memory task consisted of recalling a word list composed of negatively associated and thematically organized words. Selective processing was demonstrated in conjunction with the depressed-mood and organizational-schema induction procedures. In contrast, the arousal and neutral induction procedures did not produce selective processing of words from the list. The findings support the thesis that cognitive activity mediates the selective processing typical of mood congruence as distinct from arousal processes per se. The findings are discussed with respect to the resource allocation model and semantic network theory.

  3. Comparative aromatic hydroxylation and N-demethylation of MPTP neurotoxin and its analogs, N-methylated {beta}-carboline and isoquinoline alkaloids, by human cytochrome P450 2D6

    SciTech Connect

    Herraiz, Tomas . E-mail: therraiz@ifi.csic.es; Guillen, Hugo; Aran, Vicente J.; Idle, Jeffrey R.; Gonzalez, Frank J.

    2006-11-01

    1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxin is a chemical inducer of Parkinson's disease (PD) whereas N-methylated {beta}-carbolines and isoquinolines are naturally occurring analogues of MPTP involved in PD. This research has studied the oxidation of MPTP by human CYP2D6 (CYP2D6*1 and CYP2D6*10 allelic variants) as well as by a mixture of cytochrome P450s-resembling HLM, and the products generated compared with those afforded by human monoamine oxidase (MAO-B). MPTP was efficiently oxidized by CYP2D6 to two main products: MPTP-OH (p-hydroxylation) and PTP (N-demethylation), with turnover numbers of 10.09 min{sup -1} and K {sub m} of 79.36 {+-} 3 {mu}M (formation of MPTP-OH) and 18.95 min{sup -1} and K {sub m} 69.6 {+-} 2.2 {mu}M (PTP). Small amounts of dehydrogenated toxins MPDP{sup +} and MPP{sup +} were also detected. CYP2D6 competed with MAO-B for the oxidation of MPTP. MPTP oxidation by MAO-B to MPDP{sup +} and MPP{sup +} toxins (bioactivation) was up to 3-fold higher than CYP2D6 detoxification to PTP and MPTP-OH. Several N-methylated {beta}-carbolines and isoquinolines were screened for N-demethylation (detoxification) that was not significantly catalyzed by CYP2D6 or the P450s mixture. In contrast, various {beta}-carbolines were efficiently hydroxylated to hydroxy-{beta}-carbolines by CYP2D6. Thus, N(2)-methyl-1,2,3,4-tetrahydro-{beta}-carboline (a close MPTP analog) was highly hydroxylated to 6-hydroxy-N(2)-methyl-1,2,3,4-tetrahydro-{beta}-carboline and a corresponding 7-hydroxy-derivative. Thus, CYP2D6 could participate in the bioactivation and/or detoxification of these neuroactive compounds by an active hydroxylation pathway. The CYP2D6*1 enzymatic variant exhibited much higher metabolism of both MPTP and N(2)-methyl-1,2,3,4-tetrahydro-{beta}-carboline than the CYP2D6*10 variant, highlighting the importance of CYP2D6 polymorphism in the oxidation of these toxins. Altogether, these results suggest that CYP2D6 can play an important role

  4. Characterizing Requirements for Small Ubiquitin-like Modifier (SUMO) Modification and Binding on Base Excision Repair Activity of Thymine-DNA Glycosylase in Vivo.

    PubMed

    McLaughlin, Dylan; Coey, Christopher T; Yang, Wei-Chih; Drohat, Alexander C; Matunis, Michael J

    2016-04-22

    Thymine-DNA glycosylase (TDG) plays critical roles in DNA base excision repair and DNA demethylation. It has been proposed, based on structural studies and in vitro biochemistry, that sumoylation is required for efficient TDG enzymatic turnover following base excision. However, whether sumoylation is required for TDG activity in vivo has not previously been tested. We have developed an in vivo assay for TDG activity that takes advantage of its recently discovered role in DNA demethylation and selective recognition and repair of 5-carboxylcytosine. Using this assay, we investigated the role of sumoylation in regulating TDG activity through the use of TDG mutants defective for sumoylation and Small Ubiquitin-like Modifier (SUMO) binding and by altering TDG sumoylation through SUMO and SUMO protease overexpression experiments. Our findings indicate that sumoylation and SUMO binding are not essential for TDG-mediated excision and repair of 5-carboxylcytosine bases. Moreover, in vitro assays revealed that apurinic/apyrimidinic nuclease 1 provides nearly maximum stimulation of TDG processing of G·caC substrates. Thus, under our assay conditions, apurinic/apyrimidinic nuclease 1-mediated stimulation or other mechanisms sufficiently alleviate TDG product inhibition and promote its enzymatic turnover in vivo.

  5. Effects of Different Heat Processing on Fucoxanthin, Antioxidant Activity and Colour of Indonesian Brown Seaweeds

    NASA Astrophysics Data System (ADS)

    Susanto, Eko; Suhaeli Fahmi, A.; Winarni Agustini, Tri; Rosyadi, Septian; Dita Wardani, Ayunda

    2017-02-01

    Fucoxanthin (Fx) is major carotenoids in brown algae. It showed many health beneficial effects for oxidative stress. Fucoxanthin is lower stability which may cause problem in the application for functional food. The objective of this study was to evaluate the effects of various heat processing on Fx, antioxidant activity (IC50), total phenolic content, and colour stability of Sargassum ilicifolium. The various heat processing methods showed were not significantly affected to fucoxanthin and antioxidant activities however all treatments lower affected to brown seaweeds colour. Moreover, this study showed a useful proved in the design of brown seaweeds processing which minimize Fx, antioxidant activity and colour changes.

  6. Antioxidant Activity in Two Pearl Millet (Pennisetum typhoideum) Cultivars as Influenced by Processing

    PubMed Central

    Suma Pushparaj, Florence; Urooj, Asna

    2014-01-01

    Research on the effect of processing on the retention of bioactive components with potential antioxidant activity is gaining importance. The objective of this investigation was to evaluate the effect of various processing methods (milling, boiling, pressure cooking, roasting and germination respectively) on the antioxidant components as well as the antioxidant activities in the commonly used pearl millet cultivars—Kalukombu (K) and Maharashtra Rabi Bajra (MRB). The methanolic extracts of processed pearl millet flours were analyzed for 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, reducing power assay (RPA) and ferric reducing antioxidant power (FRAP) assays respectively. The samples were also evaluated for tannin, phytic acid and flavonoid content which was then correlated with the antioxidant activity assayed using three methods. The results indicated that the bran rich fraction showed high antioxidant activity (RPA) owing to high tannin, phytic acid and flavonoid levels. Heat treatments exhibited significantly (P ≤ 0.05) higher antioxidant activity (DPPH scavenging activity and RPA) reflecting the high flavonoid content. Processing did not have any significant effect on the FRAP activity of pearl millet. The data on the correlation coefficient suggests that DPPH radical scavenging activity and reducing power assay in the K variety was largely due to the presence of flavonoid content, however in MRB, no relationship was found between antioxidant activities and antioxidant components. PMID:26784663

  7. Control of the aeration volume in an activated sludge process for nitrogen removal.

    PubMed

    Samuelsson, P; Carlsson, B

    2002-01-01

    Biological nitrogen removal in an activated sludge process is obtained by two biological processes; nitrification and denitrification. Nitrifying bacteria need dissolved oxygen and a sufficiently large aeration volume for converting ammonium to nitrate in the wastewater. The objective of this paper is to develop an automatic control strategy for adjusting the aerated volume so that the effluent ammonium level can be kept close to a desired value despite major changes in the influent load. The strategy is based on applying exact linearization of the IAWO Activated Sludge Process Model No 1. Simulation results show that the suggested controller effectively attenuates process disturbances.

  8. Curcumin inhibits the AKT/NF-κB signaling via CpG demethylation of the promoter and restoration of NEP in the N2a cell line.

    PubMed

    Deng, Yushuang; Lu, Xi; Wang, Li; Li, Tao; Ding, Yubin; Cao, Huimin; Zhang, Yuping; Guo, Xiuming; Yu, Gang

    2014-07-01

    Curcumin (CUR), a non-toxic polyphenol from Curcuma longa, has been investigated as a potential therapy with anti-inflammatory and anti-oxidative effects for Alzheimer's disease (AD), which depicts features of chronic inflammatory environment resulting in cellular death. However, it remains largely unknown whether the anti-inflammatory effect of CUR in AD is associated with its property of CpG demethylation, which is another function of CUR with the most research interest during recent years. Neprilysin (NEP, EP24.11), a zinc-dependent metallopeptidase expressed relatively low in the brain, is emerging as a potent inhibitor of AKT/Protein Kinase B. In addition, hypermethylated promoter of NEP has been reported to be associated with decreases in NEP expression. In the present study, using bisulfite-sequencing PCR (BSP) assay, we showed that the CpG sites in NEP gene were hypermethylated both in wild-type mouse neuroblastoma N2a cells (N2a/wt) and N2a cells stably expressing human Swedish mutant amyloid precursor protein (APP) (N2a/APPswe) associated with familial early onset AD. CUR treatment induced restoration of NEP gene via CpG demethylation. This CUR-mediated upregulation of NEP expression was also concomitant with the inhibition of AKT, subsequent suppression of nuclear transcription factor-κB (NF-κB) and its downstream pro-inflammatory targets including COX-2, iNOS in N2a/APPswe cells. This study represents the first evidence on a link between CpG demethylation effect on NEP and anti-inflammation ability of CUR that may provide a novel mechanistic insight into the anti-inflammatory actions of CUR as well as new basis for using CUR as a therapeutic intervention for AD.

  9. Damage‐inducible intragenic demethylation of the human TP53 tumor suppressor gene is associated with transcription from an alternative intronic promoter

    PubMed Central

    Blackburn, James; Roden, Daniel L.; Ng, Robert; Wu, Jianmin; Bosman, Alexis

    2015-01-01

    Wild‐type TP53 exons 5–8 contain CpG dinucleotides that are prone to methylation‐dependent mutation during carcinogenesis, but the regulatory effects of methylation affecting these CpG sites are unclear. To clarify this, we first assessed site‐specific TP53 CpG methylation in normal and transformed cells. Both DNA damage and cell ageing were associated with site‐specific CpG demethylation in exon 5 accompanied by induction of a truncated TP53 isoform regulated by an adjacent intronic promoter (P2). We then synthesized novel synonymous TP53 alleles with divergent CpG content but stable encodement of the wild‐type polypeptide. Expression of CpG‐enriched TP53 constructs selectively reduced production of the full‐length transcript (P1), consistent with a causal relationship between intragenic demethylation and transcription. 450K methylation comparison of normal (TP53‐wildtype) and cancerous (TP53‐mutant) human cells and tissues revealed focal cancer‐associated declines in CpG methylation near the P1 transcription start site, accompanied by rises near the alternate exon 5 start site. These data confirm that site‐specific changes of intragenic TP53 CpG methylation are extrinsically inducible, and suggest that human cancer progression is mediated in part by dysregulation of damage‐inducible intragenic CpG demethylation that alters TP53 P1/P2 isoform expression. © 2015 The Authors. Molecular Carcinogenesis Published by Wiley Periodicals, Inc. PMID:26676339

  10. Microwave and Beam Activation of Nanostructured Catalysts for Environmentally Friendly, Energy Efficient Heavy Crude Oil Processing

    SciTech Connect

    2009-03-01

    This factsheet describes a study whose goal is initial evaluation and development of energy efficient processes which take advantage of the benefits offered by nanostructured catalysts which can be activated by microwave, RF, or radiation beams.

  11. 77 FR 60714 - Information Collection Activities: Legacy Data Verification Process (LDVP); Submitted for Office...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-04

    ... Bureau of Safety and Environmental Enforcement Information Collection Activities: Legacy Data Verification Process (LDVP); Submitted for Office of Management and Budget (OMB) Review; Comment Request ACTION... the paperwork requirements for the Notice to Lessees (NTL) on the Legacy Data Verification...

  12. Orchard factors associated with resistance and cross resistance to sterol demethylation inhibitor fungicides in populations of Venturia inaequalis from Pennsylvania.

    PubMed

    Pfeufer, Emily E; Ngugi, Henry K

    2012-03-01

    Orchard management practices, such as destroying of overwintered inoculum and limiting the number of fungicide applications, are often recommended as tactics for slowing the development of resistance to sterol demethylation-inhibitor (DMI) fungicides in populations of Venturia inaequalis. However, there is little quantitative evidence relating the use of such practices to levels of resistance in orchards. The aim of this study was to evaluate the sensitivity of V. inaequalis isolates from Pennsylvania to DMI fungicides, and to identify orchard management factors related to the incidence of resistant isolates. In total, 644 single-spore V. inaequalis cultures obtained from 20 apple orchards in 2008 or 2009 were tested for sensitivity to myclobutanil, fenbuconazole, or difenoconazole. Growers provided management history of the sampled plots. Widespread shifts toward resistance to the three fungicides were noted, with mean effective concentration for 50% inhibition (EC(50)) values of 2.136, 0.786, and 0.187 μg/ml for myclobutanil, fenbuconazole, and difenoconazole, respectively. Cross resistance to the three fungicides was documented in high correlation (Spearman's r > 0.6) between mean EC(50) values for 14 orchards. Based on a 0.5-μg/ml threshold, 66 and 26% of isolates were resistant to myclobutanil and fenbuconazole, respectively, and 22% were cross resistant to the two fungicides. A significant between-year shift toward increased resistance was noted in two of three orchards surveyed in both years. Failure to use dormant copper sprays, older trees, larger orchards, orchards with ≤10 cultivars, and application of >4 DMI sprays were positively correlated (0.0001 < P < 0.05) with the incidence of resistant isolates. Isolates from orchards with >4 DMI sprays were four times as likely to be resistant to fenbuconazole (odds ratio = 4.57; P = 0.015). Isolates from orchards without dormant copper sprays were twice as likely to be cross-shifted toward resistance to all

  13. Antioxidant activity of fresh and processed Jalapeño and Serrano peppers.

    PubMed

    Alvarez-Parrilla, Emilio; de la Rosa, Laura A; Amarowicz, Ryszard; Shahidi, Fereidoon

    2011-01-12

    In this research, total phenols, flavonoids, capsaicinoids, ascorbic acid, and antioxidant activity (ORAC, hydroxyl radical, DPPH, and TEAC assays) of fresh and processed (pickled and chipotle canned) Jalapeño and Serrano peppers were determined. All fresh and processed peppers contained capsaicin, dihydrocapsaicin, and nordihydrocapsaicin, even though the latter could be quantified only in fresh peppers. Processed peppers contained lower amounts of phytochemicals and had lower antioxidant activity, compared to fresh peppers. Good correlations between total phenols and ascorbic acid with antioxidant activity were observed. Elimination of chlorophylls by silicic acid chromatography reduced the DPPH scavenging activity of the extracts, compared to crude extracts, confirming the antioxidant activity of chlorophylls present in Jalapeño and Serrano peppers.

  14. Information entropy of activation process: Application for low-temperature fluctuations of a myoglobin molecule

    NASA Astrophysics Data System (ADS)

    Stepanov, A. V.

    2015-11-01

    Activation process for unimolecular reaction has been considered by means of radiation theory. The formulae of information entropy of activation have been derived for the Boltzmann-Arrhenius model and the activation process model (APM). The physical meaning of this entropy has been determined. It is a measure of conversion of thermal radiation energy to mechanical energy that moves atoms in a molecule during elementary activation act. It is also a measure of uncertainty of this energy conversion. The uncertainty is due to unevenness of distribution function representing the activation process. It has been shown that Arrhenius dependence is caused by the entropy change. Efficiency comparison of the two models under consideration for low-temperature fluctuations of a myoglobin molecule structure shows that the APM should be favored over the Boltzmann-Arrhenius one.

