Science.gov

Sample records for lesanded hes sellega

  1. Hypereosinophilic Syndrome (HES)

    MedlinePlus

    ... and is part of the larger group of Myeloproliferative Neoplasms. Closely related disease to HES is chronic eosinophilic ... long-term therapy. Overall experience with IFNa in myeloproliferative neoplasms is that about 25-30% of patients require ...

  2. HES6-1 and HES6-2 Function through Different Mechanisms during Neuronal Differentiation

    PubMed Central

    Vilas-Boas, Filipe; Henrique, Domingos

    2010-01-01

    Background Notch signalling plays a central role in the mechanisms regulating neuronal differentiation in the vertebrate nervous system. The transcriptional repressors encoded by Hes genes are the main effectors of this pathway, acting in neural progenitors during the lateral inhibition process to repress proneural genes and inhibit differentiation. However, Hes6 genes seem to behave differently: they are expressed in differentiating neurons and facilitate the activity of proneural genes in promoting neurogenesis. Still, the molecular mechanisms underlying this unique function of Hes6 genes are not yet understood. Methodology/Principal Findings Here, we identify two subgroups of Hes6 genes that seem conserved in most vertebrate species and characterize a novel Hes6 gene in chicken: cHes6-1. The embryonic expression pattern of cHes6-1 suggests roles for this gene in the formation of the pancreas, nervous system and in the generation of body asymmetry. We show that cHes6-1 is negatively regulated by Notch signalling in the developing embryonic spinal cord and in pancreatic progenitors, but requires Notch for the observed asymmetric expression at the lateral mesoderm. Functional studies by ectopic expression in the chick embryonic neural tube revealed that cHES6-1 up-regulates the expression of cDelta1 and cHes5 genes, in contrast with overexpression of cHES6-2, which represses the same genes. We show that this activity of cHES6-2 is dependent on its capacity to bind DNA and repress transcription, while cHES6-1 seems to function by sequestering other HES proteins and inhibit their activity as transcriptional repressors. Conclusions/Significance Our results indicate that the two chick HES6 proteins act at different phases of neuronal differentiation, contributing to the progression of neurogenesis by different mechanisms: while cHES6-2 represses the transcription of Hes genes, cHES6-1 acts later, sequestering HES proteins. Together, the two cHES6 proteins progressively

  3. HES1 in immunity and cancer.

    PubMed

    Rani, Aradhana; Greenlaw, Roseanna; Smith, Richard A; Galustian, Christine

    2016-08-01

    Hairy and enhancer of split homolog-1 (HES1) is a part of an extensive family of basic helix-loop-helix (bHLH) proteins and plays a crucial role in the control and regulation of cell cycle, proliferation, cell differentiation, survival and apoptosis in neuronal, endocrine, T-lymphocyte progenitors as well as various cancers. HES1 is a transcription factor which is regulated by the NOTCH, Hedgehog and Wnt signalling pathways. Aberrant expression of these pathways is a common feature of cancerous cells. There appears to be a fine and complicated crosstalk at the molecular level between the various signalling pathways and HES1, which contributes to its effects on the immune response and cancers such as leukaemia. Several mechanisms have been proposed, including an enhanced invasiveness and metastasis by inducing epithelial mesenchymal transition (EMT), in addition to its strict requirement for tumour cell survival. In this review, we summarize the current biology and molecular mechanisms as well as its use as a clinical target in cancer therapeutics. PMID:27066918

  4. HES6 enhances the motility of alveolar rhabdomyosarcoma cells

    SciTech Connect

    Wickramasinghe, Caroline M; Domaschenz, Renae; Amagase, Yoko; Williamson, Daniel; Missiaglia, Edoardo; Shipley, Janet; Murai, Kasumi; Jones, Philip H

    2013-01-01

    Absract: HES6, a member of the hairy-enhancer-of-split family of transcription factors, plays multiple roles in myogenesis. It is a direct target of the myogenic transcription factor MyoD and has been shown to regulate the formation of the myotome in development, myoblast cell cycle exit and the organization of the actin cytoskeleton during terminal differentiation. Here we investigate the expression and function of HES6 in rhabdomyosarcoma, a soft tissue tumor which expresses myogenic genes but fails to differentiate into muscle. We show that HES6 is expressed at high levels in the subset of alveolar rhabdomyosarcomas expressing PAX/FOXO1 fusion genes (ARMSp). Knockdown of HES6 mRNA in the ARMSp cell line RH30 reduces proliferation and cell motility. This phenotype is rescued by expression of mouse Hes6 which is insensitive to HES6 siRNA. Furthermore, expression microarray analysis indicates that the HES6 knockdown is associated with a decrease in the levels of Transgelin, (TAGLN), a regulator of the actin cytoskeleton. Knockdown of TAGLN decreases cell motility, whilst TAGLN overexpression rescues the motility defect resulting from HES6 knockdown. These findings indicate HES6 contributes to the pathogenesis of ARMSp by enhancing both proliferation and cell motility.

  5. The glucocorticoid receptor controls hepatic dyslipidemia through Hes1.

    PubMed

    Lemke, Ulrike; Krones-Herzig, Anja; Berriel Diaz, Mauricio; Narvekar, Prachiti; Ziegler, Anja; Vegiopoulos, Alexandros; Cato, Andrew C B; Bohl, Sebastian; Klingmüller, Ursula; Screaton, Robert A; Müller-Decker, Karin; Kersten, Sander; Herzig, Stephan

    2008-09-01

    Aberrant accumulation of lipids in the liver ("fatty liver" or hepatic steatosis) represents a hallmark of the metabolic syndrome and is tightly associated with obesity, type II diabetes, starvation, or glucocorticoid (GC) therapy. While fatty liver has been connected with numerous abnormalities of liver function, the molecular mechanisms of fatty liver development remain largely enigmatic. Here we show that liver-specific disruption of glucocorticoid receptor (GR) action improves the steatotic phenotype in fatty liver mouse models and leads to the induction of transcriptional repressor hairy enhancer of split 1 (Hes1) gene expression. The GR directly interferes with Hes1 promoter activity, triggering the recruitment of histone deacetylase (HDAC) activities to the Hes1 gene. Genetic restoration of hepatic Hes1 levels in steatotic animals normalizes hepatic triglyceride (TG) levels. As glucocorticoid action is increased during starvation, myotonic dystrophy, and Cushing's syndrome, the inhibition of Hes1 through the GR might explain the fatty liver phenotype in these subjects. PMID:18762022

  6. Hes-1 SUMOylation by protein inhibitor of activated STAT1 enhances the suppressing effect of Hes-1 on GADD45α expression to increase cell survival

    PubMed Central

    2014-01-01

    Background Hairy and Enhancer of split 1 (Hes-1) is a transcriptional repressor that plays an important role in neuronal differentiation and development, but post-translational modifications of Hes-1 are much less known. In the present study, we aimed to investigate whether Hes-1 could be SUMO-modified and identify the candidate SUMO acceptors on Hes-1. We also wished to examine the role of the SUMO E3 ligase protein inhibitor of activated STAT1 (PIAS1) in SUMOylation of Hes-1 and the molecular mechanism of Hes-1 SUMOylation. Further, we aimed to identify the molecular target of Hes-1 and examine how Hes-1 SUMOylation affects its molecular target to affect cell survival. Results In this study, by using HEK293T cells, we have found that Hes-1 could be SUMO-modified and Hes-1 SUMOylation was greatly enhanced by the SUMO E3 ligase PIAS1 at Lys8, Lys27 and Lys39. Furthermore, Hes-1 SUMOylation stabilized the Hes-1 protein and increased the transcriptional suppressing activity of Hes-1 on growth arrest and DNA damage-inducible protein alpha (GADD45α) expression. Overexpression of GADD45α increased, whereas knockdown of GADD45αα expression decreased cell apoptosis. In addition, H2O2 treatment increased the association between PIAS1 and Hes-1 and enhanced the SUMOylation of Hes-1 for endogenous protection. Overexpression of Hes-1 decreased H2O2-induced cell death, but this effect was blocked by transfection of the Hes-1 triple sumo-mutant (Hes-1 3KR). Overexpression of PIAS1 further facilitated the anti-apoptotic effect of Hes-1. Moreover, Hes-1 SUMOylation was independent of Hes-1 phosphorylation and vice versa. Conclusions The present results revealed, for the first time, that Hes-1 could be SUMO-modified by PIAS1 and GADD45α is a novel target of Hes-1. Further, Hes-1 SUMOylation mediates cell survival through enhanced suppression of GADD45α expression. These results revealed a novel role of Hes-1 in addition to its involvement in Notch signaling. They also

  7. Regeneration of mammalian cochlear and vestibular hair cells through Hes1/Hes5 modulation with siRNA

    PubMed Central

    Du, Xiaoping; Li, Wei; Gao, Xinsheng; West, Matthew B.; Saltzman, W. Mark; Cheng, Christopher J.; Stewart, Charles; Zheng, Jie; Cheng, Weihua; Kopke, Richard D.

    2014-01-01

    The Notch pathway is a cell signaling pathway determining initial specification and subsequent cell fate in the inner ear. Previous studies have suggested that new hair cells (HCs) can be regenerated in the inner ear by manipulating the Notch pathway. In the present study, delivery of siRNA to Hes1 and Hes5 using a transfection reagent or siRNA to Hes1 encapsulated within poly(lactide-co-glycolide acid) (PLGA) nanoparticles increased HC numbers in non-toxin treated organotypic cultures of cochleae and maculae of postnatal day 3 mouse pups. An increase in HCs was also observed in cultured cochleae and maculae of mouse pups pre-conditioned with a HC toxin (4-hydroxy-2-nonenal or neomycin) and then treated with the various siRNA formulations. Treating cochleae with siRNA to Hes1 associated with a transfection reagent or siRNA to Hes1 delivered by PLGA nanoparticles decreased Hes1 mRNA and up-regulated Atoh1 mRNA expression allowing supporting cells (SCs) to acquire a HC fate. Experiments using cochleae and maculae of p27kip1/-GFP transgenic mouse pups demonstrated that newly generated HCs trans-differentiated from SCs. Furthermore, PLGA nanoparticles are non-toxic to inner ear tissue, readily taken up by cells within the tissue of interest, and present a synthetic delivery system that is a safe alternative to viral vectors. These results indicate that when delivered using a suitable vehicle, Hes siRNAs are potential therapeutic molecules that may have the capacity to regenerate new HCs in the inner ear and possibly restore human hearing and balance function. PMID:23850665

  8. Results of NASA/NOAA HES Trade Studies

    NASA Technical Reports Server (NTRS)

    Susskind, Joel

    2011-01-01

    This slide presentation reviews the trade studies that were done for the Hyperspectral Environmental Suite (HES). The goal of the trade studies was to minimize instrument cost and risk while producing scientifically useful products. Three vendors were selected to perform the trade study, and were to conduct 11 studies, with the first study a complete wish list of things that scientists would like from GEO orbit to the 11th study which was for a Reduced Accommodation Sounder (RAS) which would still result in useful scientific products, within constrains compatible with flight on GEOS-R. The RAS's from each vendor and one other HES sounders designs are reviewed.

  9. The Hospice Environmental Survey (HES): Pilot Test of a New Measurement Instrument.

    ERIC Educational Resources Information Center

    Taylor, Jean H.; Perrill, Norman K.

    1988-01-01

    Describes development of the Hospice Environmental Survey (HES) to measure user's perception of the homelike atmosphere provided by a hospital inpatient unit called Hospice House. Presents the HES instrument, methodology, and pilot study data. (Author/NB)

  10. Dynamic reprogramming of chromatin: paradigmatic palimpsests and HES factors

    PubMed Central

    Kok, Kurtulus; Arnosti, David N.

    2015-01-01

    Temporal and spatial control of transcription in development is dictated to a great extent by transcriptional repressors. Some repressor complexes, such as Polycomp-group proteins, induce relatively long-term non-permissive states, whereas others such as hairy/enhancer of split (HES) family repressors are linked to dynamically modulated chromatin states associated with cycling expression of target genes. The mode of action and specificity of repressors involved in mediating this latter form of epigenetic control are unknown. Oscillating expression of HES repressors controlled by signaling pathways such as Notch suggests that the entire ensemble of HES–associated co-repressors and histone modifying complexes readily cycle on and off genes. Dynamic interactions between these factors and chromatin seem to be crucial in maintaining multipotency of progenitor cells, but the significance of such interactions in more differentiated cells is less well understood. We discuss here how genome-wide analyses and real-time gene expression measurements of HES regulated genes can help decipher the detailed mechanisms and biological importance of highly dynamic transcriptional switching mediated by epigenetic changes. PMID:25713582

  11. HES-Mediated Repression of Pten in Caenorhabditis elegans

    PubMed Central

    Chou, Han Ting; Vazquez, Raymarie Gomez; Wang, Kun; Campbell, Richard; Milledge, Gaolin Zheng; Walthall, Walter W.; Johnson, Casonya M.

    2015-01-01

    The hairy/enhancer-of-split (HES) group of transcription factors controls embryonic development, often by acting downstream of the Notch signaling pathway; however, little is known about postembryonic roles of these proteins. In Caenorhabditis elegans, the six proteins that make up the REF-1 family are considered to be HES orthologs that act in both Notch-dependent and Notch-independent pathways to regulate embryonic events. To further our understanding of how the REF-1 family works to coordinate postembryonic cellular events, we performed a functional characterization of the REF-1 family member, HLH-25. We show that, after embryogenesis, hlh-25 expression persists throughout every developmental stage, including dauer, into adulthood. Like animals that carry loss-of-function alleles in genes required for normal cell-cycle progression, the phenotypes of hlh-25 animals include reduced brood size, unfertilized oocytes, and abnormal gonad morphology. Using gene expression microarray, we show that the HLH-25 transcriptional network correlates with the phenotypes of hlh-25 animals and that the C. elegans Pten ortholog, daf-18, is one major hub in the network. Finally, we show that HLH-25 regulates C. elegans lifespan and dauer recovery, which correlates with a role in the transcriptional repression of daf-18 activity. Collectively, these data provide the first genetic evidence that HLH-25 may be a functional ortholog of mammalian HES1, which represses PTEN activity in mice and human cells. PMID:26438299

  12. Discovery of HeLa Cell Contamination in HES Cells

    PubMed Central

    Summerfield, Taryn L.

    2014-01-01

    Continuous cell lines are used frequently in reproductive biology research to study problems in early pregnancy events and parturition. It has been recognized for 50 years that many mammalian cell lines contain inter- or intraspecies contaminations with other cells. However, most investigators do not routinely test their culture systems for cross-contamination. The most frequent contributor to cross-contamination of cell lines is the HeLa cell isolated from an aggressive cervical adenocarcinoma. We report on the discovery of HeLa cell contamination of the human endometrial epithelial cell line HES isolated in our laboratory. Short tandem repeat analysis of 9 unique genetic loci demonstrated molecular identity between HES and HeLa cells. In addition, we verified that WISH cells, isolated originally from human amnion epithelium, were also contaminated with HeLa cells. Inasmuch as our laboratory did not culture HeLa cells at the time of HES cell derivations, the source of contamination was the WISH cell line. These data highlight the need for continued diligence in authenticating cell lines used in reproductive biology research. PMID:24520087

  13. Use of Hes1-GFP reporter mice to assess activity of the Hes1 promoter in bone cells under chronic inflammation.

    PubMed

    Zhang, Hengwei; Sun, Wen; Li, Xing; Wang, Mengmeng; Boyce, Brendan F; Hilton, Matthew J; Xing, Lianping

    2016-09-01

    Notch signaling plays a critical role in maintaining bone homeostasis partially by controlling the formation of osteoblasts from mesenchymal stem cells (MSCs). We reported that TNF activates Notch signaling in MSCs which inhibits osteoblast differentiation in TNF transgenic (TNF-Tg) mice, a mouse model of chronic inflammatory arthritis. In the current study, we used Hes1-GFP and Hes1-GFP/TNF-Tg mice to study the distribution and dynamic change of Notch active cells in normal and inflammatory bone loss and mechanisms mediating their enhanced proliferation. We found that Hes1-GFP+ cells are composed of cells expressing mesenchymal, hematopoietic and endothelial surface markers. CD45-/Hes1-GFP+ cells express high levels of mesenchymal markers and form CFU-F and CFU-ALP colonies. Expansion of CFU-F colonies is associated with a rapid increase in Hes1-GFP+ cell numbers and their GFP intensity. The GFP signal is lost when a CFU-F colony differentiates into an ALP+ osteoblast colony. TNF increases the numbers of CD45-/Hes1-GFP+ cells, which are stained negatively for osteoblast marker osteocalcin and localized adjacent to endosteal and trabecular bone surfaces. CD45-/Hes1-GFP+ cells in Hes1-GFP/TNF-Tg mice have increased BrdU incorporation and PDGFRβ levels. TNF increases the number of proliferating Hes1-GFP+ cells, which is prevented by a specific PDGFRβ inhibitor. Notch inhibition blocks TNF-mediated PDGFRβ expression and cell proliferation. Thus, TNF-induced MSC proliferation is mediated by PDGFRβ signal, which works at downstream of Notch. Hes1-GFP mice can be used to assess the activation status of Notch in bone cells. PMID:27269414

  14. Identification, characterization, and chromosomal localization of the human homolog (hES) of ES/130

    SciTech Connect

    Basson, C.T.; Morton, C.C.; MacRae, C.A.

    1996-08-01

    The chicken extracellular matrix glycoprotein ES/130 is necessary for epithelial-mesenchymal transformation in the developing hear and is also expressed in noncardiac chicken tissues such as limb and notochord. We have identified hES, the human homology of chicken ES/130. Fluorescence in situ hybridization analysis (FISH) localizes hES to human chromosome 20p11.2-p12. FISH analyses of individuals with 20p12 deletions and affected by Alagille syndrome exclude hES as a candidate gene for this disorder. Reverse transcriptase-polymerase chain reaction studies reveal that hES is expressed in both fetal and adult human tissues and that hES expression in the left ventricle is increased in the failing adult heart. Further studies will evaluate how hES mutations may relate to congenital human cardiac and skeletal anomalies as well as cardiac remodeling in the adult. 16 refs., 2 figs.

  15. Hijacking HES1: Tumors Co-opt the Anti-differentiation Strategies of Quiescent Cells

    PubMed Central

    Sang, Liyun; Roberts, James M.; Coller, Hilary A.

    2010-01-01

    Quiescent cells and tumor cells share the ability to evade irreversible cell fates. It was recently shown that the transcriptional regulator HES1 protects quiescent fibroblasts from differentiation or senescence. HES1 is highly expressed in rhabdomyosarcomas, and inhibition of HES1 restores differentiation in rhabdomyosarcoma cells. Pathways that lead to elevated HES1 levels, notch and hedgehog, are frequently upregulated in tumors. Compounds that inhibit these pathways induce differentiation and apoptosis in cancer cells and several are in clinical trials. HES1 might repress gene expression in part by recruiting histone deacetylases (HDACs). HDACs inhibit differentiation, whereas histone deacetylase inhibitors (HDACi’s) induce differentiation or apoptosis in tumors and are also showing promise as therapeutics. Small molecules that directly target HES1 itself were just identified. Thus, as we shall review in this article, therapies that target these pathways could be effective alone, in combination or in conjunction with traditional chemotherapeutics. PMID:20022559

  16. Loss of Hes1 Differentiates Sessile Serrated Adenoma/polyp from Hyperplastic Polyp

    PubMed Central

    Cui, Min; Awadallah, Amad; Liu, Wendy; Zhou, Lan; Xin, Wei

    2016-01-01

    Sessile serrated adenoma/polyp (SSA/p) is a precancerous lesion, and its differential diagnosis from hyperplastic polyp (HP) could be challenging in certain circumstances based on morphology alone. Hes1 is a downstream target of Notch signaling pathway and plays an important role in intestinal development by regulating differentiation of enterocytes. In this study, we evaluated the expression patterns of Hes1 in SSA/p and hyperplastic polyp (HP), and determine whether Hes1 immunostaining can help differentiate between these two entities. Serrated polyps with cytological dysplasia (sessile serrated adenoma with cytological dysplasia, tubular adenoma, and traditional serrated adenoma) were also studied. Hes1 is ubiquitously expressed in the nuclei of normal colon epithelial cells. The complete loss or a very weak expression of Hes1 is observed in the majority of the SSA/p in the study (58/63, 92%) compared to the normal expression of Hes1 in HP (35/35,100%). In SSA/p with cytological dysplasia, dysplastic area demonstrated cytoplasmic and/or nuclear staining for Hes1. Tubular adenoma and traditional serrated adenoma showed variability of Hes1 staining within the polyp with a mixed positive and negative staining pattern. Our study suggests that loss of Hes1 could be used as a sensitive and specific marker to differentiate SSA/p from HP, which helps the diagnosis in morphologically challenging cases. PMID:26448192

  17. Hes3 expression in the adult mouse brain is regulated during demyelination and remyelination.

    PubMed

    Toutouna, Louiza; Nikolakopoulou, Polyxeni; Poser, Steven W; Masjkur, Jimmy; Arps-Forker, Carina; Troullinaki, Maria; Grossklaus, Sylvia; Bosak, Viktoria; Friedrich, Ulrike; Ziemssen, Tjalf; Bornstein, Stefan R; Chavakis, Triantafyllos; Androutsellis-Theotokis, Andreas

    2016-07-01

    Hes3 is a component of the STAT3-Ser/Hes3 Signaling Axis controlling the growth and survival of neural stem cells and other plastic cells. Pharmacological activation of this pathway promotes neuronal rescue and behavioral recovery in models of ischemic stroke and Parkinson's disease. Here we provide initial observations implicating Hes3 in the cuprizone model of demyelination and remyelination. We focus on the subpial motor cortex of mice because we detected high Hes3 expression. This area is of interest as it is impacted both in human demyelinating diseases and in the cuprizone model. We report that Hes3 expression is reduced at peak demyelination and is partially restored within 1 week after cuprizone withdrawal. This raises the possibility of Hes3 involvement in demyelination/remyelination that may warrant additional research. Supporting a possible role of Hes3 in the maintenance of oligodendrocyte markers, a Hes3 null mouse strain shows lower levels of myelin basic protein in undamaged adult mice, compared to wild-type controls. We also present a novel method for culturing the established oligodendrocyte progenitor cell line oli-neu in a manner that maintains Hes3 expression as well as its self-renewal and differentiation potential, offering an experimental tool to study Hes3. Based upon this approach, we identify a Janus kinase inhibitor and dbcAMP as powerful inducers of Hes3 gene expression. We provide a new biomarker and cell culture method that may be of interest in demyelination/remyelination research. PMID:27018293

  18. Balanced Hydroxyethylstarch (HES 130/0.4) Impairs Kidney Function In-Vivo without Inflammation

    PubMed Central

    Schick, Martin Alexander; Baar, Wolfgang; Bruno, Raphael Romano; Wollborn, Jakob; Held, Christopher; Schneider, Reinhard; Flemming, Sven; Schlegel, Nicolas; Roewer, Norbert; Neuhaus, Winfried; Wunder, Christian

    2015-01-01

    Volume therapy is a standard procedure in daily perioperative care, and there is an ongoing discussion about the benefits of colloid resuscitation with hydroxyethylstarch (HES). In sepsis HES should be avoided due to a higher risk for acute kidney injury (AKI). Results of the usage of HES in patients without sepsis are controversial. Therefore we conducted an animal study to evaluate the impact of 6% HES 130/0.4 on kidney integrity with sepsis or under healthy conditions Sepsis was induced by standardized Colon Ascendens Stent Peritonitis (sCASP). sCASP-group as well as control group (C) remained untreated for 24 h. After 18 h sCASP+HES group (sCASP+VOL) and control+HES (C+VOL) received 50 ml/KG balanced 6% HES (VOL) 130/0.4 over 6h. After 24h kidney function was measured via Inulin- and PAH-Clearance in re-anesthetized rats, and serum urea, creatinine (crea), cystatin C and Neutrophil gelatinase-associated lipocalin (NGAL) as well as histopathology were analysed. In vitro human proximal tubule cells (PTC) were cultured +/- lipopolysaccharid (LPS) and with 0.1–4.0% VOL. Cell viability was measured with XTT-, cell toxicity with LDH-test. sCASP induced severe septic AKI demonstrated divergent results regarding renal function by clearance or creatinine measure focusing on VOL. Soleley HES (C+VOL) deteriorated renal function without sCASP. Histopathology revealed significantly derangements in all HES groups compared to control. In vitro LPS did not worsen the HES induced reduction of cell viability in PTC cells. For the first time, we demonstrated, that application of 50 ml/KG 6% HES 130/0.4 over 6 hours induced AKI without inflammation in vivo. Severity of sCASP induced septic AKI might be no longer susceptible to the way of volume expansion. PMID:26340751

  19. History of a prolific family: the Hes/Hey-related genes of the annelid Platynereis

    PubMed Central

    2014-01-01

    Background The Hes superfamily or Hes/Hey-related genes encompass a variety of metazoan-specific bHLH genes, with somewhat fuzzy phylogenetic relationships. Hes superfamily members are involved in a variety of major developmental mechanisms in metazoans, notably in neurogenesis and segmentation processes, in which they often act as direct effector genes of the Notch signaling pathway. Results We have investigated the molecular and functional evolution of the Hes superfamily in metazoans using the lophotrochozoan Platynereis dumerilii as model. Our phylogenetic analyses of more than 200 Metazoan Hes/Hey-related genes revealed the presence of five families, three of them (Hes, Hey and Helt) being pan-metazoan. Those families were likely composed of a unique representative in the last common metazoan ancestor. The evolution of the Hes family was shaped by many independent lineage specific tandem duplication events. The expression patterns of 13 of the 15 Hes/Hey-related genes in Platynereis indicate a broad functional diversification. Nevertheless, a majority of these genes are involved in two crucial developmental processes in annelids: neurogenesis and segmentation, resembling functions highlighted in other animal models. Conclusions Combining phylogenetic and expression data, our study suggests an unusual evolutionary history for the Hes superfamily. An ancestral multifunctional annelid Hes gene may have undergone multiples rounds of duplication-degeneration-complementation processes in the lineage leading to Platynereis, each gene copies ensuring their maintenance in the genome by subfunctionalisation. Similar but independent waves of duplications are at the origin of the multiplicity of Hes genes in other metazoan lineages. PMID:25250171

  20. Balanced Hydroxyethylstarch (HES 130/0.4) Impairs Kidney Function In-Vivo without Inflammation.

    PubMed

    Schick, Martin Alexander; Baar, Wolfgang; Bruno, Raphael Romano; Wollborn, Jakob; Held, Christopher; Schneider, Reinhard; Flemming, Sven; Schlegel, Nicolas; Roewer, Norbert; Neuhaus, Winfried; Wunder, Christian

    2015-01-01

    Volume therapy is a standard procedure in daily perioperative care, and there is an ongoing discussion about the benefits of colloid resuscitation with hydroxyethylstarch (HES). In sepsis HES should be avoided due to a higher risk for acute kidney injury (AKI). Results of the usage of HES in patients without sepsis are controversial. Therefore we conducted an animal study to evaluate the impact of 6% HES 130/0.4 on kidney integrity with sepsis or under healthy conditions Sepsis was induced by standardized Colon Ascendens Stent Peritonitis (sCASP). sCASP-group as well as control group (C) remained untreated for 24 h. After 18 h sCASP+HES group (sCASP+VOL) and control+HES (C+VOL) received 50 ml/KG balanced 6% HES (VOL) 130/0.4 over 6 h. After 24 h kidney function was measured via Inulin- and PAH-Clearance in re-anesthetized rats, and serum urea, creatinine (crea), cystatin C and Neutrophil gelatinase-associated lipocalin (NGAL) as well as histopathology were analysed. In vitro human proximal tubule cells (PTC) were cultured +/- lipopolysaccharid (LPS) and with 0.1-4.0% VOL. Cell viability was measured with XTT-, cell toxicity with LDH-test. sCASP induced severe septic AKI demonstrated divergent results regarding renal function by clearance or creatinine measure focusing on VOL. Soleley HES (C+VOL) deteriorated renal function without sCASP. Histopathology revealed significantly derangements in all HES groups compared to control. In vitro LPS did not worsen the HES induced reduction of cell viability in PTC cells. For the first time, we demonstrated, that application of 50 ml/KG 6% HES 130/0.4 over 6 hours induced AKI without inflammation in vivo. Severity of sCASP induced septic AKI might be no longer susceptible to the way of volume expansion. PMID:26340751

  1. HES factors regulate specific aspects of chondrogenesis and chondrocyte hypertrophy during cartilage development.

    PubMed

    Rutkowski, Timothy P; Kohn, Anat; Sharma, Deepika; Ren, Yinshi; Mirando, Anthony J; Hilton, Matthew J

    2016-06-01

    RBPjκ-dependent Notch signaling regulates multiple processes during cartilage development, including chondrogenesis, chondrocyte hypertrophy and cartilage matrix catabolism. Select members of the HES- and HEY-families of transcription factors are recognized Notch signaling targets that mediate specific aspects of Notch function during development. However, whether particular HES and HEY factors play any role(s) in the processes during cartilage development is unknown. Here, for the first time, we have developed unique in vivo genetic models and in vitro approaches demonstrating that the RBPjκ-dependent Notch targets HES1 and HES5 suppress chondrogenesis and promote the onset of chondrocyte hypertrophy. HES1 and HES5 might have some overlapping function in these processes, although only HES5 directly regulates Sox9 transcription to coordinate cartilage development. HEY1 and HEYL play no discernable role in regulating chondrogenesis or chondrocyte hypertrophy, whereas none of the HES or HEY factors appear to mediate Notch regulation of cartilage matrix catabolism. This work identifies important candidates that might function as downstream mediators of Notch signaling both during normal skeletal development and in Notch-related skeletal disorders. PMID:27160681

  2. Conjugates of methylated cyclodextrin derivatives and hydroxyethyl starch (HES): Synthesis, cytotoxicity and inclusion of anaesthetic actives

    PubMed Central

    Markenstein, Lisa; Appelt-Menzel, Antje; Metzger, Marco

    2014-01-01

    Summary The mono-6-deoxy-6-azides of 2,6-di-O-methyl-β-cyclodextrin (DIMEB) and randomly methylated-β-cyclodextrin (RAMEB) were conjugated to propargylated hydroxyethyl starch (HES) by Cu+-catalysed [2 + 3] cycloaddition. The resulting water soluble polymers showed lower critical solution temperatures (LCST) at 52.5 °C (DIMEB-HES) and 84.5 °C (RAMEB-HES), respectively. LCST phase separations could be completely avoided by the introduction of a small amount of carboxylate groups at the HES backbone. The methylated CDs conjugated to the HES backbone exhibited significantly lower cytotoxicities than the corresponding monomeric CD derivatives. Since the binding potentials of these CD conjugates were very high, they are promising candidates for new oral dosage forms of anaesthetic actives. PMID:25670977

  3. Test Report : GS battery, EPC power HES RESCU.

    SciTech Connect

    Rose, David Martin; Schenkman, Benjamin L.; Borneo, Daniel R.

    2013-10-01

    The Department of Energy Office of Electricity (DOE/OE), Sandia National Laboratories (SNL) and the Base Camp Integration Lab (BCIL) partnered together to incorporate an energy storage system into a microgrid configured Forward Operating Base to reduce the fossil fuel consumption and to ultimately save lives. Energy storage vendors will be sending their systems to SNL Energy Storage Test Pad (ESTP) for functional testing and then to the BCIL for performance evaluation. The technologies that will be tested are electro-chemical energy storage systems comprising of lead acid, lithium-ion or zinc-bromide. GS Battery and EPC Power have developed an energy storage system that utilizes zinc-bromide flow batteries to save fuel on a military microgrid. This report contains the testing results and some limited analysis of performance of the GS Battery, EPC Power HES RESCU.

  4. Hes6 is required for actin cytoskeletal organization in differentiating C2C12 myoblasts

    SciTech Connect

    Malone, Caroline M.P.; Domaschenz, Renae; Amagase, Yoko; Dunham, Ian; Murai, Kasumi; Jones, Philip H.