  15. Meditation-induced neuroplastic changes in amygdala activity during negative affective processing.

    PubMed

    Leung, Mei-Kei; Lau, Way K W; Chan, Chetwyn C H; Wong, Samuel S Y; Fung, Annis L C; Lee, Tatia M C

    2017-04-10

    Recent evidence suggests that the effects of meditation practice on affective processing and resilience have the potential to induce neuroplastic changes within the amygdala. Notably, literature speculates that meditation training may reduce amygdala activity during negative affective processing. Nonetheless, studies have thus far not verified this speculation. In this longitudinal study, participants (N = 21, 9 men) were trained in awareness-based compassion meditation (ABCM) or matched relaxation training. The effects of meditation training on amygdala activity were examined during passive viewing of affective and neutral stimuli in a non-meditative state. We found that the ABCM group exhibited significantly reduced anxiety and right amygdala activity during negative emotion processing than the relaxation group. Furthermore, ABCM participants who performed more compassion practice had stronger right amygdala activity reduction during negative emotion processing. The lower right amygdala activity after ABCM training may be associated with a general reduction in reactivity and distress. As all participants performed the emotion processing task in a non-meditative state, it appears likely that the changes in right amygdala activity are carried over from the meditation practice into the non-meditative state. These findings suggest that the distress-reducing effects of meditation practice on affective processing may transfer to ordinary states, which have important implications on stress management.

  16. Measurements of cloud condensation nuclei activity and droplet activation kinetics of wet processed regional dust samples and minerals

    NASA Astrophysics Data System (ADS)

    Kumar, P.; Sokolik, I. N.; Nenes, A.

    2011-04-01

    This study reports laboratory measurements of particle size distributions, cloud condensation nuclei (CCN) activity, and droplet activation kinetics of wet generated aerosols from clays, calcite, quartz, and desert soil samples from Northern Africa, East Asia/China, and Northern America. The dependence of critical supersaturation, sc, on particle dry diameter, Ddry, is used to characterize particle-water interactions and assess the ability of Frenkel-Halsey-Hill adsorption activation theory (FHH-AT) and Köhler theory (KT) to describe the CCN activity of the considered samples. Regional dust samples produce unimodal size distributions with particle sizes as small as 40 nm, CCN activation consistent with KT, and exhibit hygroscopicity similar to inorganic salts. Clays and minerals produce a bimodal size distribution; the CCN activity of the smaller mode is consistent with KT, while the larger mode is less hydrophilic, follows activation by FHH-AT, and displays almost identical CCN activity to dry generated dust. Ion Chromatography (IC) analysis performed on regional dust samples indicates a soluble fraction that cannot explain the CCN activity of dry or wet generated dust. A mass balance and hygroscopicity closure suggests that the small amount of ions (of low solubility compounds like calcite) present in the dry dust dissolve in the aqueous suspension during the wet generation process and give rise to the observed small hygroscopic mode. Overall these results identify an artifact that may question the atmospheric relevance of dust CCN activity studies using the wet generation method. Based on a threshold droplet growth analysis, wet generated mineral aerosols display similar activation kinetics compared to ammonium sulfate calibration aerosol. Finally, a unified CCN activity framework that accounts for concurrent effects of solute and adsorption is developed to describe the CCN activity of aged or hygroscopic dusts.

  17. 78 FR 21118 - Agency Information Collection Activities; Comment Request; Streamlined Clearance Process for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-09

    ... Agency Information Collection Activities; Comment Request; Streamlined Clearance Process for... Clearance Division, U.S. Department of Education, 400 Maryland Avenue SW., LBJ, Room 2E105, Washington, DC... Clearance Process for Discretionary Grants OMB Control Number: 1894-0001 Type of Review: An extension of...

  18. Executive Functions in Learning Processes: Do They Benefit from Physical Activity?

    ERIC Educational Resources Information Center

    Barenberg, Jonathan; Berse, Timo; Dutke, Stephan

    2011-01-01

    As executive functions play an essential role in learning processes, approaches capable of enhancing executive functioning are of particular interest to educational psychology. Recently, the hypothesis has been advanced that executive functioning may benefit from changes in neurobiological processes induced by physical activity. The present…

  19. Effect of spray drying processing parameters on the insecticidal activity of two encapsulated formulations of baculovirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this work was to evaluate the effect of spray dryer processing parameters on the process yield and insecticidal activity of baculovirus to support the development of this beneficial group of microbes as biopesticides. For each of two baculoviruses [granulovirus (GV) from Pieris rapae (L....

  20. The Relationships of Mental States and Intellectual Processes in the Learning Activities of Students

    ERIC Educational Resources Information Center

    Prokhorov, Alexander O.; Chernov, Albert V.; Yusupov, Mark G.

    2016-01-01

    Investigation of the interaction of mental states and cognitive processes in the classroom allows us to solve the problem of increasing the effectiveness of training by activating cognitive processes and management of students' mental states. This article is concerned with the most general patterns of interaction between mental state and…

  1. Prospective Elementary Mathematics Teachers' Thought Processes on a Model Eliciting Activity

    ERIC Educational Resources Information Center

    Eraslan, Ali

    2012-01-01

    Mathematical model and modeling are one of the topics that have been intensively discussed in recent years. The purpose of this study is to examine prospective elementary mathematics teachers' thought processes on a model eliciting activity and reveal difficulties or blockages in the processes. The study includes forty-five seniors taking the…

  2. Learning Computers, Speaking English: Cooperative Activities for Learning English and Basic Word Processing.

    ERIC Educational Resources Information Center

    Quann, Steve; Satin, Diana

    This textbook leads high-beginning and intermediate English-as-a-Second-Language (ESL) students through cooperative computer-based activities that combine language learning with training in basic computer skills and word processing. Each unit concentrates on a basic concept of word processing while also focusing on a grammar topic. Skills are…

  3. 75 FR 71677 - Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-24

    ... Reimbursement for Costs of Remedial Action at Active Uranium and Thorium Processing Sites AGENCY: Department of... uranium and thorium processing site licensees for reimbursement under Title X of the Energy Policy Act of... requires DOE to reimburse eligible uranium and thorium licensees for certain costs of...

  4. The Masked Semantic Priming Effect Is Task Dependent: Reconsidering the Automatic Spreading Activation Process

    ERIC Educational Resources Information Center

    de Wit, Bianca; Kinoshita, Sachiko

    2015-01-01

    Semantic priming effects are popularly explained in terms of an automatic spreading activation process, according to which the activation of a node in a semantic network spreads automatically to interconnected nodes, preactivating a semantically related word. It is expected from this account that semantic priming effects should be routinely…

  5. Plan before You Play: An Activity for Teaching the Managerial Process

    ERIC Educational Resources Information Center

    Althouse, Norm R.; Hedges, Peggy L.

    2015-01-01

    This article describes a 60-minute classroom activity using LEGO® bricks that demonstrates and reinforces the importance of the managerial process. The activity, Plan Before You Play (PBP), is targeted to introductory business classes, and differs from others in that it requires little investment or up-front planning, is easily scalable, and, with…

  6. Active Parent Consent for Health Surveys with Urban Middle School Students: Processes and Outcomes

    ERIC Educational Resources Information Center

    Secor-Turner, Molly; Sieving, Renee; Widome, Rachel; Plowman, Shari; Vanden Berk, Eric

    2010-01-01

    Background: To achieve high participation rates and a representative sample, active parent consent procedures require a significant investment of study resources. The purpose of this article is to describe processes and outcomes of utilizing active parent consent procedures with sixth-grade students from urban, ethnically diverse, economically…

  7. Process, Goal and Social Interaction Differences in Recreation: What Makes an Activity Substitutable.

    ERIC Educational Resources Information Center

    Baumgartner, Robert; Heberlein, Thomas A.

    Two recreational activities, deer hunting and goose hunting, both similar in form, are compared. It was hypothesized that the activity for which participants rated the process, the goal, and the social interaction as most important to the experience and for which participants showed the strongest family ties and social support for participation…

  8. A modified oxic-settling-anaerobic activated sludge process using gravity thickening for excess sludge reduction

    PubMed Central

    Wang, Jun; Li, Shi-Yu; Jiang, Feng; Wu, Ke; Liu, Guang-Li; Lu, Hui; Chen, Guang-Hao

    2015-01-01

    Oxic-settling-anaerobic process (OSA) was known as a cost-effective way to reduce the excess sludge production with simple upgrade of conventional activated sludge process (CAS). A low oxidation-reduction potential (ORP) level was the key factor to sludge decay and lysis in the sludge holding tank of the OSA process. However, the ORP control with nitrogen purge or chemical dosing in the OSA process would induce extra expense and complicate the operation. Hence, in this study, a sludge holding tank using gravity thickening was applied to OSA process to reduce the excess sludge production without any ORP control. Results showed that the modified OSA process not only reduced the excess sludge production effectively but also improved the sludge settleability without affected the treatment capacity. The reduction of the excess sludge production in the modified OSA process resulted from interactions among lots of factors. The key element of the process was the gravity thickening sludge holding tank. PMID:26350761

  9. A modified oxic-settling-anaerobic activated sludge process using gravity thickening for excess sludge reduction

    NASA Astrophysics Data System (ADS)

    Wang, Jun; Li, Shi-Yu; Jiang, Feng; Wu, Ke; Liu, Guang-Li; Lu, Hui; Chen, Guang-Hao

    2015-09-01

    Oxic-settling-anaerobic process (OSA) was known as a cost-effective way to reduce the excess sludge production with simple upgrade of conventional activated sludge process (CAS). A low oxidation-reduction potential (ORP) level was the key factor to sludge decay and lysis in the sludge holding tank of the OSA process. However, the ORP control with nitrogen purge or chemical dosing in the OSA process would induce extra expense and complicate the operation. Hence, in this study, a sludge holding tank using gravity thickening was applied to OSA process to reduce the excess sludge production without any ORP control. Results showed that the modified OSA process not only reduced the excess sludge production effectively but also improved the sludge settleability without affected the treatment capacity. The reduction of the excess sludge production in the modified OSA process resulted from interactions among lots of factors. The key element of the process was the gravity thickening sludge holding tank.

  10. Activity of LPO Processes in Women with Polycystic Ovarian Syndrome and Infertility.

    PubMed

    Kolesnikova, L I; Kolesnikov, S I; Darenskaya, M A; Grebenkina, L A; Nikitina, O A; Lazareva, L M; Suturina, L V; Danusevich, I N; Druzhinina, E B; Semendyaev, A A

    2017-01-01

    Specific features of LPO processes and antioxidant defense were studied in patients with polycystic ovarian syndrome (PCOS) and infertility. Changes in LPO processes in patients with PCOS were compensatory, which manifested in increased α-tocopherol and retinol concentrations and moderate decrease in superoxide dismutase activity. Intensification of prooxidant processes was found in the group of patients with infertility without PCOS. The observed changes necessitate differentiated approach to the treatment of these patients.

  11. Selective attention modulates high-frequency activity in the face-processing network.

    PubMed

    Müsch, Kathrin; Hamamé, Carlos M; Perrone-Bertolotti, Marcela; Minotti, Lorella; Kahane, Philippe; Engel, Andreas K; Lachaux, Jean-Philippe; Schneider, Till R

    2014-11-01

    Face processing depends on the orchestrated activity of a large-scale neuronal network. Its activity can be modulated by attention as a function of task demands. However, it remains largely unknown whether voluntary, endogenous attention and reflexive, exogenous attention to facial expressions equally affect all regions of the face-processing network, and whether such effects primarily modify the strength of the neuronal response, the latency, the duration, or the spectral characteristics. We exploited the good temporal and spatial resolution of intracranial electroencephalography (iEEG) and recorded from depth electrodes to uncover the fast dynamics of emotional face processing. We investigated frequency-specific responses and event-related potentials (ERP) in the ventral occipito-temporal cortex (VOTC), ventral temporal cortex (VTC), anterior insula, orbitofrontal cortex (OFC), and amygdala when facial expressions were task-relevant or task-irrelevant. All investigated regions of interest (ROI) were clearly modulated by task demands and exhibited stronger changes in stimulus-induced gamma band activity (50-150 Hz) when facial expressions were task-relevant. Observed latencies demonstrate that the activation is temporally coordinated across the network, rather than serially proceeding along a processing hierarchy. Early and sustained responses to task-relevant faces in VOTC and VTC corroborate their role for the core system of face processing, but they also occurred in the anterior insula. Strong attentional modulation in the OFC and amygdala (300 msec) suggests that the extended system of the face-processing network is only recruited if the task demands active face processing. Contrary to our expectation, we rarely observed differences between fearful and neutral faces. Our results demonstrate that activity in the face-processing network is susceptible to the deployment of selective attention. Moreover, we show that endogenous attention operates along the whole

  12. Aging process alters hippocampal and cortical secretase activities of Wistar rats.

    PubMed

    Bertoldi, Karine; Cechinel, Laura Reck; Schallenberger, Bruna; Meireles, Louisiana; Basso, Carla; Lovatel, Gisele Agustini; Bernardi, Lisiane; Lamers, Marcelo Lazzaron; Siqueira, Ionara Rodrigues

    2017-01-15

    A growing body of evidence has demonstrated amyloid plaques in aged brain; however, little attention has been given to amyloid precursor protein (APP) processing machinery during the healthy aging process. The amyloidogenic and non-amyloidogenic pathways, represented respectively by β- and α-secretases (BACE and TACE), are responsible for APP cleavage. Our working hypothesis is that the normal aging process could imbalance amyloidogenic and non-amyloidogenic pathways specifically BACE and TACE activities. Besides, although it has been showed that exercise can modulate secretase activities in Alzheimer Disease models the relationship between exercise effects and APP processing during healthy aging process is rarely studied. Our aim was to investigate the aging process and the exercise effects on cortical and hippocampal BACE and TACE activities and aversive memory performance. Young adult and aged Wistar rats were subjected to an exercise protocol (20min/day for 2 weeks) and to inhibitory avoidance task. Biochemical parameters were evaluated 1h and 18h after the last exercise session in order to verify transitory and delayed exercise effects. Aged rats exhibited impaired aversive memory and diminished cortical TACE activity. Moreover, an imbalance between TACE and BACE activities in favor of BACE activity was observed in aged brain. Moderate treadmill exercise was unable to alter secretase activities in any brain areas or time points evaluated. Our results suggest that aging-related aversive memory decline is partly linked to decreased cortical TACE activity. Additionally, an imbalance between secretase activities can be related to the higher vulnerability to neurodegenerative diseases induced by aging.

  13. Human intracranial high-frequency activity during memory processing: neural oscillations or stochastic volatility?

    PubMed

    Burke, John F; Ramayya, Ashwin G; Kahana, Michael J

    2015-04-01

    Intracranial high-frequency activity (HFA), which refers to fast fluctuations in electrophysiological recordings, increases during memory processing. Two views have emerged to explain this effect: (1) HFA reflects a synchronous signal, related to underlying gamma oscillations, that plays a mechanistic role in human memory and (2) HFA reflects an asynchronous signal that is a non-specific marker of brain activation. We review recent data supporting each of these views and conclude that HFA during memory processing is more consistent with an asynchronous signal. Memory-related HFA is therefore best conceptualized as a biomarker of neural activation that can functionally map memory with high spatial and temporal precision.

  14. Rapid regulation of sialidase activity in response to neural activity and sialic acid removal during memory processing in rat hippocampus.