    2011-07-01

    Hes6 is a member of the hairy-enhancer-of-split family of transcription factors that regulate proliferating cell fate in development and is known to be expressed in developing muscle. Here we investigate its function in myogenesis in vitro. We show that Hes6 is a direct transcriptional target of the myogenic transcription factors MyoD and Myf5, indicating that it is integral to the myogenic transcriptional program. The localization of Hes6 protein changes during differentiation, becoming predominantly nuclear. Knockdown of Hes6 mRNA levels by siRNA has no effect on cell cycle exit or induction of myosin heavy chain expression in differentiating C2C12 myoblasts, but F-actin filament formation is disrupted and both cell motility and myoblast fusion are reduced. The knockdown phenotype is rescued by expression of Hes6 cDNA resistant to siRNA. These results define a novel role for Hes6 in actin cytoskeletal dynamics in post mitotic myoblasts.

  5. MicroRNA-765 regulates neural stem cell proliferation and differentiation by modulating Hes1 expression

    PubMed Central

    Li, Siou; Zhao, Weina; Xu, Qing; Yu, Yang; Yin, Changhao

    2016-01-01

    Neural stem cells (NSCs) are multipotent, self-renewing and undifferentiated cells that have the ability to differentiate to both glial and neuronal lineages. miRNAs act a key role in regulating neuronal fate and self-renewal of NSCs. In this study, we found that ectopic expression of miR-765 promoted NSCs proliferation. Moreover, miR-765 overexpression increased the ki-67 and β-tubulin-III expression inNSCs. Overexpression of miR-765 inhibited the expression of GFAP in NSCs. Furthermore, Hes1 was identified as a direct target gene of miR-765 in NSCs. Overexpression of Hes1 decreased miR-765-induced proliferation of NSCs and inhibited NSCs differentiation to neurons in miR-765-treated NSCs. These results demonstrated that miR-765 acted a crucial role in NSCs differentiation and proliferation by inhibiting Hes1 expression. PMID:27508032

  6. GATA-1 Utilizes Ikaros and Polycomb Repressive Complex 2 To Suppress Hes1 and To Promote Erythropoiesis

    PubMed Central

    Ross, Julie; Mavoungou, Lionel; Bresnick, Emery H.

    2012-01-01

    The transcription factor Hairy Enhancer of Split 1 (HES1), a downstream effector of the Notch signaling pathway, is an important regulator of hematopoiesis. Here, we demonstrate that in primary erythroid cells, Hes1 gene expression is transiently repressed around proerythroblast stage of differentiation. Using mouse erythroleukemia cells, we found that the RNA interference (RNAi)-mediated depletion of HES1 enhances erythroid cell differentiation, suggesting that this protein opposes terminal erythroid differentiation. This is also supported by the decreased primary erythroid cell differentiation upon HES1 upregulation in Ikaros-deficient mice. A comprehensive analysis led us to determine that Ikaros favors Hes1 repression in erythroid cells by facilitating recruitment of the master regulator of erythropoiesis GATA-1 alongside FOG-1, which mediates Hes1 repression. GATA-1 is then necessary for the chromatin binding of the NuRD remodeling complex ATPase MI-2, the transcription factor GFI1B, and the histone H3K27 methyltransferase EZH2 along with Polycomb repressive complex 2. We show that EZH2 is required for the transient repression of Hes1 in erythroid cells. In aggregate, our results describe a mechanism whereby GATA-1 utilizes Ikaros and Polycomb repressive complex 2 to promote Hes1 repression as an important step in erythroid cell differentiation. PMID:22778136

  7. Hes1 Increases the Invasion Ability of Colorectal Cancer Cells via the STAT3-MMP14 Pathway

    PubMed Central

    Weng, MT; Tsao, PN; Lin, HL; Tung, CC; Change, MC; Chang, YT; Wong, JM; Wei, SC

    2015-01-01

    The Notch pathway contributes to self-renewal of tumor-initiating cell and inhibition of normal colonic epithelial cell differentiation. Deregulated expression of Notch1 and Jagged1 is observed in colorectal cancer. Hairy/enhancer of split (HES) family, the most characterized targets of Notch, involved in the development of many cancers. In this study, we explored the role of Hes1 in the tumorigenesis of colorectal cancer. Knocking down Hes1 induced CRC cell senescence and decreased the invasion ability, whereas over-expression of Hes1 increased STAT3 phosphorylation activity and up-regulated MMP14 protein level. We further explored the expression of Hes1 in human colorectal cancer and found high Hes1 mRNA expression is associated with poor prognosis in CRC patients. These findings suggest that Hes1 regulates the invasion ability through the STAT3-MMP14 pathway in CRC cells and high Hes1 expression is a predictor of poor prognosis of CRC. PMID:26650241

  8. HES6 gene is selectively overexpressed in glioma and represents an important transcriptional regulator of glioma proliferation.

    PubMed

    Haapa-Paananen, S; Kiviluoto, S; Waltari, M; Puputti, M; Mpindi, J P; Kohonen, P; Tynninen, O; Haapasalo, H; Joensuu, H; Perälä, M; Kallioniemi, O

    2012-03-01

    Malignant glioma is the most common brain tumor with 16,000 new cases diagnosed annually in the United States. We performed a systematic large-scale transcriptomics data mining study of 9783 tissue samples from the GeneSapiens database to systematically identify genes that are most glioma-specific. We searched for genes that were highly expressed in 322 glioblastoma multiforme tissue samples and 66 anaplastic astrocytomas as compared with 425 samples from histologically normal central nervous system. Transcription cofactor HES6 (hairy and enhancer of split 6) emerged as the most glioma-specific gene. Immunostaining of a tissue microarray showed HES6 expression in 335 (98.8%) out of the 339 glioma samples. HES6 was expressed in endothelial cells of the normal brain and glioma tissue. Recurrent grade 2 astrocytomas and grade 2 or 3 oligodendrogliomas showed higher levels of HES6 immunoreactivity than the corresponding primary tumors. High HES6 mRNA expression correlated with the proneural subtype that generally has a favorable outcome but is prone to recur. Functional studies suggested an important role for HES6 in supporting survival of glioma cells, as evidenced by reduction of cancer cell proliferation and migration after HES6 silencing. The biological role and consequences of HES6 silencing and overexpression was explored with genome-wide analyses, which implicated a role for HES6 in p53, c-myc and nuclear factor-κB transcriptional networks. We conclude that HES6 is important for glioma cell proliferation and migration, and may have a role in angiogenesis. PMID:21785461

  9. Premature differentiation and aberrant movement of pituitary cells lacking both Hes1 and Prop1

    PubMed Central

    Himes, Ashley D.; Raetzman, Lori T.

    2009-01-01

    In the pituitary, the transition from proliferating progenitor cell into differentiated hormone producing cell is carefully regulated in a time dependent and spatially restricted manner. We report that two targets of Notch signaling, Hes1 and Prop1, are needed to maintain progenitors within Rathke’s pouch and for the restriction of differentiated cells to the ventral pituitary. We observed ACTH and αGSU producing cells that had prematurely differentiated within Rathke’s pouch along with correlated ectopic expression of Mash1 only when both Prop1 and Hes1 were lost. We also discovered that downregulation of N-cadherin expression in cells as they transition from Rathke’s pouch to the anterior lobe appears to be essential for their movement. In the Prop1 mutant, cells are trapped in Rathke’s pouch and N-cadherin expression remains high. Also, Slug, a marker of epithelial to mesenchymal transition, is absent in the dorsal anterior lobe. When Hes1 is lost in the Prop1 mutant, N-cadherin is downregulated and cells are able to exit Rathke’s pouch but have lost their migrational cues and form ectopic foci surrounding Rathke’s pouch. Our data reveal important overlapping functions of Hes1 and Prop1 in cell differentiation and movement that are critical for pituitary organogenesis. PMID:18996108

  10. The Drosophila STUbL protein Degringolade limits HES functions during embryogenesis.

    PubMed

    Barry, Kevin C; Abed, Mona; Kenyagin, Dorit; Werwie, Timothy R; Boico, Olga; Orian, Amir; Parkhurst, Susan M

    2011-05-01

    Degringolade (Dgrn) encodes a Drosophila SUMO-targeted ubiquitin ligase (STUbL) protein similar to that of mammalian RNF4. Dgrn facilitates the ubiquitylation of the HES protein Hairy, which disrupts the repressive activity of Hairy by inhibiting the recruitment of its cofactor Groucho. We show that Hey and all HES family members, except Her, interact with Dgrn and are substrates for its E3 ubiquitin ligase activity. Dgrn displays dynamic subcellular localization, accumulates in the nucleus at times when HES family members are active and limits Hey and HES family activity during sex determination, segmentation and neurogenesis. We show that Dgrn interacts with the Notch signaling pathway by it antagonizing the activity of E(spl)-C proteins. dgrn null mutants are female sterile, producing embryos that arrest development after two or three nuclear divisions. These mutant embryos exhibit fragmented or decondensed nuclei and accumulate higher levels of SUMO-conjugated proteins, suggesting a role for Dgrn in genome stability. PMID:21486924

  11. Notch Signaling and Hes Labeling in the Normal and Drug-Damaged Organ of Corti

    PubMed Central

    Batts, Shelley A.; Shoemaker, Christopher R.; Raphael, Yehoash

    2009-01-01

    During the development of the inner ear, the Notch cell signaling pathway is responsible for the specification of the pro-sensory domain and influences cell fate decisions. It is assumed that Notch signaling ends during maturity and cannot be reinitiated to alter the fate of new or existing cells in the organ of Corti. This is in contrast to non-mammalian species which reinitiate Delta1-Notch1 signaling in response to trauma in the auditory epithelium, resulting in hair cell regeneration through transdifferentiation and/or mitosis. We report immunohistochemical data and Western protein analysis showing that in the aminoglycoside-damaged guinea pig organ of Corti, there is an increase in proteins involved in Notch activation occurring within 24 hours of a chemical hair cell lesion. The signaling response is characterized by the increased presence of Jagged1 ligand in pillar and Deiters cells, Notch1 signal in surviving supporting cell nuclei, and the absence of Jagged2 and Delta-like1. The pro-sensory bHLH protein Atoh1 was absent at all time points following an ototoxic lesion, while the repressor bHLH transcription factors Hes1 and Hes5 were detected in surviving supporting cell nuclei in the former inner and outer hair cell areas, respectively. Notch pathway proteins peaked at 2 weeks, decreased at 1 month, and nearly disappeared by 2 months. These results indicate that the mammalian auditory epithelium retains the ability to regulate Notch signaling and Notch-dependent Hes activity in response to cellular trauma and that the signaling is transient. Additionally, since Hes activity antagonizes the transcription of prosensory Atoh1, the presence of Hes after a lesion may prohibit the occurrence of transdifferentiation in the surviving supporting cells. PMID:19185606

  12. Regulation of NOTCH signaling by reciprocal inhibition of HES1 and Deltex 1 and its role in osteosarcoma invasiveness.

    PubMed

    Zhang, P; Yang, Y; Nolo, R; Zweidler-McKay, P A; Hughes, D P M

    2010-05-20

    The highly conserved NOTCH signaling pathway has many essential functions in the development of diverse cells, tissues and organs from Drosophila to humans, and dysregulated NOTCH signaling contributes to several disorders, including vascular and bone defects, as well as several cancers. Here we describe a novel mechanism of NOTCH regulation by reciprocal inhibition of two NOTCH downstream effectors: Deltex1 and HES1. This mechanism appears to regulate invasion of osteosarcoma cells, as Deltex1 blocks osteosarcoma invasiveness by downregulating NOTCH/HES1 signaling. The inhibitory effect of endogenous Deltex1 on NOTCH signaling is mediated through binding with the intracellular domain of NOTCH and ubiquitination and degradation of NOTCH receptors. Conversely, we show that the NOTCH target gene HES1 causes transcriptional inhibition of Deltex1 by directly binding to the promoter of Deltex1. An HES1 binding site is identified 400 bp upstream of the transcription start site of Deltex1. HES1-mediated repression of Deltex1 requires the C-terminal H3/H4 and WRPW domains of HES1, which associate with the TLE/Groucho corepressors. Taken together, we define a molecular mechanism regulating NOTCH signaling by reciprocal inhibition of the NOTCH target genes HES1 and Deltex1 in mammalian cells. This mechanism may have important clinical implications for targeting NOTCH signaling in osteosarcoma and other cancers. PMID:20208568

  13. Notch/HES1-mediated PARP1 activation: a cell type–specific mechanism for tumor suppression

    PubMed Central

    Kannan, Sankaranarayanan; Fang, Wendy; Song, Guangchun; Mullighan, Charles G.; Hammitt, Richard; McMurray, John

    2011-01-01

    Notch signaling plays both oncogenic and tumor suppressor roles, depending on cell type. In contrast to T-cell acute lymphoblastic leukemia (ALL), where Notch activation promotes leukemogenesis, induction of Notch signaling in B-cell ALL (B-ALL) leads to growth arrest and apoptosis. The Notch target Hairy/Enhancer of Split1 (HES1) is sufficient to reproduce this tumor suppressor phenotype in B-ALL; however, the mechanism is not yet known. We report that HES1 regulates proapoptotic signals by the novel interacting protein Poly ADP-Ribose Polymerase1 (PARP1) in a cell type–specific manner. Interaction of HES1 with PARP1 inhibits HES1 function, induces PARP1 activation, and results in PARP1 cleavage in B-ALL. HES1-induced PARP1 activation leads to self-ADP ribosylation of PARP1, consumption of nicotinamide adenine dinucleotide+, diminished adenosine triphosphate levels, and translocation of apoptosis-inducing factor from mitochondria to the nucleus, resulting in apoptosis in B-ALL but not T-cell ALL. Importantly, induction of Notch signaling by the Notch agonist peptide Delta/Serrate/Lag-2 can reproduce these events and leads to B-ALL apoptosis. The novel interaction of HES1 and PARP1 in B-ALL modulates the function of the HES1 transcriptional complex and signals through PARP1 to induce apoptosis. This mechanism shows a cell type–specific proapoptotic pathway that may lead to Notch agonist–based cancer therapeutics. PMID:21224467

  14. Hes1 promotes the IL-22-mediated antimicrobial response by enhancing STAT3-dependent transcription in human intestinal epithelial cells

    SciTech Connect

    Murano, Tatsuro; Okamoto, Ryuichi; Ito, Go; Nakata, Toru; Hibiya, Shuji; Shimizu, Hiromichi; Fujii, Satoru; Kano, Yoshihito; Mizutani, Tomohiro; Yui, Shiro; Akiyama-Morio, Junko; Nemoto, Yasuhiro; Tsuchiya, Kiichiro; Nakamura, Tetsuya; Watanabe, Mamoru

    2014-01-17

    Highlights: •Hes1 enhances IL-22-STAT3 signaling in human intestinal epithelial cells. •Hes1 enhances REG family gene induction by IL-22-STAT3 signaling. •Protein level of Hes1 restricts the response to IL-22. •Present regulation of a cytokine signal represents a new mode of Hes1 function. -- Abstract: Notch signaling plays an essential role in the proliferation and differentiation of intestinal epithelial cells (IECs). We have previously shown that Notch signaling is up-regulated in the inflamed mucosa of ulcerative colitis (UC) and thereby plays an indispensable role in tissue regeneration. Here we show that in addition to Notch signaling, STAT3 signaling is highly activated in the inflamed mucosa of UC. Forced expression of the Notch target gene Hes1 dramatically enhanced the IL-22-mediated STAT3-dependent transcription in human IECs. This enhancement of STAT3-dependent transcription was achieved by the extended phosphorylation of STAT3 by Hes1. Microarray analysis revealed that Hes1-mediated enhancement of IL-22-STAT3 signaling significantly increased the induction of genes encoding antimicrobial peptides, such as REG1A, REG3A and REG3G, in human IECs. Conversely, the reduction of Hes1 protein levels with a γ-secretase inhibitor significantly down-regulated the induction of those genes in IECs, resulting in a markedly poor response to IL-22. Our present findings identify a new role for the molecular function of Hes1 in which the protein can interact with cytokine signals and regulate the immune response of IECs.

  15. Innovation Centre Peptide and Protein Technologies HES-SO Sion: Pioneer in an Emerging Market.

    PubMed

    Heinzelmann, Elsbeth

    2016-01-01

    Peptides are small proteins typically containing a chain of up to 100 amino acids. In our bodies, they are produced every day in vast quantities and perform highly specific biological activities. As there is an increased interest in peptides for pharmaceutical applications, the HES-SO Valais-Wallis created a research group to focus on Peptide and Protein Technologies (P(2)T). PMID:27052764

  16. siRNA Targeting Hes5 Augments Hair Cell Regeneration in Aminoglycoside-damaged Mouse Utricle

    PubMed Central

    Jung, Jae Yun; Avenarius, Matt R.; Adamsky, Swetlana; Alpert, Evgenia; Feinstein, Elena; Raphael, Yehoash

    2013-01-01

    Notch signaling is active during the development of mosaic epithelial sheets and during their turnover and regeneration. After the loss of hair cells in the mosaic sheet of the vestibular sensory epithelium, new hair cells can be spontaneously generated by transdifferentiation of supporting cells. This regenerative process involves downregulation of the Hes5 gene and is known to be limited and incomplete, especially when the lesion is severe. Here, we test whether further downregulation of Hes5 gene accomplished by the use of siRNA after a severe lesion induced by an aminoglycoside in the mouse utricle can enhance the transdifferentiation of supporting cells and lead to the increased production of new hair cells. We demonstrate that Hes5 levels in the utricle decreased after the application of siRNA and that the number of hair cells in these utricles was significantly larger than following control treatment. The data suggest that siRNA technology may be useful for inducing repair and regeneration in the inner ear and that the Notch signaling pathway is a potentially useful target for specific gene expression inhibition. PMID:23439501

  17. Qingyihuaji Formula Inhibits Pancreatic Cancer and Prolongs Survival by Downregulating Hes-1 and Hey-1

    PubMed Central

    Xu, Yanli; Xu, Shan; Cai, Yueqin; Liu, Luming

    2015-01-01

    The dire prognosis of pancreatic cancer has not markedly improved during past decades. The present study was carried out to explore the effect of Qingyihuaji formula (QYHJ) on inhibiting pancreatic cancer and prolonging survival in related Notch signaling pathway. Proliferation of pancreatic cancer cells (SW1990 and PANC-1) was detected by MTT assay at 24, 48, and 72 h with exposure to various concentrations (0.08–50 mg/mL) of QYHJ water extract. Pancreatic tumor models of nude mice were divided into three groups randomly (control, QYHJ, and gemcitabine). mRNA and protein expression of Notch target genes (Hes-1, Hey-1, Hey-2, and Hey-L) in dissected tumor tissue were detected. Results showed that proliferation of SW1990 cells and PANC-1 cells was inhibited by QYHJ water extract in a dose-dependent and time-dependent manner. QYHJ effectively inhibited tumor growth and prolonged survival time in nude mice. Expression of both Hes-1 and Hey-1 was decreased significantly in QYHJ group, suggesting that Hes-1 and Hey-1 in Notch signaling pathway might be potential targets for QYHJ treatment. This research could help explain the clinical effectiveness of QYHJ and may provide advanced pancreatic cancer patients with a new therapeutic option. PMID:26783407

  18. Hes1 triggers epithelial-mesenchymal transition (EMT)-like cellular marker alterations and promotes invasion and metastasis of nasopharyngeal carcinoma by activating the PTEN/AKT pathway.

    PubMed

    Wang, Sheng-Chun; Lin, Xiao-Lin; Wang, Hui-Yan; Qin, Yu-Juan; Chen, Lin; Li, Jing; Jia, Jun-Shuang; Shen, Hong-Fen; Yang, Sheng; Xie, Rao-Ying; Wei, Fang; Gao, Fei; Rong, Xiao-Xiang; Yang, Jie; Zhao, Wen-Tao; Zhang, Ting-Ting; Shi, Jun-Wen; Yao, Kai-Tai; Luo, Wei-Ren; Sun, Yan; Xiao, Dong

    2015-11-01

    Overexpression of the transcriptional factor Hes1 (hairy and enhancer of split-1) has been observed in numerous cancers, but the precise roles of Hes1 in epithelial-mesenchymal transition (EMT), cancer invasion and metastasis remain unknown. Our current study firstly revealed that Hes1 upregulation in a cohort of human nasopharyngeal carcinoma (NPC) biopsies is significantly associated with the EMT, invasive and metastatic phenotypes of cancer. In the present study, we found that Hes1 overexpression triggered EMT-like cellular marker alterations of NPC cells, whereas knockdown of Hes1 through shRNA reversed the EMT-like phenotypes, as strongly supported by Hes1-mediated EMT in NPC clinical specimens described above. Gain-of-function and loss-of-function experiments demonstrated that Hes1 promoted the migration and invasion of NPC cells in vitro. In addition, exogenous expression of Hes1 significantly enhanced the metastatic ability of NPC cells in vivo. Chromatin immunoprecipitation (ChIP) assays showed that Hes1 inhibited PTEN expression in NPC cells through binding to PTEN promoter region. Increased Hes1 expression and decreased PTEN expression were also observed in a cohort of NPC biopsies. Additional studies demonstrated that Hes1-induced EMT-like molecular changes and increased motility and invasion of NPC cells were mediated by PTEN. Taken together, our results suggest, for what we believe is the first time, that Hes1 plays an important role in the invasion and metastasis of NPC through inhibiting PTEN expression to trigger EMT-like phenotypes. PMID:26452025

  19. Hes1 triggers epithelial-mesenchymal transition (EMT)-like cellular marker alterations and promotes invasion and metastasis of nasopharyngeal carcinoma by activating the PTEN/AKT pathway

    PubMed Central

    Qin, Yu-Juan; Chen, Lin; Li, Jing; Jia, Jun-Shuang; Shen, Hong-Fen; Yang, Sheng; Xie, Rao-Ying; Wei, Fang; Gao, Fei; Rong, Xiao-Xiang; Yang, Jie; Zhao, Wen-Tao; Zhang, Ting-Ting; Shi, Jun-Wen; Yao, Kai-Tai; Luo, Wei-Ren; Sun, Yan; Xiao, Dong

    2015-01-01

    Overexpression of the transcriptional factor Hes1 (hairy and enhancer of split-1) has been observed in numerous cancers, but the precise roles of Hes1 in epithelial-mesenchymal transition (EMT), cancer invasion and metastasis remain unknown. Our current study firstly revealed that Hes1 upregulation in a cohort of human nasopharyngeal carcinoma (NPC) biopsies is significantly associated with the EMT, invasive and metastatic phenotypes of cancer. In the present study, we found that Hes1 overexpression triggered EMT-like cellular marker alterations of NPC cells, whereas knockdown of Hes1 through shRNA reversed the EMT-like phenotypes, as strongly supported by Hes1-mediated EMT in NPC clinical specimens described above. Gain-of-function and loss-of-function experiments demonstrated that Hes1 promoted the migration and invasion of NPC cells in vitro. In addition, exogenous expression of Hes1 significantly enhanced the metastatic ability of NPC cells in vivo. Chromatin immunoprecipitation (ChIP) assays showed that Hes1 inhibited PTEN expression in NPC cells through binding to PTEN promoter region. Increased Hes1 expression and decreased PTEN expression were also observed in a cohort of NPC biopsies. Additional studies demonstrated that Hes1-induced EMT-like molecular changes and increased motility and invasion of NPC cells were mediated by PTEN. Taken together, our results suggest, for what we believe is the first time, that Hes1 plays an important role in the invasion and metastasis of NPC through inhibiting PTEN expression to trigger EMT-like phenotypes. PMID:26452025

  20. ZNF70, a novel ILDR2-interacting protein, contributes to the regulation of HES1 gene expression.

    PubMed

    Watanabe, Kazuhisa; Nakayama, Kazuhiro; Ohta, Satoshi; Tago, Kenji; Boonvisut, Supichaya; Millings, Elizabeth J; Fischer, Stuart G; LeDuc, Charles A; Leibel, Rudolph L; Iwamoto, Sadahiko

    2016-09-01

    A diabetes susceptibility gene, immunoglobulin-like domain containing receptor 2 (Ildr2), encodes a transmembrane protein localized to the endoplasmic reticulum membrane that is closely related to hepatic lipid metabolism. The livers of ob/ob mice in which Ildr2 is transiently overexpressed are relieved of hepatic steatosis. However, the molecular mechanisms through which ILDR2 affects these changes in hepatic lipid metabolism remain unknown. This study aimed to identify ILDR2-interacting proteins to further elucidate the molecular mechanisms underlying the role of ILDR2 in lipid homeostasis. We purified ILDR2-containing protein complexes using tandem affinity purification tagging and identified ZNF70, a member of the Kruppel C2H2-type zinc finger protein family, as a novel ILDR2-interacting protein. We demonstrated that ZNF70 interacts with ZFP64 and activates HES1 transcription by binding to the HES1 promoter. In addition, HES1 gene expression is increased in ILDR2-knockdown HepG2 cells, in which ZNF70 is translocated from the cytoplasm to the nucleus, suggesting that ZNF70 migration to the nucleus after dissociating from the ILDR2-ZNF70 complex activates HES1 transcription. These results support a novel link between ILDR2 and HES1 gene expression and suggest that ILDR2 is involved in a novel pathway in hepatic steatosis. PMID:27353377

  1. miR-381 Regulates Neural Stem Cell Proliferation and Differentiation via Regulating Hes1 Expression

    PubMed Central

    Liu, Baoquan; Yang, Chunxiao; Nie, Xuedan; Wang, Xiaokun; Zheng, Jiaolin; Wang, Yue; Zhu, Yulan

    2015-01-01

    Neural stem cells are self-renewing, multipotent and undifferentiated precursors that retain the capacity for differentiation into both glial (astrocytes and oligodendrocytes) and neuronal lineages. Neural stem cells offer cell-based therapies for neurological disorders such as Alzheimer's disease, Parkinson's disease, Huntington's disease and spinal cord injuries. However, their cellular behavior is poorly understood. MicroRNAs (miRNAs) are a class of small noncoding RNAs involved in cell development, proliferation and differentiation through regulating gene expression at post-transcriptional level. The role of miR–381 in the development of neural stem cells remains unknown. In this study, we showed that overexpression of miR–381 promoted neural stem cells proliferation. It induced the neural stem cells differentiation to neurons and inhibited their differentiation to astrocytes. Furthermore, we identified HES1 as a direct target of miR–381 in neural stem cells. Moreover, re-expression of HES1 impaired miR-381-induced promotion of neural stem cells proliferation and induce neural stem cells differentiation to neurons. In conclusion, miR–381 played important role in neural stem cells proliferation and differentiation. PMID:26431046

  2. Inflammatory Kidney and Liver Tissue Response to Different Hydroxyethylstarch (HES) Preparations in a Rat Model of Early Sepsis

    PubMed Central

    Schimmer, Ralph C.; Urner, Martin; Voigtsberger, Stefanie; Booy, Christa; Roth Z’Graggen, Birgit; Beck-Schimmer, Beatrice; Schläpfer, Martin

    2016-01-01

    Background Tissue hypoperfusion and inflammation in sepsis can lead to organ failure including kidney and liver. In sepsis, mortality of acute kidney injury increases by more than 50%. Which type of volume replacement should be used is still an ongoing debate. We investigated the effect of different volume strategies on inflammatory mediators in kidney and liver in an early sepsis model. Material and Methods Adult male Wistar rats were subjected to sepsis by cecal ligation and puncture (CLP) and assigned to three fluid replenishment groups. Animals received 30mL/kg of Ringer’s lactate (RL) for 2h, thereafter RL (75mL/kg), hydroxyethyl starch (HES) balanced (25mL/kg), containing malate and acetate, or HES saline (25mL/kg) for another 2h. Kidney and liver tissue was assessed for inflammation. In vitro rat endothelial cells were exposed to RL, HES balanced or HES saline for 2h, followed by stimulation with tumor necrosis factor-α (TNF-α) for another 4h. Alternatively, cells were exposed to malate, acetate or a mixture of malate and acetate, reflecting the according concentration of these substances in HES balanced. Pro-inflammatory cytokines were determined in cell supernatants. Results Cytokine mRNA in kidney and liver was increased in CLP animals treated with HES balanced compared to RL, but not after application of HES saline. MCP-1 was 3.5fold (95% CI: 1.3, 5.6) (p<0.01) and TNF-α 2.3fold (95% CI: 1.2, 3.3) (p<0.001) upregulated in the kidney. Corresponding results were seen in liver tissue. TNF-α-stimulated endothelial cells co-exposed to RL expressed 3529±1040pg/mL MCP-1 and 59±23pg/mL CINC-1 protein. These cytokines increased by 2358pg/mL (95% CI: 1511, 3204) (p<0.001) and 29pg/ml (95% CI: 14, 45) (p<0.01) respectively when exposed to HES balanced instead. However, no further upregulation was observed with HES saline. PBS supplemented with acetate increased MCP-1 by 1325pg/mL (95% CI: 741, 1909) (p<0.001) and CINC-1 by 24pg/mL (95% CI: 9, 38) (p<0

  3. Association between perinatal methylation of the neuronal differentiation regulator HES1 and later childhood neurocognitive function and behaviour

    PubMed Central

    Lillycrop, Karen A; Costello, Paula M; Teh, Ai Ling; Murray, Robert J; Clarke-Harris, Rebecca; Barton, Sheila J; Garratt, Emma S; Ngo, Sherry; Sheppard, Allan M; Wong, Johnny; Dogra, Shaillay; Burdge, Graham C; Cooper, Cyrus; Inskip, Hazel M; Gale, Catharine R; Gluckman, Peter D; Harvey, Nicholas C; Chong, Yap-Seng; Yap, Fabian; Meaney, Michael J; Rifkin-Graboi, Anne; Holbrook, Joanna D; Godfrey, Keith M

    2015-01-01

    Background Early life environments induce long-term changes in neurocognitive development and behaviour. In animal models, early environmental cues affect neuropsychological phenotypes via epigenetic processes but, as yet, there is little direct evidence for such mechanisms in humans. Method We examined the relation between DNA methylation at birth and child neuropsychological outcomes in two culturally diverse populations using a genome-wide methylation analysis and validation by pyrosequencing. Results Within the UK Southampton Women’s Survey (SWS) we first identified 41 differentially methylated regions of interest (DMROI) at birth associated with child’s full-scale IQ at age 4 years. Associations between HES1 DMROI methylation and later cognitive function were confirmed by pyrosequencing in 175 SWS children. Consistent with these findings, higher HES1 methylation was associated with higher executive memory function in a second independent group of 200 SWS 7-year-olds. Finally, we examined a pathway for this relationship within a Singaporean cohort (n = 108). Here, HES1 DMROI methylation predicted differences in early infant behaviour, known to be associated with academic success. In vitro, methylation of HES1 inhibited ETS transcription factor binding, suggesting a functional role of this site. Conclusions Thus, our findings suggest that perinatal epigenetic processes mark later neurocognitive function and behaviour, providing support for a role of epigenetic processes in mediating the long-term consequences of early life environment on cognitive development. PMID:25906782

  4. STAT3-Ser/Hes3 Signaling: A New Molecular Component of the Neuroendocrine System?

    PubMed

    Nikolakopoulou, P; Poser, S W; Masjkur, J; Fernandez Rubin de Celis, M; Toutouna, L; Andoniadou, C L; McKay, R D; Chrousos, G; Ehrhart-Bornstein, M; Bornstein, S R; Androutsellis-Theotokis, A

    2016-02-01

    The endocrine system involves communication among different tissues in distinct organs, including the pancreas and components of the Hypothalamic-Pituitary-Adrenal Axis. The molecular mechanisms underlying these complex interactions are a subject of intense study as they may hold clues for the progression and treatment of a variety of metabolic and degenerative diseases. A plethora of signaling pathways, activated by hormones and other endocrine factors have been implicated in this communication. Recent advances in the stem cell field introduce a new level of complexity: adult progenitor cells appear to utilize distinct signaling pathways than the more mature cells in the tissue they co-reside. It is therefore important to elucidate the signal transduction requirements of adult progenitor cells in addition to those of mature cells. Recent evidence suggests that a common non-canonical signaling pathway regulates adult progenitors in several different tissues, rendering it as a potentially valuable starting point to explore their biology. The STAT3-Ser/Hes3 Signaling Axis was first identified as a major regulator of neural stem cells and, subsequently, cancer stem cells. In the endocrine/neuroendocrine system, this pathway operates on several levels, regulating other types of plastic cells: (a) it regulates pancreatic islet cell function and insulin release; (b) insulin in turn activates the pathway in broadly distributed neural progenitors and possibly also hypothalamic tanycytes, cells with important roles in the control of the adrenal gland; (c) adrenal progenitors themselves operate this pathway. The STAT3-Ser/Hes3 Signaling Axis therefore deserves additional research in the context of endocrinology. PMID:26783739

  5. Prolyl isomerase Pin1 and protein kinase HIPK2 cooperate to promote cortical neurogenesis by suppressing Groucho/TLE:Hes1-mediated inhibition of neuronal differentiation.

    PubMed

    Ciarapica, R; Methot, L; Tang, Y; Lo, R; Dali, R; Buscarlet, M; Locatelli, F; del Sal, G; Rota, R; Stifani, S

    2014-02-01

    The Groucho/transducin-like Enhancer of split 1 (Gro/TLE1):Hes1 transcriptional repression complex acts in cerebral cortical neural progenitor cells to inhibit neuronal differentiation. The molecular mechanisms that regulate the anti-neurogenic function of the Gro/TLE1:Hes1 complex during cortical neurogenesis remain to be defined. Here we show that prolyl isomerase Pin1 (peptidyl-prolyl cis-trans isomerase NIMA-interacting 1) and homeodomain-interacting protein kinase 2 (HIPK2) are expressed in cortical neural progenitor cells and form a complex that interacts with the Gro/TLE1:Hes1 complex. This association depends on the enzymatic activities of both HIPK2 and Pin1, as well as on the association of Gro/TLE1 with Hes1, but is independent of the previously described Hes1-activated phosphorylation of Gro/TLE1. Interaction with the Pin1:HIPK2 complex results in Gro/TLE1 hyperphosphorylation and weakens both the transcriptional repression activity and the anti-neurogenic function of the Gro/TLE1:Hes1 complex. These results provide evidence that HIPK2 and Pin1 work together to promote cortical neurogenesis, at least in part, by suppressing Gro/TLE1:Hes1-mediated inhibition of neuronal differentiation. PMID:24270405

  6. Germ Cell Nuclear Factor (GCNF) Represses Oct4 Expression and Globally Modulates Gene Expression in Human Embryonic Stem (hES) Cells*

    PubMed Central

    Wang, Hongran; Wang, Xiaohong; Xu, Xueping; Kyba, Michael; Cooney, Austin J.

    2016-01-01

    Oct4 is considered a key transcription factor for pluripotent stem cell self-renewal. It binds to specific regions within target genes to regulate their expression and is downregulated upon induction of differentiation of pluripotent stem cells; however, the mechanisms that regulate the levels of human Oct4 expression remain poorly understood. Here we show that expression of human Oct4 is directly repressed by germ cell nuclear factor (GCNF), an orphan nuclear receptor, in hES cells. Knockdown of GCNF by siRNA resulted in maintenance of Oct4 expression during RA-induced hES cell differentiation. While overexpression of GCNF promoted repression of Oct4 expression in both undifferentiated and differentiated hES cells. The level of Oct4 repression was dependent on the level of GCNF expression in a dose-dependent manner. mRNA microarray analysis demonstrated that overexpression of GCNF globally regulates gene expression in undifferentiated and differentiated hES cells. Within the group of altered genes, GCNF down-regulated 36% of the genes, and up-regulated 64% in undifferentiated hES cells. In addition, GCNF also showed a regulatory gene pattern that is different from RA treatment during hES cell differentiation. These findings increase our understanding of the mechanisms that maintain hES cell pluripotency and regulate gene expression during the differentiation process. PMID:26769970

  7. Correlation of Notch1/Hes1 Genes Expression Levels in Egyptian Paediatric Patients with Newly Diagnosed and Persistent Primary Immune(Idiopathic) Thrombocytopenic Purpura.

    PubMed

    Gawdat, Rania Mohsen; Hammam, Amira Ahmed; Ezzat, Dina Ahmed

    2016-09-01

    Notch signalling is involved in the development of several autoimmune diseases, one of such diseases is ITP. The aim of this study was to investigate and compare the expression levels of Notch1 receptor and its target Hes1 gene in Egyptian paediatric ITP patients. Real-time quantitative reverse transcriptase polymerase chain reaction was used to analyse the expression levels of Notch1 and Hes1 in 42 children with primary ITP (22 newly diagnosed and 20 persistent) cases. Twenty age and sex matched non-ITP controls were included. The expression levels of Notch1 were higher in newly diagnosed and persistent cases than controls with high statistical significant difference (P value < 0.001, P < 0.001) respectively, similarly as regards the expression levels of HES1 (P value < 0.001, P < 0.007) respectively. A significant positive correlation was found between Notch1 and Hes1 expression levels in newly diagnosed cases (r = 0.587, P value = 0.004). There was an association between levels of both genes in most of ITP patients but Hes1 was markedly elevated than Notch1 in few cases. High expression levels of Notch1/Hes1 indicated the important role of Notch signalling in both newly diagnosed and persistent ITP. High expression levels of Hes1 than Notch1 may shed light on its value as a therapeutic target for future research in ITP. PMID:27429531

  8. MicroRNA-9 promotes proliferation of leukemia cells in adult CD34-positive acute myeloid leukemia with normal karyotype by downregulation of Hes1.

    PubMed

    Tian, Chen; You, M James; Yu, Yong; Zhu, Lei; Zheng, Guoguang; Zhang, Yizhuo

    2016-06-01

    Acute myeloid leukemia (AML) is a group of heterogeneous hematopoietic malignancies sustained by leukemic stem cells (LSCs) that can resist treatment. Previously, we found that low expression of Hes1 was a poor prognostic factor for AML. However, the activation status of Hes1 and its regulation in LSCs and leukemic progenitors (LPs) as well as normal hematopoietic stem cells (HSCs) in Hes1-low AML patients have not been elucidated. In this study, the expression of Hes1 in LSCs and LPs was analyzed in adult CD34(+) Hes1-low AML with normal karyotype and the upstream microRNA (miRNA) regulators were screened. Our results showed that the level of either Hes1 or p21 was lower in LSCs or LPs than in HSCs whereas the level of miR-9 was highest in LPs and lowest in HSCs. An inverse correlation was observed in the expression of Hes1 and miR-9. Furthermore, we validated miR-9 as one of the regulators of Hes1 by reporter gene analysis. Knockdown of miR-9 by lentivirus infection suppressed the proliferation of AML cells by the induction of G0 arrest and apoptosis in vitro. Moreover, knockdown of miR-9 resulted in decreased circulating leukemic cell counts in peripheral blood and bone marrow, attenuated splenomegaly, and prolonged survival in a xenotransplant mouse model. Our results indicate that the miR-9 plays an important role in supporting AML cell growth and survival by downregulation of Hes1 and that miR-9 has potential as a therapeutic target for treating AML. PMID:26678889

  9. Posterior-anterior gradient of zebrafish hes6 expression in the presomitic mesoderm is established by the combinatorial functions of the downstream enhancer and 3'UTR.

    PubMed

    Kawamura, Akinori; Ovara, Hiroki; Ooka, Yuko; Kinoshita, Hirofumi; Hoshikawa, Miki; Nakajo, Kenji; Yokota, Daisuke; Fujino, Yuuri; Higashijima, Shin-ichi; Takada, Shinji; Yamasu, Kyo

    2016-01-15

    In vertebrates, the periodic formation of somites from the presomitic mesoderm (PSM) is driven by the molecular oscillator known as the segmentation clock. The hairy-related gene, hes6/her13.2, functions as a hub by dimerizing with other oscillators of the segmentation clock in zebrafish embryos. Although hes6 exhibits a posterior-anterior expression gradient in the posterior PSM with a peak at the tailbud, the detailed mechanisms underlying this unique expression pattern have not yet been clarified. By establishing several transgenic lines, we found that the transcriptional regulatory region downstream of hes6 in combination with the hes6 3'UTR recapitulates the endogenous gradient of hes6 mRNA expression. This downstream region, which we termed the PT enhancer, possessed several putative binding sites for the T-box and Ets transcription factors that were required for the regulatory activity. Indeed, the T-box transcription factor (Tbx16) and Ets transcription factor (Pea3) bound specifically to the putative binding sites and regulated the enhancer activity in zebrafish embryos. In addition, the 3'UTR of hes6 is required for recapitulation of the endogenous mRNA expression. Furthermore, the PT enhancer with the 3'UTR of hes6 responded to the inhibition of retinoic acid synthesis and fibroblast growth factor signaling in a manner similar to endogenous hes6. The results showed that transcriptional regulation by the T-box and Ets transcription factors, combined with the mRNA stability given by the 3'UTR, is responsible for the unique expression gradient of hes6 mRNA in the posterior PSM of zebrafish embryos. PMID:26596999

  10. Incredibly Versatile Microbial Fuel Cells Innovative Ideas at HES-SO Valais-Wallis for Solving Topical Problems.

    PubMed

    Heinzelmann, Elsbeth

    2016-01-01

    At HES-SO Valais-Wallis, Prof. Fabian Fischer is specialized in microbial fuel cells for novel applications that meet the challenge of producing renewable energies. He and his team possess a unique expertise in bioelectric energy vector generation, phosphate extraction (CHIMIA 2015, 69, 296) and the testing of antimicrobial surfaces. Let's take a look behind the scenes of the Institute of Life Technologies in Sion. PMID:27363382

  11. Whole Genome Sequencing Identifies a Missense Mutation in HES7 Associated with Short Tails in Asian Domestic Cats

    PubMed Central

    Xu, Xiao; Sun, Xin; Hu, Xue-Song; Zhuang, Yan; Liu, Yue-Chen; Meng, Hao; Miao, Lin; Yu, He; Luo, Shu-Jin

    2016-01-01

    Domestic cats exhibit abundant variations in tail morphology and serve as an excellent model to study the development and evolution of vertebrate tails. Cats with shortened and kinked tails were first recorded in the Malayan archipelago by Charles Darwin in 1868 and remain quite common today in Southeast and East Asia. To elucidate the genetic basis of short tails in Asian cats, we built a pedigree of 13 cats segregating at the trait with a founder from southern China and performed linkage mapping based on whole genome sequencing data from the pedigree. The short-tailed trait was mapped to a 5.6 Mb region of Chr E1, within which the substitution c. 5T > C in the somite segmentation-related gene HES7 was identified as the causal mutation resulting in a missense change (p.V2A). Validation in 245 unrelated cats confirmed the correlation between HES7-c. 5T > C and Chinese short-tailed feral cats as well as the Japanese Bobtail breed, indicating a common genetic basis of the two. In addition, some of our sampled kinked-tailed cats could not be explained by either HES7 or the Manx-related T-box, suggesting at least three independent events in the evolution of domestic cats giving rise to short-tailed traits. PMID:27560986

  12. Notch-HES1 signaling axis controls hemato-endothelial fate decisions of human embryonic and induced pluripotent stem cells.

    PubMed

    Lee, Jung Bok; Werbowetski-Ogilvie, Tamra E; Lee, Jong-Hee; McIntyre, Brendan A S; Schnerch, Angelique; Hong, Seok-Ho; Park, In-Hyun; Daley, George Q; Bernstein, Irwin D; Bhatia, Mickie

    2013-08-15

    Notch signaling regulates several cellular processes including cell fate decisions and proliferation in both invertebrates and mice. However, comparatively less is known about the role of Notch during early human development. Here, we examined the function of Notch signaling during hematopoietic lineage specification from human pluripotent stem cells of both embryonic and adult fibroblast origin. Using immobilized Notch ligands and small interfering RNA to Notch receptors we have demonstrated that Notch1, but not Notch2, activation induced hairy and enhancer of split 1 (HES1) expression and generation of committed hematopoietic progenitors. Using gain- and loss-of-function approaches, this was shown to be attributed to Notch-signaling regulation through HES1, which dictated cell fate decisions from bipotent precursors either to the endothelial or hematopoietic lineages at the clonal level. Our study reveals a previously unappreciated role for the Notch pathway during early human hematopoiesis, whereby Notch signaling via HES1 represents a toggle switch of hematopoietic vs endothelial fate specification. PMID:23733337

  13. Whole Genome Sequencing Identifies a Missense Mutation in HES7 Associated with Short Tails in Asian Domestic Cats.

    PubMed

    Xu, Xiao; Sun, Xin; Hu, Xue-Song; Zhuang, Yan; Liu, Yue-Chen; Meng, Hao; Miao, Lin; Yu, He; Luo, Shu-Jin

    2016-01-01

    Domestic cats exhibit abundant variations in tail morphology and serve as an excellent model to study the development and evolution of vertebrate tails. Cats with shortened and kinked tails were first recorded in the Malayan archipelago by Charles Darwin in 1868 and remain quite common today in Southeast and East Asia. To elucidate the genetic basis of short tails in Asian cats, we built a pedigree of 13 cats segregating at the trait with a founder from southern China and performed linkage mapping based on whole genome sequencing data from the pedigree. The short-tailed trait was mapped to a 5.6 Mb region of Chr E1, within which the substitution c. 5T > C in the somite segmentation-related gene HES7 was identified as the causal mutation resulting in a missense change (p.V2A). Validation in 245 unrelated cats confirmed the correlation between HES7-c. 5T > C and Chinese short-tailed feral cats as well as the Japanese Bobtail breed, indicating a common genetic basis of the two. In addition, some of our sampled kinked-tailed cats could not be explained by either HES7 or the Manx-related T-box, suggesting at least three independent events in the evolution of domestic cats giving rise to short-tailed traits. PMID:27560986

  14. Hydroxyethyl Starch (HES 130/0.4) Impairs Intestinal Barrier Integrity and Metabolic Function: Findings from a Mouse Model of the Isolated Perfused Small Intestine

    PubMed Central

    Dombrowsky, Heike; Zitta, Karina; Bein, Berthold; Krause, Thorsten; Goldmann, Torsten; Frerichs, Inez; Steinfath, Markus; Weiler, Norbert; Albrecht, Martin

    2015-01-01

    Background The application of hydroxyethyl starch (HES) for volume resuscitation is controversially discussed and clinical studies have suggested adverse effects of HES substitution, leading to increased patient mortality. Although, the intestine is of high clinical relevance and plays a crucial role in sepsis and inflammation, information about the effects of HES on intestinal function and barrier integrity is very scarce. We therefore evaluated the effects of clinically relevant concentrations of HES on intestinal function and barrier integrity employing an isolated perfused model of the mouse small intestine. Methods An isolated perfused model of the mouse small intestine was established and intestines were vascularly perfused with a modified Krebs-Henseleit buffer containing 3% Albumin (N=7) or 3% HES (130/0.4; N=7). Intestinal metabolic function (galactose uptake, lactate-to-pyruvate ratio), edema formation (wet-to-dry weight ratio), morphology (histological and electron microscopical analysis), fluid shifts within the vascular, lymphatic and luminal compartments, as well as endothelial and epithelial barrier permeability (FITC-dextran translocation) were evaluated in both groups. Results Compared to the Albumin group, HES perfusion did not significantly change the wet-to-dry weight ratio and lactate-to-pyruvate ratio. However, perfusing the small intestine with 3% HES resulted in a significant loss of vascular fluid (p<0.01), an increased fluid accumulation in the intestinal lumen (p<0.001), an enhanced translocation of FITC-dextran from the vascular to the luminal compartment (p<0.001) and a significantly impaired intestinal galactose uptake (p<0.001). Morphologically, these findings were associated with an aggregation of intracellular vacuoles within the intestinal epithelial cells and enlarged intercellular spaces. Conclusion A vascular perfusion with 3% HES impairs the endothelial and epithelial barrier integrity as well as metabolic function of the small

  15. Regulation of the Notch target gene Hes-1 by TGF{alpha} induced Ras/MAPK signaling in human neuroblastoma cells

    SciTech Connect

    Stockhausen, Marie-Therese; Sjoelund, Jonas; Axelson, Hakan . E-mail: hakan.axelson@molmed.mas.lu.se

    2005-10-15

    Ras and Notch signaling have recently been shown to cooperate in the maintenance of neoplastic transformation. Here, we show that TGF{alpha}, a known activator of Ras signaling, can drive cell proliferation and at the same time induce the expression of the Notch target Hes-1 in the neuroblastoma cell line SK-N-BE(2)c. The up-regulation of Hes-1 occurred both at the transcriptional and protein levels and by use of EGFR and MEK inhibitors we could show that the Hes-1 response was dependent on activation of the MAP kinase ERK. Blocking Notch activation by {gamma}-secretase inhibition did not profoundly affect the Hes-1 levels, neither in untreated nor in TGF{alpha} treated cells. The up-regulation of Hes-1 was associated with down-regulation of its pro-neuronal target gene Hash-1. Taken together, these results show that TGF{alpha} is a potent mitogen of neuroblastoma cells and suggest a connection between activation of ERK and Hes-1, thus providing a link between the Ras and Notch signaling pathways.

  16. Hes1 promotes cell proliferation and migration by activating Bmi-1 and PTEN/Akt/GSK3β pathway in human colon cancer.

    PubMed

    Gao, Fei; Huang, Wei; Zhang, YuQin; Tang, ShaoHui; Zheng, Lin; Ma, Feng; Wang, YiMing; Tang, Hui; Li, Xin

    2015-11-17

    Hes1 is a transcription factor that influences cell proliferation and differentiation. However, the effect of Hes1 on invasiveness and the underlying mechanism remain unknown. In the current study, we found that Hes1 suppressed cell apoptosis, promoted cell growth, induced EMT phenotype and cytoskeleton reconstruction, and enhanced the metastatic potential of colon cancer cells in vitro and in vivo. Furthermore, we indicated that Bmi-1 mediated Hes1-induced cell proliferation and migration, downregulated PTEN and activated the Akt/GSK3β pathway, consequently induced EMT and cytoskeleton reconstruction, ultimately leading to enhanced invasiveness of cancer cells. In addition, we also found that both Hes1 and Bmi-1 could directly regulate PTEN by associating at the PTEN locus, and played important roles in regulating PTEN/Akt/GSK3β pathway. Our results provide functional and mechanistic links between Hes1 and Bmi-1/PTEN/Akt/GSK3β signaling in the development and progression of colon cancer. PMID:26452029

  17. Hes1 promotes cell proliferation and migration by activating Bmi-1 and PTEN/Akt/GSK3β pathway in human colon cancer

    PubMed Central

    Gao, Fei; Huang, Wei; Zhang, YuQin; Tang, ShaoHui; Zheng, Lin; Ma, Feng; Wang, YiMing; Tang, Hui; Li, Xin

    2015-01-01

    Hes1 is a transcription factor that influences cell proliferation and differentiation. However, the effect of Hes1 on invasiveness and the underlying mechanism remain unknown. In the current study, we found that Hes1 suppressed cell apoptosis, promoted cell growth, induced EMT phenotype and cytoskeleton reconstruction, and enhanced the metastatic potential of colon cancer cells in vitro and in vivo. Furthermore, we indicated that Bmi-1 mediated Hes1-induced cell proliferation and migration, downregulated PTEN and activated the Akt/GSK3β pathway, consequently induced EMT and cytoskeleton reconstruction, ultimately leading to enhanced invasiveness of cancer cells. In addition, we also found that both Hes1 and Bmi-1 could directly regulate PTEN by associating at the PTEN locus, and played important roles in regulating PTEN/Akt/GSK3β pathway. Our results provide functional and mechanistic links between Hes1 and Bmi-1/PTEN/Akt/GSK3β signaling in the development and progression of colon cancer. PMID:26452029

  18. Canine Disorder Mirrors Human Disease: Exonic Deletion in HES7 Causes Autosomal Recessive Spondylocostal Dysostosis in Miniature Schnauzer Dogs

    PubMed Central

    Willet, Cali E.; Makara, Mariano; Reppas, George; Tsoukalas, George; Malik, Richard; Haase, Bianca; Wade, Claire M.

    2015-01-01

    Spondylocostal dysostosis is a congenital disorder of the axial skeleton documented in human families from diverse racial backgrounds. The condition is characterised by truncal shortening, extensive hemivertebrae and rib anomalies including malalignment, fusion and reduction in number. Mutations in the Notch signalling pathway genes DLL3, MESP2, LFNG, HES7 and TBX6 have been associated with this defect. In this study, spondylocostal dysostosis in an outbred family of miniature schnauzer dogs is described. Computed tomography demonstrated that the condition mirrors the skeletal defects observed in human cases, but unlike most human cases, the affected dogs were stillborn or died shortly after birth. Through gene mapping and whole genome sequencing, we identified a single-base deletion in the coding region of HES7. The frameshift mutation causes loss of functional domains essential for the oscillatory transcriptional autorepression of HES7 during somitogenesis. A restriction fragment length polymorphism test was applied within the immediate family and supported a highly penetrant autosomal recessive mode of inheritance. The mutation was not observed in wider testing of 117 randomly sampled adult miniature schnauzer and six adult standard schnauzer dogs; providing a significance of association of Praw = 4.759e-36 (genome-wide significant). Despite this apparently low frequency in the Australian population, the allele may be globally distributed based on its presence in two unrelated sires from geographically distant locations. While isolated hemivertebrae have been observed in a small number of other dog breeds, this is the first clinical and genetic diagnosis of spontaneously occurring spondylocostal dysostosis in a non-human mammal and offers an excellent model in which to study this devastating human disorder. The genetic test can be utilized by dog breeders to select away from the disease and avoid unnecessary neonatal losses. PMID:25659135

  19. Canine disorder mirrors human disease: exonic deletion in HES7 causes autosomal recessive spondylocostal dysostosis in miniature Schnauzer dogs.

    PubMed

    Willet, Cali E; Makara, Mariano; Reppas, George; Tsoukalas, George; Malik, Richard; Haase, Bianca; Wade, Claire M

    2015-01-01

    Spondylocostal dysostosis is a congenital disorder of the axial skeleton documented in human families from diverse racial backgrounds. The condition is characterised by truncal shortening, extensive hemivertebrae and rib anomalies including malalignment, fusion and reduction in number. Mutations in the Notch signalling pathway genes DLL3, MESP2, LFNG, HES7 and TBX6 have been associated with this defect. In this study, spondylocostal dysostosis in an outbred family of miniature schnauzer dogs is described. Computed tomography demonstrated that the condition mirrors the skeletal defects observed in human cases, but unlike most human cases, the affected dogs were stillborn or died shortly after birth. Through gene mapping and whole genome sequencing, we identified a single-base deletion in the coding region of HES7. The frameshift mutation causes loss of functional domains essential for the oscillatory transcriptional autorepression of HES7 during somitogenesis. A restriction fragment length polymorphism test was applied within the immediate family and supported a highly penetrant autosomal recessive mode of inheritance. The mutation was not observed in wider testing of 117 randomly sampled adult miniature schnauzer and six adult standard schnauzer dogs; providing a significance of association of Praw = 4.759e-36 (genome-wide significant). Despite this apparently low frequency in the Australian population, the allele may be globally distributed based on its presence in two unrelated sires from geographically distant locations. While isolated hemivertebrae have been observed in a small number of other dog breeds, this is the first clinical and genetic diagnosis of spontaneously occurring spondylocostal dysostosis in a non-human mammal and offers an excellent model in which to study this devastating human disorder. The genetic test can be utilized by dog breeders to select away from the disease and avoid unnecessary neonatal losses. PMID:25659135

  20. Biosorption at the HES-SO Valais-Wallis Sion Looking for Partners to Implement Innovative Technologies.

    PubMed

    Heinzelmann, Elsbeth

    2016-01-01

    Water pollution contributes to a number of health problems and disorders in humans, and dramatically affects wild animals and the environment as a whole. Biosorption is a sound and sustainable way to remove pollutants like metals, dyes, phenols, pesticides or drug metabolites from liquid effluents. But despite active research worldwide, technical limitations still impede its widespread use. At HES-SO Valais-Wallis in Sion, Prof. Simon Crelier is working on the development of efficient, economically viable and scalable biosorption processes. PMID:27198820

  1. METHOTREXATE AND MYOTREXATE INDUCE APOPTOSIS IN HUMAN MYOMA FIBROBLASTS (T hES CELL LINE) VIA MITOCHONDRIAL PATHWAY.

    PubMed

    Kastratović, Tatjana; Arsenijević, Slobodan; Matović, Zoran; Mitrović, Marina; Nikolić, Ivana; Milosavljević, Zoran; Protrka, Zoran; Šorak, Marija; Đurić, Janko

    2015-01-01

    Uterine leiomyomas (fibroids) are the most common benign tumors in women of reproductive age. Although the local application of low doses of methotrexate (MTX) is used as an effective treatment of the myomas, myotrexate could be a promising new drug. This study investigated the cytotoxic and apoptotic effects of both MTX and myotrexate in human fibroblasts derived from the uterine fibroids (T hES cell line). The myotrexate adduct is an aqueous solution of MTX and L-arginine. Cells were treated with a graded concentrations of both MTX and myothrexate (0.1-16 µM) for 24 h. The cytotoxicity was assayed by MTT test, apoptosis was evaluated by Annexin V-FITC assay and their possible role in apoptosis was determined by immnu- flourescence. Both MTX and myotrexate induced apoptosis in T hES cells in a dose dependent manner (p < 0.001). Myotrexate significantly increased the percentage of AnnexinV positive cells, BAX/Bcl-2 ratio and subsequent caspase-3 activation compared to the MTX treated cells (p < 0.05). Both MTX or myotrexate treatment showed a diffuse staining of cytochrome c indicating its release from mitochondria to the cytosol, suggesting that their mechanisms of action most likely involves the mitochondrial apoptotic pathway. PMID:26642654

  2. Aberrant expression of Notch1, HES1, and DTX1 genes in glioblastoma formalin-fixed paraffin-embedded tissues.

    PubMed

    Narayanappa, Rajeswari; Rout, Pritilata; Aithal, Madhuri G S; Chand, Ashis Kumar

    2016-05-01

    Glioblastoma is the most common malignant brain tumor accounting for more than 54 % of all gliomas. Despite aggressive treatments, median survival remains less than 1 year. This might be due to the unavailability of effective molecular diagnostic markers and targeted therapy. Thus, it is essential to discover molecular mechanisms underlying disease by identifying dysregulated pathways involved in tumorigenesis. Notch signaling is one such pathway which plays an important role in determining cell fates. Since it is found to play a critical role in many cancers, we investigated the role of Notch genes in glioblastoma with an aim to identify biomarkers that can improve diagnosis. Using real-time PCR, we assessed the expression of Notch genes including receptors (Notch1, Notch2, Notch3, and Notch4), ligands (JAG1, JAG2, and DLL3), downstream targets (HES1 and HEY2), regulator Deltex1 (DTX1), inhibitor NUMB along with transcriptional co-activator MAML1, and a component of gamma-secretase complex APH1A in 15 formalin-fixed paraffin-embedded (FFPE) patient samples. Relative quantification was done by the 2(-ΔΔCt) method; the data are presented as fold change in gene expression normalized to an internal control gene and relative to the calibrator. The data revealed aberrant expression of Notch genes in glioblastoma compared to normal brain. More than 85 % of samples showed high Notch1 (P = 0.0397) gene expression and low HES1 (P = 0.011) and DTX1 (P = 0.0001) gene expression. Our results clearly show aberrant expression of Notch genes in glioblastoma which can be used as putative biomarkers together with histopathological observation to improve diagnosis, therapeutic strategies, and patient prognosis. PMID:26662803

  3. Effect of rapid set binder on early strength and permeability of HES latex modified road repair pre-packed concrete

    NASA Astrophysics Data System (ADS)

    Han, J. W.; Lee, S. K.; Yu, C.; Park, C. G.

    2015-12-01

    The early strength development characteristics and permeability resistance of high early strength (HES) pre-packed road repair concrete incorporating a rapid-set binder material were evaluated for emergency repairs to road pavement. The rapid-set binder is a mixture of rapid-set cement and silica sands whose fluidity improves with the addition of styrene butadiene latex (latex). The resulting mixture has a compressive strength of 21 MPa or higher and a flexural strength of greater than 3.5 MPa after 4 hours, the maximum curing age allowed for emergency repair materials. This study examines the strength development properties and permeability resistance of HES latex-modified pre-packed road repair concrete using a rapid- set binder as a function of the latex-to-binder mixing ratio at values of 0.40, 0.33, 0.29 and 0.25. Both early strength development properties and permeability resistance increased as the ratio of latex to rapid-set binder decreased. The mixture showed a compressive strength of 21 MPa or higher after 4 hours, which is the design standard of emergency repair concrete, only when this ratio was 0.29 or lower. A flexural strength of 3.5 MPa or greater was observed after hours only when this ratio was 0.33 or lower. The standard for permeability resistance, less than 2,000 C of chloride after 7 days of curing, was satisfied by all ratios. The ratio of latex to rapid-set binder satisfying all of the conditions for an emergency road repair material was 0.29 or less.

  4. Gamma-secretase Inhibitor Prevents Proliferation and Migration of Ductus Arteriosus Smooth Muscle Cells through the Notch3-HES1/2/5 Pathway

    PubMed Central

    Wu, Jiunn-Ren; Yeh, Jwu-Lai; Liou, Shu-Fen; Dai, Zen-Kong; Wu, Bin-Nan; Hsu, Jong-Hau

    2016-01-01

    Patent ductus arteriosus (PDA) can cause morbidity and mortality in neonates. Vascular remodeling, characterized by proliferation and migration of smooth muscle cells (SMCs), is an essential process for postnatal DA closure. Notch signaling is an important mediator of vascular remodelling but its role in DA is unkonwn. We investigated the effects and underlying mechanisms of γ-secretase inhibitor DAPT, a Notch signaling inhibitor on angiotensin II (Ang II)-induced proliferation and migration of DASMCs. Proliferation and migration of DASMCs cultured from neonatal Wistar rats were induced by Ang II, with or without DAPT pre-treatment. In addition, potential underlying mechanisms including cell cycle progression, Ca2+ influx, reactive oxygen species (ROS) production, signal transduction of MAPK and Akt, and Notch receptor with its target gene pathway were examined. We found that DAPT inhibited Ang II-induced DASMCs proliferation and migration dose dependently. DAPT also arrested the cell cycle progression in the G0/G1-phase, and attenuated calcium overload and ROS production caused by Ang II. Moreover, DAPT inhibited nuclear translocation of Notch3 receptor intracellular domain, with decreased expression of its down-stream genes including HES1, HES2 and HES5. Finally, Ang II-activated ERK1/2, JNK and Akt were also counteracted by DAPT. In conclusion, DAPT inhibits Ang II-induced DASMCs proliferation and migration. These effects are potentially mediated by decreased calcium influx, reduced ROS production, and down-regulation of ERK1/2, JNK and Akt, through the Notch3-HES1/2/5 pathway. Therefore, Notch signaling has a role in DA remodeling and may provide a target pathway for therapeutic intervention of PDA. PMID:27570480

  5. Notch-Hes-1 axis controls TLR7-mediated autophagic death of macrophage via induction of P62 in mice with lupus.

    PubMed

    Li, Xiaojing; Liu, Fei; Zhang, Xuefang; Shi, Guoping; Ren, Jing; Ji, Jianjian; Ding, Liang; Fan, Hongye; Dou, Huan; Hou, Yayi

    2016-01-01

    The increased death of macrophages has been considered as a pathogenic factor for systemic lupus erythematosus (SLE), and dysfunction of autophagy may contribute to improper cell death. However, the effect of autophagy on macrophage during the pathogenesis of SLE is still unclear. Here we found that the death rate and autophagy level of macrophages significantly increased in MRL/lpr lupus-prone mice. Activation of toll-like receptor 7 (TLR7) triggered macrophage death in an autophagy-dependent but caspase-independent way in vitro. Moreover, P62/SQSTM1 is thought to have an essential role in selective autophagy. We also demonstrated that P62/SQSTM1 was required for TLR7-induced autophagy, and knockdown of P62 suppressed R848-induced cell death and LC3II protein accumulation. As an important mediator for cell-cell communication, Notch signaling is responsible for cell-fate decisions. Our results showed that activation of TLR7 also upregulated the expression of Notch1, especially its downstream target gene Hairy and enhancer of split 1 (Hes-1) in macrophages. Of note, we found that Hes-1, as a transcriptional factor, controlled TLR7-induced autophagy by regulating P62 expression. Furthermore, to confirm the above results in vivo, TLR7 agonist imiquimod (IMQ)-induced lupus mouse model was prepared. Splenic macrophages from IMQ-treated mice exhibited increased autophagy and cell death as well as enhanced expressions of Notch1 and Hes-1. Our results indicate that Notch1-Hes-1 signaling controls TLR7-induced autophagic death of macrophage via regulation of P62 in mice with lupus. PMID:27537524

  6. Gamma-secretase Inhibitor Prevents Proliferation and Migration of Ductus Arteriosus Smooth Muscle Cells through the Notch3-HES1/2/5 Pathway.