    PubMed

    Minami, Akira; Meguro, Yuko; Ishibashi, Sayaka; Ishii, Ami; Shiratori, Mako; Sai, Saki; Horii, Yuuki; Shimizu, Hirotaka; Fukumoto, Hokuto; Shimba, Sumika; Taguchi, Risa; Takahashi, Tadanobu; Otsubo, Tadamune; Ikeda, Kiyoshi; Suzuki, Takashi

    2017-04-07

    Sialidase cleaves sialic acids on the extracellular cell surface as well as inside the cell and is necessary for normal long-term potentiation (LTP) at mossy fiber-CA3 pyramidal cell synapses and for hippocampus-dependent spatial memory. Here, we investigated in detail the role of sialidase in memory processing. Sialidase activity measured with 4-methylumbelliferyl-α-d-N-acetylneuraminic acid (4MU-Neu5Ac) or 5-bromo-4-chloroindol-3-yl-α-d-N-acetylneuraminic acid (X-Neu5Ac) and Fast Red Violet LB was increased by high-K(+)-induced membrane depolarization. Sialidase activity was also increased by chemical LTP induction with forskolin and activation of BDNF signaling, non-NMDA receptors, or NMDA receptors. The increase in sialidase activity with neural excitation appears to be caused not by secreted sialidase or by an increase in sialidase expression but by a change in the subcellular localization of sialidase. Astrocytes as well as neurons are also involved in the neural activity-dependent increase in sialidase activity. Sialidase activity visualized with a benzothiazolylphenol-based sialic acid derivative (BTP3-Neu5Ac), a highly sensitive histochemical imaging probe for sialidase activity, at the CA3 stratum lucidum of rat acute hippocampal slices was immediately increased in response to LTP-inducible high-frequency stimulation on a time scale of seconds. To obtain direct evidence for sialic acid removal on the extracellular cell surface during neural excitation, the extracellular free sialic acid level in the hippocampus was monitored using in vivo microdialysis. The free sialic acid level was increased by high-K(+)-induced membrane depolarization. Desialylation also occurred during hippocampus-dependent memory formation in a contextual fear-conditioning paradigm. Our results show that neural activity-dependent desialylation by sialidase may be involved in hippocampal memory processing.

  15. Preparation of sulfurized powdered activated carbon from waste tires using an innovative compositive impregnation process.

    PubMed

    Yuan, Chung-Shin; Lin, Hsun-Yu; Wu, Chun-Hsin; Liu, Ming-Han; Hung, Chung-Hsuang

    2004-07-01

    The objective of this study is to develop an innovative compositive impregnation process for preparing sulfurized powdered activated carbon (PAC) from waste tires. An experimental apparatus, including a pyrolysis and activation system and a sulfur (S) impregnation system, was designed and applied to produce sulfurized PAC with a high specific surface area. Experimental tests involved the pyrolysis, activation, and sulfurization of waste tires. Waste-tire-derived PAC (WPAC) was initially produced in the pyrolysis and activation system. Experimental results indicated that the Brunauer-Emmett-Teller (BET) surface area of WPAC increased, and the average pore radius of WPAC decreased, as water feed rate and activation time increased. In this study, a conventional direct impregnation process was used to prepare the sulfurized PAC by impregnating WPAC with sodium sulfide (Na2S) solution. Furthermore, an innovative compositive impregnation process was developed and then compared with the conventional direct impregnation process. Experimental results showed that the compositive impregnation process produced the sulfurized WPAC with high BET surface area and a high S content. A maximum BET surface area of 886 m2/g and the S content of 2.61% by mass were obtained at 900 degrees C and at the S feed ratio of 2160 mg Na2S/g C. However, the direct impregnation process led to a BET surface area of sulfurized WPAC that decreased significantly as the S content increased.

  16. Epileptiform activity in the mouse visual cortex interferes with cortical processing in connected areas

    PubMed Central

    Petrucco, L.; Pracucci, E.; Brondi, M.; Ratto, G. M.; Landi, S.

    2017-01-01

    Epileptiform activity is associated with impairment of brain function even in absence of seizures, as demonstrated by failures in various testing paradigm in presence of hypersynchronous interictal spikes (ISs). Clinical evidence suggests that cognitive deficits might be directly caused by the anomalous activity rather than by its underlying etiology. Indeed, we seek to understand whether ISs interfere with neuronal processing in connected areas not directly participating in the hypersynchronous activity in an acute model of epilepsy. Here we cause focal ISs in the visual cortex of anesthetized mice and we determine that, even if ISs do not invade the opposite hemisphere, the local field potential is subtly disrupted with a modulation of firing probability imposed by the contralateral IS activity. Finally, we find that visual processing is altered depending on the temporal relationship between ISs and stimulus presentation. We conclude that focal ISs interact with normal cortical dynamics far from the epileptic focus, disrupting endogenous oscillatory rhythms and affecting information processing. PMID:28071688

  17. Pilot and full scale applications of sulfur-based autotrophic denitrification process for nitrate removal from activated sludge process effluent.

    PubMed

    Sahinkaya, Erkan; Kilic, Adem; Duygulu, Bahadir

    2014-09-01

    Sulfur-based autotrophic denitrification of nitrified activated sludge process effluent was studied in pilot and full scale column bioreactors. Three identical pilot scale column bioreactors packed with varying sulfur/lime-stone ratios (1/1-3/1) were setup in a local wastewater treatment plant and the performances were compared under varying loading conditions for long-term operation. Complete denitrification was obtained in all pilot bioreactors even at nitrate loading of 10 mg NO3(-)-N/(L.h). When the temperature decreased to 10 °C during the winter time at loading of 18 mg NO3(-)-N/(L.h), denitrification efficiency decreased to 60-70% and the bioreactor with S/L ratio of 1/1 gave slightly better performance. A full scale sulfur-based autotrophic denitrification process with a S/L ratio of 1/1 was set up for the denitrification of an activated sludge process effluent with a flow rate of 40 m(3)/d. Almost complete denitrification was attained with a nitrate loading rate of 6.25 mg NO3(-)-N/(L.h).

  18. Induction of a specific CD8+ T-cell response to cancer/testis antigens by demethylating pre-treatment against osteosarcoma.

    PubMed

    Li, Binghao; Zhu, Xiaobing; Sun, Lingling; Yuan, Li; Zhang, Jian; Li, Hengyuan; Ye, Zhaoming

    2014-11-15

    Conventional non-surgical therapeutic regimens against osteosarcoma are subject to chemoresistance and tumor relapse, and immunotherapy may be promising for this tumor. However, it's hard to find satisfactory epitopes for immunotherapy against osteosarcoma. Cancer/testis antigens (CTAs), such as MAGE-A family and NY-ESO-1, the potential antigens that almost exclusively express in tumor cells and immune-privileged sites, have been found expressed in osteosarcoma also. Nevertheless, the expression of CTAs is downregulated in many tumors, constraining the application of immunotherapy. In this article, we demonstrate that the expression of MAGE-A family and NY-ESO-1 in osteosarcoma cells can be upregulated following treatment with demethylating agent 5-aza-2'-deoxycytidine and consequently induces a CTA specific CD8+ T-cell response against osteosarcoma in vitro and in vivo. The in vivo imaging was realized by using luciferase-transfected HOS cells and DiR labeled T-cells in severely combined immunodeficiency mouse models. Cytotoxic T cells specifically recognizing MAGE-A family and NY-ESO-1 clustered at the tumor site in mice pre-treated with DAC and resulted in tumor growth suppression, while it was not observed in mice without DAC pre-treatment. This study is important for more targeted therapeutic approaches and suggests that adoptive immunotherapy, combined with demethylating treatment, has the potential for non-surgical therapeutic strategy against osteosarcoma.

  19. Growth-inducing effects of argon plasma on soybean sprouts via the regulation of demethylation levels of energy metabolism-related genes

    PubMed Central

    Zhang, Jiao Jiao; Jo, Jin Oh; Huynh, Do Luong; Mongre, Raj Kumar; Ghosh, Mrinmoy; Singh, Amit Kumar; Lee, Sang Baek; Mok, Young Sun; Hyuk, Park; Jeong, Dong Kee

    2017-01-01

    This study was conducted to determine the effects of argon plasma on the growth of soybean [Glycine max (L.) Merr.] sprouts and investigate the regulation mechanism of energy metabolism. The germination and growth characteristics were modified by argon plasma at different potentials and exposure durations. Upon investigation, plasma treatment at 22.1 kV for 12 s maximized the germination and seedling growth of soybean, increasing the concentrations of soluble protein, antioxidant enzymes, and adenosine triphosphate (ATP) as well as up-regulating ATP a1, ATP a2, ATP b1, ATP b2, ATP b3, target of rapamycin (TOR), growth-regulating factor (GRF) 1–6, down-regulating ATP MI25 mRNA expression, and increasing the demethylation levels of the sequenced region of ATP a1, ATP b1, TOR, GRF 5, and GRF 6 of 6-day-old soybean sprouts. These observations indicate that argon plasma promotes soybean seed germination and sprout growth by regulating the demethylation levels of ATP, TOR, and GRF. PMID:28167819

  20. Growth-inducing effects of argon plasma on soybean sprouts via the regulation of demethylation levels of energy metabolism-related genes

    NASA Astrophysics Data System (ADS)

    Zhang, Jiao Jiao; Jo, Jin Oh; Huynh, Do Luong; Mongre, Raj Kumar; Ghosh, Mrinmoy; Singh, Amit Kumar; Lee, Sang Baek; Mok, Young Sun; Hyuk, Park; Jeong, Dong Kee

    2017-02-01

    This study was conducted to determine the effects of argon plasma on the growth of soybean [Glycine max (L.) Merr.] sprouts and investigate the regulation mechanism of energy metabolism. The germination and growth characteristics were modified by argon plasma at different potentials and exposure durations. Upon investigation, plasma treatment at 22.1 kV for 12 s maximized the germination and seedling growth of soybean, increasing the concentrations of soluble protein, antioxidant enzymes, and adenosine triphosphate (ATP) as well as up-regulating ATP a1, ATP a2, ATP b1, ATP b2, ATP b3, target of rapamycin (TOR), growth-regulating factor (GRF) 1–6, down-regulating ATP MI25 mRNA expression, and increasing the demethylation levels of the sequenced region of ATP a1, ATP b1, TOR, GRF 5, and GRF 6 of 6-day-old soybean sprouts. These observations indicate that argon plasma promotes soybean seed germination and sprout growth by regulating the demethylation levels of ATP, TOR, and GRF.

  1. Demethylated HSATII DNA and HSATII RNA Foci Sequester PRC1 and MeCP2 into Cancer-Specific Nuclear Bodies.

    PubMed

    Hall, Lisa L; Byron, Meg; Carone, Dawn M; Whitfield, Troy W; Pouliot, Gayle P; Fischer, Andrew; Jones, Peter; Lawrence, Jeanne B

    2017-03-21

    This study reveals that high-copy satellite II (HSATII) sequences in the human genome can bind and impact distribution of chromatin regulatory proteins and that this goes awry in cancer. In many cancers, master regulatory proteins form two types of cancer-specific nuclear bodies, caused by locus-specific deregulation of HSATII. DNA demethylation at the 1q12 mega-satellite, common in cancer, causes PRC1 aggregation into prominent Cancer-Associated Polycomb (CAP) bodies. These loci remain silent, whereas HSATII loci with reduced PRC1 become derepressed, reflecting imbalanced distribution of UbH2A on these and other PcG-regulated loci. Large nuclear foci of HSATII RNA form and sequester copious MeCP2 into Cancer-Associated Satellite Transcript (CAST) bodies. Hence, HSATII DNA and RNA have an exceptional capacity to act as molecular sponges and sequester chromatin regulatory proteins into abnormal nuclear bodies in cancer. The compartmentalization of regulatory proteins within nuclear structure, triggered by demethylation of "junk" repeats, raises the possibility that this contributes to further compromise of the epigenome and neoplastic progression.

  2. Main contribution of the cytochrome P450 isoenzyme 1A2 (CYP1A2) to N-demethylation and 5-sulfoxidation of the phenothiazine neuroleptic chlorpromazine in human liver--A comparison with other phenothiazines.

    PubMed

    Wójcikowski, Jacek; Boksa, Jan; Daniel, Władysława A

    2010-10-15

    The aim of the present study was to identify cytochrome P450 (CYP) isoenzymes involved in the 5-sulfoxidation, mono-N-demethylation and di-N-demethylation of the aliphatic-type phenothiazine neuroleptic chlorpromazine in human liver. Experiments were performed in vitro using cDNA-expressed human CYP isoforms (Supersomes 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4), liver microsomes from different donors and CYP-selective inhibitors. The obtained results indicate that CYP1A2 is the only CYP isoform that catalyzes the mono-N-demethylation and di-N-demethylation of chlorpromazine (100%) and is the main isoform responsible for chlorpromazine 5-sulfoxidation (64%) at a therapeutic concentration of the drug (10 microM). CYP3A4 contributes to a lesser degree to chlorpromazine 5-sulfoxidation (34%). The role of CYP2B6, CYP2C19 and CYP2D6 in catalyzing of the latter reaction is negligible (0.1-2%). Similar results were obtained at a higher, non-therapeutic concentration of the drug (100 microM); however, the contribution of CYP1A2 to chlorpromazine mono-N-demethylation was noticeably lower (75%), mostly in favour of CYP2C19 and CYP3A4 (about 12% each). The obtained results indicate that the catalysis of chlorpromazine N-demethylation and 5-sulfoxidation in humans exhibits a stricter CYP1A2 preference compared to the previously tested phenothiazines (promazine, perazine, and thioridazine). Hence pharmacokinetic interactions involving chlorpromazine and CYP1A2 substrates and inhibitors are likely to occur. Considering strong dopaminergic D(2), noradrenergic alpha(1) and cholinergic M(1) receptor blocking properties of chlorpromazine and some of its metabolites, as well as their serious side effects, the obtained results may be of pharmacological and clinical importance.

  3. Idaho Chemical Processing Plant low-activity waste grout stabilization development program FY-97 status report

    SciTech Connect

    Herbst, A.K.; Marshall, D.W.; McCray, J.A.

    1998-02-01

    The general purpose of the Grout Development Program is to solidify and stabilize the liquid low-activity wastes (LAW) generated at the Idaho Chemical Processing Plant (ICPP). It is anticipated that LAW will be produced from the following: (1) chemical separation of the tank farm high-activity sodium-bearing waste, (2) retrieval, dissolution, and chemical separation of the aluminum, zirconium, and sodium calcines, (3) facility decontamination processes, and (4) process equipment waste. Grout formulation studies for sodium-bearing LAW, including decontamination and process equipment waste, continued this fiscal year. A second task was to develop a grout formulation to solidify potential process residual heels in the tank farm vessels when the vessels are closed.

  4. The influence of spontaneous activity on stimulus processing in primary visual cortex.

    PubMed

    Schölvinck, M L; Friston, K J; Rees, G

    2012-02-01

    Spontaneous activity in the resting human brain has been studied extensively; however, how such activity affects the local processing of a sensory stimulus is relatively unknown. Here, we examined the impact of spontaneous activity in primary visual cortex on neuronal and behavioural responses to a simple visual stimulus, using functional MRI. Stimulus-evoked responses remained essentially unchanged by spontaneous fluctuations, combining with them in a largely linear fashion (i.e., with little evidence for an interaction). However, interactions between spontaneous fluctuations and stimulus-evoked responses were evident behaviourally; high levels of spontaneous activity tended to be associated with increased stimulus detection at perceptual threshold. Our results extend those found in studies of spontaneous fluctuations in motor cortex and higher order visual areas, and suggest a fundamental role for spontaneous activity in stimulus processing.