    PubMed

    Wu, Jiunn-Ren; Yeh, Jwu-Lai; Liou, Shu-Fen; Dai, Zen-Kong; Wu, Bin-Nan; Hsu, Jong-Hau

    2016-01-01

    Patent ductus arteriosus (PDA) can cause morbidity and mortality in neonates. Vascular remodeling, characterized by proliferation and migration of smooth muscle cells (SMCs), is an essential process for postnatal DA closure. Notch signaling is an important mediator of vascular remodelling but its role in DA is unkonwn. We investigated the effects and underlying mechanisms of γ-secretase inhibitor DAPT, a Notch signaling inhibitor on angiotensin II (Ang II)-induced proliferation and migration of DASMCs. Proliferation and migration of DASMCs cultured from neonatal Wistar rats were induced by Ang II, with or without DAPT pre-treatment. In addition, potential underlying mechanisms including cell cycle progression, Ca(2+) influx, reactive oxygen species (ROS) production, signal transduction of MAPK and Akt, and Notch receptor with its target gene pathway were examined. We found that DAPT inhibited Ang II-induced DASMCs proliferation and migration dose dependently. DAPT also arrested the cell cycle progression in the G0/G1-phase, and attenuated calcium overload and ROS production caused by Ang II. Moreover, DAPT inhibited nuclear translocation of Notch3 receptor intracellular domain, with decreased expression of its down-stream genes including HES1, HES2 and HES5. Finally, Ang II-activated ERK1/2, JNK and Akt were also counteracted by DAPT. In conclusion, DAPT inhibits Ang II-induced DASMCs proliferation and migration. These effects are potentially mediated by decreased calcium influx, reduced ROS production, and down-regulation of ERK1/2, JNK and Akt, through the Notch3-HES1/2/5 pathway. Therefore, Notch signaling has a role in DA remodeling and may provide a target pathway for therapeutic intervention of PDA. PMID:27570480

  7. Fbw7 Repression by Hes5 Creates a Feedback Loop That Modulates Notch-Mediated Intestinal and Neural Stem Cell Fate Decisions

    PubMed Central

    Tendeng, Christian; Clurman, Bruce E.; Lewis, Julian; Behrens, Axel

    2013-01-01

    FBW7 is a crucial component of an SCF-type E3 ubiquitin ligase, which mediates degradation of an array of different target proteins. The Fbw7 locus comprises three different isoforms, each with its own promoter and each suspected to have a distinct set of substrates. Most FBW7 targets have important functions in developmental processes and oncogenesis, including Notch proteins, which are functionally important substrates of SCF(Fbw7). Notch signalling controls a plethora of cell differentiation decisions in a wide range of species. A prominent role of this signalling pathway is that of mediating lateral inhibition, a process where exchange of signals that repress Notch ligand production amplifies initial differences in Notch activation levels between neighbouring cells, resulting in unequal cell differentiation decisions. Here we show that the downstream Notch signalling effector HES5 directly represses transcription of the E3 ligase Fbw7β, thereby directly bearing on the process of lateral inhibition. Fbw7Δ/+ heterozygous mice showed haploinsufficiency for Notch degradation causing impaired intestinal progenitor cell and neural stem cell differentiation. Notably, concomitant inactivation of Hes5 rescued both phenotypes and restored normal stem cell differentiation potential. In silico modelling suggests that the NICD/HES5/FBW7β positive feedback loop underlies Fbw7 haploinsufficiency. Thus repression of Fbw7β transcription by Notch signalling is an essential mechanism that is coupled to and required for the correct specification of cell fates induced by lateral inhibition. PMID:23776410

  8. Notch-Hes pathway mediates the impaired osteogenic differentiation of bone marrow mesenchymal stromal cells from myelodysplastic syndromes patients through the down-regulation of Runx2

    PubMed Central

    Fei, Chengming; Guo, Juan; Zhao, Youshan; Gu, Shucheng; Zhao, Sida; Li, Xiao; Chang, Chunkang

    2015-01-01

    Previous studies have demonstrated that bone marrow mesenchymal stromal cells (BMMSCs) from patients with myelodysplastic syndromes (MDS) display defective proliferative potential and impaired osteogenic differentiation ability. However, the underlying mechanisms are unclear. In the present study, the impaired osteogenic differentiation potential of BMMSCs was found in cases with RARS (83.3%), RCMD (75.0%), RAEB I (44.4%), RAEB II (40%). We also observed that MDS-BMMSCs with impaired osteogenic differentiation potential exhibited accelerate senescence and decreased hematopoietic supporting function. Further, we found that an abnormal activation of Notch-Hes signaling pathway in MDS-BMMSCs. By overexpression of Notch intracellular domain (NICD) in BMMSCs from healthy donors, we confirmed that Notch signaling pathway negatively regulated BMMSCs osteogenesis through inhibition of Runx2 transcriptional activity. Importantly, treatment with DAPT, a γ-secretase inhibitor of Notch signaling reversed the osteogenic differentiation in MDS-BMMSCs. Collectively, we provide evidence that activation of Notch-Hes signaling pathway is involved in the impaired osteogenic differentiation of MDS-BMMSCs and support the concept of a primary BMMSCs defect that might have a contributory effect in MDS pathogenesis. PMID:26692937

  9. Prostate tumor OVerexpressed-1 (PTOV1) down-regulates HES1 and HEY1 notch targets genes and promotes prostate cancer progression

    PubMed Central

    2014-01-01

    Background PTOV1 is an adaptor protein with functions in diverse processes, including gene transcription and protein translation, whose overexpression is associated with a higher proliferation index and tumor grade in prostate cancer (PC) and other neoplasms. Here we report its interaction with the Notch pathway and its involvement in PC progression. Methods Stable PTOV1 knockdown or overexpression were performed by lentiviral transduction. Protein interactions were analyzed by co-immunoprecipitation, pull-down and/or immunofluorescence. Endogenous gene expression was analyzed by real time RT-PCR and/or Western blotting. Exogenous promoter activities were studied by luciferase assays. Gene promoter interactions were analyzed by chromatin immunoprecipitation assays (ChIP). In vivo studies were performed in the Drosophila melanogaster wing, the SCID-Beige mouse model, and human prostate cancer tissues and metastasis. The Excel package was used for statistical analysis. Results Knockdown of PTOV1 in prostate epithelial cells and HaCaT skin keratinocytes caused the upregulation, and overexpression of PTOV1 the downregulation, of the Notch target genes HEY1 and HES1, suggesting that PTOV1 counteracts Notch signaling. Under conditions of inactive Notch signaling, endogenous PTOV1 associated with the HEY1 and HES1 promoters, together with components of the Notch repressor complex. Conversely, expression of active Notch1 provoked the dismissal of PTOV1 from these promoters. The antagonist role of PTOV1 on Notch activity was corroborated in the Drosophila melanogaster wing, where human PTOV1 exacerbated Notch deletion mutant phenotypes and suppressed the effects of constitutively active Notch. PTOV1 was required for optimal in vitro invasiveness and anchorage-independent growth of PC-3 cells, activities counteracted by Notch, and for their efficient growth and metastatic spread in vivo. In prostate tumors, the overexpression of PTOV1 was associated with decreased expression

  10. Quality of water used during cage cultivation of rainbow trout (Oncorhynchus mykiss) in Bereket HES IV Dam Lake (Muğla, Turkey).

    PubMed

    Ozdemir, Nedim; Demirak, Ahmet; Keskin, Feyyaz

    2014-12-01

    A thorough investigation of the impact of rainbow trout (Oncorhynchus mykiss) cultivation on surface water quality in the area known as Bereket HES IV Dam Lake was conducted. Water samples were collected from October 2009 to June 2010 from four stations in the Dam Lake and analyzed for water temperature, pH, dissolved oxygen, electrical conductivity, nitrite nitrogen, nitrate nitrogen, and orthophosphate. Surface water quality was then evaluated based on the comparison of samples collected from three stations located near fish cages to those collected from a reference station outside the cultivation area as well as by the comparison with standards specified in the Water Pollution Registration Act. According to the Water Pollution Registration Act, the surface water quality of the Dam Lake was class I. Additionally, there were no significant differences in water quality within the Dam Lake among any of the sampling stations, including the reference station. Overall, these findings indicate that cage cultivation of rainbow trout may have a negative impact on the entire Dam Lake. PMID:25204896

  11. Selection of cell fate in the organ of Corti involves the integration of Hes/Hey signaling at the Atoh1 promoter.

    PubMed

    Abdolazimi, Yassan; Stojanova, Zlatka; Segil, Neil

    2016-03-01

    Determination of cell fate within the prosensory domain of the developing cochlear duct relies on the temporal and spatial regulation of the bHLH transcription factor Atoh1. Auditory hair cells and supporting cells arise in a wave of differentiation that patterns them into discrete rows mediated by Notch-dependent lateral inhibition. However, the mechanism responsible for selecting sensory cells from within the prosensory competence domain remains poorly understood. We show in mice that rather than being upregulated in rows of cells, Atoh1 is subject to transcriptional activation in groups of prosensory cells, and that highly conserved sites for Hes/Hey repressor binding in the Atoh1 promoter are needed to select the hair cell and supporting cell fate. During perinatal supporting cell transdifferentiation, which is a model of hair cell regeneration, we show that derepression is sufficient to induce Atoh1 expression, suggesting a mechanism for priming the 3' Atoh1 autoregulatory enhancer needed for hair cell expression. PMID:26932672

  12. Short curcumin treatment modulates oxidative stress, arginase activity, aberrant crypt foci, and TGF-β1 and HES-1 transcripts in 1,2-dimethylhydrazine-colon carcinogenesis in mice.

    PubMed

    Bounaama, Abdelkader; Djerdjouri, Bahia; Laroche-Clary, Audrey; Le Morvan, Valérie; Robert, Jacques

    2012-12-16

    This study investigated the effect of short curcumin treatment, a natural antioxidant on 1,2-dimethylhydrazine (DMH)-induced aberrant crypt foci (ACF) in mice. The incidence of aberrant crypt foci (ACF) was 100%, with 54 ± 6 per colon, 10 weeks after the first DMH injection and reached 67 ± 12 per colon after 12 weeks. A high level of undifferentiated goblet cells and a weak apoptotic activity were shown in dysplastic ACF. The morphological alterations of colonic mucosa were associated to severe oxidative stress ratio with 43% increase in malondialdehyde vs. 36% decrease in GSH. DMH also increased inducible nitric synthase (iNOS) mRNA transcripts (250%), nitrites level (240%) and arginase activity (296%), leading to nitrosative stress and cell proliferation. Curcumin treatment, starting at week 10 post-DMH injection for 14 days, reduced the number of ACF (40%), iNOS expression (25%) and arginase activity (73%), and improved redox status by approximately 46%, compared to DMH-treated mice. Moreover, curcumin induced apoptosis of dysplastic ACF cells without restoring goblet cells differentiation. Interestingly, curcumin induced a parallel increase in TGF-β1 and HES-1 transcripts (42% and 26%, respectively). In conclusion, the protective effect of curcumin was driven by the reduction of arginase activity and nitrosative stress. The up regulation of TGF-β1 and HES-1 expression by curcumin suggests for the first time, a potential interplay between these signalling pathways in the chemoprotective mechanism of curcumin. PMID:22982865

  13. Methylprednisolone inhibits the proliferation and affects the differentiation of rat spinal cord-derived neural progenitor cells cultured in low oxygen conditions by inhibiting HIF-1α and Hes1 in vitro.

    PubMed

    Wang, Wenhao; Wang, Peng; Li, Shiyuan; Yang, Jiewen; Liang, Xinjun; Tang, Yong; Li, Yuxi; Yang, Rui; Wu, Yanfeng; Shen, Huiyong

    2014-09-01

    Although there is much controversy over the use of methylprednisolone (MP), it is one of the main drugs used in the treatment of acute spinal cord injury (SCI). The induction of the proliferation and differentiation of endogenous neural progenitor cells (NPCs) is considered a promising mode of treatment for SCI. However, the effects of MP on spinal cord-derived endogenous NPCs in a low oxygen enviroment remain to be delineated. Thus, the aim of this study was to investigate the potential effects of MP on NPCs cultured under low oxygen conditions in vitro and to elucidate the molecular mechanisms involved. Fetal rat spinal cord-derived NPCs were harvested and divided into 4 groups: 2 groups of cells cultured under normal oxygen conditions and treated with or without MP, and 2 groups incubated in 3% O2 (low oxygen) treated in a similar manner. We found that MP induced suppressive effects on NPC proliferation even under low oxygen conditions (3% O2). The proportion of nestin-positive NPCs decreased from 51.8±2.46% to 36.17±3.55% following the addition of MP and decreased more significantly to 27.20±2.68% in the cells cultured in 3% O2. In addition, a smaller number of glial fibrillary acidic protein (GFAP)-positive cells and a greater number of microtubule-associated protein 2 (MAP2)-positive cells was observed following the addition of MP under both normal (normoxic) and low oxygen (hypoxic) conditions. In response to MP treatment, hypoxia-inducible factor-1α (HIF-1α) and the Notch signaling pathway downstream protein, Hes1, but not the upstream Notch-1 intracelluar domain (NICD), were inhibited. After blocking NICD with a γ-secretase inhibitor (DAPT) MP still inhibited the expression of Hes1. Our results provide insight into the molecular mechanisms responsible for the MP-induced inhibition of proliferation and its effects on differentiation and suggest that HIF-1α and Hes1 play an important role in this effect. PMID:24992925

  14. Technical Education in 2-Year Colleges. HES Survey Number 17.

    ERIC Educational Resources Information Center

    Burton, Lawrence; Celebuski, Carin A.

    Based on a January 1993 survey of 347 two-year colleges nationwide, this report describes aspects of engineering technology and science technology education and training in the nation's two-year colleges. Following introductory materials, the first chapter estimates the number of colleges offering engineering and science programs, reviews types of…

  15. Hematology Expert System (HES) For Tonsillectomy/Adenoidectomy Patients

    NASA Astrophysics Data System (ADS)

    Pizzi, Nicolino J.; Kapoor, Sandhya; Gerrard, Jon M.

    1989-03-01

    The purpose of this expert system is to assess a predisposition to bleeding in a patient undergoing a tonsillectomy and/or adenoidectomy as may occur with patients who have certain blood conditions such as hemophilia and von Willebrand's disease. This goal is achieved by establishing a correlation between the patients' responses to a medical questionnaire and the relative quantities of blood lost during the operation.

  16. Determination of Physical Parameter Estimates for Metal-Poor Stars from the HK and HES Surveys

    NASA Astrophysics Data System (ADS)

    Beers, Timothy C.; Lee, Y.; Placco, V.; Carollo, D.; Christlieb, N.; Fiorenza, S.

    2013-06-01

    Over the past quarter century, well over ten thousand metal-poor star candidates selected during the course of the HK survey of Beers, Preston, and Shectman and the Hamburg/ESO survey of Christlieb and collaborators have had medium-resolution (R ~ 2500-3000) spectroscopy obtained, using a host of telescopes and spectrographs. During that time, we have refined the analysis tools at our disposal, and collected broadband photometry for many of these stars. Here we report on an application of the recently refined non-SEGUE Stellar Parameter Pipeline (n-SSPP) to obtain estimates of effective temperatures (Teff), surface gravities (log g), and metallicities ([Fe/H]) for many of these stars. In the case of some of the follow-up spectra, the wavelength coverage and S/N ratios also permit the determination of [C/Fe] and [alpha/Fe] ratios. Ongoing detailed analyses of the chemistry and kinematics of these stars will be described.

  17. Hydrogen Energy Storage (HES) and Power-to-Gas Economic Analysis; NREL (National Renewable Energy Laboratory)

    SciTech Connect

    Eichman, Joshua

    2015-07-30

    This presentation summarizes opportunities for hydrogen energy storage and power-to-gas and presents the results of a market analysis performed by the National Renewable Energy Laboratory to quantify the value of energy storage. Hydrogen energy storage and power-to-gas systems have the ability to integrate multiple energy sectors including electricity, transportation, and industrial. On account of the flexibility of hydrogen systems, there are a variety of potential system configurations. Each configuration will provide different value to the owner, customers and grid system operator. This presentation provides an economic comparison of hydrogen storage, power-to-gas and conventional storage systems. The total cost is compared to the revenue with participation in a variety of markets to assess the economic competitiveness. It is found that the sale of hydrogen for transportation or industrial use greatly increases competitiveness. Electrolyzers operating as demand response devices (i.e., selling hydrogen and grid services) are economically competitive, while hydrogen storage that inputs electricity and outputs only electricity have an unfavorable business case. Additionally, tighter integration with the grid provides greater revenue (e.g., energy, ancillary service and capacity markets are explored). Lastly, additional hours of storage capacity is not necessarily more competitive in current energy and ancillary service markets and electricity markets will require new mechanisms to appropriately compensate long duration storage devices.

  18. Hydrogen Energy Storage (HES) Activities at NREL; NREL (National Renewable Energy Laboratory)

    SciTech Connect

    Eichman, J.

    2015-04-21

    This presentation provides an overview of hydrogen and energy storage, including hydrogen storage pathways and international power-to-gas activities, and summarizes the National Renewable Energy Laboratory's hydrogen energy storage activities and results.

  19. Transition metal HE`s - VII ACP. Progress report, August 1971--October 1971

    SciTech Connect

    Clink, G.L.

    1998-12-31

    Physical and physicochemical properties of hexaamminechromium III perchlorate (ACP) were investigated to determine its potentiality for HE and/or HE component application. Physicochemical properties obtained through investigation of isothermally aged ACP under conditions of various system and boundary restrictions (open and closed; 50 and 80 C; 0.65 and 0.85 g/cc) show the material to be substantially stable under these simulated isothermal storage conditions from a standpoint of infrared and DTA thermal pattern behavior and chemical analysis of aged residues. Spark sensitivity was 0.25 joules at 5 kv (by LASL type test) and impact sensitivity was about 23 and 14 cm on sandpaper (12A) and steel (12B), respectively.

  20. Low-energy R -matrix fits for the 6Li(d ,α ) 4HeS factor

    NASA Astrophysics Data System (ADS)

    Grineviciute, J.; Lamia, L.; Mukhamedzhanov, A. M.; Spitaleri, C.; La Cognata, M.

    2015-01-01

    Background: Information about the 6Li(d ,α ) 4He reaction rates of astrophysical interest can be obtained by extrapolating direct data to lower energies, or by indirect methods. The indirect Trojan horse method, as well as various R -matrix and polynomial fits to direct data, estimate electron screening energies much larger than the adiabatic limit. Calculations that include the subthreshold resonance estimate smaller screening energies. Purpose: Obtain the 6Li(d,α ) 4He reaction R -matrix parameters and the bare astrophysical S factor for energies relevant to the stellar plasmas by fitting R -matrix formulas for the subthreshold resonances to the S -factor data above 60 keV. Methods: The bare S factor is calculated using the single- and the two-level R -matrix formulas for the closest to the threshold 0+ and 2+ subthreshold states at 22.2 ,20.2 , and 20.1 MeV. The electron screening potential U e is then obtained by fitting it as a single parameter to the low-energy data. The calculations are also done by fitting U e simultaneously with other parameters. Results: The low-energy S factor is dominated by the 2+ subthreshold resonance at 22.2 MeV. The influence of the other two subthreshold states is small. The resultant electron screening is smaller than the adiabatic value. The fits that neglect the electron screening above 60 keV produce a significantly smaller electron screening potential. The calculations show a large ambiguity associated with a choice of the initial channel radius. Conclusions: The R -matrix fits do not show a significantly larger Ue than predicted by the atomic physics models. The R -matrix best fit provides U e=149.5 eV and Sb(0 ) =21.7 MeV b.

  1. "HES Presidential Address": Confessions of a Positivist--How Foucault Led Me to a Meta-Narrative about School Desegregation

    ERIC Educational Resources Information Center

    Hutcheson, Philo

    2012-01-01

    This address derives from the intellectual contributions of young scholars and doctoral students, in faded memory of the author's life as a doctoral student and young scholar. This address has three purposes: (1) to define school desegregation; (2) to place--albeit briefly--that definition within the larger context of the literature on school…

  2. A Combination of Culture Conditions and Gene Expression Analysis Can Be Used to Investigate and Predict hES Cell Differentiation Potential towards Male Gonadal Cells

    PubMed Central

    Kjartansdóttir, Kristín Rós; Reda, Ahmed; Panula, Sarita; Day, Kelly; Hultenby, Kjell; Söder, Olle; Hovatta, Outi; Stukenborg, Jan-Bernd

    2015-01-01

    Human embryonic stem cell differentiation towards various cell types belonging to ecto-, endo- and mesodermal cell lineages has been demonstrated, with high efficiency rates using standardized differentiation protocols. However, germ cell differentiation from human embryonic stem cells has been very inefficient so far. Even though the influence of various growth factors has been evaluated, the gene expression of different cell lines in relation to their differentiation potential has not yet been extensively examined. In this study, the potential of three male human embryonic stem cell lines to differentiate towards male gonadal cells was explored by analysing their gene expression profiles. The human embryonic stem cell lines were cultured for 14 days as monolayers on supporting human foreskin fibroblasts or as spheres in suspension, and were differentiated using BMP7, or spontaneous differentiation by omitting exogenous FGF2. TLDA analysis revealed that in the undifferentiated state, these cell lines have diverse mRNA profiles and exhibit significantly different potentials for differentiation towards the cell types present in the male gonads. This potential was associated with important factors directing the fate of the male primordial germ cells in vivo to form gonocytes, such as SOX17 or genes involved in the NODAL/ACTIVIN pathway, for example. Stimulation with BMP7 in suspension culture resulted in up-regulation of cytoplasmic SOX9 protein expression in all three lines. The observation that human embryonic stem cells differentiate towards germ and somatic cells after spontaneous and BMP7-induced stimulation in suspension emphasizes the important role of somatic cells in germ cell differentiation in vitro. PMID:26630562

  3. A Combination of Culture Conditions and Gene Expression Analysis Can Be Used to Investigate and Predict hES Cell Differentiation Potential towards Male Gonadal Cells.

    PubMed

    Kjartansdóttir, Kristín Rós; Reda, Ahmed; Panula, Sarita; Day, Kelly; Hultenby, Kjell; Söder, Olle; Hovatta, Outi; Stukenborg, Jan-Bernd

    2015-01-01

    Human embryonic stem cell differentiation towards various cell types belonging to ecto-, endo- and mesodermal cell lineages has been demonstrated, with high efficiency rates using standardized differentiation protocols. However, germ cell differentiation from human embryonic stem cells has been very inefficient so far. Even though the influence of various growth factors has been evaluated, the gene expression of different cell lines in relation to their differentiation potential has not yet been extensively examined. In this study, the potential of three male human embryonic stem cell lines to differentiate towards male gonadal cells was explored by analysing their gene expression profiles. The human embryonic stem cell lines were cultured for 14 days as monolayers on supporting human foreskin fibroblasts or as spheres in suspension, and were differentiated using BMP7, or spontaneous differentiation by omitting exogenous FGF2. TLDA analysis revealed that in the undifferentiated state, these cell lines have diverse mRNA profiles and exhibit significantly different potentials for differentiation towards the cell types present in the male gonads. This potential was associated with important factors directing the fate of the male primordial germ cells in vivo to form gonocytes, such as SOX17 or genes involved in the NODAL/ACTIVIN pathway, for example. Stimulation with BMP7 in suspension culture resulted in up-regulation of cytoplasmic SOX9 protein expression in all three lines. The observation that human embryonic stem cells differentiate towards germ and somatic cells after spontaneous and BMP7-induced stimulation in suspension emphasizes the important role of somatic cells in germ cell differentiation in vitro. PMID:26630562

  4. Guiding the morphogenesis of dissociated newborn mouse retinal cells and hES cell-derived retinal cells by soft lithography-patterned microchannel PLGA scaffolds

    PubMed Central

    McUsic, Andrew C.; Lamba, Deepak A.; Reh, Thomas A.

    2011-01-01

    Embryonic stem (ES) cell-derived photoreceptors are a promising cell source for enhanced in vitro models of retinal degenerative diseases, but the more differentiated characteristics of retinal cells do not typically develop in dissociated cell cultures. Therefore, we have reconstructed organized retinal tissue by seeding dissociated cells into an array of aligned units that more faithfully mimics the retina. We solvent-processed poly(lactic-co-glycolic acid) (PLGA) into a microchannel scaffold format to achieve this geometric constraint. We compared the effect of PLGA concentration on channel morphology and, along with other culture conditions, on the infiltration of dissociated newborn mouse retinal cells into the channels. Culturing scaffolds at the gas-liquid interface with low serum media increased infiltrated rod photoreceptor viability 18-fold over submerged, high serum cultures when evaluated after seven days. Rod photoreceptors and Müller glia aligned processes parallel to the microchannel walls. Otx2+ and Pax6+ subpopulations recapitulated lamination behavior. Further, we constructed scaffold/retinal pigment epithelium (RPE) co-cultures and observed rods extending rhodopsin-positive processes toward RPE cells, mimicking normal rod polarization and morphology. Finally, human embryonic stem cell-derived photoreceptors exhibited infiltration and morphological characteristics similar to mouse retinal cells inside the scaffolds. These findings constitute an important advance in generating tissue-level retinal models from dissociated cells for use as drug screening platforms and in regenerative medicine. PMID:22115999

  5. Peripheral Neuropathy Associated withHypereosinophilic Syndrome

    PubMed Central

    Lee, Kyung Ho; Kim, Jung Eun

    2008-01-01

    The idiopathic hypereosinophilic syndrome (HES) represents a leukoproliferative disorder, characterized by unexplained prolonged eosinophilia (>6 months) and evidence of specific organ damage. So far, the peripheral neuropathy associated with skin manifestations of HES has not been reported in the dermatologic literature although the incidence of peripheral neuropathy after HES ranges from 6~52%. Herein, we report the peripheral neuropathy associated with HES, documented by clinical, histopathological, and electrodiagnostic criteria. PMID:27303181

  6. Higher Education for Sustainability: Developing a Comprehensive Research Agenda

    ERIC Educational Resources Information Center

    Wright, Tarah

    2007-01-01

    From 27 to 29 October 2005, 35 experts in higher education for sustainability (HES) representing 17 countries, gathered in Halifax, Nova Scotia. This consultation represents the first gathering of HES researchers in Canada, and brought Canadian and international researchers together to further intellectual understanding of HES research and to…

  7. Role of ICT in Shaping the Future of Pakistani Higher Education System

    ERIC Educational Resources Information Center

    Shaikh, Zaffar Ahmed; Khoja, Shakeel Ahmed

    2011-01-01

    This study examined the challenges faced by the Pakistani higher education system (HES) in integrating information and communication technology (ICT); it aimed at understanding ICT needs, measuring the increase in ICT demand, determining the relationship between ICT and HES performance, and understanding how the HES copes with the challenges of…

  8. HYPEREOSINOPHILIC SYNDROME: CUTANEOUS INVOLVEMENT AS THE SOLE MANIFESTATION

    PubMed Central

    Sundaramurthi, Vidya lakshmi; Prabhavathy, D; Somasundaram, S V; Wahab, Afthab Jameela

    2011-01-01

    Hypereosinophilic syndrome (HES) encompasses a group of leukoproliferative disorders with variable involvement of the internal organs. More than half of all patients have cutaneous involvement. In a minority of the reported cases, skin involvement has been the only manifestation of HES . We report one such rare case of HES, with cutaneous involvement as the sole manifestation. PMID:21572808

  9. Raman-based detection of hydroxyethyl starch in kidney allograft biopsies as a potential marker of allograft quality in kidney transplant recipients.

    PubMed

    Vuiblet, Vincent; Fere, Michael; Bankole, Ezechiel; Wynckel, Alain; Gobinet, Cyril; Birembaut, Philippe; Piot, Olivier; Rieu, Philippe

    2016-01-01

    In brain-dead donor resuscitation, hydroxyethyl starch (HES) use has been associated with presence of osmotic-nephrosis-like lesions in kidney transplant recipients. Our aim was to determine whether the presence of HES in protocol renal graft biopsies at three months (M3) after transplantation is associated with renal graft quality. According to the HES administered to the donor during the procurement procedure, two groups of patients were defined according graft exposition to HES: HES group, (N = 20) and control group (N = 6). Detection and relative quantification of HES was performed by Raman spectroscopy microimaging on M3 protocol renal graft biopsies. Statistical analyses were used to investigate the association between Raman data and graft characteristics. HES spectral signal was revealed negative in the control group, whereas it was positive in 40% of biopsies from the HES group. In the HES group, a stronger HES signal was associated with a lower risk of graft failure measured by the Kidney Donor Risk Index (KDRI) and was correlated with the allograft kidney function. Thus, HES accumulation in donor kidney, as probed by Raman biophotonic technique, is correlated with the quality of donor kidney and consequently the graft renal function and graft survival. PMID:27608775

  10. Raman-based detection of hydroxyethyl starch in kidney allograft biopsies as a potential marker of allograft quality in kidney transplant recipients

    PubMed Central

    Vuiblet, Vincent; Fere, Michael; Bankole, Ezechiel; Wynckel, Alain; Gobinet, Cyril; Birembaut, Philippe; Piot, Olivier; Rieu, Philippe

    2016-01-01

    In brain-dead donor resuscitation, hydroxyethyl starch (HES) use has been associated with presence of osmotic-nephrosis-like lesions in kidney transplant recipients. Our aim was to determine whether the presence of HES in protocol renal graft biopsies at three months (M3) after transplantation is associated with renal graft quality. According to the HES administered to the donor during the procurement procedure, two groups of patients were defined according graft exposition to HES: HES group, (N = 20) and control group (N = 6). Detection and relative quantification of HES was performed by Raman spectroscopy microimaging on M3 protocol renal graft biopsies. Statistical analyses were used to investigate the association between Raman data and graft characteristics. HES spectral signal was revealed negative in the control group, whereas it was positive in 40% of biopsies from the HES group. In the HES group, a stronger HES signal was associated with a lower risk of graft failure measured by the Kidney Donor Risk Index (KDRI) and was correlated with the allograft kidney function. Thus, HES accumulation in donor kidney, as probed by Raman biophotonic technique, is correlated with the quality of donor kidney and consequently the graft renal function and graft survival. PMID:27608775

  11. Identifying developmental toxicity pathways for a subset of ToxCast chemicals using human embryonic stem cells and metabolomics

    EPA Science Inventory

    Metabolomics analysis was performed on the supernatant of human embryonic stem (hES) cell cultures exposed to a blinded subset of 11 chemicals selected from the chemical library of EPA's ToxCast™ chemical screening and prioritization research project. Metabolites from hES cultur...

  12. 75 FR 6658 - Science Advisory Board Staff Office; Notification of a Public Teleconference of the Health...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-10

    ...The Environmental Protection Agency (EPA) Science Advisory Board (SAB) Staff Office announces a public teleconference of the Health Effects Subcommittee (HES) of the Advisory Council on Clean Air Compliance Analysis (Council). The HES, supplemented with additional members from the Council, will discuss its draft peer review report on EPA's health benefits analyses and uncertainty analyses in......

  13. Renal thrombotic microangiopathy and FIP1L1/PDGFRα-associated myeloproliferative variant of hypereosinophilic syndrome

    PubMed Central

    Langlois, Anne Lyse; Shehwaro, Nathalie; Rondet, Claire; Benbrik, Youssef; Maloum, Karim; Gueutin, Victor; Rouvier, Philippe; Izzedine, Hassane

    2013-01-01

    We report a case of renal thrombotic microangiopathy (TMA) in a myeloproliferative variant of hypereosinophilic syndrome (HES) in a 24-year-old man which resolved with imatinib therapy. This is one of a few cases in the literature to date describing TMA in HES, suggesting that the pathogenesis of thrombosis is at least in part related to damage from activated eosinophils. PMID:27293571

  14. The Word from a Walrus: Five Decades of the History of Education Society

    ERIC Educational Resources Information Center

    Urban, Wayne J.