  5. Modelling of activation processes for GR-280 graphite at Ignalina NPP.

    PubMed

    Smaizys, Arturas; Narkunas, Ernestas; Poskas, Povilas

    2005-01-01

    Ignalina Nuclear Power Plant (INPP) operates two RBMK-1500 water-cooled graphite-moderated channel-type power reactors. The total mass of graphite in the cores of both units at INPP is about 3600 tons. Modelling of activation processes in the reactor's structural materials is necessary for decommissioning planning, because large amounts of activated structural materials (graphite, stainless steel, concrete, etc.) should be managed as radioactive waste. Knowledge of radiological characteristics and a radioactive inventory of irradiated materials are essential in planning of the decommissioning processes. The purpose of this work was to perform conservative neutron activation analysis for decommissioning purposes of INPP. ORIGEN computer code was used for the calculations. Activity levels were calculated for different nuclides present in the graphite and estimates were made how these activity levels depend on irradiation time, neutron flux value and other parameters. Obtained results were compared with the data available from other investigations for GR-280 graphite.

  6. Comparison of natural estrogen removal efficiency in the conventional activated sludge process and the oxidation ditch process.

    PubMed

    Hashimoto, T; Onda, K; Nakamura, Y; Tada, K; Miya, A; Murakami, T

    2007-05-01

    The presence of natural estrogens, 17beta-estradiol (E2), estrone (E1) and estriol (E3), as well as estrogenic activity in wastewater influents and secondary effluents were investigated in 20 full-scale wastewater treatment plants in Japan. In all of the influent samples, natural estrogens were detected at concentrations above the minimum limits of detection (0.5ng/L). The concentrations of natural estrogens detected in the effluent of oxidation ditch plants were generally lower than previously reported values. On the other hand, in the conventional activated sludge plants, increments of E1 during biological treatment were frequently observed although E2 and E3 were removed effectively in the process. The removal rates of natural estrogens or estrogenic activity show no observed statistical relationship with the solids retention time (SRT) and the hydraulic retention time (HRT). However, the plants with high SRT or HRT generally showed high and stable removal of both natural estrogens and estrogenic activity.

  7. Relative Contributions of Specific Activity Histories and Spontaneous Processes to Size Remodeling of Glutamatergic Synapses

    PubMed Central

    Dvorkin, Roman; Ziv, Noam E.

    2016-01-01

    The idea that synaptic properties are defined by specific pre- and postsynaptic activity histories is one of the oldest and most influential tenets of contemporary neuroscience. Recent studies also indicate, however, that synaptic properties often change spontaneously, even in the absence of specific activity patterns or any activity whatsoever. What, then, are the relative contributions of activity history-dependent and activity history-independent processes to changes synapses undergo? To compare the relative contributions of these processes, we imaged, in spontaneously active networks of cortical neurons, glutamatergic synapses formed between the same axons and neurons or dendrites under the assumption that their similar activity histories should result in similar size changes over timescales of days. The size covariance of such commonly innervated (CI) synapses was then compared to that of synapses formed by different axons (non-CI synapses) that differed in their activity histories. We found that the size covariance of CI synapses was greater than that of non-CI synapses; yet overall size covariance of CI synapses was rather modest. Moreover, momentary and time-averaged sizes of CI synapses correlated rather poorly, in perfect agreement with published electron microscopy-based measurements of mouse cortex synapses. A conservative estimate suggested that ~40% of the observed size remodeling was attributable to specific activity histories, whereas ~10% and ~50% were attributable to cell-wide and spontaneous, synapse-autonomous processes, respectively. These findings demonstrate that histories of naturally occurring activity patterns can direct glutamatergic synapse remodeling but also suggest that the contributions of spontaneous, possibly stochastic, processes are at least as great. PMID:27776122

  8. Role of invertase activity in processing quality of potatoes: Effect of storage temperature and duration.

    PubMed

    Bandana; Sharma, Vineet; Singh, Brajesh; Raigond, Pinky; Kaushik, S K

    2016-03-01

    Invertase activity and processing attributes of three potato cultivars were studied to find the reason for deterioration of processing quality during their prolonged storage in commercial cold stores (4°C) as compared to elevated temperature storage (12 ± 0.5°C), with CIPC {Isopropyl-N-(3-Cholorophenyl) carbamate}. Lower storage temperature (4°C) tended to be more effective in increasing invertase activity of potato tubers than elevated temperature. Non-processing cultivar viz., Kufri Pukhraj resulted in accumulation of more invertase activity than relatively two processing cultivars. Kufri Chipsona-1 and Kufri Chipsona-3 at 12 ± 0.5°C possessed basal invertase activity ranging from 39.3 to 79.8 and 54.1 to 93.8 (pmoles hexose h⁻¹ g⁻¹ f.wt.) respectively, during two years. Total invertase activity at 4°C increased abruptly and remained high from 30 to 60 days of storage. The activity progressively reached 90.6 to 106.6 and 81.4 to 101.3 during both the years respectively, after 60 days of storage to that observed initially. Reducing sugar content increased from 23.3 to 105.7 and 389.0 to 1138.2 (mg 100g⁻¹ f.wt.) after 90 days of storage at 12 ± 0.5°C and 4°C, respectively. Studies concluded that basal and total invertase, were responsible for cold-induced sweetening and resulted in deterioration of processing quality of potatoes during storage at 4°C. Since this activity is low at 12 ± 0.5°C, the processing traits remained acceptable to industry and consumers.

  9. Left ventral occipitotemporal activation during orthographic and semantic processing of auditory words.

    PubMed

    Ludersdorfer, Philipp; Wimmer, Heinz; Richlan, Fabio; Schurz, Matthias; Hutzler, Florian; Kronbichler, Martin

    2016-01-01

    The present fMRI study investigated the hypothesis that activation of the left ventral occipitotemporal cortex (vOT) in response to auditory words can be attributed to lexical orthographic rather than lexico-semantic processing. To this end, we presented auditory words in both an orthographic ("three or four letter word?") and a semantic ("living or nonliving?") task. In addition, a auditory control condition presented tones in a pitch evaluation task. The results showed that the left vOT exhibited higher activation for orthographic relative to semantic processing of auditory words with a peak in the posterior part of vOT. Comparisons to the auditory control condition revealed that orthographic processing of auditory words elicited activation in a large vOT cluster. In contrast, activation for semantic processing was only weak and restricted to the middle part vOT. We interpret our findings as speaking for orthographic processing in left vOT. In particular, we suggest that activation in left middle vOT can be attributed to accessing orthographic whole-word representations. While activation of such representations was experimentally ascertained in the orthographic task, it might have also occurred automatically in the semantic task. Activation in the more posterior vOT region, on the other hand, may reflect the generation of explicit images of word-specific letter sequences required by the orthographic but not the semantic task. In addition, based on cross-modal suppression, the finding of marked deactivations in response to the auditory tones is taken to reflect the visual nature of representations and processes in left vOT.

  10. Retinoic acid dependent histone 3 demethylation of the clustered HOX genes during neural differentiation of human embryonic stem cells.

    PubMed

    Shahhoseini, Maryam; Taghizadeh, Zeinab; Hatami, Maryam; Baharvand, Hossein

    2013-04-01

    Gene activation of HOX clusters is an early event in embryonic development. These genes are highly expressed and active in the vertebrate nervous system. Based on the presence of retinoic acid response elements (RAREs) in the regulatory region of many of the HOX genes, it is deduced that retinoic acid (RA) can influence epigenetic regulation and consequently the expression pattern of HOX during RA-induced differentiation of embryonic model systems. In this investigation, the expression level as well as the epigenetic regulation of several HOX genes of the 4 A-D clusters was analyzed in human embryonic stem cells, and also through their neural induction, in the presence and absence of RA. Expression analysis data significantly showed increased mRNA levels of all examined HOX genes in the presence of RA. Epigenetic analysis of the HOX gene regulatory regions also showed a significant decrease in methylation of histone H3K27 parallel to an absolute preferential incorporation of the demethylase UTX rather than JMJD3 in RA-induced neural differentiated cells. This finding clearly showed the functional role of UTX in epigenetic alteration of HOX clusters during RA-induced neural differentiation; the activity could not be detectable for the demethylase JMJD3 during this developmental process.

  11. Patterns of frontoparietal activation as a marker for unsuccessful visuospatial processing in healthy aging.

    PubMed

    Drag, Lauren L; Light, Sharee N; Langenecker, Scott A; Hazlett, Kathleen E; Wilde, Elisabeth A; Welsh, Robert; Steinberg, Brett A; Bieliauskas, Linas A

    2016-09-01

    Visuospatial abilities are sensitive to age-related decline, although the neural basis for this decline (and its everyday behavioral correlates) is as yet poorly understood. fMRI was employed to examine age-related differences in patterns of functional activation that underlie changes in visuospatial processing. All participants completed a brief neuropsychological battery and also a figure ground task (FGT) assessing visuospatial processing while fMRI was recorded. Participants included 16 healthy older adults (OA; aged 69-82 years) and 16 healthy younger adults (YA; aged 20-35 years). We examined age-related differences in behavioral performance on the FGT in relation to patterns of fMRI activation. OA demonstrated reduced performance on the FGT task and showed increased activation of supramarginal parietal cortex as well as increased activation of frontal and temporal regions compared to their younger counterparts. Performance on the FGT related to increased supramarginal gyrus activity and increased medial prefrontal activity in OAs, but not YAs. Our results are consistent with an anterior-posterior compensation model. Successful FGT performance requires the perception and integration of multiple stimuli and thus it is plausible that healthy aging may be accompanied by changes in visuospatial processing that mimic a subtle form of dorsal simultanagnosia. Overall, decreased visuospatial processing in OA relates to an altered frontoparietal neurobiological signature that may contribute to the general phenomenon of increasingly fragmented execution of behavior associated with normal aging.

  12. Characterizing the effect of cytochrome P450 (CYP) 2C8, CYP2C9, and CYP2D6 genetic polymorphisms on stereoselective N-demethylation of fluoxetine.

    PubMed

    Wang, Zhangting; Wang, Shengjia; Huang, Minmin; Hu, Haihong; Yu, Lushan; Zeng, Su

    2014-03-01

    Fluoxetine (FLX) is one of the most widely prescribed selective serotonin reuptake inhibitors. Although FLX is used as racemate in the clinic, the clinical pharmacokinetics of FLX and its N-demethylation metabolite norfluoxetine (NFLX) show obvious cytochrome P450 (CYP) polymorphism dependency and exhibit marked stereoselectivity. However, the kinetic profiles of CYP variants to FLX remain unclear. In the present study, some variants of human CYP2C8, CYP2C9, and CYP2D6 were first expressed in insect cells, and their catalytic roles with respect to FLX enantiomers were then investigated. CYP2C8.4 and CYP2C9.10 showed significantly lower activity and CYP2C8.3 showed significantly higher activity toward both R- and S-FLX compared with the wildtype, while CYP2C9.3, CYP2C9.13, and CYP2C9.16 showed significantly lower activity only toward R-FLX. Five CYP2C9 variants and CYP2D6.1 exhibited significantly stereoselective kinetic profiles prior to R-FLX, and CYP2C8.3 showed a slight stereoselectivity. Interestingly, obvious substrate inhibition was observed in the CYP2C9 wildtype and its three variants only in the case of R-FLX. Together, these findings suggest that CYP2C9 and CYP2D6 polymorphism may play an important role in the clearance of FLX and also in the stereoselective kinetic profiles of FLX enantiomers.

  13. Antioxidant activity of processed table beets (Beta vulgaris var, conditiva) and green beans (Phaseolus vulgaris L.).

    PubMed

    Jiratanan, Thudnatkorn; Liu, Rui Hai

    2004-05-05

    It has been shown that thermal processing of tomatoes and sweet corn results in increased antioxidant activities despite the loss of vitamin C. Until now, it is unclear whether this positive effect of thermal processing occurs with all crop produce. Therefore, analysis of a root vegetable (beets) and of a legume (green beans) was undertaken to address this question. Antioxidant activity of beets processed under typical commercial processing conditions remained constant despite an 8% loss of vitamin C, a 60% loss of color, and 30% loss of dietary folate. There was a slight but significant 5% increase in phenolic content of processed beets. In contrast, vitamin C and dietary folate content of green beans remained constant, whereas a 32% reduction in phenolic compounds occurred after typical commercial processing conditions. The antioxidant activity of green beans was reduced by 20%. These findings along with previous works suggest that the effects of thermal processing vary with the respective produce crop type. It also reinforces the concept that optimal health benefits may be achieved when a wide variety of plant foods (fruits, vegetables and whole grains) and preparation methods are incorporated into the diet.

  14. Evaluation of aeration energy saving in two modified activated sludge processes.

    PubMed

    Lee, Ingyu; Lim, Honglae; Jung, Byunghun; Colosimo, Mark F; Kim, Hyunook

    2015-12-01

    A variety of modified activated sludge processes are widely used in wastewater treatment plants (WWTPs) for removing organics and nutrients (N and P). Since energy consumption in aeration basin accounts for the major part of the overall energy usage in WWTPs, efforts have been made to find ways to reduce aeration energy. In this study, two modified activated sludge processes in a pilot scale designed for nutrient removal were evaluated for the extent of energy saving: (1) ABA(2) process - adjusting air on/off period (i.e., with a temporal change); and (2) MB-A(2)O process - changing volume ratio of aerobic tank to anoxic tank (i.e., with a spatial change). For the 1st process, the air on/off period was fixed at 60min/45min with aerobic fraction being 0.57, while for the 2nd process, the aerobic/anoxic volume ratio was reduced from 0.58 to 0.42. The results demonstrate that the effluent COD, TN, NH4(+) and TP concentrations are acceptable while reduced aeration time/volume certainly saves significant energy consumption. To the best of our knowledge, this is 1st attempt to reduce the aeration period or aeration volume to save the aeration energy in these two modified activated sludge processes. The implication of these observations is further discussed.

  15. Granular activated carbon promoted ozonation of a food-processing secondary effluent.

    PubMed

    Alvarez, Pedro M; Pocostales, J Pablo; Beltrán, Fernando J

    2011-01-30

    This paper reports on the application of a simultaneous combination of ozone and a granular activated carbon (O(3)/GAC) as a tertiary treatment of a wastewater generated from the activity of various food-processing industries. Prior to the O(3)/GAC treatment, the wastewater was subjected to conventional primary and secondary treatments in a full-scale wastewater treatment plant (WWTP). The effluent from the WWTP presented high organic load (COD>500 mg/l and TOC>150 mg/l), which could be much reduced by the O(3)/GAC treatment. Results from the O(3)/GAC experiments were compared with those obtained in single ozonation, single adsorption onto GAC and sequential O(3)-GAC adsorption experiments. While single processes and the sequential one showed limited capacity to remove organic matter for the food-processing effluent (COD removal <40%), the simultaneous O(3)/GAC process led to decreases of COD up to 82% at the conditions here applied. The combined process also improved the ozone consumption, which decreased from about 19 g O(3)/g TOC (single ozonation process) to 8.2-10.7 g O(3)/g TOC (O(3)/GAC process). The reusability of the GAC throughout a series of consecutive O(3)/GAC experiments was studied with no apparent loss of activity for a neutral GAC (PZC = 6.7) but for a basic GAC (PZC = 9.1).

  16. In vitro Anti-Thrombotic Activity of Extracts from Blacklip Abalone (Haliotis rubra) Processing Waste

    PubMed Central

    Suleria, Hafiz Ansar Rasul; Hines, Barney M.; Addepalli, Rama; Chen, Wei; Masci, Paul; Gobe, Glenda; Osborne, Simone A.

    2016-01-01

    Waste generated from the processing of marine organisms for food represents an underutilized resource that has the potential to provide bioactive molecules with pharmaceutical applications. Some of these molecules have known anti-thrombotic and anti-coagulant activities and are being investigated as alternatives to common anti-thrombotic drugs, like heparin and warfarin that have serious side effects. In the current study, extracts prepared from blacklip abalone (Haliotis rubra) processing waste, using food grade enzymes papain and bromelain, were found to contain sulphated polysaccharide with anti-thrombotic activity. Extracts were found to be enriched with sulphated polysaccharides and assessed for anti-thrombotic activity in vitro through heparin cofactor-II (HCII)-mediated inhibition of thrombin. More than 60% thrombin inhibition was observed in response to 100 μg/mL sulphated polysaccharides. Anti-thrombotic potential was further assessed as anti-coagulant activity in plasma and blood, using prothrombin time (PT), activated partial thromboplastin time (aPTT), and thromboelastography (TEG). All abalone extracts had significant activity compared with saline control. Anion exchange chromatography was used to separate extracts into fractions with enhanced anti-thrombotic activity, improving HCII-mediated thrombin inhibition, PT and aPTT almost 2-fold. Overall this study identifies an alternative source of anti-thrombotic molecules that can be easily processed offering alternatives to current anti-thrombotic agents like heparin. PMID:28042854

  17. Start-up of the Anammox process from the conventional activated sludge in a membrane bioreactor.

    PubMed

    Wang, Tao; Zhang, Hanmin; Yang, Fenglin; Liu, Sitong; Fu, Zhimin; Chen, Huihui

    2009-05-01

    A lab-scale membrane bioreactor (MBR) was used to start-up the anaerobic ammonium oxidation (Anammox) process from the conventional activated sludge for 2 months. Results indicated the MBR could be a novel and suitable system for start-up of the Anammox process. The Anammox activity appeared after 16 days operation, and the average removal efficiencies of ammonia and nitrite were both over 90% in the end. A final specific Anammox activity of 0.35 g NH(4)(+)-N+NO(2)(-)-N (gVSS *d)(-1) was obtained. Fluorescence in situ hybridization (FISH) analysis confirmed the existence of Anammox bacteria and aerobic ammonia oxidizing bacteria. On the basis of results on MBR performance and FISH analysis, it was proposed that the start-up process was essentially a microbial community succession under man-made disturbance, and a climax community with Anammox bacteria as the dominant population was finally established.

  18. Coal liquefaction process streams characterization and evaluation. Volume 1, Base program activities

    SciTech Connect

    Robbins, G.A.; Brandes, S.D.; Winschel, R.A.; Burke, F.P.

    1994-05-01

    This 4.5-year project consisted of routine analytical support to DOE`s direct liquefaction process development effort (the Base Program), and an extensive effort to develop, demonstrate, and apply new analytical methods for the characterization of liquefaction process streams (the Participants Program). The objective of the Base Program was to support the on-going DOE direct coal liquefaction process development program. Feed, process, and product samples were used to assess process operations, product quality, and the effects of process variables, and to direct future testing. The primary objective of the Participants Program was to identify and demonstrate analytical methods for use in support of liquefaction process development, and in so doing, provide a bridge between process design, and development, and operation and analytical chemistry. To achieve this objective, novel analytical methods were evaluated for application to direct coal liquefaction-derived materials. CONSOL teamed with 24 research groups in the program. Well-defined and characterized samples of coal liquefaction process-derived materials were provided to each group. CONSOL made an evaluation of each analytical technique. During the performance of this project, we obtained analyses on samples from numerous process development and research programs and we evaluated a variety of analytical techniques for their usefulness in supporting liquefaction process development. Because of the diverse nature of this program, we provide here an annotated bibliography of the technical reports, publications, and formal presentations that resulted from this program to serve as a comprehensive summary of contract activities.

  19. Mechanistic investigation of industrial wastewater naphthenic acids removal using granular activated carbon (GAC) biofilm based processes.

    PubMed

    Islam, Md Shahinoor; Zhang, Yanyan; McPhedran, Kerry N; Liu, Yang; Gamal El-Din, Mohamed

    2016-01-15

    Naphthenic acids (NAs) found in oil sands process-affected waters (OSPW) have known environmental toxicity and are resistant to conventional wastewater treatments. The granular activated carbon (GAC) biofilm treatment process has been shown to effectively treat OSPW NAs via combined adsorption/biodegradation processes despite the lack of research investigating their individual contributions. Presently, the NAs removals due to the individual processes of adsorption and biodegradation in OSPW bioreactors were determined using sodium azide to inhibit biodegradation. For raw OSPW, after 28 days biodegradation and adsorption contributed 14% and 63% of NA removal, respectively. For ozonated OSPW, biodegradation removed 18% of NAs while adsorption reduced NAs by 73%. Microbial community 454-pyrosequencing of bioreactor matrices indicated the importance of biodegradation given the diverse carbon degrading families including Acidobacteriaceae, Ectothiorhodospiraceae, and Comamonadaceae. Overall, results highlight the ability to determine specific processes of NAs removals in the combined treatment process in the presence of diverse bacteria metabolic groups found in GAC bioreactors.

  20. Activation processes on GaAs photocathode by different currents of oxygen source

    NASA Astrophysics Data System (ADS)

    Miao, Zhuang; Shi, Feng; Cheng, Hongchang; Wang, Shufei; Zhang, Xiaohui; Yuan, Yuan; Chen, Chang

    2015-04-01

    In order to know the influence of activation processes on GaAs photocathodes, three GaAs samples were activated by a fixed current of cesium source and different currents of oxygen source. The current of caesium source is same during activation to ensure initial adsorption of caesium quantum is similar, which is the base to show the difference during alternation activation of caesium and oxygen. Analysed with the activation data, it is indicated that Cs-to-O current ratio of 1.07 is the optimum ratio to obtain higher sensitivity and better stability. According to double dipole model, stable and uniform double dipole layers of GaAs-O-Cs:Cs-O-Cs are formed and negative electron affinity is achieved on GaAs surface by activation with cesium and oxygen. The analytical result is just coincident with the model. Thus there is an efficient technological method to improve sensitivity and stability of GaAs photocathode.

  1. Activation of bean (Phaseolus vulgaris) [alpha]-amylase inhibitor requires proteolytic processing of the proprotein

    SciTech Connect

    Pueyo, J.J.; Hunt, D.C.; Chrispeels, M.J. )

    1993-04-01

    Seeds of the common bean (Phaseolus vulgaris) contain a plant defense protein that inhibits the [alpha]-amylases of mammals and insects. This [alpha]-amylase inhibitor ([alpha]Al) is synthesized as a proprotein on the endoplasmic reticulum and is proteolytically processed after arrival in the protein storage vacuoles to polypeptides of relative molecular weight (M[sub r]) 15,000 to 18,000. The authors report two types of evidence that proteolytic processing is linked to activation of the inhibitory activity. First, by surveying seed extracts of wild accessions of P. vulgaris and other species in the genus Phaseolus, they found that antibodies to [alpha]Al recognize large (M[sub r] 30,000-35,000) polypeptides as well as typical [alpha]Al processing products (M[sub r] 15,000-18,000). [alpha]Al activity was found in all extracts that had the typical [alpha]Al processed polypeptides, but was absent from seed extracts that lacked such polypeptides. Second, they made a mutant [alpha]Al in which asparagine-77 is changed to aspartic acid-77. This mutation slows down the proteolytic processing of pro-[alpha]Al when the gene is expressed in tobacco. When pro-[alpha]Al was separated from mature [alpha]Al by gel filtration, pro-[alpha]Al was found not to have [alpha]-amylase inhibitory activity. The authors interpret these results to mean that formation of the active inhibitor is causally related to proteolytic processing of the proprotein. They suggest that the polypeptide cleavage removes a conformation constraint on the precursor to produce the biochemically active molecule. 43 refs., 5 figs., 1 tab.

  2. Overview of materials processing in space activity at Marshall Space Flight Center

    NASA Technical Reports Server (NTRS)

    Williams, J. R.; Chassay, R. P.; Moore, W. W.; Ruff, R. C.; Yates, I. C.

    1984-01-01

    An overview of activities involving the Space Transportation System (STS), now in the operational phase, and results of some of the current space experiments, as well as future research opportunities in microgravity environment, are presented. The experiments of the Materials Processing in Space Program flown on the STS, such as bioseparation processes, isoelectric focusing, solidification and crystal growth processes, containerless processes, and the Materials Experiment Assembly experiments are discussed. Special consideration is given to the experiments to be flown aboard the Spacelab 3 module, the Fluids Experiments System, and the Vapor Crystal Growth System. Ground-based test facilities and planned space research facilities, as well as the nature of the commercialization activities, are briefly explained.

  3. Effects of Cu on metabolisms and enzyme activities of microbial communities in the process of composting.

    PubMed

    Guo, Xingliang; Gu, Jie; Gao, Hua; Qin, Qingjun; Chen, Zhixue; Shao, Li; Chen, Lin; Li, Hailong; Zhang, Weijuan; Chen, Shengnan; Liu, Jiang

    2012-03-01

    With the compost matrix of pig manure, wheat straw, and spent mushroom substrate, and then inoculated with the Compound Microbe Preparation, the study investigated the effects of the heavy metal Cu on the process of composting. Biolog EcoPlate™ test revealed that at a low content, Cu could improve the capacities of microbial communities to transform and exploit carbon sources in the form of polymer, thus speeding up the decomposition of agricultural wastes, and at a high content, Cu presented inhibiting effect on microbial communities to exploit complex macromolecular carbon sources, thus extending the decomposition of agricultural wastes. Enzyme activity testing showed that at a low content, Cu presented enzyme activity-activating effect at the early period of composting and inhibiting effect in the late period of composting, and at a high content, Cu presented enzyme activity-inhibiting effects through the process of composting.

  4. Enzyme activity as an indicator of soil-rehabilitation processes at a zinc and lead ore mining and processing area.

    PubMed

    Ciarkowska, Krystyna; Sołek-Podwika, Katarzyna; Wieczorek, Jerzy

    2014-01-01

    The activities of soil enzymes in relation to the changes occurring in the soil on a degraded area in southern Poland after zinc and lead mining were analyzed. An evaluation of the usefulness of urease and invertase activities for estimating the progress of the rehabilitation processes in degraded soil was performed. The data show that the soil samples differed significantly in organic carbon (0.68-104.0 g kg(-1)) and total nitrogen (0.03-8.64 g kg(-1)) content in their surface horizons. All of the soil samples (apart from one covered with forest) had very high total concentrations of zinc (4050-10,884 mg kg(-1)), lead (959-6661 mg kg(-1)) and cadmium (24.4-174.3 mg kg(-1)) in their surface horizons, and similar concentrations in their deeper horizons. Nevertheless, the amounts of the soluble forms of the above-mentioned heavy metals were quite low and they accounted for only a small percentage of the total concentrations: 1.4% for Zn, 0.01% for Pb and 2.6% for Cd. Urease activities were ranked as follows: soil from flotation settler (0.88-1.78 μg N-NH4(+) 2h(-1) g(-1))activity (2.14-5.73 μg N-NH4(+) 2h(-1) g(-1)). Invertase activities were similar in soil that was undisturbed by mining and in soil from old slag heaps, ranging from 20.5 to 77.1mg of the inverted sugar, but they were much lower in soil from the flotation settler (0.12-6.95 mg of the inverted sugar). The results demonstrated that heavy pollution with Zn, Pb and Cd slightly decreased the activities of urease and invertase. It is thought that it resulted from the enzyme reactions occurring in slightly acidic or alkaline soil conditions. Under such conditions, heavy metals occur mainly in insoluble forms. The activities of these enzymes are strongly dependent on the content and decomposition of organic matter in the soil.

  5. [Combination of two signals of danger--a principal cause of activation of chronic inflammatory processes].

    PubMed

    Lebedev, K A

    2012-01-01

    Chronic inflammatory processes have long current during which there is a change of remission by an aggravation of disease. Until recently was considered, that occurrence and activation of chronic inflammatory process is caused by one signal of danger. Most often it are served with microorganisms. The sum of our end literary data shows that activation is connected with accumulation in the center of an inflammation of macrophages and their hyperactivation as a result of action of two signals of danger--microorganisms and xenobiotics.

  6. Activity of processes on the visible surfaces of Solar System bodies

    NASA Astrophysics Data System (ADS)

    Vidmachenko, A. P.

    2016-10-01

    We consider the physical processes on the surfaces of Solar System bodies, which lead to visible changes in their reflective characteristics. It is shown that each body in the Solar system has a set of chemical elements and their compounds, converting of which indicates significant activity in such a significant temperature change range from 700 K (for Mercury) to 30 K for Pluto. That is, all objects in the Solar system show a significant activity. However, they are very individual for the list and the type of the processes that take place on each body in the Solar system.

  7. Effect of process conditions on recovery of protein activity after freezing and freeze-drying.

    PubMed

    Jiang, S; Nail, S L

    1998-05-01

    The objective of this research was to gain a better understanding of the degree to which recovery of activity of model proteins after freeze-drying can be maximized by manipulation of freeze-dry process conditions in the absence of protective solutes. Catalase, beta-galactosidase and lactate dehydrogenase (LDH) were used as model proteins. All of the three proteins exhibited a concentration-dependent loss of activity after freezing, with significantly higher recovery at higher concentration. The freezing method and the type of buffer were also important, with sodium phosphate buffer and freezing by immersion of vials in liquid nitrogen associated with the lowest recovery of activity. Differential scanning calorimetry was predictive of the onset of collapse during freeze-drying only for beta-galactosidase. For the other proteins, either no Tg' transition was observed, or the apparent glass transition did not correlate with the microscopically-observed collapse temperature. The time course of activity loss for beta-galactosidase and LDH was compared during freeze-drying under conditions which produced collapse of the dried matrix and conditions which produced retention of microstructure in the dried solid. Recovery of activity decreased continuously during primary drying, with no sharp drop in recovery of activity associated with the onset of collapse. The most important drying process variable affecting recovery of activity was residual moisture level, with a dramatic drop in activity recovery associated with residual moisture levels less than about 10%.

  8. Mid-fusiform activation during object discrimination reflects the process of differentiating structural descriptions.

    PubMed

    Liu, Xun; Steinmetz, Nicholas A; Farley, Alison B; Smith, Charles D; Joseph, Jane E

    2008-09-01

    The present study explored constraints on mid-fusiform activation during object discrimination. In three experiments, participants performed a matching task on simple line configurations, nameable objects, three dimensional (3-D) shapes, and colors. Significant bilateral mid-fusiform activation emerged when participants matched objects and 3-D shapes, as compared to when they matched two-dimensional (2-D) line configurations and colors, indicating that the mid-fusiform is engaged more strongly for processing structural descriptions (e.g., comparing 3-D volumetric shape) than perceptual descriptions (e.g., comparing 2-D or color information). In two of the experiments, the same mid-fusiform regions were also modulated by the degree of structural similarity between stimuli, implicating a role for the mid-fusiform in fine differentiation of similar visual object representations. Importantly, however, this process of fine differentiation occurred at the level of structural, but not perceptual, descriptions. Moreover, mid-fusiform activity was more robust when participants matched shape compared to color information using the identical stimuli, indicating that activity in the mid-fusiform gyrus is not driven by specific stimulus properties, but rather by the process of distinguishing stimuli based on shape information. Taken together, these findings further clarify the nature of object processing in the mid-fusiform gyrus. This region is engaged specifically in structural differentiation, a critical component process of object recognition and categorization.