    2010-01-01

    This essay is an attempt at an institutional history of the History of Education Society (HES), from its inception in 1960 to the present day. Many of the intellectual currents and cross-currents, as well as the "History of Education Quarterly" (HEQ), the journal of the HES in which these intellectual movements were featured, are slighted in this…

  15. National Innovation and Knowledge Performance: The Role of Higher Education Teaching and Training

    ERIC Educational Resources Information Center

    Saad, Mohammed; Guermat, Cherif; Brodie, Lisa

    2015-01-01

    This paper investigates the role of the higher education system (HES) in the production of national innovation. We focus on the issue of institutional diversity of HESs and its impact on national innovation systems. We identify four key HES characteristics and hypothesize their influence on the production of national innovation. The empirical…

  16. General model of a cascade of reactions with time delays: Global stability analysis

    NASA Astrophysics Data System (ADS)

    Bodnar, Marek

    2015-07-01

    The problem considered in this paper consists of a cascade of reactions with discrete as well as distributed delays, which arose in the context of Hes1 gene expression. For the abstract general model sufficient conditions for global stability are presented. Then the abstract result is applied to the Hes1 model.

  17. Nucleofection Mediates High-Efficiency Stable Gene Knockdown and Transgene Expression in Human Embryonic Stem Cells

    PubMed Central

    Hohenstein, Kristi A.; Pyle, April D.; Chern, Jing Yi; Lock, Leslie F.; Donovan, Peter J.

    2013-01-01

    High-efficiency genetic modification of human embryonic stem (hES) cells would enable manipulation of gene activity, routine gene targeting, and development of new human disease models and treatments. Chemical transfection, nucleofection, and electroporation of hES cells result in low transfection efficiencies. Viral transduction is efficient but has significant drawbacks. Here we describe techniques to transiently and stably express transgenes in hES cells with high efficiency using a widely available vector system. The technique combines nucleofection of single hES cells with improved methods to select hES cells at clonal density. As validation, we reduced Oct4 and Nanog expression using siRNAs and shRNA vectors in hES cells. Furthermore, we derived many hES cell clones with either stably reduced alkaline phosphatase activity or stably overexpressed green fluorescent protein. These clones retained stem cell characteristics (normal karyotype, stem cell marker expression, self-renewal, and pluripotency). These studies will accelerate efforts to interrogate gene function and define the parameters that control growth and differentiation of hES cells. PMID:18323409

  18. A hyperbolic-equation system approach for magnetized electron fluids in quasi-neutral plasmas

    SciTech Connect

    Kawashima, Rei; Komurasaki, Kimiya; Schönherr, Tony

    2015-03-01

    A new approach using a hyperbolic-equation system (HES) is proposed to solve for the electron fluids in quasi-neutral plasmas. The HES approach avoids treatments of cross-diffusion terms which cause numerical instabilities in conventional approaches using an elliptic equation (EE). A test calculation reveals that the HES approach can robustly solve problems of strong magnetic confinement by using an upwind method. The computation time of the HES approach is compared with that of the EE approach in terms of the size of the problem and the strength of magnetic confinement. The results indicate that the HES approach can be used to solve problems in a simple structured mesh without increasing computational time compared to the EE approach and that it features fast convergence in conditions of strong magnetic confinement.

  19. Assessment of coagulation with 6% hydroxyethyl starch 130/0.4 in cesarean section

    PubMed Central

    Oh, Chung-Sik; Kim, Seong-Hyop; Kim, Duk-Kyung; Lim, Jeong-Ae; Woo, Nam-Sik

    2012-01-01

    Background Third-generation hydroxyethyl starch (HES) solutions have been developed to minimize negative effects on hemostasis. In normal pregnancy, the coagulation activity increases, reaching a maximum around term. This study examined the effects of hemodilution with HES 130/0.4 (6%) on blood coagulation in parturients in vivo and in vitro. Methods Forty parturients scheduled for cesarean sections were assigned randomly to receive either 500 or 1,000 ml of HES 130/0.4 (6%). Rotation thromboelastometry (ROTEM®) measurements were performed before and after administering HES 130/0.4 (6%). In addition, blood samples obtained from 20 randomly selected parturients were diluted 10% to 40% using HES 130/0.4 (6%), and ROTEM® measurements were performed before and after dilution. The changes from baseline and the effects of dilution were analyzed by ROTEM® parameters. Results Infusions of 500 or 1,000 ml of HES 130/0.4 (6%) in the parturients altered the clot formation time, α angle, and maximal clot firmness, although all remained within normal ranges. HES 130/0.4 (6%) affected in vitro blood coagulation in parturients' blood containing 10, 20, 30, and 40% HES. The clotting time was prolonged at each dilution percentage, but remained within the normal range. Other parameters showed an impairment of the coagulation system. Conclusions Blood coagulation in parturients may be compromised at high dilution ratios of HES 130/0.4 (6%) to blood. Nevertheless, the infusion of 1,000 ml of HES 130/0.4 (6%) in normal parturients did not significantly affect blood coagulation. PMID:22558500

  20. Effect of perioperative crystalloid or colloid fluid therapy on hemorrhage, coagulation competence, and outcome

    PubMed Central

    Rasmussen, Kirsten C.; Secher, Niels H.; Pedersen, Tom

    2016-01-01

    Abstract Background: A meta-analysis concerning perioperative coagulation competence, hemorrhage, and outcome was conducted including the use of hydroxyethyl starches (HESs), dextran, or albumin versus administration of a crystalloid as control to assess the efficacy and safety of colloids and crystalloids for fluid administration during major elective surgery. Surgery was restricted to cardiovascular and noncardiovascular surgery, and HESs were stratified to HES 130/0.4 and HES 200/0.5. Methods: We searched Cochrane Central Register of Controlled Trials, MEDLINE, ISI Web of Science, EMBASE, conference proceedings, reference lists, and databases of ongoing trials. Results: Thirty one primary clinical randomized controlled trials included 2287 patients undergoing major surgery from January 2000 to August 2015. The perioperative changes in coagulation competence were measured by thromboelastography (TEG) maximum amplitude (MA) in 9 studies administering crystalloids versus HES and in 4 studies administering albumin versus HES. All studies but 1 disclosed increased reduction in TEG-MA following HES administration (P = 0.0001 and 0.0002). The total loss of blood was reported in 17 studies in which crystalloids were compared to HES and 12 studies reported increased blood loss after administration of HES (P < 0.003). When administering albumin versus HES, 6 studies reported reduced hemorrhage associated with albumin administration (P = 0.005). Reoperation was not significantly reduced by the use of crystalloids, but may be more frequent after HESs compared to albumin (P < 0.03). In this analysis, more patients admitted to administration of HESs were exposed to decrease coagulation competence, compared to perioperative crystalloids and albumin administration. Conclusion: This stratified meta-analysis showed that increased blood loss was found in noncardiovascular surgery among patients receiving HES compared with crystalloids, followed by a marked

  1. Hemodynamic effects of 6% hydroxyethyl starch infusion in sevoflurane-anesthetized thoroughbred horses.

    PubMed

    Ohta, Minoru; Kurimoto, Shinjiro; Tokushige, Hirotaka; Kuroda, Taisuke; Ishikawa, Yuhiro

    2013-07-31

    To determine hemodynamic effects of hydroxyethyl starch (HES) infusion during anesthesia in horses, incremental doses of 6% HES were administered to 6 healthy Thoroughbred horses. Anesthesia was induced with xylazine, guaifenesin and thiopental and maintained with sevoflurane at 2.8% of end-tidal concentration in all horses. The horses were positioned in right lateral recumbency and administered 3 intravenous dose of 6% HES (5 ml/kg) over 15 min with 15-min intervals in addition to constant infusion of lactated Ringer's solution at 10 ml/kg/hr. Hemodynamic parameters were measured before and every 15 min until 90 min after the administration of 6% HES. There was no significant change in heart rate and arterial blood pressures throughout the experiment. The HES administration produced significant increases in mean right atrial pressure, stroke volume, cardiac output (CO) and decrease in systemic vascular resistance (SVR) in a dose-dependent manner. There was no significant change in electrolytes (Na(+), K(+), Cl(-)) throughout the experiment, however, packed cell volume, hemoglobin concentration, and total protein and albumin concentrations decreased in a dose-dependent manner following the HES administration. In conclusion, the HES administration provides a dose-dependent increase in CO, but has no impact upon arterial blood pressures due to a simultaneous decrease in SVR. PMID:23411483

  2. Roles of bHLH genes in neural stem cell differentiation

    SciTech Connect

    Kageyama, Ryoichiro . E-mail: rkageyam@virus.kyoto-u.ac.jp; Ohtsuka, Toshiyuki; Hatakeyama, Jun; Ohsawa, Ryosuke

    2005-06-10

    Neural stem cells change their characteristics over time during development: they initially proliferate only and then give rise to neurons first and glial cells later. In the absence of the repressor-type basic helix-loop-helix (bHLH) genes Hes1, Hes3 and Hes5, neural stem cells do not proliferate sufficiently but prematurely differentiate into neurons and become depleted without making the later born cell types such as astrocytes and ependymal cells. Thus, Hes genes are essential for maintenance of neural stem cells to make cells not only in correct numbers but also in full diversity. Hes genes antagonize the activator-type bHLH genes, which include Mash1, Math and Neurogenin. The activator-type bHLH genes promote the neuronal fate determination and induce expression of Notch ligands such as Delta. These ligands activate Notch signaling and upregulate Hes1 and Hes5 expression in neighboring cells, thereby maintaining these cells undifferentiated. Thus, the activator-type and repressor-type bHLH genes regulate each other, allowing only subsets of cells to undergo differentiation while keeping others to stay neural stem cells. This regulation is essential for generation of complex brain structures of appropriate size, shape and cell arrangement.

  3. Hexane extract from Sargassum serratifolium inhibits the cell proliferation and metastatic ability of human glioblastoma U87MG cells.

    PubMed

    Kang, Chang-Won; Park, Min-Seok; Kim, Nan-Hee; Lee, Ji-Hyun; Oh, Chul-Woong; Kim, Hyeung-Rak; Kim, Gun-Do

    2015-11-01

    The present study is the first to demonstrate the anticancer effects of a hexane extract from the brown algae Sargassum serratifolium (HES) on human cancer cell lines, including glioblastoma U87MG, cervical cancer HeLa and gastric cancer MKN-28 cells, as well as liver cancer SK-HEP 1 cells. Among these cancer cell lines, U87MG cells were most sensitive to the cell death induced by HES. HES exhibited a cytotoxic effect on U87MG cells at concentrations of 14-16 µg/ml, yet an effect was not observed in human embryonic kidney HEK293 cells. The antiproliferative effects of HES were regulated by inhibition of the MAPK/ERK signaling pathway which plays a pivotal role in the proliferation of glioblastoma U87MG cells. In addition, treatment with HES led to cell morphological changes and cell cytoskeleton degradation through regulation of actin dynamic signaling. Furthermore, migration and invasion of the U87MG cells were inhibited by HES via suppression of matrix metalloproteinase (MMP)-2 and -9 expression. Thus, our results suggest that HES is a potential therapeutic agent which has anticancer effects on glioblastoma. PMID:26323587

  4. Changes in the molecular composition of circulating hydroxyethyl starch

    PubMed Central

    Farrow, S. P.; Hall, M.; Ricketts, C. R.

    1970-01-01

    1. Hydroxyethyl starch (HES) of molecular weight 450,000 has been extensively investigated as a plasma volume expander and HES of molecular weight 60,000 has been proposed for use as a cryoprotective agent in the freezing for preservation of red blood cells. In these applications considerable amounts of HES may be injected intravenously and it is therefore desirable to understand its behaviour in vivo. 2. After intravenous injection into rabbits, plasma levels of HES and clinical dextrans were determined using anthrone. HES persisted longer in the circulation than did the dextrans. 3. HES was recovered from the bloodstream of rabbits and shown by gel filtration to be of a narrower molecular size distribution than the injected material. Smaller molecules were removed, presumably by filtration at the glomerulus. Larger molecules may have been removed by cells of the reticuloendothelial system or possibly through the action of amylase. 4. The amylase in saliva was shown to break down HES into molecules of intermediate size with very little production of maltose units. PMID:5441787

  5. Suppression of type 2 immunity and allergic airway inflammation by secreted products of the helminth Heligmosomoides polygyrus

    PubMed Central

    McSorley, Henry J; O'Gorman, Mary T; Blair, Natalie; Sutherland, Tara E; Filbey, Kara J; Maizels, Rick M

    2016-01-01

    Summary Allergic asthma is less prevalent in countries with parasitic helminth infections, and mice infected with parasites such as Heligmosomoides polygyrus are protected from allergic airway inflammation. To establish whether suppression of allergy could be mediated by soluble products of this helminth, we tested H. polygyrus excretory-secretory (HES) material for its ability to impair allergic inflammation. When HES was added to sensitising doses of ovalbumin, the subsequent allergic airway response was suppressed, with ablated cell infiltration, a lower ratio of effector (CD4+CD25+Foxp3−) to regulatory (CD4+Foxp3+) T (Treg) cells, and reduced Th1, Th2 and Th17 cytokine production. HES exposure reduced IL-5 responses and eosinophilia, abolished IgE production, and inhibited the type 2 innate molecules arginase-1 and RELM-α. Although HES contains a TGF-β-like activity, similar effects in modulating allergy were not observed when administering mammalian TGF-β alone. HES also protected previously sensitised mice, suppressing recruitment of eosinophils to the airways when given at challenge, but no change in Th or Treg cell populations was apparent. Because heat-treatment of HES did not impair suppression at sensitisation, but compromised its ability to suppress at challenge, we propose that HES contains distinct heat-stable and heat-labile immunomodulatory molecules which modulate pro-allergic adaptive and innate cell populations. PMID:22706967

  6. A triple-blinded randomized trial comparing the hemostatic effects of large-dose 10% hydroxyethyl starch 264/0.45 versus 5% albumin during major reconstructive surgery.

    PubMed

    Arellano, Ramiro; Gan, Bing Siang; Salpeter, Mary Jane; Yeo, Erik; McCluskey, Stuart; Pinto, Ruxandra; Irish, Jonathan; Ross, Douglas C; Doyle, D John; Parkin, John; Brown, Dale; Rotstein, Lorne; Witterick, Ian; Matthews, Wayne; Yoo, John; Neligan, Peter C; Gullane, Pat; Lampe, Howard

    2005-06-01

    In Canada, hydroxyethyl starch 264/0.45 (HES 264/0.45; molar weight 264 kDa, molar substitution 0.45) has largely replaced albumin as the colloidal fluid of choice for perioperative intravascular volume expansion. The maximum recommended dose of HES 264/0.45 is 28 mL/kg; however, there are no clinical data supporting this limit. In this study we compared the hemostatic effects of HES 264/0.45 versus 5% albumin in doses up to 45 mL/kg over 24 h during major reconstructive head and neck surgery. Fifty patients were randomized to receive HES 264/0.45 or 5% human albumin from the induction of anesthesia until 24 h thereafter. Both albumin and HES 264/0.45 effectively maintained physiologic variables in the perioperative and postoperative periods. The partial thromboplastin time and international normalized ratio were significantly increased in the HES 264/0.45 group compared with the albumin group after infusion of 30 mL/kg and 45 mL/kg (P < 0.05). Factor VIII activity and von Willebrand factor level were significantly reduced in the HES 264/0.45 group compared with the albumin group after infusion of 15 mL/kg, 30 mL/kg, and 45 mL/kg (P < 0.05). Significantly more subjects in the HES 264/0.45 group received allogeneic red blood cell transfusions (P < 0.02). We conclude that HES 264/0.45 infusions >30 mL/kg over 24 h impair coagulation to a greater extent than albumin, possibly leading to more allogeneic transfusions. PMID:15920225

  7. How to cross immunogenetic hurdles to human embryonic stem cell transplantation.

    PubMed

    de Rham, Casimir; Villard, Jean

    2011-11-01

    Implantation of human embryonic stem cells (hES), derived progenitors or mature cells derived from hES has great therapeutic potential for many diseases. If hES would come from genetically unrelated individuals, it would be probably rejected by the immune system of the recipient. Blood groups, MHC and minor antigens are the immunogenetic hurdles that have to be crossed for successful transplantation. Autologous transplantation with adult stem cells would be the best approach but several elements argue against this option. Classical immunosuppression, depleting antibody, induction of tolerance and stem cell banking are alternative methods that could be proposed to limit the risk of rejection. PMID:21461719

  8. Idiopathic Hypereosinophilic Syndrome With Cutaneous Manifestations and Flame Figures: A Spectrum of Eosinophilic Dermatoses Whose Features Overlap With Wells' Syndrome

    PubMed Central

    Kiracofe, Elizabeth A.; Clark, Lindsey N.; Gru, Alejandro A.

    2015-01-01

    Importance: Wells syndrome (WS) (eosinophilic cellulitis) is an uncommon eosinophilic dermatitis that has been rarely described in association with, but distinct from, hypereosinophilic syndrome (HES). Observations: We report a case of an eosinophilic dermatosis with flame figures in association with idiopathic HES, manifested by inflammatory myocarditis, asthma, and peripheral blood eosinophilia. Conclusions and Relevance: The diagnoses of WS and HES, rather than being distinct findings, may represent 2 entities on a spectrum of hypereosinophilic diseases. The diagnosis of WS should be made with caution and should prompt a thorough investigation that includes a work-up for a systemic eosinophilic disorder. PMID:25839890

  9. 26 CFR 1.121-3 - Reduced maximum exclusion for taxpayers failing to meet certain requirements.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... section 121(c)(2). Example 4. B, who has chronic asthma, purchases a house in Minnesota in 2003 that he...'s asthma symptoms. In 2004 B sells his house and moves to Arizona to relieve his asthma...

  10. 26 CFR 1.121-3 - Reduced maximum exclusion for taxpayers failing to meet certain requirements.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... section 121(c)(2). Example 4. B, who has chronic asthma, purchases a house in Minnesota in 2003 that he...'s asthma symptoms. In 2004 B sells his house and moves to Arizona to relieve his asthma...

  11. 26 CFR 1.121-3 - Reduced maximum exclusion for taxpayers failing to meet certain requirements.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... section 121(c)(2). Example 4. B, who has chronic asthma, purchases a house in Minnesota in 2003 that he...'s asthma symptoms. In 2004 B sells his house and moves to Arizona to relieve his asthma...

  12. 26 CFR 1.121-3 - Reduced maximum exclusion for taxpayers failing to meet certain requirements.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... section 121(c)(2). Example 4. B, who has chronic asthma, purchases a house in Minnesota in 2003 that he...'s asthma symptoms. In 2004 B sells his house and moves to Arizona to relieve his asthma...

  13. 26 CFR 1.121-3 - Reduced maximum exclusion for taxpayers failing to meet certain requirements.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... section 121(c)(2). Example 4. B, who has chronic asthma, purchases a house in Minnesota in 2003 that he...'s asthma symptoms. In 2004 B sells his house and moves to Arizona to relieve his asthma...

  14. Hydrogen Energy Storage: Grid and Transportation Services (Technical Report)

    SciTech Connect

    Not Available

    2015-02-01

    Proceedings of an expert workshop convened by the U.S. Department of Energy and Industry Canada, and hosted by the National Renewable Energy Laboratory and the California Air Resources Board, May 14-15, 2014, in Sacramento, California, to address the topic of hydrogen energy storage (HES). HES systems provide multiple opportunities to increase the resilience and improve the economics of energy sup supply systems underlying the electric grid, gas pipeline systems, and transportation fuels. This is especially the case when considering particular social goals and market drivers, such as reducing carbon emissions, increasing reliability of supply, and reducing consumption of conventional petroleum fuels. This report compiles feedback collected during the workshop, which focused on policy and regulatory issues related to HES systems. Report sections include an introduction to HES pathways, market demand, and the "smart gas" concept; an overview of the workshop structure; and summary results from panel presentations and breakout groups.

  15. Hybrid Energy System Modeling in Modelica

    SciTech Connect

    William R. Binder; Christiaan J. J. Paredis; Humberto E. Garcia

    2014-03-01

    In this paper, a Hybrid Energy System (HES) configuration is modeled in Modelica. Hybrid Energy Systems (HES) have as their defining characteristic the use of one or more energy inputs, combined with the potential for multiple energy outputs. Compared to traditional energy systems, HES provide additional operational flexibility so that high variability in both energy production and consumption levels can be absorbed more effectively. This is particularly important when including renewable energy sources, whose output levels are inherently variable, determined by nature. The specific HES configuration modeled in this paper include two energy inputs: a nuclear plant, and a series of wind turbines. In addition, the system produces two energy outputs: electricity and synthetic fuel. The models are verified through simulations of the individual components, and the system as a whole. The simulations are performed for a range of component sizes, operating conditions, and control schemes.

  16. The Louisville Twin Study: Developmental Synchronies in Behavior.

    ERIC Educational Resources Information Center

    Wilson, Ronald S.

    1983-01-01

    Results show individual differences in intelligence progressively stabilizing by school age, with each child following a distinct pattern of mental development. Monozygotic twins became increasingly concordant over an Hes and were closely matched on developmental trends. (Author/RH)

  17. Evaluation and differential diagnosis of marked, persistent eosinophilia

    PubMed Central

    Mejia, Rojelio; Nutman, Thomas B.

    2012-01-01

    High grade eosinophilia in patients can be a diagnostic dilemma, as the etiologies are extensive and varied. Hypereosinophilic syndromes (HES) are a group of heterogeneous disorders, many of which remain ill-defined. By definition, HES must be distinguished from other disorders with persistently elevated eosinophilia with a defined cause. Although marked eosinophilia worldwide is most commonly caused by helminth (worm) infections, non-infectious causes must be sought including drug reactions, malignancies, and immunologic, inflammatory and allergic diseases. PMID:22449625

  18. Influence of Two Colloidal Extracorporeal Primes on Coagulation of Cardiac Surgical Patients: A Prospectively Randomized Open-Label Pilot Trial.

    PubMed

    Bethlehem, Irene; Wierda, Korry; Visser, Cornelis; Jekel, Lilian; Koopmans, Matty; Kuiper, Michael A

    2014-12-01

    The search for the ideal priming fluid continues as more evidence is discovered about side effects of volume expanders. With the availability of modern, balanced hydroxyethyl starch (HES) solutions with less side effects than former HES solutions, we considered to replace our gelatin- (modified gelatin) based extracorporeal circuit prime for a HES (130/.42) prime. Therefore, we studied the influence of two colloidal priming fluids on postoperative coagulation in patients undergoing cardiac surgery. The primary endpoint was to compare clot formation time between the HES group and the gelatin group with rotational thromboelastometry (ROTEM). Additionally we compared colloid osmotic pressure and fluid balance of both groups. Forty patients, undergoing elective first time coronary artery bypass grafting or single-valve surgery, were included in this prospectively randomized open-label pilot study. Laboratory data and ROTEM data were collected and analyzed for differences between the two groups. ROTEM data show significantly more prolongation in Extem clot formation time and significant more decrease in Extem alpha in the HES group. Fibtem maximum clot firmness was significantly smaller in the HES group; this was consistent with fibrinogen concentration measurement, which decreased more in the HES group than in the gelatin group and recovered more over time in the gelatin group. We found no significant difference in colloid. osmotic pressure between the groups. In this trial, HES (130/.42) impairs coagulation significantly more compared with gelatin. These differences in influence on coagulation did not lead to a difference in blood loss or fluid balance, so clinical relevance could not be proven. PMID:26357798

  19. Derivation and characterization of human ESC-derived mesenchymal stem cells.

    PubMed

    Lai, Ruenn Chai; Choo, Andre; Lim, Sai Kiang

    2011-01-01

    Mesenchymal stem cells (MSCs) are multipotent stem cells that have been isolated from numerous sources including human embryonic stem cells (hES). Derivation from hES is unique in that hES must be differentiated. In our hands, trypsinizing hES into single cells and plating them on gelatin coated plates in a DMEM medium supplemented with serum replacement media and FGF2 with either PDGF AB or EGF will induce differentiation of hES and selectively enhance the survival of MSCs over hES. Repeated passaging by trypsinization results in a highly enriched MSC culture. Enriched MSC cultures can be further purified to homogeneity by limiting dilution or FACS sorting for a CD105+ or CD73+ and CD24- cell population. The resulting hES-MSCs fulfill the ISCT minimal defining criteria for human MSCs, namely adherence to plastic, a surface antigen expression profile of CD29+, CD44+, CD49a+ CD49e+, CD73+, CD105+, CD166+, CD34-, CD45-, and a differentiation potential that includes adipogenesis, osteogenesis, and chondrogenesis. Finally, hES-MSCs can be extensively and stably propagated. This method of deriving hES-MSCs without the need for a xenogeneic feeder and use of animal serum could be used to derive clinically compliant MSCs from hESCs. PMID:21431516

  20. Effect of hydroxyethyl starch on blood glucose levels

    PubMed Central

    Shim, Soo Bin; Choi, Woo Young

    2016-01-01

    Background Hydroxyethyl starch (HES), a commonly used resuscitation fluid, has the property to induce hyperglycemia as it contains large ethyl starch, which can be metabolized to produce glucose. We evaluated the effect of 6% HES-130 on the blood glucose levels in non-diabetic patients undergoing surgery under spinal anesthesia. Methods Patients scheduled to undergo elective lower limb surgery were enrolled. Fifty-eight patients were divided into two groups according to the type of the main intravascular fluid used before spinal anesthesia (Group LR: lactated Ringer's solution, n = 30 vs. Group HES: 6% hydroxyethyl starch 130/0.4, n = 28). Blood glucose levels were measured at the following time points: 0 (baseline), 20 min (T1), 1 h (T2), 2 h (T3), 4 h (T4), and 6 h (T6). Results Mean blood glucose levels at T5 in the LR group and T4, T5 in the HES group, increased significantly compared to baseline. There were no significant changes in the serial differences of mean blood glucose levels from baseline between the two groups. Conclusions Administration of 6% HES-130 increased blood glucose levels within the physiologic limits, but the degree of glucose increase was not greater than that caused by administration of lactated Ringer's solution. In conclusion, we did not find evidence that 6% HES-130 induces hyperglycemia in non-diabetic patients. PMID:27482311

  1. Health Education Specialists' Knowledge, Attitudes, and Perceptions of the Patient Protection and Affordable Care Act.

    PubMed

    Strong, Jessica; Hanson, Carl L; Magnusson, Brianna; Neiger, Brad

    2016-03-01

    The changing landscape of health care as a result of the Patient Protection and Affordable Care Act (ACA) may provide new opportunities for health education specialists (HES). The purpose of this study was to survey HES in the United States on their knowledge and attitudes of the ACA and assess their perceptions of job growth under the law. A random sample of 220 (36% response rate) certified HES completed a 53-item cross sectional survey administered online through Qualtrics. Findings were compared to public opinion on health care reform. HES are highly favorable of the law (70%) compared to the general public (23%). A total of 85% of respondents were able to list a provision of the ACA, and most (81%) thought the ACA would be successful at increasing insured Americans. Over half (64.6%) believe job opportunities will increase. Those who viewed the law favorably were significantly more likely to score better on a knowledge scale related to the ACA. HES understand publicized provisions but are uncertain about common myths and specific provisions related to Title IV, "Prevention of Chronic Disease and Improving Public Health." Directed and continuing education to HES regarding the ACA is warranted. PMID:26272884

  2. Hybrid entanglement concentration assisted with single coherent state

    NASA Astrophysics Data System (ADS)

    Rui, Guo; Lan, Zhou; Shi-Pu, Gu; Xing-Fu, Wang; Yu-Bo, Sheng

    2016-03-01

    Hybrid entangled state (HES) is a new type of entanglement, which combines the advantages of an entangled polarization state and an entangled coherent state. HES is widely discussed in the applications of quantum communication and computation. In this paper, we propose three entanglement concentration protocols (ECPs) for Bell-type HES, W-type HES, and cluster-type HES, respectively. After performing these ECPs, we can obtain the maximally entangled HES with some success probability. All the ECPs exploit the single coherent state to complete the concentration. These protocols are based on the linear optics, which are feasible in future experiments. Project supported by the National Natural Science Foundation of China (Grant Nos. 11474168 and 61401222), the Natural Science Foundation of Jiangsu Province, China (Grant No. BK20151502), the Qing Lan Project in Jiangsu Province, China, the Natural Science Foundation of Jiangsu Higher Education Institutions, China (Grant No. 15KJA120002), and the Priority Academic Development Program of Jiangsu Higher Education Institutions, China.

  3. Hypoglycemic effect of a hot-water extract from defatted sesame (Sesamum indicum L.) seed on the blood glucose level in genetically diabetic KK-Ay mice.

    PubMed

    Takeuchi, H; Mooi, L Y; Inagaki, Y; He, P

    2001-10-01

    Genetically diabetic (type II) KK-Ay mice, male and 5 weeks of age, were divided into one group of 12 mice that were fed on a basal (BAS) diet and three groups of 6 mice each that were fed on the test diets for 4 weeks. Each test diet contained 4.0% of the hot-water extract (HES) from defatted sesame (Sesamum indicum L.) seed, 1.4% of the water eluent fraction (WFH) of HES or 0.7% of the methanol eluent fraction (MFH) of HES from a glass column packed with HP-20 resin. At the end of the feeding period, the BAS group was divided into the MAL and MALH groups which were respectively force-fed with 1 ml per mouse of a 20% maltose solution in water with or without 4.0% HES. The plasma glucose concentration and amount of urinary excreted glucose were lower from the HES and MFH diets than from the BAS and WFH diets. The levels of plasma glucose and serum insulin were lower in the MALH group than in the MAL group. These results indicate that HES and MFH had a reductive effect on the plasma glucose concentration of KK-Ay mice, and this effect is suggested to have been caused by the delayed glucose absorption. PMID:11758931

  4. The effects of haemodilution with hydroxyethyl starch 130/0.4 solution on coagulation as assessed by thromboelastography and platelet receptor function studies in vitro.