  9. Study the left prefrontal cortex activity of Chinese children with dyslexia in phonological processing by NIRS

    NASA Astrophysics Data System (ADS)

    Zhang, Zhili; Li, Ting; Zheng, Yi; Luo, Qingming; Song, Ranran; Gong, Hui

    2006-02-01

    Developmental dyslexia, a kind of prevalent psychological disease, represents that dyslexic children have unexpected difficulties in phonological processing and recognition test of Chinese characters. Some functional imaging technologies, such as fMRI and PET, have been used to study the brain activities of the children with dyslexia whose first language is English. In this paper, a portable, 16-channel, continuous-wave (CW) NIRS instrument was used to monitor the concentration changes of each hemoglobin species when Chinese children did the task of phonological processing and recognition test. The NIRS recorded the hemodynamic changes in the left prefrontal cortex of the children. 20 dyslexia-reading children (10~12 years old) and 20 normal-reading children took part in the phonological processing of Chinese characters including the phonological awareness section and the phonological decoding section. During the phonological awareness section, the changed concentration of deoxy-hemoglobin in dyslexia-reading children were significantly higher (p<0.05) than normal-reading children in the left ventrolateral prefrontal cortex (VLPFC). While in the phonological decoding section, both normal and dyslexic reading children had more activity in the left VLPFC, but only normal-reading children had activity in the left middorsal prefrontal cortex. In conclusion, both dyslexic and normal-reading children have activity in the left prefrontal cortex, but the degree and the areas of the prefrontal cortex activity are different between them when they did phonological processing.

  10. Right Occipital Cortex Activation Correlates with Superior Odor Processing Performance in the Early Blind

    PubMed Central

    Grandin, Cécile B.; Dricot, Laurence; Plaza, Paula; Lerens, Elodie; Rombaux, Philippe; De Volder, Anne G.

    2013-01-01

    Using functional magnetic resonance imaging (fMRI) in ten early blind humans, we found robust occipital activation during two odor-processing tasks (discrimination or categorization of fruit and flower odors), as well as during control auditory-verbal conditions (discrimination or categorization of fruit and flower names). We also found evidence for reorganization and specialization of the ventral part of the occipital cortex, with dissociation according to stimulus modality: the right fusiform gyrus was most activated during olfactory conditions while part of the left ventral lateral occipital complex showed a preference for auditory-verbal processing. Only little occipital activation was found in sighted subjects, but the same right-olfactory/left-auditory-verbal hemispheric lateralization was found overall in their brain. This difference between the groups was mirrored by superior performance of the blind in various odor-processing tasks. Moreover, the level of right fusiform gyrus activation during the olfactory conditions was highly correlated with individual scores in a variety of odor recognition tests, indicating that the additional occipital activation may play a functional role in odor processing. PMID:23967263

  11. Assessment of MSFCs Process for the Development and Activation of Space Act Agreement

    NASA Technical Reports Server (NTRS)

    Daugherty, Rachel

    2014-01-01

    Space Act Agreements (SAAs) are contractual agreements that NASA utilizes to form partnerships with researchers, industry, and academia to stimulate cutting-edge innovation within the science and technology communities. center dot This study assessed the current SAA development and activation process at Marshall Space Flight Center (MSFC) to determine if improvements could be implemented to increase productivity, decrease time to activation, and improve the quality of deliverables.

  12. Effects of human activities on the ecological processes of river biofilms in a highly urbanized river

    NASA Astrophysics Data System (ADS)

    Hung, R.; Li, M.

    2013-12-01

    Many anthropogenic disturbances and their effects of aquatic ecosystem are difficult to quantify in urbanized rivers. In past, specific taxa analysis of community structure was a common approach in river health monitoring studies. However, it is still difficult to understand stream ecosystem integrity without considering ecosystem processes. The complex species composition and metabolism of a river biofilm have the capacity to interact and/or modulate their surrounding environment. Because of their short life cycles, species richness, and worldwide distribution, structure and function of river biofilm communities are sensitive to change in environmental conditions. Therefore, biofilms are widely used as early warning systems of water pollution for water quality monitoring studies. In this study, we used river biofilms as a bioindicator by examining their extracellular enzyme activities and photosynthesis efficiency to understand human activities on the ecological processes of river ecosystem in a highly urbanized river. We sampled four sites along the Keelung River, Taiwan, based on different intensities of anthropogenic disturbances including water pollution index, population densities, land use types and types of stream habitats. Two study sites are heavily influenced by human activities and the others are not. The activities of extracellular enzymes within the biofilm play an important function for organic matter decomposition and nutrient cycling. We measured seven extracellular enzyme activities (β-d-glucosidase, phosphatase, leucine-aminopeptidase, sulfatase, peroxidase, polyphenol oxidase, and esterase) to examine specific enzyme activity changes at four study sites monthly. In addition, relative proportion of each extracellular enzyme activity on total enzyme activities was calculated in order to examine the relationship between functional biofilm profiles and different urban intensities. Among four study sites, leucine-aminopeptidase and esterase

  13. Clinical process analysis and activity-based costing at a heart center.

    PubMed

    Ridderstolpe, Lisa; Johansson, Andreas; Skau, Tommy; Rutberg, Hans; Ahlfeldt, Hans

    2002-08-01

    Cost studies, productivity, efficiency, and quality of care measures, the links between resources and patient outcomes, are fundamental issues for hospital management today. This paper describes the implementation of a model for process analysis and activity-based costing (ABC)/management at a Heart Center in Sweden as a tool for administrative cost information, strategic decision-making, quality improvement, and cost reduction. A commercial software package (QPR) containing two interrelated parts, "ProcessGuide and CostControl," was used. All processes at the Heart Center were mapped and graphically outlined. Processes and activities such as health care procedures, research, and education were identified together with their causal relationship to costs and products/services. The construction of the ABC model in CostControl was time-consuming. However, after the ABC/management system was created, it opened the way for new possibilities including process and activity analysis, simulation, and price calculations. Cost analysis showed large variations in the cost obtained for individual patients undergoing coronary artery bypass grafting (CABG) surgery. We conclude that a process-based costing system is applicable and has the potential to be useful in hospital management.

  14. Mechanism of the activation process for the formation of a surface-conduction electron-emitter

    NASA Astrophysics Data System (ADS)

    Tsukamoto, Takeo; Okuda, Masahiro; Arai, Yutaka; Miyata, Hirokatsu

    2016-01-01

    The major role of the chemical reaction between a silica substrate and deposited carbon in the activation process for the formation of a surface-conduction electron emitter (SCE) is investigated. The SCE emits electrons by the tunneling effect when an electric field is applied across a nanoscale gap. The nanogap is spontaneously formed by the activation process, wherein a pulse voltage is applied between a pair of electrodes, which are separated by a narrow gap inside a vacuum chamber in the presence of hydrocarbons. At the gap, two elemental processes compete; the deposition of carbon by the electron-induced decomposition of hydrocarbons and the consumption of carbon by reaction with the silica substrate. The balance of the dynamics of the two processes, which simply depends on the temperature at the gap, is responsible for the spontaneous determination of the width of the nanogap. The calculation based on the model that involves the two competitive processes agrees with the experimental results on the activation process.

  15. Passive and active processes in visuo-spatial memory: double dissociation in developmental learning disabilities.

    PubMed

    Cornoldi, C; Rigoni, F; Venneri, A; Vecchi, T

    2000-01-01

    The distinction between passive and active visuo-spatial memory has been useful to interpret various pattern of deficits reported in individual differences studies. However, this interpretation raises the issue of task difficulty, since active tasks could be failed simply because more complex and the corresponding deficit could reflect a reduced capacity of the system. We describe two children with Nonverbal Learning Disability whose performance provides evidence of a dissociation between passive and active memory processes. One of the children showed a selective impairment in passive tasks and performed flawlessly in active tasks, whereas the second child displayed the opposite pattern. These data suggest that a qualitative difference between passive and active processes does exist and that differences in performance do not reflect a lower/higher level of task difficulty. Further, these data underlie the importance of formulating theoretical models of visuo-spatial memory including both material-related (i.e., visual vs spatial) and process-related (i.e., passive vs active) distinctions.

  16. Impact of thermal processing and the Maillard reaction on the basophil activation of hazelnut allergic patients.

    PubMed

    Cucu, Tatiana; De Meulenaer, Bruno; Bridts, Chris; Devreese, Bart; Ebo, Didier

    2012-05-01

    Food allergy, an abnormal immunological response due to sensitization to a food component, has become an important health problem, especially in industrialized countries. The aim of this study was to investigate the impact of thermal processing and glycation on the basophil activation by hazelnut proteins using a basophil activation test. Patients with systemic allergic reactions (SR; n=6) to hazelnut as well as patients with an isolated oral allergy syndrome (OAS; n=4) were investigated. Thermal processing of hazelnut proteins either in the presence or absence of wheat proteins did not result in major changes in the stimulatory activity of the basophils for patients with SR or OAS. For the patients with OAS, incubation of hazelnut proteins with glucose led to complete depletion of the stimulatory activity of the basophils. An increase in stimulatory activity of the basophils for two out of six patients with SR was observed. For the other four patients slight or complete abolition of the stimulatory activity was observed. These results indicate that some patients with SR to hazelnut are at risk when exposed to hazelnut proteins, even in processed foods.

  17. DNA Demethylation Rescues the Impaired Osteogenic Differentiation Ability of Human Periodontal Ligament Stem Cells in High Glucose

    PubMed Central

    Liu, Zhi; Chen, Tian; Sun, Wenhua; Yuan, Zongyi; Yu, Mei; Chen, Guoqing; Guo, Weihua; Xiao, Jingang; Tian, Weidong

    2016-01-01

    Diabetes mellitus, characterized by abnormally high blood glucose levels, gives rise to impaired bone remodeling. In response to high glucose (HG), the attenuated osteogenic differentiation capacity of human periodontal ligament stem cells (hPDLSCs) is associated with the loss of alveolar bone. Recently, DNA methylation was reported to affect osteogenic differentiation of stem cells in pathological states. However, the intrinsic mechanism linking DNA methylation to osteogenic differentiation ability in the presence of HG is still unclear. In this study, we found that diabetic rats with increased DNA methylation levels in periodontal ligaments exhibited reduced bone mass and density. In vitro application of 5-aza-2′-deoxycytidine (5-aza-dC), a DNA methyltransferase inhibitor, to decrease DNA methylation levels in hPDLSCs, rescued the osteogenic differentiation capacity of hPDLSCs under HG conditions. Moreover, we demonstrated that the canonical Wnt signaling pathway was activated during this process and, under HG circumstances, the 5-aza-dC-rescued osteogenic differentiation capacity was blocked by Dickkopf-1, an effective antagonist of the canonical Wnt signaling pathway. Taken together, these results demonstrate for the first time that suppression of DNA methylation is able to facilitate the osteogenic differentiation capacity of hPDLSCs exposed to HG, through activation of the canonical Wnt signaling pathway. PMID:27273319

  18. Two C4-sterol methyl oxidases (Erg25) catalyse ergosterol intermediate demethylation and impact environmental stress adaptation in Aspergillus fumigatus

    PubMed Central

    Blosser, Sara J.; Merriman, Brittney; Grahl, Nora; Chung, Dawoon

    2014-01-01

    The human pathogen Aspergillus fumigatus adapts to stress encountered in the mammalian host as part of its ability to cause disease. The transcription factor SrbA plays a significant role in this process by regulating genes involved in hypoxia and low-iron adaptation, antifungal drug responses and virulence. SrbA is a direct transcriptional regulator of genes encoding key enzymes in the ergosterol biosynthesis pathway, including erg25A and erg25B, and ΔsrbA accumulates C4-methyl sterols, suggesting a loss of Erg25 activity [C4-sterol methyl oxidase (SMO)]. Characterization of the two genes encoding SMOs in Aspergillus fumigatus revealed that both serve as functional C4-demethylases, with Erg25A serving in a primary role, as Δerg25A accumulates more C4-methyl sterol intermediates than Δerg25B. Single deletion of these SMOs revealed alterations in canonical ergosterol biosynthesis, indicating that ergosterol may be produced in an alternative fashion in the absence of SMO activity. A Δerg25A strain displayed moderate susceptibility to hypoxia and the endoplasmic reticulum stress-inducing agent DTT, but was not required for virulence in murine or insect models of invasive aspergillosis. Inducing expression of erg25A partially restored the hypoxia growth defect of ΔsrbA. These findings implicated Aspergillus fumigatus SMOs in the maintenance of canonical ergosterol biosynthesis and indicated an overall involvement in the fungal stress response. PMID:25107308

  19. Deep processing activates the medial temporal lobe in young but not in old adults.

    PubMed

    Daselaar, Sander M; Veltman, Dick J; Rombouts, Serge A R B; Raaijmakers, Jeroen G W; Jonker, Cees

    2003-11-01

    Age-related impairments in episodic memory have been related to a deficiency in semantic processing, based on the finding that elderly adults typically benefit less than young adults from deep, semantic as opposed to shallow, nonsemantic processing of study items. In the present study, we tested the hypothesis that elderly adults are not able to perform certain cognitive operations under deep processing conditions. We further hypothesised that this inability does not involve regions commonly associated with lexical/semantic retrieval processes, but rather involves a dysfunction of the medial temporal lobe (MTL) memory system. To this end, we used functional MRI on rather extensive groups of young and elderly adults to compare brain activity patterns obtained during a deep (living/nonliving) and a shallow (uppercase/lowercase) classification task. Common activity in relation to semantic classification was observed in regions that have been previously related to semantic retrieval, including mainly left-lateralised activity in the inferior prefrontal, middle temporal, and middle frontal/anterior cingulate gyrus. Although the young adults showed more activity in some of these areas, the finding of mainly overlapping activation patterns during semantic classification supports the idea that lexical/semantic retrieval processes are still intact in elderly adults. This received further support by the finding that both groups showed similar behavioural performances as well on the deep and shallow classification tasks. Importantly, though, the young revealed significantly more activity than the elderly adults in the left anterior hippocampus during deep relative to shallow classification. This finding is in line with the idea that age-related impairments in episodic encoding are, at least partly, due to an under-recruitment of the medial temporal lobe memory system.

  20. Kinetic behaviour of zymogen activation processes in the presence of an inhibitor.

    PubMed Central

    Varón, R; Manjabacas, M C; García-Moreno, M; Valero, E; Garcia-Canovas, F

    1993-01-01

    A global kinetic analysis of a general zymogen activation model, where not only the activating but also the activated enzyme suffer an irreversible inhibition is presented. A reaction in which the enzyme acts upon a substrate is coupled to monitor the process. In addition, we determined the corresponding kinetic equations for a number of particular cases of the general model studied. Finally, a kinetic data analysis and a procedure to discriminate among the different mechanisms considered, which are based on the kinetic equations obtained, are suggested. PMID:8452535

  1. Activity of fuel batches processed through Hanford separations plants, 1944 through 1989

    SciTech Connect

    Watrous, R.A.; Wootan, D.W.

    1997-07-29

    This document provides a printout of the ``Fuel Activity Database`` (version U6) generated by the Hanford DKPRO code and transmitted to the Los Alamos National Laboratory for input to their ``Hanford Defined Waste`` model of waste tank inventories. This fuel activity file consists of 1,276 records--each record representing the activity associated with a batch of spent reactor fuel processed by month (or shorter period) through individual Hanford separations plants between 1944 and 1989. Each record gives the curies for 46 key radionuclides, decayed to a common reference date of January 1, 1994.

  2. Community Vision and Interagency Alignment: A Community Planning Process to Promote Active Transportation.

    PubMed

    DeGregory, Sarah Timmins; Chaudhury, Nupur; Kennedy, Patrick; Noyes, Philip; Maybank, Aletha

    2016-04-01

    In 2010, the Brooklyn Active Transportation Community Planning Initiative launched in 2 New York City neighborhoods. Over a 2-year planning period, residents participated in surveys, school and community forums, neighborhood street assessments, and activation events-activities that highlighted the need for safer streets locally. Consensus among residents and key multisectoral stakeholders, including city agencies and community-based organizations, was garnered in support of a planned expansion of bicycling infrastructure. The process of building on community assets and applying a collective impact approach yielded changes in the built environment, attracted new partners and resources, and helped to restore a sense of power among residents.

  3. Treatment of coke-oven wastewater with the powdered activated carbon-contact stabilization activated sludge process. Final report

    SciTech Connect

    Suidan, M.T.; Deady, M.A.; Gee, C.S.

    1983-11-01

    The objective of the study was to determine optimum parameters for the operation of an innovative process train used in the treatment of coke-over wastewater. The treatment process train consisted of a contact-stabilization activated sludge system with powdered activated carbon (PAC) addition, followed by activated sludge nitrification, followed by denitrification in an anoxic filter. The control and operating parameters evaluated during the study were: (a) the average mixed-liquor PAC concentration maintained in the contact-stabilization system, (b) the solids retention time practiced in the contact-stabilization system, and (c) the hydraulic detention time maintained in the contact aeration tank. Three identical treatement process trains were constructed and employed in this study. The coke-oven wastewater used for this investigation was fed to the treatment units at 30% strength. The first part of the study was devoted to determining the interactions between the mixed liquor PAC concentration and the solids retention time in the contact-stabilization tanks. Results showed that optimum overall system performance is attainable when the highest sludge age (30 day) and highest mixed liquor PAC concentration were practiced. During the second phase of the study, all three systems were operated at a 30 day solids retention time while different detention times of 1, 2/3 and 1/3 day were evaluated in the contact tank. PAC addition rates were maintained at the former levels and, consequently, reduced contact times entailed higher mixed liquor carbon concentrations. Once again, the system receiving the highest PAC addition rate of PAC exhibited the best overall performance. This system exhibited no deterioration in process performance as a result of decreased contact detention time. 72 references, 41 figures, 24 tables.

  4. Proteolytic Processing and Activation of Clostridium perfringens Epsilon Toxin by Caprine Small Intestinal Contents

    PubMed Central

    Freedman, John C.; Li, Jihong; Uzal, Francisco A.

    2014-01-01

    ABSTRACT Epsilon toxin (ETX), a pore-forming toxin produced by type B and D strains of Clostridium perfringens, mediates severe enterotoxemia in livestock and possibly plays a role in human disease. During enterotoxemia, the nearly inactive ETX prototoxin is produced in the intestines but then must be activated by proteolytic processing. The current study sought to examine ETX prototoxin processing and activation ex vivo using the intestinal contents of a goat, a natural host species for ETX-mediated disease. First, this study showed that the prototoxin has a KEIS N-terminal sequence with a molecular mass of 33,054 Da. When the activation of ETX prototoxin ex vivo by goat small intestinal contents was assessed by SDS-PAGE, the prototoxin was processed in a stepwise fashion into an ~27-kDa band or higher-molecular-mass material that could be toxin oligomers. Purified ETX corresponding to the ~27-kDa band was cytotoxic. When it was biochemically characterized by mass spectrometry, the copresence of three ETX species, each with different C-terminal residues, was identified in the purified ~27-kDa ETX preparation. Cytotoxicity of each of the three ETX species was then demonstrated using recombinant DNA approaches. Serine protease inhibitors blocked the initial proteotoxin processing, while carboxypeptidase inhibitors blocked further processing events. Taken together, this study provides important new insights indicating that, in the intestinal lumen, serine protease (including trypsin and possibly chymotrypsin) initiates the processing of the prototoxin but other proteases, including carboxypeptidases, then process the prototoxin into multiple active and stable species. PMID:25336460

  5. Activities of the Institute of Chemical Processing of Coal at Zabrze

    SciTech Connect

    Dreszer, K.

    1995-12-31

    The Institute of Chemical Processing of Coal at Zabrze was established in 1955. The works on carbochemical technologies have been, therefore, carried out at the Institute for 40 years. The targets of the Institute`s activities are research, scientific and developing works regarding a sensible utilization of fuels via their processing into more refined forms, safe environment, highly efficient use of energy carriers and technological products of special quality. The Institute of Chemical Processing of Coal has been dealing with the following: optimized use of home hard coals; improvement of classic coal coking technologies, processing and utilization of volatile coking products; production technologies of low emission rate fuels for communal management; analyses of coal processing technologies; new technologies aimed at increasing the efficiency of coal utilization for energy-generating purposes, especially in industry and studies on the ecological aspects of these processes; production technologies of sorbents and carbon activating agents and technologies of the utilization; rationalization of water and wastes management in the metallurgical and chemical industries in connection with removal of pollution especially dangerous to the environment from wastes; utilization technologies of refined materials (electrode cokes, binders, impregnating agents) for making electrodes, refractories and new generation construction carbon materials; production technologies of high quality bituminous and bituminous and resin coating, anti-corrosive and insulation materials; environmentally friendly utilization technologies for power station, mine and other wastes, and dedusting processes in industrial gas streams.

  6. Innovative Application of Mechanical Activation for Rare Earth Elements Recovering: Process Optimization and Mechanism Exploration

    PubMed Central

    Tan, Quanyin; Deng, Chao; Li, Jinhui

    2016-01-01

    With the rapidly expanding use of fluorescent lamps (FLs) and increasing interest in conservation and sustainable utilization of critical metals such as rare earth elements (REEs), the recovering of REEs from phosphors in waste FLs is becoming a critical environmental and economic issue. To effectively recycle REEs with metallurgical methods, mechanical activation by ball milling was introduced to pretreat the waste phosphors. This current study put the emphasis on the mechanical activation and leaching processes for REEs, and explored the feasibility of the method from both theoretical and practical standpoints. Results showed physicochemical changes of structural destruction and particle size reduction after mechanical activation, leading to the easy dissolution of REEs in the activated samples. Under optimal conditions, dissolution yields of 89.4%, 93.1% and 94.6% for Tb, Eu and Y, respectively, were achieved from activated waste phosphors using hydrochloric acid as the dissolution agent. The shrinking core model proved to be the most applicable for the leaching procedure, with an apparent activation energy of 10.96 ± 2.79 kJ/mol. This novel process indicates that mechanical activation is an efficient method for recovering REEs from waste phosphors, and it has promising potential for REE recovery with low cost and high efficiency. PMID:26819083

  7. Asymmetrical Processing of Olfactory Input in the Piriform Cortex Mediates "Activation" of the Avian Navigation Circuitry.

    PubMed

    Jorge, Paulo E; Marques, Paulo A M; Pinto, Belmiro V; Phillips, John B

    2016-08-11

    The role of odors in the long-distance navigation of birds has elicited intense debate for more than half a century. Failure to resolve many of the issues fueling this debate is due at least in part to the absence of controls for a variety of non-specific effects that odors have on the navigational process. The present experiments were carried out to investigate whether the olfactory inputs are involved only in "activation" of neuronal circuitry involved in navigation or are also playing a role in providing directional information. Experienced adult pigeons were exposed to controlled olfactory stimuli during different segments of the journey (release site vs. displacement + release site). Protein levels of IEGs (immediate early genes used to mark synaptic activity) were analyzed in areas within the olfactory/navigation avian circuitry. The results indicate that 1) exposure to natural odors at the release site (and not before) elicit greater activation across brain regions than exposure to filtered air, artificial odors, and natural odors along the entire outward journey (from home to the release site, inclusive); 2) activation of the piriform cortex in terms of odor discrimination is lateralized; 3) activation of the navigation circuitry is achieved by means of lateralized activation of piriform cortex neurons. Altogether, the findings provide the first direct evidence that activation of the avian navigation circuitry is mediated by asymmetrical processing of olfactory input occurring in the right piriform cortex.

  8. Innovative Application of Mechanical Activation for Rare Earth Elements Recovering: Process Optimization and Mechanism Exploration

    NASA Astrophysics Data System (ADS)

    Tan, Quanyin; Deng, Chao; Li, Jinhui

    2016-01-01

    With the rapidly expanding use of fluorescent lamps (FLs) and increasing interest in conservation and sustainable utilization of critical metals such as rare earth elements (REEs), the recovering of REEs from phosphors in waste FLs is becoming a critical environmental and economic issue. To effectively recycle REEs with metallurgical methods, mechanical activation by ball milling was introduced to pretreat the waste phosphors. This current study put the emphasis on the mechanical activation and leaching processes for REEs, and explored the feasibility of the method from both theoretical and practical standpoints. Results showed physicochemical changes of structural destruction and particle size reduction after mechanical activation, leading to the easy dissolution of REEs in the activated samples. Under optimal conditions, dissolution yields of 89.4%, 93.1% and 94.6% for Tb, Eu and Y, respectively, were achieved from activated waste phosphors using hydrochloric acid as the dissolution agent. The shrinking core model proved to be the most applicable for the leaching procedure, with an apparent activation energy of 10.96 ± 2.79 kJ/mol. This novel process indicates that mechanical activation is an efficient method for recovering REEs from waste phosphors, and it has promising potential for REE recovery with low cost and high efficiency.

  9. Temporal dynamics of activation of thematic and functional knowledge during conceptual processing of manipulable artifacts.

    PubMed

    Kalénine, Solène; Mirman, Daniel; Middleton, Erica L; Buxbaum, Laurel J

    2012-09-01

    The current research aimed at specifying the activation time course of different types of semantic information during object conceptual processing and the effect of context on this time course. We distinguished between thematic and functional knowledge and the specificity of functional similarity. Two experiments were conducted with healthy older adults using eye tracking in a word-to-picture matching task. The time course of gaze fixations was used to assess activation of distractor objects during the identification of manipulable artifact targets (e.g., broom). Distractors were (a) thematically related (e.g., dustpan), (b) related by a specific function (e.g., vacuum cleaner), or (c) related by a general function (e.g., sponge). Growth curve analyses were used to assess competition effects when target words were presented in isolation (Experiment 1) and embedded in contextual sentences of different generality levels (Experiment 2). In the absence of context, there was earlier and shorter lasting activation of thematically related as compared to functionally related objects. The time course difference was more pronounced for general functions than specific functions. When contexts were provided, functional similarities that were congruent with context generality level increased in salience with earlier activation of those objects. Context had little impact on thematic activation time course. These data demonstrate that processing a single manipulable artifact concept implicitly activates thematic and functional knowledge with different time courses and that context speeds activation of context-congruent functional similarity.

  10. Innovative Application of Mechanical Activation for Rare Earth Elements Recovering: Process Optimization and Mechanism Exploration.

    PubMed

    Tan, Quanyin; Deng, Chao; Li, Jinhui

    2016-01-28

    With the rapidly expanding use of fluorescent lamps (FLs) and increasing interest in conservation and sustainable utilization of critical metals such as rare earth elements (REEs), the recovering of REEs from phosphors in waste FLs is becoming a critical environmental and economic issue. To effectively recycle REEs with metallurgical methods, mechanical activation by ball milling was introduced to pretreat the waste phosphors. This current study put the emphasis on the mechanical activation and leaching processes for REEs, and explored the feasibility of the method from both theoretical and practical standpoints. Results showed physicochemical changes of structural destruction and particle size reduction after mechanical activation, leading to the easy dissolution of REEs in the activated samples. Under optimal conditions, dissolution yields of 89.4%, 93.1% and 94.6% for Tb, Eu and Y, respectively, were achieved from activated waste phosphors using hydrochloric acid as the dissolution agent. The shrinking core model proved to be the most applicable for the leaching procedure, with an apparent activation energy of 10.96 ± 2.79 kJ/mol. This novel process indicates that mechanical activation is an efficient method for recovering REEs from waste phosphors, and it has promising potential for REE recovery with low cost and high efficiency.

  11. Enhancing the NASA Expendable Launch Vehicle Payload Safety Review Process Through Program Activities

    NASA Technical Reports Server (NTRS)

    Palo, Thomas E.

    2007-01-01

    The safety review process for NASA spacecraft flown on Expendable Launch Vehicles (ELVs) has been guided by NASA-STD 8719.8, Expendable Launch Vehicle Payload Safety Review Process Standard. The standard focused primarily on the safety approval required to begin pre-launch processing at the launch site. Subsequent changes in the contractual, technical, and operational aspects of payload processing, combined with lessons-learned supported a need for the reassessment of the standard. This has resulted in the formation of a NASA ELV Payload Safety Program. This program has been working to address the programmatic issues that will enhance and supplement the existing process, while continuing to ensure the safety of ELV payload activities.

  12. The process for the activation of frog epidermis pro-tyrosinase.

    PubMed Central

    Peñafiel, R; Galindo, J D; Pedreño, E; Lozano, J A

    1982-01-01

    1. Purified pro-tyrosinase from epidermis of the frog Rana esculenta ridibunda can be activated in vitro by several proteinases (trypsin, alpha-chymotrypsin, Pronase) and by light. 2. Both pro-tyrosinase and tyrosinase are composed of a single type of subunit having pI 7.2 and approximate molecular weights 68000 and 62000 respectively. A peptide of low molecular weight is released as a consequence of the proteolytic activation. Pro-tyrosinase and tyrosinase have different quaternary structures, the proenzyme being a dimer of Mr approx. 115000 and the enzyme a tetramer of Mr approx. 210 000. 3. The activation process was affected by several agents (L-3,4-dihydroxyphenylalanine, urea, formamide) that prevented, partially or totally, the activation of pro-tyrosinase. 4. The activation of pro-tyrosinase seems to be the result of a cleavage of the polypeptide chain that determines changes in tertiary or quaternary structure. PMID:6814426

  13. Activated Biological Filters (ABF Towers). Student Manual. Biological Treatment Process Control.

    ERIC Educational Resources Information Center

    Wooley, John F.

    This student manual contains textual material for a two-lesson unit on activated bio-filters (ABF). The first lesson (the sewage treatment plant) examines those process units that are unique to the ABF system. The lesson includes a review of the structural components of the ABF system and their functions and a discussion of several operational…

  14. Fantasy Activity and the Televiewing Event: Considerations for an Information Processing Construct of Involvement.

    ERIC Educational Resources Information Center

    Lindlof, Thomas R.

    The similarities between television viewing and fantasy activity (daydreaming, reverie, mind-wandering, internal dialogue) more than warrant the building of a theoretical construct, especially in the context of recent empirical research on television viewing consequences. A construct of the television viewing process, based on cognitive theories…

  15. Operational Control Procedures for the Activated Sludge Process, Part III-A: Calculation Procedures.

    ERIC Educational Resources Information Center

    West, Alfred W.