    PubMed

    Williams, P; Yang, K; Kershaw, G; Wong, G; Dunkley, S; Kam, P C A

    2015-11-01

    This study evaluated the effects of haemodilution with either 6% hydroxyethyl starch (HES) 130/0.4 (Voluven(®)) or 0.9% normal saline (NS) on blood coagulation in vitro. Haemodilution with 6% HES 130/0.4 impaired coagulation, as indicated by the changes in thromboelastographic parameters k-time, α-angle and maximum amplitude. Light transmission aggregometry and multiple electrode aggregometry demonstrated that impaired platelet receptor function occurred only at high levels of haemodilution (40%) with both fluids, but there was no significant difference between the two fluids (P=0.05). The thromboelastographic functional fibrinogen assay showed that the fibrinogen component of clot strength was significantly impaired with haemodilution with HES 130/0.4 compared with haemodilution with NS (whole blood [14.4 ± 4.6 mm] versus 40% HES dilution [3.7 ± 1.9], [P=0.001]; versus 40% NS dilution [10.4 ± 4.6], [P=0.129]). These findings suggest that there is little difference between HES or NS in relation to coagulation or platelet function during minor or moderate haemodilution, but at high levels of haemodilution with HES, fibrinogen activity is more impaired compared with NS. PMID:26603798

  5. Rates of self-harm presenting to general hospitals: a comparison of data from the Multicentre Study of Self-Harm in England and Hospital Episode Statistics

    PubMed Central

    Turnbull, Pauline; Hawton, Keith; Geulayov, Galit; Waters, Keith; Ness, Jennifer; Townsend, Ellen; Khundakar, Kazem; Kapur, Nav

    2016-01-01

    Objective Rates of hospital presentation for self-harm in England were compared using different national and local data sources. Design The study was descriptive and compared bespoke data collection methods for recording self-harm presentations to hospital with routinely collected hospital data. Setting Local area data on self-harm from the 3 centres of the Multicentre Study of Self-harm in England (Oxford, Manchester and Derby) were used along with national and local routinely collected data on self-harm admissions and emergency department attendances from Hospital Episode Statistics (HES). Primary outcome Rate ratios were calculated to compare rates of self-harm generated using different data sources nationally and locally (between 2010 and 2012) and rates of hospital presentations for self-harm were plotted over time (between 2003 and 2012), based on different data sources. Results The total number of self-harm episodes between 2010 and 2012 was 13 547 based on Multicentre Study data, 9600 based on HES emergency department data and 8096 based on HES admission data. Nationally, routine HES data underestimated overall rates of self-harm by approximately 60% compared with rates based on Multicentre Study data (rate ratio for HES emergency department data, 0.41 (95% CI 0.35 to 0.49); rate ratio for HES admission data, 0.42 (95% CI 0.36 to 0.49)). Direct local area comparisons confirmed an overall underascertainment in the HES data, although the difference varied between centres. There was a general increase in self-harm over time according to HES data which contrasted with a fall and then a rise in the Multicentre Study data. Conclusions There was a consistent underestimation of presentations for self-harm recorded by HES emergency department data, and fluctuations in year-on-year figures. HES admission data appeared more reliable but missed non-admitted episodes. Routinely collected data may miss important trends in self-harm and cannot be used in isolation as

  6. Efficient derivation of Chinese human embryonic stem cell lines from frozen embryos.

    PubMed

    Li, Chunliang; Yang, Ying; Lu, Xiaowei; Sun, Yijuan; Gu, Junjie; Feng, Yun; Jin, Ying

    2010-04-01

    Human embryonic stem (hES) cells are pluripotent cells derived from the inner cell mass of blastocysts. Their unique properties of self-renewal and pluripotency make them an attractive tool for basic research as well as a potential cell resource for therapy. However, each hES cell line demonstrates different identity. It is desirable to obtain more fully characterized hES cell lines with newly developed technologies associated with hES cell culture. Here, we report our experience of efficient derivation of three new Chinese hES cell lines (SHhES2, SHhES3, and SHhES4) from in vitro fertilization discarded embryos donated by women with polycystic ovary syndrome. These cell lines were derived under conditions minimizing exposure to animal components and maintained at an undifferentiated state for long-term culture. They retained a normal karyotype and expressed ALP, OCT4, SOX2, SSEA-4, TRA-1-60 and TRA-1-81. RT-PCR analysis also revealed high expression levels of pluripotency markers such as OCT4, LEFTY A, SOX2, TDGF-1, THY1, FGF4, NANOG, and REX1. When suspended in low-attachment culture dishes, embryoid bodies formed and were comprised of various differentiated cell types from all three embryonic germ layers. However, well-shaped teratomas were only harvested from line SHhES2, not from SHhES3 and SHhES4, indicating that the differentiation ability in vivo differs among the three cell lines. Collectively, the three new hES cell lines were established and fully characterized. The effort paves the way toward generating hES cell lines without contamination by animal components. All of these cell lines are available by contact Ying Jin at yjin@sibs.ac.cn. PMID:20186511

  7. Enhancement of Cell Recovery for Dissociated Human Embryonic Stem Cells After Cryopreservation

    PubMed Central

    Xu, Xia; Cowley, Sally; Flaim, Christopher J; James, William; Seymour, Lenard W; Cui, Zhanfeng

    2010-01-01

    Due to widespread applications of human embryonic stem (hES) cells, it is essential to establish effective protocols for cryopreservation and subsequent culture of hES cells to improve cell recovery. We have developed a new protocol for cryopreservation of dissociated hES cells and subsequent culture. We examined the effects of new formula of freezing solution containing 7.5% dimethylsulfoxide (DMSO) (v/v %) and 2.5% polyethylene glycol (PEG) (w/v %) on cell survival and recovery of hES cells after cryopreservation, and further investigated the role of the combination of Rho-associated kinase (ROCK) inhibitor and p53 inhibitor on cell recovery during the subsequent culture. Compared with the conventional slow-freezing method which uses 10% DMSO as a freezing solution and then cultured in the presence of ROCK inhibitor at the first day of culture, we found out that hES cell recovery was significantly enhanced by around 30 % (P < 0.05) by the new freezing solution. Moreover, at the first day of post-thaw culture, the presence of 10 μM ROCK inhibitor (Y-27632) and 1 μM pifithrin-μ together further significantly improved cell recovery by around 20% (P < 0.05) either for feeder-dependent or feeder-independent culture. hES cells remained their undifferentiated status after using this novel protocol for cryopreservation and subsequent culture. Furthermore, this protocol is a scalable cryopreservation method for handling large quantities of hES cells. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010 PMID:20014103

  8. From Observation to Information: Data-Driven Understanding of on Farm Yield Variation

    PubMed Central

    Jiménez, Daniel; Dorado, Hugo; Cock, James; Prager, Steven D.; Delerce, Sylvain; Grillon, Alexandre; Andrade Bejarano, Mercedes; Benavides, Hector; Jarvis, Andy

    2016-01-01

    Agriculture research uses “recommendation domains” to develop and transfer crop management practices adapted to specific contexts. The scale of recommendation domains is large when compared to individual production sites and often encompasses less environmental variation than farmers manage. Farmers constantly observe crop response to management practices at a field scale. These observations are of little use for other farms if the site and the weather are not described. The value of information obtained from farmers’ experiences and controlled experiments is enhanced when the circumstances under which it was generated are characterized within the conceptual framework of a recommendation domain, this latter defined by Non-Controllable Factors (NCFs). Controllable Factors (CFs) refer to those which farmers manage. Using a combination of expert guidance and a multi-stage analytic process, we evaluated the interplay of CFs and NCFs on plantain productivity in farmers’ fields. Data were obtained from multiple sources, including farmers. Experts identified candidate variables likely to influence yields. The influence of the candidate variables on yields was tested through conditional forests analysis. Factor analysis then clustered harvests produced under similar NCFs, into Homologous Events (HEs). The relationship between NCFs, CFs and productivity in intercropped plantain were analyzed with mixed models. Inclusion of HEs increased the explanatory power of models. Low median yields in monocropping coupled with the occasional high yields within most HEs indicated that most of these farmers were not using practices that exploited the yield potential of those HEs. Varieties grown by farmers were associated with particular HEs. This indicates that farmers do adapt their management to the particular conditions of their HEs. Our observations confirm that the definition of HEs as recommendation domains at a small-scale is valid, and that the effectiveness of distinct

  9. From Observation to Information: Data-Driven Understanding of on Farm Yield Variation.

    PubMed

    Jiménez, Daniel; Dorado, Hugo; Cock, James; Prager, Steven D; Delerce, Sylvain; Grillon, Alexandre; Andrade Bejarano, Mercedes; Benavides, Hector; Jarvis, Andy

    2016-01-01

    Agriculture research uses "recommendation domains" to develop and transfer crop management practices adapted to specific contexts. The scale of recommendation domains is large when compared to individual production sites and often encompasses less environmental variation than farmers manage. Farmers constantly observe crop response to management practices at a field scale. These observations are of little use for other farms if the site and the weather are not described. The value of information obtained from farmers' experiences and controlled experiments is enhanced when the circumstances under which it was generated are characterized within the conceptual framework of a recommendation domain, this latter defined by Non-Controllable Factors (NCFs). Controllable Factors (CFs) refer to those which farmers manage. Using a combination of expert guidance and a multi-stage analytic process, we evaluated the interplay of CFs and NCFs on plantain productivity in farmers' fields. Data were obtained from multiple sources, including farmers. Experts identified candidate variables likely to influence yields. The influence of the candidate variables on yields was tested through conditional forests analysis. Factor analysis then clustered harvests produced under similar NCFs, into Homologous Events (HEs). The relationship between NCFs, CFs and productivity in intercropped plantain were analyzed with mixed models. Inclusion of HEs increased the explanatory power of models. Low median yields in monocropping coupled with the occasional high yields within most HEs indicated that most of these farmers were not using practices that exploited the yield potential of those HEs. Varieties grown by farmers were associated with particular HEs. This indicates that farmers do adapt their management to the particular conditions of their HEs. Our observations confirm that the definition of HEs as recommendation domains at a small-scale is valid, and that the effectiveness of distinct

  10. Mixing properties of lyophilized protein systems: a spectroscopic and calorimetric study.

    PubMed

    Katayama, Derrick S; Carpenter, John F; Menard, Kevin P; Manning, Mark Cornell; Randolph, Theodore W

    2009-09-01

    The purpose of this study was to investigate the solid-state properties of lyophilized formulations of protein (ribonuclease A) containing sucrose or trehalose across a wide range of compositions, both in the presence or absence of hydroxyethylstarch (HES). Infrared spectroscopy reveals that the protein forms hydrogen bonds to sugars (sucrose or trehalose) as water is removed from the sample. The strength and/or number of hydrogen bonds in dried samples increase as the weight fraction of sugar increases. Significant deviations of glass transition temperatures (T(g)'s) from those predicted by free volume theory are seen in both protein-sugar systems. The behavior can be explained by formation of protein-sugar hydrogen bonds at the expense of self-interactions between the sugars. Attractive interactions between lyophilized ribonuclease A and HES were detected spectroscopically and from thermodynamic analysis of T(g) values, contrary to the view that HES is sterically hindered from interacting with the protein surface. Sucrose-HES interactions were much less favorable than trehalose-HES interactions, suggesting that phase separation in sugar/HES/protein mixtures would be more likely in the presence of sucrose than trehalose. Finally, the thermodynamics of mixing were investigated using differential scanning calorimetry (DSC) providing some of the first data for such solid protein sugar formulations with and without HES. In nearly all samples, positive excess enthalpy, excess entropy and excess free energy were observed, with the excess free energy being greater for samples containing sucrose rather than trehalose. Analysis of Flory-Huggins chi parameters suggests that phase separation between protein and excipients may be thermodynamically favored in these dried solid preparations. PMID:18623211

  11. In vitro and in vivo analyses of human embryonic stem cell-derived dopamine neurons.

    PubMed

    Park, Chang-Hwan; Minn, Yang-Ki; Lee, Ji-Yeon; Choi, Dong Ho; Chang, Mi-Yoon; Shim, Jae-Won; Ko, Ji-Yun; Koh, Hyun-Chul; Kang, Min Jeong; Kang, Jin Sun; Rhie, Duck-Joo; Lee, Yong-Sung; Son, Hyeon; Moon, Shin Yong; Kim, Kwang-Soo; Lee, Sang-Hun

    2005-03-01

    Human embryonic stem (hES) cells, due to their capacity of multipotency and self-renewal, may serve as a valuable experimental tool for human developmental biology and may provide an unlimited cell source for cell replacement therapy. The purpose of this study was to assess the developmental potential of hES cells to replace the selectively lost midbrain dopamine (DA) neurons in Parkinson's disease. Here, we report the development of an in vitro differentiation protocol to derive an enriched population of midbrain DA neurons from hES cells. Neural induction of hES cells co-cultured with stromal cells, followed by expansion of the resulting neural precursor cells, efficiently generated DA neurons with concomitant expression of transcriptional factors related to midbrain DA development, such as Pax2, En1 (Engrailed-1), Nurr1, and Lmx1b. Using our procedure, the majority of differentiated hES cells (> 95%) contained neuronal or neural precursor markers and a high percentage (> 40%) of TuJ1+ neurons was tyrosine hydroxylase (TH)+, while none of them expressed the undifferentiated ES cell marker, Oct 3/4. Furthermore, hES cell-derived DA neurons demonstrated functionality in vitro, releasing DA in response to KCl-induced depolarization and reuptake of DA. Finally, transplantation of hES-derived DA neurons into the striatum of hemi-parkinsonian rats failed to result in improvement of their behavioral deficits as determined by amphetamine-induced rotation and step-adjustment. Immunohistochemical analyses of grafted brains revealed that abundant hES-derived cells (human nuclei+ cells) survived in the grafts, but none of them were TH+. Therefore, unlike those from mouse ES cells, hES cell-derived DA neurons either do not survive or their DA phenotype is unstable when grafted into rodent brains. PMID:15715675

  12. Enhancement of cell recovery for dissociated human embryonic stem cells after cryopreservation.

    PubMed

    Xu, Xia; Cowley, Sally; Flaim, Christopher J; James, William; Seymour, Lenard W; Cui, Zhanfeng

    2010-01-01

    Due to widespread applications of human embryonic stem (hES) cells, it is essential to establish effective protocols for cryopreservation and subsequent culture of hES cells to improve cell recovery. We have developed a new protocol for cryopreservation of dissociated hES cells and subsequent culture. We examined the effects of new formula of freezing solution containing 7.5% dimethylsulfoxide (DMSO) (v/v %) and 2.5% polyethylene glycol (PEG) (w/v %) on cell survival and recovery of hES cells after cryopreservation, and further investigated the role of the combination of Rho-associated kinase (ROCK) inhibitor and p53 inhibitor on cell recovery during the subsequent culture. Compared with the conventional slow-freezing method which uses 10% DMSO as a freezing solution and then cultured in the presence of ROCK inhibitor at the first day of culture, we found out that hES cell recovery was significantly enhanced by around 30 % (P < 0.05) by the new freezing solution. Moreover, at the first day of post-thaw culture, the presence of 10 microM ROCK inhibitor (Y-27632) and 1 microM pifithrin-mu together further significantly improved cell recovery by around 20% (P < 0.05) either for feeder-dependent or feeder-independent culture. hES cells remained their undifferentiated status after using this novel protocol for cryopreservation and subsequent culture. Furthermore, this protocol is a scalable cryopreservation method for handling large quantities of hES cells. PMID:20014103

  13. The stellar content of the Hamburg/ESO survey. IV. Selection of candidate metal-poor stars

    NASA Astrophysics Data System (ADS)

    Christlieb, N.; Schörck, T.; Frebel, A.; Beers, T. C.; Wisotzki, L.; Reimers, D.

    2008-06-01

    We present the quantitative methods used for selecting candidate metal-poor stars in the Hamburg/ESO objective-prism survey (HES). The selection is based on the strength of the Ca II K line, B-V colors (both measured directly from the digital HES spectra), as well as J-K colors from the 2 Micron All Sky Survey. The KP index for Ca II K can be measured from the HES spectra with an accuracy of 1.0 Å, and a calibration of the HES B-V colors, using CCD photometry, yields a 1-σ uncertainty of 0.07 mag for stars in the color range 0.3 < B-V < 1.4. These accuracies make it possible to reliably reject stars with [Fe/H] > -2.0 without sacrificing completeness at the lowest metallicities. A test of the selection using 1121 stars of the HK survey of Beers, Preston, and Shectman present on HES plates suggests that the completeness at [Fe/H] < -3.5 is close to 100% and that, at the same time, the contamination of the candidate sample with false positives is low: 50% of all stars with [Fe/H] > -2.5 and 97% of all stars with [Fe/H] > -2.0 are rejected. The selection was applied to 379 HES fields, covering a nominal area of 8853 deg2 of the southern high Galactic latitude sky. The candidate sample consists of 20 271 stars in the magnitude range 10 ≲ B ≲ 18. A comparison of the magnitude distribution with that of the HK survey shows that the magnitude limit of the HES sample is about 2 mag fainter. Taking the overlap of the sky areas covered by both surveys into account, it follows that the survey volume for metal-poor stars has been increased by the HES by about a factor of 10 with respect to the HK survey. We have already identified several very rare objects with the HES, including, e.g., the three most heavy-element deficient stars currently known. Based on observations collected at the European Southern Observatory, Chile (Proposal ID 145.B-0009). Tables A.1 and A.2 are only available in electronic form at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or

  14. Dynamic Analysis of Hybrid Energy Systems under Flexible Operation and Variable Renewable Generation -- Part I: Dynamic Performance Analysis and Part II: Dynamic Cost

    SciTech Connect

    Humberto E. Garcia; Amit Mohanty; Wen-Chiao Lin; Robert S. Cherry

    2013-04-01

    Dynamic analysis of hybrid energy systems (HES) under flexible operation and variable renewable generation is considered in order to better understand various challenges and opportunities associated with the high system variability arising from the integration of renewable energy into the power grid. Unique consequences are addressed by devising advanced HES solutions in which multiple forms of energy commodities, such as electricity and chemical products, may be exchanged. Dynamic models of various unit operations are developed and integrated within two different HES options. One HES option, termed traditional, produces electricity only and consists of a primary heat generator (PHG) (e.g., a small modular reactor), a steam turbine generator, a wind farm, and a battery storage. The other HES option, termed advanced, includes not only the components present in the traditional option but also a chemical plant complex to repurpose excess energy for non-electricity services, such as for the production of chemical goods (e.g., transportation fuel). In either case, a given HES is connected to the power grid at a point of common coupling and requested to deliver a certain electricity generation profile as dictated by a regional power grid operator based on a predicted demand curve. Dynamic analysis of these highly-coupled HES are performed to identify their key dynamical properties and limitations and to prescribe solutions for best managing and mitigating the high variability introduced from incorporating renewable energy into the energy mix. A comparative dynamic cost analysis is also conducted to determine best HES options. The cost function includes a set of metrics for computing fixed costs, such as fixed operations and maintenance (O&M) and overnight capital costs, and also variable operational costs, such as cost of variability, variable O&M cost, and cost of environmental impact, together with revenues. Assuming different options for implementing PHG (e

  15. Evidence for bystander signalling between human trophoblast cells and human embryonic stem cells.

    PubMed

    Jones, Anna J; Gokhale, Paul J; Allison, Thomas F; Sampson, Barry; Athwal, Sharan; Grant, Simon; Andrews, Peter W; Allen, Nicholas D; Case, C Patrick

    2015-01-01

    Maternal exposure during pregnancy to toxins can occasionally lead to miscarriage and malformation. It is currently thought that toxins pass through the placental barrier, albeit bi-layered in the first trimester, and damage the fetus directly, albeit at low concentration. Here we examined the responses of human embryonic stem (hES) cells in tissue culture to two metals at low concentration. We compared direct exposures with indirect exposures across a bi-layered model of the placenta cell barrier. Direct exposure caused increased DNA damage without apoptosis or a loss of cell number but with some evidence of altered differentiation. Indirect exposure caused increased DNA damage and apoptosis but without loss of pluripotency. This was not caused by metal ions passing through the barrier. Instead the hES cells responded to signalling molecules (including TNF-α) secreted by the barrier cells. This mechanism was dependent on connexin 43 mediated intercellular 'bystander signalling' both within and between the trophoblast barrier and the hES colonies. These results highlight key differences between direct and indirect exposure of hES cells across a trophoblast barrier to metal toxins. It offers a theoretical possibility that an indirectly mediated toxicity of hES cells might have biological relevance to fetal development. PMID:26170169

  16. Evidence for bystander signalling between human trophoblast cells and human embryonic stem cells

    PubMed Central

    Jones, Anna J; Gokhale, Paul J; Allison, Thomas F; Sampson, Barry; Athwal, Sharan; Grant, Simon; Andrews, Peter W; Allen, Nicholas D; Patrick Case, C

    2015-01-01

    Maternal exposure during pregnancy to toxins can occasionally lead to miscarriage and malformation. It is currently thought that toxins pass through the placental barrier, albeit bi-layered in the first trimester, and damage the fetus directly, albeit at low concentration. Here we examined the responses of human embryonic stem (hES) cells in tissue culture to two metals at low concentration. We compared direct exposures with indirect exposures across a bi-layered model of the placenta cell barrier. Direct exposure caused increased DNA damage without apoptosis or a loss of cell number but with some evidence of altered differentiation. Indirect exposure caused increased DNA damage and apoptosis but without loss of pluripotency. This was not caused by metal ions passing through the barrier. Instead the hES cells responded to signalling molecules (including TNF-α) secreted by the barrier cells. This mechanism was dependent on connexin 43 mediated intercellular ‘bystander signalling’ both within and between the trophoblast barrier and the hES colonies. These results highlight key differences between direct and indirect exposure of hES cells across a trophoblast barrier to metal toxins. It offers a theoretical possibility that an indirectly mediated toxicity of hES cells might have biological relevance to fetal development. PMID:26170169

  17. Profiling TRA-1-81 antigen distribution on a human embryonic stem cell

    SciTech Connect

    Qiu Dengli; Xiang Jialing; Li Zhaoxia; Krishnamoorthy, Aparna; Chen Liaohai; Wang Rong

    2008-05-02

    Human embryonic stem (hES) cells hold great promise in regenerative medicine. Although hES cells have unlimited self-renewal potential, they tend to differentiate spontaneously in culture. TRA-1-81 is a biomarker of undifferentiated hES cells. Quantitative characterization of TRA-1-81 expression level in a single cell helps capture the 'turn-on' signal and understand the mechanism of early differentiation. Here, we report on our examination of TRA-1-81 distribution and association on a hES cell membrane using an atomic force microscope (AFM). Our results suggest that aggregated distribution of TRA-1-81 antigen is characteristic for undifferentiated hES cells. We also evaluated the TRA-1-81 expression level at {approx}17,800 epitopes and {approx}700 epitopes per cell on an undifferentiated cell and a spontaneously differentiated cell, respectively. The method in this study can be adapted in examining other surface proteins on various cell types, thus providing a general tool for investigating protein distribution and association at the single cell level.

  18. A Defined, Controlled Culture System for Primary Bovine Chromaffin Progenitors Reveals Novel Biomarkers and Modulators

    PubMed Central

    Masjkur, Jimmy; Levenfus, Ian; Lange, Sven; Arps-Forker, Carina; Poser, Steve; Qin, Nan; Vukicevic, Vladimir; Chavakis, Triantafyllos; Eisenhofer, Graeme; Bornstein, Stefan R.; Ehrhart-Bornstein, Monika

    2014-01-01

    We present a method to efficiently culture primary chromaffin progenitors from the adult bovine adrenal medulla in a defined, serum-free monolayer system. Tissue is dissociated and plated for expansion under support by the mitogen basic fibroblast growth factor (bFGF). The cultures, although not homogenous, contain a subpopulation of cells expressing the neural stem cell marker Hes3 that also propagate. In addition, Hes3 is also expressed in the adult adrenal medulla from where the tissue is taken. Differentiation is induced by bFGF withdrawal and switching to Neurobasal medium containing B27. Following differentiation, Hes3 expression is lost, and cells acquire morphologies and biomarker expression patterns of chromaffin cells and dopaminergic neurons. We tested the effect of different treatments that we previously showed regulate Hes3 expression and cell number in cultures of fetal and adult rodent neural stem cells. Treatment of the cultures with a combination of Delta4, Angiopoietin2, and a Janus kinase inhibitor increases cell number during the expansion phase without significantly affecting catecholamine content levels. Treatment with cholera toxin does not significantly affect cell number but reduces the ratio of epinephrine to norepinephrine content and increases the dopamine content relative to total catecholamines. These data suggest that this defined culture system can be used for target identification in drug discovery programs and that the transcription factor Hes3 may serve as a new biomarker of putative adrenomedullary chromaffin progenitor cells. PMID:24855275

  19. EMMPRIN Is Secreted by Human Uterine Epithelial Cells in Microvesicles and Stimulates Metalloproteinase Production by Human Uterine Fibroblast Cells

    PubMed Central

    Dayger, C. A.; Mehrotra, P.; Belton, R. J.; Nowak, R. A.

    2012-01-01

    Endometrial remodeling is a physiological process involved in the gynecological disease, endometriosis. Tissue remodeling is directed by uterine fibroblast production of matrix metalloproteinases (MMPs). Several MMPs are regulated directly by the protein extracellular matrix metalloproteinase inducer (EMMPRIN) and also by proinflammatory cytokines such as interleukin (IL)1-α/β. We hypothesized that human uterine epithelial cells (HESs) secrete intact EMMPRIN to stimulate MMPs. Microvesicles from HES cell-conditioned medium (CM) expressed intact EMMPRIN protein. Treatment of HES cells with estradiol or phorbyl 12-myristate-13-acetate increased the release of EMMPRIN-containing microvesicles. The HES CM stimulated MMP-1, -2, and -3 messenger RNA levels in human uterine fibroblasts (HUFs) and EMMPRIN immunodepletion from HES-cell concentrated CM reduced MMP stimulation (P < .05). Treatment of HUF cells with low concentrations of IL-1β/α stimulated MMP production (P < .05). These results indicate that HES cells regulate MMP production by HUF cells by secretion of EMMPRIN, in response to ovarian hormones, proinflammatory cytokines as well as activation of protein kinase C. PMID:22729071

  20. Lovastatin Decreases the Expression of CD133 and Influences the Differentiation Potential of Human Embryonic Stem Cells

    PubMed Central

    Kallas-Kivi, Ade

    2016-01-01

    The lipophilic statin lovastatin decreases cholesterol synthesis and is a safe and effective treatment for the prevention of cardiovascular diseases. Growing evidence points at antitumor potential of lovastatin. Therefore, understanding the molecular mechanism of lovastatin function in different cell types is critical to effective therapy design. In this study, we investigated the effects of lovastatin on the differentiation potential of human embryonic stem (hES) cells (H9 cell line). Multiparameter flow cytometric assay was used to detect changes in the expression of transcription factors characteristic of hES cells. We found that lovastatin treatment delayed NANOG downregulation during ectodermal and endodermal differentiation. Likewise, expression of ectodermal (SOX1 and OTX2) and endodermal (GATA4 and FOXA2) markers was higher in treated cells. Exposure of hES cells to lovastatin led to a minor decrease in the expression of SSEA-3 and a significant reduction in CD133 expression. Treated cells also formed fewer embryoid bodies than control cells. By analyzing hES with and without CD133, we discovered that CD133 expression is required for proper formation of embryoid bodies. In conclusion, lovastatin reduced the heterogeneity of hES cells and impaired their differentiation potential. PMID:27247576

  1. Topology and dynamics of the zebrafish segmentation clock core circuit.

    PubMed

    Schröter, Christian; Ares, Saúl; Morelli, Luis G; Isakova, Alina; Hens, Korneel; Soroldoni, Daniele; Gajewski, Martin; Jülicher, Frank; Maerkl, Sebastian J; Deplancke, Bart; Oates, Andrew C

    2012-01-01

    During vertebrate embryogenesis, the rhythmic and sequential segmentation of the body axis is regulated by an oscillating genetic network termed the segmentation clock. We describe a new dynamic model for the core pace-making circuit of the zebrafish segmentation clock based on a systematic biochemical investigation of the network's topology and precise measurements of somitogenesis dynamics in novel genetic mutants. We show that the core pace-making circuit consists of two distinct negative feedback loops, one with Her1 homodimers and the other with Her7:Hes6 heterodimers, operating in parallel. To explain the observed single and double mutant phenotypes of her1, her7, and hes6 mutant embryos in our dynamic model, we postulate that the availability and effective stability of the dimers with DNA binding activity is controlled in a "dimer cloud" that contains all possible dimeric combinations between the three factors. This feature of our model predicts that Hes6 protein levels should oscillate despite constant hes6 mRNA production, which we confirm experimentally using novel Hes6 antibodies. The control of the circuit's dynamics by a population of dimers with and without DNA binding activity is a new principle for the segmentation clock and may be relevant to other biological clocks and transcriptional regulatory networks. PMID:22911291

  2. Hydroxyethyl starch-10-hydroxy camptothecin conjugate: synthesis, pharmacokinetics, cytotoxicity and pharmacodynamics research.

    PubMed

    Li, Guofei; Cai, Cuifang; Qi, Yingjie; Tang, Xing

    2016-01-01

    10-hydroxy camptothecin (10-HCPT) is an antitumor agent effective in the treatment of several solid tumors but its use is hampered by poor water solubility, low lactone stability, short plasma half-life and dose-limiting toxicity. In this study, 10-HCPT-hydroxyethyl starch (HES) conjugate was prepared to overcome these limits of 10-HCPT. The solubility of 10-HCPT conjugate was 0.72 mg/ml, about 100 times to free 10-HCPT. The 10-HCPT conjugate showed good sustained release effect in phosphate-buffered saline (PBS), rat plasma and liver homogenate. Meanwhile, 10-HCPT-HES conjugate achieved much lower IC50 and higher cytotoxicity effects than the free 10-HCPT on Hep-3B liver cancer cells. The pharmacokinetics results of 10-HCPT-HES conjugate demonstrated that the biological half-life of 10-HCPT was increased from 10 min to 4.38 h and the bioavailability was 40 times higher than the commercial 10-HCPT injection. The pharmacodynamics results indicated that 10-HCPT-HES conjugate had a better antitumor efficiency against nude mouse with Hep-3B tumor than the commercial 10-HCPT injection, and the inhibition ratio of tumor was 83.9 and 27.8%, respectively, at the same administration dosage. These findings suggest that 10-HCPT-HES conjugate is a promising drug delivery system providing improved long circulating effect, greater stability and better antitumor effect. PMID:24833272

  3. Hydroxyethyl starch conjugates for improving the stability, pharmacokinetic behavior and antitumor activity of 10-hydroxy camptothecin.

    PubMed

    Li, Guofei; Li, Yan; Tang, Yilin; Zhang, Yu; Zhang, Yan; Yin, Tian; Xu, Hui; Cai, Cuifang; Tang, Xing

    2014-08-25

    10-Hydroxy camptothecin (10-HCPT)-hydroxyethyl starch (HES) conjugates were prepared to improve the water solubility, prolong the half-life in plasma and increase the antitumor efficacy of 10-HCPT, and the structures of the conjugates were confirmed by NMR and infrared spectroscopy. The 10-HCPT conjugates showed good sustained release effect in phosphate-buffered saline (PBS), rat plasma and liver homogenate. Meanwhile, 10-HCPT-HES conjugates achieved much lower IC50 and higher cytotoxicity effects than the free 10-HCPT on Hep-3B and SMMC-7721 cell lines. The pharmacokinetics results of 10-HCPT-HES conjugates demonstrated that the biological half-life of 10-HCPT was increased from 10 min to 2.94 h and 3.76 h, respectively, in comparison with the commercial 10-HCPT injection. The pharmacodynamics results indicated that 10-HCPT-HES conjugate had a better antitumor efficiency against nude mouse with Hep-3B tumor than the commercial 10-HCPT injection, and the inhibition ratio of tumor was 78.3% and 31.5%, respectively, at the dose of 1.0 mg/kg. These findings suggest that 10-HCPT-HES conjugate is a promising drug delivery system providing improved long circulating effect, greater stability and better antitumor effect. PMID:24861941

  4. Hesperidin Inhibits Inflammatory Response Induced by Aeromonas hydrophila Infection and Alters CD4+/CD8+ T Cell Ratio

    PubMed Central

    Abuelsaad, Abdelaziz S. A.; Allam, Gamal; Al-Solumani, Adnan A. A.

    2014-01-01

    Background. Aeromonas hydrophila is an opportunistic bacterial pathogen that is associated with a number of human diseases. Hesperidin (HES) has been reported to exert antioxidant and anti-inflammatory activities. Objectives. The aim of this study was to investigate the potential effect of HES treatment on inflammatory response induced by A. hydrophila infection in murine. Methods. A. hydrophila-infected mice were treated with HES at 250 mg/kg b.wt./week for 4 consecutive weeks. Phagocytosis, reactive oxygen species production, CD4+/CD8+ T cell ratio, and CD14 expression on intestinal infiltrating monocytes were evaluated. The expression of E-selectin and intercellular adhesion molecule 1 on stimulated HUVECs and RAW macrophage was evaluated. Results. Percentage of CD4+ T cells in the intestinal tissues of infected treated mice was highly significantly increased; however, phagocytic index, ROS production, CD8+ T cells percentage, and CD14 expression on monocytes were significantly reduced. On the other hand, HES significantly inhibited A-LPS- and A-ECP-induced E-selectin and ICAM-1 expression on HUVECs and ICAM-1 expression on RAW macrophage. Conclusion. Present data indicated that HES has a potential role in the suppression of inflammatory response induced by A. hydrophila toxins through downmodulation of ROS production and CD14 and adhesion molecules expression, as well as increase of CD4+/CD8+ cell ratio. PMID:24891765

  5. US hydropower resource assessment for Colorado

    SciTech Connect

    Francfort, J.E.

    1994-05-01

    The US Department of Energy is developing an estimate of the hydropower development potential in this country. Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE, menu-driven software application. HES allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the State of Colorado.

  6. U.S. Hydropower Resource Assessment Final Report

    SciTech Connect

    Conner, Alison M.; Francfort, James E.; Rinehart, Ben N.

    1998-12-01

    To provide a more accurate assessment of the domestic undeveloped hydropower capacity, the U.S. Department of Energy’s Hydropower Program developed a computer model, Hydropower Evaluation Software (HES). HES allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental, legal, and institutional attributes present, and generate reports based on these suitability factors. This report describes the development of HES, its data requirements, and its application to each state assessment; in addition, it summarizes the data derivation process and data for the states. Modeling of the undeveloped hydropower resources in the United States, based on environmental, legal, and institutional constraints, has identified 5,677 sites that have a total undeveloped capacity of about 30,000 megawatts.