    This is the second in a series of documents developed by the National Training and Operational Technology Center describing operational control procedures for the activated sludge process used in wastewater treatment. This document deals exclusively with the calculation procedures, including simplified mixing formulas, aeration tank…

  16. Collaborative Design Processes: An Active and Reflective Learning Course in Multidisciplinary Collaboration.

    ERIC Educational Resources Information Center

    O'Brien, William J.; Soibelman, Lucio; Elvin, George

    2003-01-01

    In a capstone course, graduate students from two universities participated in collaborative design in the architectural, engineering, and construction industries in multidisciplinary teams via the Internet. Students also developed process designs to integrate technology into multidisciplinary teamwork, combining active and reflective learning.…

  17. FATE OF WATER SOLUBLE AZO DYES IN THE ACTIVATED SLUDGE PROCESS

    EPA Science Inventory

    The objective of this study was to determine the partitioning of water soluble azo dyes in the activated sludge process (ASP). Azo dyes are of concern because some of the dyes, dye precursors , and/or their degradation products such as aromatic amines (which are also dye precurso...

  18. The Effect of Process Writing Activities on the Writing Skills of Prospective Turkish Teachers

    ERIC Educational Resources Information Center

    Dilidüzgün, Sükran

    2013-01-01

    Problem statement: Writing an essay is a most difficult creative work and consequently requires detailed instruction. There are in fact two types of instruction that contribute to the development of writing skills: Reading activities analysing texts in content and schematic structure to find out how they are composed and process writing…

  19. Processes and Outcomes of Joint Activity with E-Books for Promoting Kindergarteners' Emergent Literacy

    ERIC Educational Resources Information Center

    Shamir, Adina

    2009-01-01

    This research investigated the effects of an educational electronic book (e-book) on low socioeconomic status (SES) kindergarteners' emergent literacy while focusing on the relationship between process and outcomes during joint learning. The sample (96 kindergarteners, aged five to six) was randomly assigned to experimental (e-book activation) and…

  20. Brain Activation during Semantic Processing in Autism Spectrum Disorders via Functional Magnetic Resonance Imaging

    ERIC Educational Resources Information Center

    Harris, Gordon J.; Chabris, Christopher F.; Clark, Jill; Urban, Trinity; Aharon, Itzhak; Steele, Shelley; McGrath, Lauren; Condouris, Karen; Tager-Flusberg, Helen

    2006-01-01

    Language and communication deficits are core features of autism spectrum disorders (ASD), even in high-functioning adults with ASD. This study investigated brain activation patterns using functional magnetic resonance imaging in right-handed adult males with ASD and a control group, matched on age, handedness, and verbal IQ. Semantic processing in…

  1. Active in-database processing to support ambient assisted living systems.

    PubMed

    de Morais, Wagner O; Lundström, Jens; Wickström, Nicholas

    2014-08-12

    As an alternative to the existing software architectures that underpin the development of smart homes and ambient assisted living (AAL) systems, this work presents a database-centric architecture that takes advantage of active databases and in-database processing. Current platforms supporting AAL systems use database management systems (DBMSs) exclusively for data storage. Active databases employ database triggers to detect and react to events taking place inside or outside of the database. DBMSs can be extended with stored procedures and functions that enable in-database processing. This means that the data processing is integrated and performed within the DBMS. The feasibility and flexibility of the proposed approach were demonstrated with the implementation of three distinct AAL services. The active database was used to detect bed-exits and to discover common room transitions and deviations during the night. In-database machine learning methods were used to model early night behaviors. Consequently, active in-database processing avoids transferring sensitive data outside the database, and this improves performance, security and privacy. Furthermore, centralizing the computation into the DBMS facilitates code reuse, adaptation and maintenance. These are important system properties that take into account the evolving heterogeneity of users, their needs and the devices that are characteristic of smart homes and AAL systems. Therefore, DBMSs can provide capabilities to address requirements for scalability, security, privacy, dependability and personalization in applications of smart environments in healthcare.

  2. Abstracts from the Cognitive Processes of Children Engaged in Music Activity Conference.

    ERIC Educational Resources Information Center

    Bulletin of the Council for Research in Music Education, 1999

    1999-01-01

    Contains abstracts of the papers that were presented at the Cognitive Processes of Children Engaged in Musical Activity Conference (Champaign-Urbana, IL, June 3-5, 1999). Covers topics such as children's melodic improvisations, toddlers' kinesthetic reactions to music, mother-infant play, a theory of multiple musical intelligences, and reflective…

  3. Operational Control Procedures for the Activated Sludge Process, Part I - Observations, Part II - Control Tests.

    ERIC Educational Resources Information Center

    West, Alfred W.

    This is the first in a series of documents developed by the National Training and Operational Technology Center describing operational control procedures for the activated sludge process used in wastewater treatment. Part I of this document deals with physical observations which should be performed during each routine control test. Part II…

  4. Specific and Nonspecific Neural Activity during Selective Processing of Visual Representations in Working Memory

    ERIC Educational Resources Information Center

    Oh, Hwamee; Leung, Hoi-Chung

    2010-01-01

    In this fMRI study, we investigated prefrontal cortex (PFC) and visual association regions during selective information processing. We recorded behavioral responses and neural activity during a delayed recognition task with a cue presented during the delay period. A specific cue ("Face" or "Scene") was used to indicate which one of the two…

  5. A Process of Environmental Education Communication through Community Cultural Activity Area

    ERIC Educational Resources Information Center

    Wongpaibool, Duangporn; Rawang, Wee; Supapongpichate, Ratchanont; Pichayapibool, Pataraboon

    2016-01-01

    The purpose of this research was: 1. To investigate social context, environment, way of life and community culture. 2. To gather the views and opinions regarding environmental conservation and restoration. 3. To synthesize a process of environmental education communication based on community cultural activity area. 4. To evaluate the efficacy of…

  6. Relatedness Proportion Effects in Semantic Categorization: Reconsidering the Automatic Spreading Activation Process

    ERIC Educational Resources Information Center

    de Wit, Bianca; Kinoshita, Sachiko

    2014-01-01

    Semantic priming effects at a short prime-target stimulus onset asynchrony are commonly explained in terms of an automatic spreading activation process. According to this view, the proportion of related trials should have no impact on the size of the semantic priming effect. Using a semantic categorization task ("Is this a living…

  7. NREL Develops Accelerated Sample Activation Process for Hydrogen Storage Materials (Fact Sheet)

    SciTech Connect

    Not Available

    2010-12-01

    This fact sheet describes NREL's accomplishments in developing a new sample activation process that reduces the time to prepare samples for measurement of hydrogen storage from several days to five minutes and provides more uniform samples. Work was performed by NREL's Chemical and Materials Science Center.

  8. Rhythms of Dialogue and Referential Activity: Implicit Process across Procedural and Verbal Realms

    ERIC Educational Resources Information Center

    Ritter, Michael S.

    2009-01-01

    This work examines the relationship between implicit procedural and implicit verbal processes as they occur in natural adult conversation. Theoretical insights and empirical findings are rooted in a move towards integration of Bucci's "Referential Activity" (RA) and "Multiple Code" perspectives and Beebe and Jaffe's…

  9. Really Writing! Ready-To-Use Writing Process Activities for the Elementary Grades. 2nd Edition

    ERIC Educational Resources Information Center

    Sunflower, Cherlyn

    2005-01-01

    The second edition of "Really Writing!" provides 40 real-life writing activities designed to capture the attention of young authors (grades 2 through 6) who are just learning the composing process. This book is also a resource for teaching advanced writers who are ready to experiment with a variety of writing and speaking genres. Each of the…

  10. Working through: In-Session Processes that Promote Between-Session Thoughts and Activities

    ERIC Educational Resources Information Center

    Owen, Jesse; Quirk, Kelley; Hilsenroth, Mark J.; Rodolfa, Emil

    2012-01-01

    This study examined whether clients' ratings of the working alliance as well as their perception of cognitive-behavioral (CB) and psychodynamic-interpersonal (PI) techniques (delivered by therapists who used both) were associated with clients' intersession processes (i.e., their thoughts about therapy and therapeutic activity between sessions).…

  11. 77 FR 28401 - Information Collection Activities: Legacy Data Verification Process (LDVP); Submitted for Office...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-14

    ... Bureau of Safety and Environmental Enforcement Information Collection Activities: Legacy Data Verification Process (LDVP); Submitted for Office of Management and Budget (OMB) Review; Comment Request ACTION... comments on a collection of information that we will submit to the Office of Management and Budget...

  12. A Joint Learning Activity in Process Control and Distance Collaboration between Future Engineers and Technicians

    ERIC Educational Resources Information Center

    Deschênes, Jean-Sebastien; Barka, Noureddine; Michaud, Mario; Paradis, Denis; Brousseau, Jean

    2013-01-01

    A joint learning activity in process control is presented, in the context of a distance collaboration between engineering and technical-level students, in a similar fashion as current practices in the industry involving distance coordination and troubleshooting. The necessary infrastructure and the setup used are first detailed, followed by a…

  13. Active In-Database Processing to Support Ambient Assisted Living Systems

    PubMed Central

    de Morais, Wagner O.; Lundström, Jens; Wickström, Nicholas

    2014-01-01

    As an alternative to the existing software architectures that underpin the development of smart homes and ambient assisted living (AAL) systems, this work presents a database-centric architecture that takes advantage of active databases and in-database processing. Current platforms supporting AAL systems use database management systems (DBMSs) exclusively for data storage. Active databases employ database triggers to detect and react to events taking place inside or outside of the database. DBMSs can be extended with stored procedures and functions that enable in-database processing. This means that the data processing is integrated and performed within the DBMS. The feasibility and flexibility of the proposed approach were demonstrated with the implementation of three distinct AAL services. The active database was used to detect bed-exits and to discover common room transitions and deviations during the night. In-database machine learning methods were used to model early night behaviors. Consequently, active in-database processing avoids transferring sensitive data outside the database, and this improves performance, security and privacy. Furthermore, centralizing the computation into the DBMS facilitates code reuse, adaptation and maintenance. These are important system properties that take into account the evolving heterogeneity of users, their needs and the devices that are characteristic of smart homes and AAL systems. Therefore, DBMSs can provide capabilities to address requirements for scalability, security, privacy, dependability and personalization in applications of smart environments in healthcare. PMID:25120164

  14. Analyzing the Learning Process of an Online Role-Playing Discussion Activity

    ERIC Educational Resources Information Center

    Hou, Huei-Tse

    2012-01-01

    Instructional activities based on online discussion strategies have gained prevalence in recent years. Within this context, a crucial research topic is to design innovative and appropriate online discussion strategies that assist learners in attaining a deeper level of interaction and higher cognitive skills. By analyzing the process of online…

  15. Hypoxia-Inducible Histone Lysine Demethylases: Impact on the Aging Process and Age-Related Diseases

    PubMed Central

    Salminen, Antero; Kaarniranta, Kai; Kauppinen, Anu

    2016-01-01

    Hypoxia is an environmental stress at high altitude and underground conditions but it is also present in many chronic age-related diseases, where blood flow into tissues is impaired. The oxygen-sensing system stimulates gene expression protecting tissues against hypoxic insults. Hypoxia stabilizes the expression of hypoxia-inducible transcription factor-1α (HIF-1α), which controls the expression of hundreds of survival genes related to e.g. enhanced energy metabolism and autophagy. Moreover, many stress-related signaling mechanisms, such as oxidative stress and energy metabolic disturbances, as well as the signaling cascades via ceramide, mTOR, NF-κB, and TGF-β pathways, can also induce the expression of HIF-1α protein to facilitate cell survival in normoxia. Hypoxia is linked to prominent epigenetic changes in chromatin landscape. Screening studies have indicated that the stabilization of HIF-1α increases the expression of distinct histone lysine demethylases (KDM). HIF-1α stimulates the expression of KDM3A, KDM4B, KDM4C, and KDM6B, which enhance gene transcription by demethylating H3K9 and H3K27 sites (repressive epigenetic marks). In addition, HIF-1α induces the expression of KDM2B and KDM5B, which repress transcription by demethylating H3K4me2,3 sites (activating marks). Hypoxia-inducible KDMs support locally the gene transcription induced by HIF-1α, although they can also control genome-wide chromatin landscape, especially KDMs which demethylate H3K9 and H3K27 sites. These epigenetic marks have important role in the control of heterochromatin segments and 3D folding of chromosomes, as well as the genetic loci regulating cell type commitment, proliferation, and cellular senescence, e.g. the INK4 box. A chronic stimulation of HIF-1α can provoke tissue fibrosis and cellular senescence, which both are increasingly present with aging and age-related diseases. We will review the regulation of HIF-1α-dependent induction of KDMs and clarify their role in

  16. Sampling frequency affects the processing of Actigraph raw acceleration data to activity counts.

    PubMed

    Brønd, Jan Christian; Arvidsson, Daniel

    2016-02-01

    ActiGraph acceleration data are processed through several steps (including band-pass filtering to attenuate unwanted signal frequencies) to generate the activity counts commonly used in physical activity research. We performed three experiments to investigate the effect of sampling frequency on the generation of activity counts. Ideal acceleration signals were produced in the MATLAB software. Thereafter, ActiGraph GT3X+ monitors were spun in a mechanical setup. Finally, 20 subjects performed walking and running wearing GT3X+ monitors. Acceleration data from all experiments were collected with different sampling frequencies, and activity counts were generated with the ActiLife software. With the default 30-Hz (or 60-Hz, 90-Hz) sampling frequency, the generation of activity counts was performed as intended with 50% attenuation of acceleration signals with a frequency of 2.5 Hz by the signal frequency band-pass filter. Frequencies above 5 Hz were eliminated totally. However, with other sampling frequencies, acceleration signals above 5 Hz escaped the band-pass filter to a varied degree and contributed to additional activity counts. Similar results were found for the spinning of the GT3X+ monitors, although the amount of activity counts generated was less, indicating that raw data stored in the GT3X+ monitor is processed. Between 600 and 1,600 more counts per minute were generated with the sampling frequencies 40 and 100 Hz compared with 30 Hz during running. Sampling frequency affects the processing of ActiGraph acceleration data to activity counts. Researchers need to be aware of this error when selecting sampling frequencies other than the default 30 Hz.

  17. Prefrontal Brain Activation During Emotional Processing: A Functional Near Infrared Spectroscopy Study (fNIRS)

    PubMed Central

    Glotzbach, Evelyn; Mühlberger, Andreas; Gschwendtner, Kathrin; Fallgatter, Andreas J; Pauli, Paul; Herrmann, Martin J

    2011-01-01

    The limbic system and especially the amygdala have been identified as key structures in emotion induction and regulation. Recently research has additionally focused on the influence of prefrontal areas on emotion processing in the limbic system and the amygdala. Results from fMRI studies indicate that the prefrontal cortex (PFC) is involved not only in emotion induction but also in emotion regulation. However, studies using fNIRS only report prefrontal brain activation during emotion induction. So far it lacks the attempt to compare emotion induction and emotion regulation with regard to prefrontal activation measured with fNIRS, to exclude the possibility that the reported prefrontal brain activation in fNIRS studies are mainly caused by automatic emotion regulation processes. Therefore this work tried to distinguish emotion induction from regulation via fNIRS of the prefrontal cortex. 20 healthy women viewed neutral pictures as a baseline condition, fearful pictures as induction condition and reappraised fearful pictures as regulation condition in randomized order. As predicted, the view-fearful condition led to higher arousal ratings than the view-neutral condition with the reappraise-fearful condition in between. For the fNIRS results the induction condition showed an activation of the bilateral PFC compared to the baseline condition (viewing neutral). The regulation condition showed an activation only of the left PFC compared to the baseline condition, although the direct comparison between induction and regulation condition revealed no significant difference in brain activation. Therefore our study underscores the results of previous fNIRS studies showing prefrontal brain activation during emotion induction and rejects the hypothesis that this prefrontal brain activation might only be a result of automatic emotion regulation processes. PMID:21673974

  18. Active content determination of pharmaceutical tablets using near infrared spectroscopy as Process Analytical Technology tool.

    PubMed