  7. U.S. Hydropower Resource Assessment Final Report

    SciTech Connect

    A. M. Conner; J. E. Francfort; B. N. Rinehart

    1998-12-01

    To provide a more accurate assessment of the domestic undeveloped hydropower capacity, the US Department of Energy's Hydropower Program developed a computer model, Hydropower Evaluation Software (HES). HES allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental, legal, and institutional attributes present, and generate reports based on these suitability factors. This report describes the development of HES, its data requirements, and its application to each state assessment; in addition, it summarizes the data derivation process and data for the states. Modeling of the undeveloped hydropower resources in the US, based on environmental, legal, and institutional constraints, has identified 5,677 sites that have a total undeveloped capacity of about 30,000 megawatts.

  8. Nuclear Hybrid Energy Systems - Regional Studies. West Texas and Northeastern Arizona

    SciTech Connect

    Garcia, Humberto E.; Chen, Jun; Kim, Jong S.; McKellar, Michael G.; Deason, Wesley R.; Vilim, Richard B.; Bragg-Sitton, Shannon M.; Boardman, Richard D.

    2015-04-01

    The primary objective of this study is to conduct a preliminary dynamic analysis of two realistic hybrid energy systems (HES) including a nuclear reactor as the main baseload heat generator (denoted as nuclear HES or nuclear hybrid energy systems [NHES]) and to assess the local (e.g., HES owners) and system (e.g., the electric grid) benefits attainable by the application of NHES in scenarios with multiple commodity production and high penetration of renewable energy. It is performed for regional cases - not generic examples - based on available resources, existing infrastructure, and markets within the selected regions. This study also briefly addresses the computational capabilities developed to conduct such analyses, reviews technical gaps, and suggests some research paths forward.

  9. Nuclear Hybrid Energy Systems Regional Studies: West Texas & Northeastern Arizona

    SciTech Connect

    Garcia, Humberto E.; Chen, Jun; Kim, Jong Suk; McKellar, Michael George; Deason, Wesley R; Richard B. Vilim; Bragg-Sitton, Shannon M.; Boardman, Richard D.

    2015-04-01

    The primary objective of this study is to conduct a preliminary dynamic analysis of two realistic hybrid energy systems (HES) including a nuclear reactor as the main baseload heat generator (denoted as nuclear HES or nuclear hybrid energy systems [[NHES]) and to assess the local (e.g., HES owners) and system (e.g., the electric grid) benefits attainable by the application of NHES in scenarios with multiple commodity production and high penetration of renewable energy. It is performed for regional cases not generic examples based on available resources, existing infrastructure, and markets within the selected regions. This study also briefly addresses the computational capabilities developed to conduct such analyses, reviews technical gaps, and suggests some research paths forward.

  10. [Human embryonic stem cell research in Germany. The scientific reviewing of applications for the import and use of human embryonic stem cells].

    PubMed

    Kiechle, M

    2008-09-01

    From July 2002 to May 2008, 36 applications for the import and use of human embryonic stem cells (hES) were reviewed by the German Central Ethics Committee for Stem Cell Research (ZES). A flood of applications anticipated by opponents to human embryonic stem cell research has not occurred since the enactment of the German Stem Cell Act in 2002. On the contrary, German hES cell research is below international average in terms of project numbers. The current restrictions for using hES cells in Germany might be causative for the opinion that this type of research is not considered to be very promising. This could hold true especially for research aiming at clinical applications. Consequently, potential research goals of premium importance, especially those of potential clinical relevance, could be seriously jeopardized. PMID:18769900

  11. An Optimization Framework for Dynamic Hybrid Energy Systems

    SciTech Connect

    Wenbo Du; Humberto E Garcia; Christiaan J.J. Paredis

    2014-03-01

    A computational framework for the efficient analysis and optimization of dynamic hybrid energy systems (HES) is developed. A microgrid system with multiple inputs and multiple outputs (MIMO) is modeled using the Modelica language in the Dymola environment. The optimization loop is implemented in MATLAB, with the FMI Toolbox serving as the interface between the computational platforms. Two characteristic optimization problems are selected to demonstrate the methodology and gain insight into the system performance. The first is an unconstrained optimization problem that optimizes the dynamic properties of the battery, reactor and generator to minimize variability in the HES. The second problem takes operating and capital costs into consideration by imposing linear and nonlinear constraints on the design variables. The preliminary optimization results obtained in this study provide an essential step towards the development of a comprehensive framework for designing HES.

  12. Alternative sources of pluripotent stem cells: scientific solutions to an ethical dilemma.

    PubMed

    Rao, Mahendra; Condic, Maureen L

    2008-02-01

    Stem cell researchers in the United States have faced a quagmire of uncertainty due to multiple factors: the ethical divide over the use of embryos for research, the lack of clarity in federal guidelines governing this research, the restrictive patent situation surrounding the generation of new human embryonic stem (HES) cell lines; and the limits on types of research eligible for federal funding. In this commentary, we describe how recent advances in derivation of hES cell-like lines may allow at least some of these uncertainties to be resolved. More importantly, we suggest that the derivation of hES cell-like lines by morally acceptable methods would not only avoid the corrosive effects of a protracted ethical debate over stem cell research, but would also allow U.S. researchers to access federal funds and compete on a more level international playing field. PMID:18271697

  13. Hypereosinophilic syndrome presenting with multiple organ infiltration and deep venous thrombosis

    PubMed Central

    Gao, Su-jun; Wei, Wei; Chen, Jiang-tao; Tan, Ye-hui; Yu, Cheng-bao; Litzow, Mark Robert; Liu, Qiu-ju

    2016-01-01

    Abstract Background: Hypereosinophilic syndrome (HES) can be fatal, particularly when eosinophils infiltrate vital organs and/or if extensive thrombosis develops. However there are no standard recommendations for the use of anticoagulant therapy of HES in the setting of thrombosis. Methods: We herein present a case of a 46-year-old female who presented with marked peripheral eosinophilia with symptoms of multi-organ infiltration and extensive deep venous thrombosis (DVT). In this case, evaluation was carried out before the diagnosis was established, and timely standard-dose corticosteroids combined with a new oral anticoagulant (NOAC) therapy were carried out. Results: These measures resulted in a rapid response and long-term disease control. Conclusion: Although there are no data to support which anticoagulant is preferred in this setting, this case indicates that the new oral anticoagulants may play an important role in the treatment of thrombosis in HES. PMID:27583887

  14. Characterization of dynamics in complex lyophilized formulations: II. Analysis of density variations in terms of glass dynamics and comparisons with global mobility, fast dynamics, and Positron Annihilation Lifetime Spectroscopy (PALS)

    PubMed Central

    Chieng, Norman; Cicerone, Marcus T.; Zhong, Qin; Liu, Ming; Pikal, Michael J.

    2013-01-01

    Amorphous HES/disaccharide (trehalose or sucrose) formulations, with and without added polyols (glycerol and sorbitol) and disaccharide formulations of human growth hormone (hGH), were prepared by freeze drying and characterized with particular interest in methodology for using high precision density measurements to evaluate free volume changes and a focus on comparisons between “free volume” changes obtained from analysis of density data, fast dynamics (local mobility), and PALS characterization of “free volume” hole size. Density measurements were performed using a helium gas pycnometer, and fast dynamics was characterized using incoherent neutron scattering spectrometer. Addition of sucrose and trehalose to hGH decreases free volume in the system with sucrose marginally more effective than trehalose, consistent with superior pharmaceutical stability of sucrose hGH formulations well below Tg relative to trehalose. We find that density data may be analyzed in terms of free volume changes by evaluation of volume changes on mixing and calculation of apparent specific volumes from the densities. Addition of sucrose to HES decreases free volume, but the effect of trehalose is not detectable above experimental error. Addition of sorbitol or glycerol to HES/trehalose base formulations appears to significantly decrease free volume, consistent with the positive impact of such additions on pharmaceutical stability (i.e., degradation) in the glassy state. Free volume changes, evaluated from density data, fast dynamics amplitude of local motion, and PALS hole size data generally are in qualitative agreement for the HES/disaccharide systems studied. All predict decreasing molecular mobility as disaccharides are added to HES. Global mobility as measured by enthalpy relaxation times, increases as disaccharides, particularly sucrose, are added to HES. PMID:23623797

  15. The Implementation of an Integrated Information System for Substance Use Screening in General Medical Settings

    PubMed Central

    Sorensen-Alawad, A.; Carney, B.L.; Persand, I.; Cruz, A.; Botticelli, M.; Pressman, K.; Adams, W.G.; Brolin, M.; Alford, D.P.

    2014-01-01

    Summary The Massachusetts Screening, Brief Intervention and Referral to Treatment (MASBIRT) Program, a substance use screening program in general medical settings, created a web-based, point-of-care (POC), application – the MASBIRT Portal (the “Portal”) to meet program goals. Objectives We report on development and implementation of the Portal. Methods Five year program process outcomes recorded by an independent evaluator and an anonymous survey of Health Educator’s (HEs) adoption, perceptions and Portal use with a modified version of the Technology Readiness Index are described. [8] Specific management team members, selected based on their roles in program leadership, development and implementation of the Portal and supervision of HEs, participated in semi-structured, qualitative interviews. Results At the conclusion of the program 73% (24/33) of the HEs completed a survey on their experience using the Portal. HEs reported that the Portal made recording screening information easy (96%); improved planning their workday (83%); facilitated POC data collection (84%); decreased time dedicated to data entry (100%); and improved job satisfaction (59%). The top two barriers to use were “no or limited wireless connectivity” (46%) and “the tablet was too heavy/bulky to carry” (29%). Qualitative management team interviews identified strategies for successful HIT implementation: importance of engaging HEs in outlining specifications and workflow needs, collaborative testing prior to implementation and clear agreement on data collection purpose, quality requirements and staff roles. Discussion Overall, HEs perceived the Portal favorably with regard to time saving ability and improved workflow. Lessons learned included identifying core requirements early during system development and need for managers to institute and enforce consistent behavioral work norms. Conclusion Barriers and HEs’ views of technology impacted the utilization of the MASBIRT Portal

  16. Hyperbranched Polyglycerol as a Colloid in Cold Organ Preservation Solutions

    PubMed Central

    Gao, Sihai; Guan, Qiunong; Chafeeva, Irina; Brooks, Donald E.; Nguan, Christopher Y. C.; Kizhakkedathu, Jayachandran N.; Du, Caigan

    2015-01-01

    Hydroxyethyl starch (HES) is a common colloid in organ preservation solutions, such as in University of Wisconsin (UW) solution, for preventing graft interstitial edema and cell swelling during cold preservation of donor organs. However, HES has undesirable characteristics, such as high viscosity, causing kidney injury and aggregation of erythrocytes. Hyperbranched polyglycerol (HPG) is a branched compact polymer that has low intrinsic viscosity. This study investigated HPG (MW-0.5 to 119 kDa) as a potential alternative to HES for cold organ preservation. HPG was synthesized by ring-opening multibranching polymerization of glycidol. Both rat myocardiocytes and human endothelial cells were used as an in vitro model, and heart transplantation in mice as an in vivo model. Tissue damage or cell death was determined by both biochemical and histological analysis. HPG polymers were more compact with relatively low polydispersity index than HES in UW solution. Cold preservation of mouse hearts ex vivo in HPG solutions reduced organ damage in comparison to those in HES-based UW solution. Both size and concentration of HPGs contributed to the protection of the donor organs; 1 kDa HPG at 3 wt% solution was superior to HES-based UW solution and other HPGs. Heart transplants preserved with HPG solution (1 kDa, 3%) as compared with those with UW solution had a better functional recovery, less tissue injury and neutrophil infiltration in syngeneic recipients, and survived longer in allogeneic recipients. In cultured myocardiocytes or endothelial cells, significantly more cells survived after cold preservation with the HPG solution than those with the UW solution, which was positively correlated with the maintenance of intracellular adenosine triphosphate and cell membrane fluidity. In conclusion, HPG solution significantly enhanced the protection of hearts or cells during cold storage, suggesting that HPG is a promising colloid for the cold storage of donor organs and cells in

  17. Hyperbranched polyglycerol as a colloid in cold organ preservation solutions.

    PubMed

    Gao, Sihai; Guan, Qiunong; Chafeeva, Irina; Brooks, Donald E; Nguan, Christopher Y C; Kizhakkedathu, Jayachandran N; Du, Caigan

    2015-01-01

    Hydroxyethyl starch (HES) is a common colloid in organ preservation solutions, such as in University of Wisconsin (UW) solution, for preventing graft interstitial edema and cell swelling during cold preservation of donor organs. However, HES has undesirable characteristics, such as high viscosity, causing kidney injury and aggregation of erythrocytes. Hyperbranched polyglycerol (HPG) is a branched compact polymer that has low intrinsic viscosity. This study investigated HPG (MW-0.5 to 119 kDa) as a potential alternative to HES for cold organ preservation. HPG was synthesized by ring-opening multibranching polymerization of glycidol. Both rat myocardiocytes and human endothelial cells were used as an in vitro model, and heart transplantation in mice as an in vivo model. Tissue damage or cell death was determined by both biochemical and histological analysis. HPG polymers were more compact with relatively low polydispersity index than HES in UW solution. Cold preservation of mouse hearts ex vivo in HPG solutions reduced organ damage in comparison to those in HES-based UW solution. Both size and concentration of HPGs contributed to the protection of the donor organs; 1 kDa HPG at 3 wt% solution was superior to HES-based UW solution and other HPGs. Heart transplants preserved with HPG solution (1 kDa, 3%) as compared with those with UW solution had a better functional recovery, less tissue injury and neutrophil infiltration in syngeneic recipients, and survived longer in allogeneic recipients. In cultured myocardiocytes or endothelial cells, significantly more cells survived after cold preservation with the HPG solution than those with the UW solution, which was positively correlated with the maintenance of intracellular adenosine triphosphate and cell membrane fluidity. In conclusion, HPG solution significantly enhanced the protection of hearts or cells during cold storage, suggesting that HPG is a promising colloid for the cold storage of donor organs and cells in

  18. Validating the use of Hospital Episode Statistics data and comparison of costing methodologies for economic evaluation: an end-of-life case study from the Cluster randomised triAl of PSA testing for Prostate cancer (CAP)

    PubMed Central

    Thorn, Joanna C; Turner, Emma L; Hounsome, Luke; Walsh, Eleanor; Down, Liz; Verne, Julia; Donovan, Jenny L; Neal, David E; Hamdy, Freddie C; Martin, Richard M; Noble, Sian M

    2016-01-01

    Objectives To evaluate the accuracy of routine data for costing inpatient resource use in a large clinical trial and to investigate costing methodologies. Design Final-year inpatient cost profiles were derived using (1) data extracted from medical records mapped to the National Health Service (NHS) reference costs via service codes and (2) Hospital Episode Statistics (HES) data using NHS reference costs. Trust finance departments were consulted to obtain costs for comparison purposes. Setting 7 UK secondary care centres. Population A subsample of 292 men identified as having died at least a year after being diagnosed with prostate cancer in Cluster randomised triAl of PSA testing for Prostate cancer (CAP), a long-running trial to evaluate the effectiveness and cost-effectiveness of prostate-specific antigen (PSA) testing. Results Both inpatient cost profiles showed a rise in costs in the months leading up to death, and were broadly similar. The difference in mean inpatient costs was £899, with HES data yielding ∼8% lower costs than medical record data (differences compatible with chance, p=0.3). Events were missing from both data sets. 11 men (3.8%) had events identified in HES that were all missing from medical record review, while 7 men (2.4%) had events identified in medical record review that were all missing from HES. The response from finance departments to requests for cost data was poor: only 3 of 7 departments returned adequate data sets within 6 months. Conclusions Using HES routine data coupled with NHS reference costs resulted in mean annual inpatient costs that were very similar to those derived via medical record review; therefore, routinely available data can be used as the primary method of costing resource use in large clinical trials. Neither HES nor medical record review represent gold standards of data collection. Requesting cost data from finance departments is impractical for large clinical trials. Trial registration number ISRCTN92187251

  19. Modeling the zebrafish segmentation clock's gene regulatory network constrained by expression data suggests evolutionary transitions between oscillating and nonoscillating transcription.

    PubMed

    Schwendinger-Schreck, Jamie; Kang, Yuan; Holley, Scott A

    2014-06-01

    During segmentation of vertebrate embryos, somites form in accordance with a periodic pattern established by the segmentation clock. In the zebrafish (Danio rerio), the segmentation clock includes six hairy/enhancer of split-related (her/hes) genes, five of which oscillate due to negative autofeedback. The nonoscillating gene hes6 forms the hub of a network of 10 Her/Hes protein dimers, which includes 7 DNA-binding dimers and 4 weak or non-DNA-binding dimers. The balance of dimer species is critical for segmentation clock function, and loss-of-function studies suggest that the her genes have both unique and redundant functions within the clock. However, the precise regulatory interactions underlying the negative feedback loop are unknown. Here, we combine quantitative experimental data, in silico modeling, and a global optimization algorithm to identify a gene regulatory network (GRN) designed to fit measured transcriptional responses to gene knockdown. Surprisingly, we find that hes6, the clock gene that does not oscillate, responds to negative feedback. Consistent with prior in silico analyses, we find that variation in transcription, translation, and degradation rates can mediate the gain and loss of oscillatory behavior for genes regulated by negative feedback. Extending our study, we found that transcription of the nonoscillating Fgf pathway gene sef responds to her/hes perturbation similarly to oscillating her genes. These observations suggest a more extensive underlying regulatory similarity between the zebrafish segmentation clock and the mouse and chick segmentation clocks, which exhibit oscillations of her/hes genes as well as numerous other Notch, Fgf, and Wnt pathway genes. PMID:24663100

  20. From Dynamic Expression Patterns to Boundary Formation in the Presomitic Mesoderm

    PubMed Central

    Tiedemann, Hendrik B.; Schneltzer, Elida; Zeiser, Stefan; Hoesel, Bastian; Beckers, Johannes; Przemeck, Gerhard K. H.; de Angelis, Martin Hrabě

    2012-01-01

    The segmentation of the vertebrate body is laid down during early embryogenesis. The formation of signaling gradients, the periodic expression of genes of the Notch-, Fgf- and Wnt-pathways and their interplay in the unsegmented presomitic mesoderm (PSM) precedes the rhythmic budding of nascent somites at its anterior end, which later develops into epithelialized structures, the somites. Although many in silico models describing partial aspects of somitogenesis already exist, simulations of a complete causal chain from gene expression in the growth zone via the interaction of multiple cells to segmentation are rare. Here, we present an enhanced gene regulatory network (GRN) for mice in a simulation program that models the growing PSM by many virtual cells and integrates WNT3A and FGF8 gradient formation, periodic gene expression and Delta/Notch signaling. Assuming Hes7 as core of the somitogenesis clock and LFNG as modulator, we postulate a negative feedback of HES7 on Dll1 leading to an oscillating Dll1 expression as seen in vivo. Furthermore, we are able to simulate the experimentally observed wave of activated NOTCH (NICD) as a result of the interactions in the GRN. We esteem our model as robust for a wide range of parameter values with the Hes7 mRNA and protein decays exerting a strong influence on the core oscillator. Moreover, our model predicts interference between Hes1 and HES7 oscillators when their intrinsic frequencies differ. In conclusion, we have built a comprehensive model of somitogenesis with HES7 as core oscillator that is able to reproduce many experimentally observed data in mice. PMID:22761566

  1. Comparison of Albumin, Hydroxyethyl Starch and Ringer Lactate Solution as Priming Fluid for Cardiopulmonary Bypass in Paediatric Cardiac Surgery

    PubMed Central

    Prajapati, Mrugesh; Solanki, Atul; Pandya, Himani

    2016-01-01

    Introduction In paediatric cardiac surgery, there is still not any information with regard to the best choice of priming fluids for Cardiopulmonary Bypass (CPB). Albumin, Hydroxyethyl Starch (HES) & ringer lactate are equally used, but each has its advantages & disadvantages. Albumin & HES had better fluid balance which affect outcome in paediatric cardiac surgery significantly. Aim To compare priming solution containing albumin, hydroxyethyl starch and ringer lactate during elective open-heart surgery in paediatrics aged up to 3 years. Materials and Methods All patients were managed by standardized institution protocol and were randomly distributed into three groups based on the priming solution which is used in the CPB Circuit and having 35 patients in each group. Group A: Receive albumin 10 ml/kg in priming solution, Group B: Receive Hydroxyethyl starch (HES130/0.4) 6% 20ml/kg in priming solution, Group C: Receive ringer lactate priming solution. Primary outcome variable included perioperative haemoglobin, total protein, colloid osmotic pressure, platelets, fluid balance, urine output, post-operative blood loss, blood products usage, renal & liver function, extubation time, ICU stay & outcome. Results Patients receiving albumin had higher perioperative platelet count, total protein level & colloid osmotic pressure, lesser post-operative blood loss & blood products requirement. Patients receiving HES had lower level of platelets postoperatively than ringer lactate group but not associated with increase blood loss. HES did not affect renal function & haemostasis in this dose. Patients receiving ringer lactate had positive fluid balance intraoperatively. All three groups have similar effect on renal & liver function, urine output, time to extubation, ICU stay & outcome. Conclusion We conclude that albumin is expensive but better prime as maintain haemostasis, colloid oncotic pressure & reduced blood product requirement. HES will not hamper haemostasis & renal

  2. Modeling the Zebrafish Segmentation Clock’s Gene Regulatory Network Constrained by Expression Data Suggests Evolutionary Transitions Between Oscillating and Nonoscillating Transcription

    PubMed Central

    Schwendinger-Schreck, Jamie; Kang, Yuan; Holley, Scott A.

    2014-01-01

    During segmentation of vertebrate embryos, somites form in accordance with a periodic pattern established by the segmentation clock. In the zebrafish (Danio rerio), the segmentation clock includes six hairy/enhancer of split-related (her/hes) genes, five of which oscillate due to negative autofeedback. The nonoscillating gene hes6 forms the hub of a network of 10 Her/Hes protein dimers, which includes 7 DNA-binding dimers and 4 weak or non-DNA-binding dimers. The balance of dimer species is critical for segmentation clock function, and loss-of-function studies suggest that the her genes have both unique and redundant functions within the clock. However, the precise regulatory interactions underlying the negative feedback loop are unknown. Here, we combine quantitative experimental data, in silico modeling, and a global optimization algorithm to identify a gene regulatory network (GRN) designed to fit measured transcriptional responses to gene knockdown. Surprisingly, we find that hes6, the clock gene that does not oscillate, responds to negative feedback. Consistent with prior in silico analyses, we find that variation in transcription, translation, and degradation rates can mediate the gain and loss of oscillatory behavior for genes regulated by negative feedback. Extending our study, we found that transcription of the nonoscillating Fgf pathway gene sef responds to her/hes perturbation similarly to oscillating her genes. These observations suggest a more extensive underlying regulatory similarity between the zebrafish segmentation clock and the mouse and chick segmentation clocks, which exhibit oscillations of her/hes genes as well as numerous other Notch, Fgf, and Wnt pathway genes. PMID:24663100

  3. US hydropower resource assessment for Hawaii

    SciTech Connect

    Francfort, J.E.

    1996-09-01

    US DOE is developing an estimate of the undeveloped hydropower potential in US. The Hydropower Evaluation Software (HES) is a computer model developed by INEL for this purpose. HES measures the undeveloped hydropower resources available in US, using uniform criteria for measurement. The software was tested using hydropower information and data provided by Southwestern Power Administration. It is a menu-driven program that allows the PC user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes, and generate reports. This report describes the resource assessment results for the State of Hawaii.

  4. Renal involvement in idiopathic hypereosinophic syndrome

    PubMed Central

    Shehwaro, Nathalie; Langlois, Anne Lyse; Gueutin, Victor; Izzedine, Hassane

    2013-01-01

    The hypereosinophilic syndromes (HESs) are a group of disorders marked by the sustained overproduction of eosinophils, in which eosinophilic infiltration and mediator release cause damage to multiple organs. In idiopathic HES, the underlying cause of hypereosinophilia (HE) remains unknown despite thorough aetiological work-up. Kidney disease is thought to be rare in HES. Renal manifestations described include eosinophilic interstitial nephritis, various types of glomerulopathies, thrombotic microangiopathy (TMA) and electrolyte disturbances. The diagnosis must be made in time, because a recovery of renal function can be obtained if treatment is initiated promptly. PMID:26064485

  5. A Case of Erythroderma Secondary to Hypereosinophilia

    PubMed Central

    Abdulsalam, Mohammed Shafi; Ghanta, Hari Chandana; Menon, Maya; Jacob, Sheba S.K.

    2016-01-01

    Hypereosinophilic syndrome (HES) is a myeloproliferative disorder characterised by marked peripheral eosinophilia and end organ damage attributable to eosinophilia without secondary cause. Early recognition and treatment are essential to prevent morbidity and mortality. Cytoreduction with Steroids, Hydroxyurea and Imatinib are the main stay of treatment. Molecular studies like Fip1-like-1 fused with platelet derived growth factor receptor alpha (FIP1L1-PDGFRα) etc., are recommended in view of therapeutic implication. In this paper we report a rare case of HES developing in a lady 6 months after surgical removal of lymphangioma of spleen, which in itself is rare.

  6. [A propose to suspend the use of hydroxyethyl starch for fluid resuscitation in shock phase of severe burns].

    PubMed

    Luo, Gao-xing; Peng, Yi-zhi; Wu, Jun

    2013-10-01

    Based on the result of randomized controlled trials and meta-analysis recently, the infusion of hydroxyethyl starch (HES) was not shown to over match routine crystalline solution in exerting resuscitation effect against hypovolemia of patients with burn shock, severe systematic infection, or other critical conditions, on the other hand, it may induce renal toxicity and other toxic and side effects. Since the pathological mechanism underlying hypovolemia during shock phase after burn is similar to that of severe systemic infection, we propose to suspend the use of HES for fluid resuscitation during the shock phase of severe burn until further elucidation. PMID:24359998

  7. Moderate lead poisoning: trends in blood lead levels in unchelated children.

    PubMed Central

    Markowitz, M E; Bijur, P E; Ruff, H A; Balbi, K; Rosen, J F

    1996-01-01

    The appropriate clinical management of children who are moderately poisoned with lead (Pb) is under active investigation. To determine the pattern of change in blood Pb (BPb) levels in the absence of chelation therapy, we followed moderately Pb-poisoned children (initial blood Pb levels 1.21-2.66 mumol/l or 25-55 micrograms/dl) for 6 months with repeated BPb level measurements. Chelation therapy was not administered because all the children had negative lead mobilization tests indicating limited response to the chelating agent, calcium disodium edetate (CaNa2EDTA). Eligible children received the following interventions: notification of the health department to remediate lead hazards; reinforced educational efforts about the toxicity sources and treatment of Pb during 10 clinic and 3 home visits; and iron therapy for children with ferritin levels less than 16 micrograms/l. To quantify the lead paint hazards in the home, we combined a visual rating of the surfaces (intact to peeling) with an X-ray fluorescence (XRF) measurement of the lead content of the painted surface. The sum of these assessments is termed the home environmental score (HES). Data were analyzed from 79 children. BPb levels declined by 27%, on average, over 6 months. HES was correlated with BPb at enrollment, but neither the initial nor later HES measurements predicted BPb at other time points. The HES was highest at enrollment and declined by 50% and 75% at the second and third home visits, respectively. However, only a minority of the children (20%) achieved an HES of 0, indicating no lead paint hazards at home. Despite some ongoing Pb exposure, a parallel fall in BPb levels was observed in subgroups of children with either initially low or high HES (above or below the median HES of 37). Iron status did not account for the change in BPb levels. These data provide evidence that our measure, the HES, is quantifiably related to BPb levels in children, that this correlation is significant only prior

  8. Value-Driven Design and Sensitivity Analysis of Hybrid Energy Systems using Surrogate Modeling

    SciTech Connect

    Wenbo Du; Humberto E. Garcia; William R. Binder; Christiaan J. J. Paredis

    2001-10-01

    A surrogate modeling and analysis methodology is applied to study dynamic hybrid energy systems (HES). The effect of battery size on the smoothing of variability in renewable energy generation is investigated. Global sensitivity indices calculated using surrogate models show the relative sensitivity of system variability to dynamic properties of key components. A value maximization approach is used to consider the tradeoff between system variability and required battery size. Results are found to be highly sensitive to the renewable power profile considered, demonstrating the importance of accurate renewable resource modeling and prediction. The documented computational framework and preliminary results represent an important step towards a comprehensive methodology for HES evaluation, design, and optimization.

  9. Efficiency of 7.2% hypertonic saline hydroxyethyl starch 200/0.5 versus mannitol 15% in the treatment of increased intracranial pressure in neurosurgical patients – a randomized clinical trial [ISRCTN62699180

    PubMed Central

    Harutjunyan, Lilit; Holz, Carsten; Rieger, Andreas; Menzel, Matthias; Grond, Stefan; Soukup, Jens

    2005-01-01

    Introduction This prospective randomized clinical study investigated the efficacy and safety of 7.2% hypertonic saline hydroxyethyl starch 200/0.5 (7.2% NaCl/HES 200/0.5) in comparison with 15% mannitol in the treatment of increased intracranial pressure (ICP). Methods Forty neurosurgical patients at risk of increased ICP were randomized to receive either 7.2% NaCl/HES 200/0.5 or 15% mannitol at a defined infusion rate, which was stopped when ICP was < 15 mmHg. Results Of the 40 patients, 17 patients received 7.2% NaCl/HES 200/0.5 and 15 received mannitol 15%. In eight patients, ICP did not exceed 20 mmHg so treatment was not necessary. Both drugs decreased ICP below 15 mmHg (p < 0.0001); 7.2% NaCl/HES 200/0.5 within 6.0 (1.2–15.0) min (all results are presented as median (minimum-maximum range)) and mannitol within 8.7 (4.2–19.9) min (p < 0.0002). 7.2% NaCl/HES 200/0.5 caused a greater decrease in ICP than mannitol (57% vs 48%; p < 0.01). The cerebral perfusion pressure was increased from 60 (39–78) mmHg to 72 (54–85) mmHg by infusion with 7.2% NaCl/HES 200/0.5 (p < 0.0001) and from 61 (47–71) mmHg to 70 (50–79) mmHg with mannitol (p < 0.0001). The mean arterial pressure was increased by 3.7% during the infusion of 7.2% NaCl/HES 200/0.5 but was not altered by mannitol. There were no clinically relevant effects on electrolyte concentrations and osmolarity in the blood. The mean effective dose to achieve an ICP below 15 mmHg was 1.4 (0.3–3.1) ml/kg for 7.2% NaCl/HES 200/0.5 and 1.8 (0.45–6.5) ml/kg for mannitol (p < 0.05). Conclusion 7.2% NaCl/HES 200/0.5 is more effective than mannitol 15% in the treatment of increased ICP. A dose of 1.4 ml/kg of 7.2% NaCl/HES 200/0.5 can be recommended as effective and safe. The advantage of 7.2% NaCl/HES 200/0.5 might be explained by local osmotic effects, because there were no clinically relevant differences in hemodynamic clinical chemistry parameters. PMID:16277715

  10. [Comparison of the selected methods of cord processing for transplantation purposes].

    PubMed

    Ołdak, T; Machaj, E K; Gajkowska, A; Kruszewski, M; Kłos, M; Szczecina, R; Czajkowski, K; Kuczyńska-Sicińska, J; Pojda, Z

    2000-09-01

    Human umbilical cord blood (UCB) has been successfully used as a source of allogeneic hematopoietic cells for transplantation. Banking of the UCB requires its volume reduction to decrease storage space, costs and volume of infused DMSO. In order to select an optimal method for volume reduction we compared several methods of cord blood processing, namely buffy coat centrifugation, red cell lysis, hydroxyethyl starch (HES)-, methylcellulose- and gelatin-sedimentations. The viability of cells and the recoveries of total white blood cells, mononuclear cells and CD34+ cells was evaluated. We also compared the efficacy of red cells depletion from the original UCB sample. Buffy coat centrifugation, red cell lysis, HES, gelatin or methylcellulose resulted in high mononuclear cell recoveries, whereas high hematopoietic cell recovery was observed only after HES sedimentation and buffy coat processing. The HES sedimentation procedure compared to buffy coat processing is more time and labor consuming and resulted in higher red blood cell and platelets depletion. Both methods can be recommended as a method at choice for the umbilical cord blood processing before banking. PMID:11083012

  11. POPULATION EXPOSURES TO PARTICULATE MATTER: A COMPARISON OF EXPOSURE MODEL PREDICTIONS AND MEASUREMENT DATA

    EPA Science Inventory

    The US EPA National Exposure Research Laboratory (NERL) is currently developing an integrated human exposure source-to-dose modeling system (HES2D). This modeling system will incorporate models that use a probabilistic approach to predict population exposures to environmental ...

  12. Leukemia inhibitory factor (LIF) enhances MAP2 + and HUC/D + neurons and influences neurite extension during differentiation of neural progenitors derived from human embryonic stem cells.

    EPA Science Inventory

    Leukemia Inhibitory Factor (L1F), a member of the Interleukin 6 cytokine family, has a role in differentiation of Human Neural Progenitor (hNP) cells in vitro. hNP cells, derived from Human Embryonic Stem (hES) cells, have an unlimited capacity for self-renewal in monolayer cultu...

  13. Nonviral Reprogramming of Human Wharton's Jelly Cells Reveals Differences Between ATOH1 Homologues.

    PubMed

    Mellott, Adam J; Devarajan, Keerthana; Shinogle, Heather E; Moore, David S; Talata, Zsolt; Laurence, Jennifer S; Forrest, M Laird; Noji, Sumihare; Tanaka, Eiji; Staecker, Hinrich; Detamore, Michael S

    2015-06-01

    The transcription factor atonal homolog 1 (ATOH1) has multiple homologues that are functionally conserved across species and is responsible for the generation of sensory hair cells. To evaluate potential functional differences between homologues, human and mouse ATOH1 (HATH1 and MATH-1, respectively) were nonvirally delivered to human Wharton's jelly cells (hWJCs) for the first time. Delivery of HATH1 to hWJCs demonstrated superior expression of inner ear hair cell markers and characteristics than delivery of MATH-1. Inhibition of HES1 and HES5 signaling further increased the atonal effect. Transfection of hWJCs with HATH1 DNA, HES1 siRNA, and HES5 siRNA displayed positive identification of key hair cell and support cell markers found in the cochlea, as well as a variety of cell shapes, sizes, and features not native to hair cells, suggesting the need for further examination of other cell types induced by HATH1 expression. In the first side-by-side evaluation of HATH1 and MATH-1 in human cells, substantial differences were observed, suggesting that the two atonal homologues may not be interchangeable in human cells, and artificial expression of HATH1 in hWJCs requires further study. In the future, this line of research may lead to engineered systems that would allow for evaluation of drug ototoxicity or potentially even direct therapeutic use. PMID:25760435

  14. Spectroscopic investigation of Lambda hypernuclei in the wide mass region using the (e, e-prime K+) reaction

    SciTech Connect

    Satoshi N. Nakamura

    2011-09-01

    The third generation spectroscopic study of {Lambda} hypernuclei using (e,e'K{sup +}) reaction (JLab E05-115) was performed at JLab Hall-C in 2009. The experiment introduced the newly developed high-resolution electron spectrometer (HES) with the existing high-resolution kaon spectrometer (HKS). Experimental configuration, conditions, spectrometer designs and current status of analysis are presented.

  15. Mineral nutrient recovery from pyrolysis systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bioenergy crops such as high-energy sorghum (HES), bioenergy rice, corn stover, and switchgrass can be thermo-chemically converted by pyrolysis to produce bio-oil, synthesis gas from non-condensable gases, and biochar. The biochar fraction can be recycled back to the production field to improve soil...

  16. Hesperidin metabolite hesperetin-7-O-glucuronide, but not hesperetin-3'-O-glucuronide, exerts hypotensive, vasodilatory, and anti-inflammatory activities.

    PubMed

    Yamamoto, Masaki; Jokura, Hiroko; Hashizume, Koujiro; Ominami, Hideo; Shibuya, Yusuke; Suzuki, Atsushi; Hase, Tadashi; Shimotoyodome, Akira

    2013-09-01

    Orally ingested hesperidin (HES) is hydrolyzed into hesperetin in the gastrointestinal tract and conjugated during absorption. Hesperetin conjugates are the main circulating metabolites in human and rat plasma. We previously reported that glucosyl hesperidin (GHES), a water-soluble HES derivative, prevents hypertension via improvement of endothelial dysfunction in spontaneously hypertensive rats (SHRs). Although these hesperetin conjugates seem to be responsible for hypotensive and endothelium-dependent vasodilatory activities of dietary GHES, little is known about the mechanisms of action of these conjugated metabolites. Therefore, the aim of the present study was to investigate the effects of hesperetin-7-O-β-d-glucuronide (HPT7G) and hesperetin-3'-O-β-d-glucuronide (HPT3'G), which are the predominant HES metabolites in rat plasma, on blood pressure and endothelial function. Intravenous administration of HPT7G (5 mg kg(-1)) decreased blood pressure in anesthetized SHRs. HPT7G enhanced endothelium-dependent vasodilation in response to acetylcholine, but had no effect on endothelium-independent vasodilation in response to sodium nitroprusside (SNP) in aortas isolated from SHRs. HPT7G decreased hydrogen peroxide-induced intracellular adhesion molecule-1 and monocyte chemoattractant protein-1 mRNA expression in rat aortic endothelial cells. In contrast, HPT3'G had little effect on these parameters. In conclusion, HPT7G exerted hypotensive, vasodilatory and anti-inflammatory activities, similar to hesperetin and these effects are associated, in part, with the activity of GHES and HES to improve hypertension and endothelial dysfunction. PMID:23831969

  17. Enterprise Curriculum for Creative Industries Students: An Exploration of Current Attitudes and Issues

    ERIC Educational Resources Information Center

    Carey, Charlotte; Naudin, Annette

    2006-01-01

    Purpose: This paper seeks to report on the current state and attitudes towards Enterprise Curriculum within higher education (HE) for the creative industries sector. It is based on preliminary findings from the Creative Enterprise Conference (2006) held at UCE Birmingham, which examined the role of HEs in developing future entrepreneurs in this…

  18. The Elementary and Secondary Education Act at Fifty: A Changing Federal Role in American Education

    ERIC Educational Resources Information Center

    Nelson, Adam R.

    2016-01-01

    For this first "History of Education Quarterly Policy Forum," we invited participants in the special Plenary Session at the 2015 Annual Meeting of the History of Education Society (HES) in St. Louis to publish their remarks on the historical significance of the Elementary and Secondary Education Act (ESEA) at fifty. Organized and…

  19. Nonviral Reprogramming of Human Wharton's Jelly Cells Reveals Differences Between ATOH1 Homologues

    PubMed Central

    Mellott, Adam J.; Devarajan, Keerthana; Shinogle, Heather E.; Moore, David S.; Talata, Zsolt; Laurence, Jennifer S.; Forrest, M. Laird; Noji, Sumihare; Tanaka, Eiji; Staecker, Hinrich

    2015-01-01

    The transcription factor atonal homolog 1 (ATOH1) has multiple homologues that are functionally conserved across species and is responsible for the generation of sensory hair cells. To evaluate potential functional differences between homologues, human and mouse ATOH1 (HATH1 and MATH-1, respectively) were nonvirally delivered to human Wharton's jelly cells (hWJCs) for the first time. Delivery of HATH1 to hWJCs demonstrated superior expression of inner ear hair cell markers and characteristics than delivery of MATH-1. Inhibition of HES1 and HES5 signaling further increased the atonal effect. Transfection of hWJCs with HATH1 DNA, HES1 siRNA, and HES5 siRNA displayed positive identification of key hair cell and support cell markers found in the cochlea, as well as a variety of cell shapes, sizes, and features not native to hair cells, suggesting the need for further examination of other cell types induced by HATH1 expression. In the first side-by-side evaluation of HATH1 and MATH-1 in human cells, substantial differences were observed, suggesting that the two atonal homologues may not be interchangeable in human cells, and artificial expression of HATH1 in hWJCs requires further study. In the future, this line of research may lead to engineered systems that would allow for evaluation of drug ototoxicity or potentially even direct therapeutic use. PMID:25760435

  20. Reading and Arithmetic Achievement Among Youths 12-17 Years as Measured by the Wide Range Achievement Test. Vital and Health Statistics Series 11, No. 136.

    ERIC Educational Resources Information Center

    Hitchcock, Dale C.; Pinder, Glenn B.

    National estimates of school achievement as measured by the reading and arithmetic subtests of the Wide Range Achievement Test (WRAT) for the noninstitutionalized population of the United States aged 12-17 years are presented. Data were obtained in the Health Examination Survey (HES) of 1966-70. In the survey a probability sample of 7,514 youths…

  1. Country's Competitiveness and Sustainability in the Context of the Higher Education System Reforms

    ERIC Educational Resources Information Center

    Jermolajeva, Elita; Aleksejeva, Ludmila

    2013-01-01

    The accumulation of knowledge and its use have become important factors that promote economic development as they contribute to a country's competitiveness in the global economy. The basic significance of research is obtained by defining new approaches in the organisation, function and efficiency of the higher education system (HES) by emphasising…

  2. 75 FR 6740 - Self-Regulatory Organizations; NYSE Arca, Inc.; Notice of Filing and Immediate Effectiveness of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-10

    .... 146 ERTS Electronic Arts Inc. 207 TCK Teck Resources Ltd. 121 EWJ iShares MSCI Japan Index Fund. 196... Act Release No. 60711 (September 23, 2009), 74 FR 49419 (September 28, 2009) (order approving SR... Holdings PLC. 204 WFR MEMC Electronic Materials Inc. 197 HES Hess Corp. 115 WFT Weatherford...

  3. 75 FR 6756 - Self-Regulatory Organizations; NYSE Arca, Inc.; Notice of Filing and Immediate Effectiveness of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-10

    ... Energy Nemours & Co. Inc. 146 ERTS Electronic Arts 207 TCK Teck Resources Inc. Ltd. 121 EWJ iShares MSCI... Act Release No. 60711 (September 23, 2009), 74 FR 49419 (September 28, 2009) (order approving SR... United Technologies Corp. 170 HBC HSBC Holdings PLC. 204 WFR MEMC Electronic Materials Inc. 197 HES...

  4. Trends in the human embryonic stem cell patent field.

    PubMed

    Karlsson, Ulrika; Hyllner, Johan; Runeberg, Kristina

    2007-01-01

    The successful derivation of human embryonic stem (hES) cell lines in late 1990s marks the birth of a new era in biomedical research. In the USA, this landmark invention is protected by granted composition-of-matter patents. In addition to these patents, several others have been granted on further development of hES cell research, such as on differentiated cell types and in vitro and in vivo use aspects. In Europe, there is presently no consensus pertaining to the patentability of hES cells, and all patent applications pending at the European patent office are therefore awaiting a principal decision by the Enlarged Board of Appeal. The authors argue that it will be of importance to the stem cell industry that patents are granted on inventions downstream in the value chain, e.g on specialised cell types derived from hES cells and different drug discovery applications. Patents and patent applications on such inventions for the three germ layers ectoderm/neuro, endoderm/hepato and mesoderm/cardio have been examined. The number of patents increased in the period 2001 to 2006 for all three lineages with ectoderm/neuro as the most patent intensive field. There where 9-13 times more US patent applications filed related to the three lineages compared to in Europe. PMID:19076035

  5. High Speed Video for Airborne Instrumentation Application

    NASA Technical Reports Server (NTRS)

    Tseng, Ting; Reaves, Matthew; Mauldin, Kendall

    2006-01-01

    A flight-worthy high speed color video system has been developed. Extensive system development and ground and environmental. testing hes yielded a flight qualified High Speed Video System (HSVS), This HSVS was initially used on the F-15B #836 for the Lifting Insulating Foam Trajectory (LIFT) project.

  6. Applying GIS to Develop a Web-Based Spatial-Person-Temporal History Educational System

    ERIC Educational Resources Information Center

    Lo, Jia-Jiunn; Chang, Chuen-Jung; Tu, Hsiao-Han; Yeh, Shiou-Wen

    2009-01-01

    Developing interactive history learning materials to facilitate historical thinking is one of the challenges in history teaching and learning. This study developed a web-based history educational system, which has used the acronym HES-SPATO (history educational system based on SPATO), to increase the understandability of history learning…

  7. Developing Research Priorities with a Cohort of Higher Education for Sustainability Experts

    ERIC Educational Resources Information Center

    Wright, Tarah S. A.

    2007-01-01

    Purpose: The purpose of this paper is to examine the results of a Delphi exercise used at the Halifax Consultation in which 35 experts representing 17 countries gathered to develop research priorities for the emerging field of higher education for sustainability (HES). Design/methodology/approach: The Delphi technique was used to elicit the…

  8. Suppression of OVA-alum induced allergy by Heligmosomoides polygyrus products is MyD88-, TRIF-, regulatory T- and B cell-independent, but is associated with reduced innate lymphoid cell activation.

    PubMed

    McSorley, Henry J; Blair, Natalie F; Robertson, Elaine; Maizels, Rick M

    2015-11-01

    The murine intestinal nematode Heligmosomoides polygyrus exerts multiple immunomodulatory effects in the host, including the suppression of allergic inflammation in mice sensitized to allergen presented with alum adjuvant. Similar suppression is attained by co-administration of H. polygyrus excretory/secretory products (HES) with the sensitizing dose of ovalbumin (OVA) in alum. We investigated the mechanism of suppression by HES in this model, and found it was maintained in MyD88xTRIF-deficient mice, implying no role for helminth- or host-derived TLR ligands, or IL-1 family cytokines that signal in a MyD88- or TRIF-dependent manner. We also found suppression was unchanged in µMT mice, which lack B2 B cells, and that suppression was not abrogated when regulatory T cells were depleted in Foxp3.LuciDTR-4 mice. However, reduced IL-5 production was seen in the first 12 h after injection of OVA-alum when HES was co-administered, associated with reduced activation of IL-5(+) and IL-13(+) group 2 innate lymphoid cells. Thus, the suppressive effects of HES on alum-mediated OVA sensitization are reflected in the very earliest innate response to allergen exposure in vivo. PMID:25728231

  9. Highly efficient transient gene expression and gene targeting in primate embryonic stem cells with helper-dependent adenoviral vectors

    PubMed Central

    Suzuki, Keiichiro; Mitsui, Kaoru; Aizawa, Emi; Hasegawa, Kouichi; Kawase, Eihachiro; Yamagishi, Toshiyuki; Shimizu, Yoshihiko; Suemori, Hirofumi; Nakatsuji, Norio; Mitani, Kohnosuke

    2008-01-01

    Human embryonic stem (hES) cells are regarded as a potentially unlimited source of cellular materials for regenerative medicine. For biological studies and clinical applications using primate ES cells, the development of a general strategy to obtain efficient gene delivery and genetic manipulation, especially gene targeting via homologous recombination (HR), would be of paramount importance. However, unlike mouse ES (mES) cells, efficient strategies for transient gene delivery and HR in hES cells have not been established. Here, we report that helper-dependent adenoviral vectors (HDAdVs) were able to transfer genes in hES and cynomolgus monkey (Macaca fasicularis) ES (cES) cells efficiently. Without losing the undifferentiated state of the ES cells, transient gene transfer efficiency was ≈100%. Using HDAdVs with homology arms, approximately one out of 10 chromosomal integrations of the vector was via HR, whereas the rate was only ≈1% with other gene delivery methods. Furthermore, in combination with negative selection, ≈45% of chromosomal integrations of the vector were targeted integrations, indicating that HDAdVs would be a powerful tool for genetic manipulation in hES cells and potentially in other types of human stem cells, such as induced pluripotent stem (iPS) cells. PMID:18768795

  10. Oscillatory control of Delta-like1 in cell interactions regulates dynamic gene expression and tissue morphogenesis

    PubMed Central

    Shimojo, Hiromi; Isomura, Akihiro; Ohtsuka, Toshiyuki; Kori, Hiroshi; Miyachi, Hitoshi; Kageyama, Ryoichiro

    2016-01-01

    Notch signaling regulates tissue morphogenesis through cell–cell interactions. The Notch effectors Hes1 and Hes7 are expressed in an oscillatory manner and regulate developmental processes such as neurogenesis and somitogenesis, respectively. Expression of the mRNA for the mouse Notch ligand Delta-like1 (Dll1) is also oscillatory. However, the dynamics of Dll1 protein expression are controversial, and their functional significance is unknown. Here, we developed a live-imaging system and found that Dll1 protein expression oscillated in neural progenitors and presomitic mesoderm cells. Notably, when Dll1 expression was accelerated or delayed by shortening or elongating the Dll1 gene, Dll1 oscillations became severely dampened or quenched at intermediate levels, as modeled mathematically. Under this condition, Hes1 and Hes7 oscillations were also dampened. In the presomitic mesoderm, steady Dll1 expression led to severe fusion of somites and their derivatives, such as vertebrae and ribs. In the developing brain, steady Dll1 expression inhibited proliferation of neural progenitors and accelerated neurogenesis, whereas optogenetic induction of Dll1 oscillation efficiently maintained neural progenitors. These results indicate that the appropriate timing of Dll1 expression is critical for the oscillatory networks and suggest the functional significance of oscillatory cell–cell interactions in tissue morphogenesis. PMID:26728556

  11. Physicochemical properties and responses in microcirculation of native tapioca starch-based plasma expander.

    PubMed

    Chatpun, Surapong; Meesane, Jirut; Rujirojindakul, Pairaya

    2016-02-01

    Plasma expanders (PEs) such as hydroxyethyl strach are widely used for volume replacement. The plantation and production of tapioca in Thailand is abundant which may provide a new source for PEs starch with novel properties. This work investigated the properties and circulatory effects of native tapioca starch-based PE (TPE). Various formulations of mixture between native tapioca starch and 0.9% sodium chloride solution were prepared and characterized in order to obtain the proper physicochemical and rheological properties. About 1% concentration by weight per volume of TPE was compared with 6% hydroxyethyl starch 130/0.4 in 0.9% sodium chloride (HES130/0.4) using an acute hemodilution by 40% of blood volume in an animal protocol. TPE had higher turbidity and viscosity but lower colloid osmotic pressure compared with HES 130/0.4. The in vivo study demonstrated that Golden Syrian hamsters hemodiluted with TPE maintained a mean arterial blood pressure and no significant difference compared to HES 130/0.4. The arterial vasodilation and functional capillary density in the animals hemodiluted with TPE had higher values than in the animals hemodiluted with HES 130/0.4. Although the in vivo study reported positive results using this native tapioca starch-based PE, the product needs work to improve some of its physiochemical properties. PMID:25892486

  12. microRNA input into a neural ultradian oscillator controls emergence and timing of alternative cell states

    PubMed Central

    Goodfellow, Marc; Phillips, Nicholas E.; Manning, Cerys; Galla, Tobias; Papalopulu, Nancy

    2014-01-01

    Progenitor maintenance, timed differentiation and the potential to enter quiescence are three fundamental processes that underlie the development of any organ system. In the nervous system, progenitor cells show short-period oscillations in the expression of the transcriptional repressor Hes1, while neurons and quiescent progenitors show stable low and high levels of Hes1, respectively. Here we use experimental data to develop a mathematical model of the double-negative interaction between Hes1 and a microRNA, miR-9, with the aim of understanding how cells transition from one state to another. We show that the input of miR-9 into the Hes1 oscillator tunes its oscillatory dynamics, and endows the system with bistability and the ability to measure time to differentiation. Our results suggest that a relatively simple and widespread network of cross-repressive interactions provides a unifying framework for progenitor maintenance, the timing of differentiation and the emergence of alternative cell states. PMID:24595054

  13. INVADING AND METASTASIZING CARDIAC HEMANGIOENDOTHELIAL NEOPLASMS IN A COHORT OF THE FISH RIVULUS MARMORATUS: UNUSUALLY HIGH PREVALENCE, HISTOPATHOLOGY, AND POSSIBLE ETIOLOGIES

    EPA Science Inventory

    An unusually high, unprecedented prevalence of cardiac hemangioendotheliosarcomas (HES) and hemangioendotheliomas (HE) was found in a cohort of the small, teleost fish Rivulus marmoratus. he neoplasms occurred in 50/220 fish (22.7%) used in a carcinogenicity study of butyrated hy...

  14. Characterization and physiological effect of tapioca maltodextrin colloid plasma expander in hemorrhagic shock and resuscitation model.

    PubMed

    Chatpun, Surapong; Sawanyawisuth, Kittisak; Wansuksri, Rungtiva; Piyachomkwan, Kuakoon

    2016-05-01

    Plasma expanders (PEs) are administered fluids to replace blood volume when massive blood loss has occured. Maltodextrin from tapioca starch was selected as a study candidate to prepare a colloid PE due to an uncomplicated production process. The formulations of mixture between tapioca maltodextrin and 0.9 % sodium chloride solution were prepared and then characterized. This was to investigate the effects of a dextrose equivalent (DE) and the concentration on the physical properties. Storage stability of each formulation was also determined and compared with clinically used PE [6 % hydroxyethyl starch (HES), 130/0.4]. The effects on the circulatory system in hamsters with hemorrhagic shock and resuscitation using prepared PE were also investigated. The results showed that low DE value led to high retrogradation, turbidity and viscosity but low colloid osmotic pressure and poor solubility. Among the prepared solutions, tapioca maltodextrin with DE6 at 10 % w/v concentration had comparable properties with 6 % HES 130/0.4. Animals resuscitated with 10 % DE6 PE had improved mean arterial blood pressure similar to those resuscitated with 6 % HES 130/0.4. However, several parameters in animals resuscitated with 10 % DE6 PE were lower than those resuscitated with 6 % HES 130/0.4, i.e., heart rate, functional capillary density. Therefore, if using tapioca maltodextrin for PE, some properties have to be considered and efficiently optimized. PMID:27015687

  15. The use of hydroxy-ethyl starch sedimentation for autologous buffy coat preparation.

    PubMed

    Gannon, Timothy; McConnell, Kristen; Riley, Jeffrey

    2005-09-01

    The use of hydroxy-ethyl starch (HES) has been used in the cord blood banking industry for the separation of blood into its individual components. The focus of this study is to examine whether the use of HES is feasible in obtaining a buffy coat for the use in the construction of platelet gels. Blood was collected from seven canines into anticoagulant citrate dextrose at a ratio of 1:8. A sample of whole blood was used to obtain initial cell counts and a base line Thrombelastograph (TEG). 6% Hespan was added to the whole blood and allowed to sediment for 60 minutes. Cell counts and TEGs were performed on the resulting layers: red blood cells (RBCs), buffy coat (BC), and plasma. The results of this study (see Table 1) suggest that the gravity sedimentation of whole blood with HES can significantly reduce the RBCs in the BC and plasma layers while increasing the platelet count in the plasma layer. These data also suggest that HES does not affect the clotting capabilities of the BC and plasma layers. PMID:16350387

  16. Context dependence between subdomains in the DNA binding interface of the I-CreI homing endonuclease

    PubMed Central

    Grizot, Sylvestre; Duclert, Aymeric; Thomas, Séverine; Duchateau, Philippe; Pâques, Frédéric

    2011-01-01

    Homing endonucleases (HE) have emerged as precise tools for achieving gene targeting events. Redesigned HEs with tailored specificities can be used to cleave new sequences, thereby considerably expanding the number of targetable genes and loci. With HEs, as well as with other protein scaffolds, context dependence of DNA/protein interaction patterns remains one of the major limitations for rational engineering of new DNA binders. Previous studies have shown strong crosstalk between different residues and regions of the DNA binding interface. To investigate this phenomenon, we systematically combined mutations from three groups of amino acids in the DNA binding regions of the I-CreI HE. Our results confirm that important crosstalk occurs throughout this interface in I-CreI. Detailed analysis of success rates identified a nearest-neighbour effect, with a more pronounced level of dependence between adjacent regions. Taken together, these data suggest that combinatorial engineering does not necessarily require the identification of separable functional or structural regions, and that groups of amino acids provide acceptable building blocks that can be assembled, overcoming the context dependency of the DNA binding interface. Furthermore, the present work describes a sequential method to engineer tailored HEs, wherein three contiguous regions are individually mutated and assembled to create HEs with engineered specificity. PMID:21482539

  17. Automated embolus identification using a rule-based expert system.

    PubMed

    Fan, L; Evans, D H; Naylor, A R

    2001-08-01

    Transcranial Doppler ultrasound (US) can be used to detect microemboli in the cerebral circulation, but is still limited because it usually relies on "human experts" (HEs) to identify signals corresponding to embolic events. The purpose of this study was to develop an automatic system that could replace the HE and, thus, make the technique more widely applicable and, potentially, more reliable. An expert system, based around a digital signal-processing board, analysed Doppler signal patterns in both the time domain and frequency domain. The system was trained and tested on Doppler signals recorded during the dissection and recovery phases of carotid endarterectomy. It was tested with 74 separate 2.5-min recordings that contained at least 575 artefacts in addition to 253 s of diathermy interference. The results were compared with the results obtained by three HEs. Using a "gold-standard" that classified any event detected by the majority of HEs as an embolus, the automatic system displayed a sensitivity of 94.7% and a specificity of 95.1% for 1151 candidate events 7 dB or more above the clutter (signal-to-clutter ratio, SCR, > or = 7 dB), and 89.6% and 95.3%, respectively, for 2098 candidate events with SCR > or = 5 dB. The system had a very similar performance to individual HEs for SCR > or = 7dB, and was only marginally worse for SCR > or = 5 dB. PMID:11527593

  18. Eosinophilic Granulomatosis With Polyangiitis With Thrombotic Microangiopathy: Is Simultaneous Systemic Lupus Erythematosus Associated With Clinical Manifestations?: A Case Report and Review of the Literature.

    PubMed

    Fukui, Shoichi; Iwamoto, Naoki; Tsuji, Sosuke; Umeda, Masataka; Nishino, Ayako; Nakashima, Yoshikazu; Suzuki, Takahisa; Horai, Yoshiro; Koga, Tomohiro; Kawashiri, Shin-Ya; Ichinose, Kunihiro; Hirai, Yasuko; Tamai, Mami; Nakamura, Hideki; Origuchi, Tomoki; Kinoshita, Naóe; Kawakami, Atsushi

    2015-11-01

    Eosinophilic granulomatosis with polyangiitis (EGPA) is one of the antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, which is characterized by vasculitis of the small to medium-sized vessels. On the contrary, thrombotic microangiopathy (TMA) is a life-threatening condition which can cause ischemic organ injury. Although several case reports have described patients with TMA associated with ANCA-associated vasculitis except for EGPA, there are no previous case reports of EGPA associated with TMA.A 71-year-old Japanese man was diagnosed with EGPA based on his asthma, eosinophilia, lung opacity, refractory sinusitis, and positive myeloperoxidase-ANCA. He was also diagnosed with TMA based on peripheral schizocytes and hemolytic anemia. We performed plasmapheresis and started high-dose corticosteroid therapy; thereafter, he improved promptly. His case also fulfilled the classification criteria of systemic lupus erythematosus (SLE) based on the pleural effusion, renal disorder, anemia, thrombocytopenia, positive antidouble-stranded DNA antibody, and low complement. Elements of SLE were thought to affect his clinical course.We reviewed 11 patients with EGPA or hypereosinophilic syndrome (HES) associated with SLE, including our case. Patients with EGPA or HES associated with SLE had more heart complications than patients with simple EGPA or simple HES did. Patients with EGPA or HES associated with SLE had more pleural effusion than patients with simple SLE did.Clinical manifestations of eosinophilia with SLE or SLE with eosinophilia may differ from simple SLE or simple eosinophilia. PMID:26559264

  19. Cyclin D2 and the CDK substrate p220(NPAT) are required for self-renewal of human embryonic stem cells.

    PubMed

    Becker, Klaus A; Ghule, Prachi N; Lian, Jane B; Stein, Janet L; van Wijnen, Andre J; Stein, Gary S

    2010-02-01

    Self-renewal of pluripotent human embryonic stem (hES) cells utilizes an abbreviated cell cycle that bypasses E2F/pRB-dependent growth control. We investigated whether self-renewal is alternatively regulated by cyclin/CDK phosphorylation of the p220(NPAT)/HiNF-P complex to activate histone gene expression at the G1/S phase transition. We show that cyclin D2 is prominently expressed in pluripotent hES cells, but cyclin D1 eclipses cyclin D2 during differentiation. Depletion of cyclin D2 or p220(NPAT) causes a cell cycle defect in G1 reflected by diminished phosphorylation of p220(NPAT), decreased cell cycle dependent histone H4 expression and reduced S phase progression. Thus, cyclin D2 and p220(NPAT) are principal cell cycle regulators that determine competency for self-renewal in pluripotent hES cells. While pRB/E2F checkpoint control is relinquished in human ES cells, fidelity of physiological regulation is secured by cyclin D2 dependent activation of the p220(NPAT)/HiNF-P mechanism that may explain perpetual proliferation of hES cells without transformation or tumorigenesis. PMID:19890848

  20. Alternatives to the Fish Early Life-Stage Test: A Research Strategy for Discovering and Annotating Adverse Outcome Pathways During Fish Development

    EPA Science Inventory

    The OECD 210 fish early life]stage (FELS) test is the primary guideline test used to estimate chronic fish toxicity, as well as support ecological risk assessments and chemical management programs around the world. As a step toward developing alternatives to the FELS test, a HES...

  1. Effect of haemodilution, acidosis, and hypothermia on the activity of recombinant factor VIIa (NovoSeven®)

    PubMed Central

    Viuff, D.; Lauritzen, B.; Pusateri, A. E.; Andersen, S.; Rojkjaer, R.; Johansson, P. I.

    2008-01-01

    Background A range of plasma volume expanders is used clinically, often in settings where haemostasis may already be impaired. The haemostatic agent, recombinant activated factor VII (rFVIIa, NovoSeven®), may be used to improve haemostasis but potential interactions with different volume expanders are poorly understood. Methods Clot formation was measured by thromboelastography (TEG) using blood from healthy volunteers. In vitro effects of rFVIIa with haemodilution, acidosis, and hypothermia were examined. Conditions were induced by dilution with NaCl (0.9%), lactated Ringer's solution, albumin 5%, or hydroxyethyl starch (HES) solutions [MW (molecular weight) 130–670 kDa]; by adjusting pH to 6.8 with 1 M HEPES (N-2-hydroxyethylpiperazine-N′-2-ethanesulphonic acid) buffer; or by reducing temperature to 32°C. We also studied the effect of low vs high MW HES (MW 200 vs 600 kDa) and rFVIIa on in vivo bleeding time (BT) in rabbits. Results Haemodilution progressively altered TEG parameters. rFVIIa improved TEG parameters in the presence of acidosis, hypothermia or 20% haemodilution (P<0.05). At 40% haemodilution, the rFVIIa effect was diminished particularly with high MW HES. In vivo, rFVIIa shortened the BT (P<0.05) with low but not high MW HES. Conclusions Efficacy of rFVIIa was affected by the degree of haemodilution and type of volume expander, but not by acidosis or hypothermia. PMID:18565966

  2. Investigating Students' Ideas about Buoyancy and the Influence of Haptic Feedback

    ERIC Educational Resources Information Center

    Minogue, James; Borland, David

    2016-01-01

    While haptics (simulated touch) represents a potential breakthrough technology for science teaching and learning, there is relatively little research into its differential impact in the context of teaching and learning. This paper describes the testing of a haptically enhanced simulation (HES) for learning about buoyancy. Despite a lifetime of…

  3. Evaluation of 1066 ToxCast Chemicals in a human stem cell assay for developmental toxicity (SOT)

    EPA Science Inventory

    To increase the diversity of assays used to assess potential developmental toxicity, the ToxCast chemical library was screened in the Stemina devTOX quickPREDICT assay using human embryonic stem (hES) cells. A model for predicting teratogenicity was based on a training set of 23 ...

  4. THE CONTRIBUTION OF AMBIENT PM2.5 TO TOTAL PERSONAL EXPOSURES: RESULTS FROM A POPULATION EXPOSURE MODEL FOR PHILADELPHIA, PA

    EPA Science Inventory

    The US EPA National Exposure Research Laboratory (NERL) is currently developing an integrated human exposure source-to-dose modeling system (HES2D). This modeling system will incorporate population exposure modules that use a probabilistic approach to predict population exposu...

  5. Computational enzyme design

    NASA Astrophysics Data System (ADS)

    Bolon, Daniel N.

    2002-08-01

    The long-term objective of computational enzyme design is the ability to generate efficient protein catalysts for any chemical reaction. This thesis develops and experimentally validates a general computational approach for the design of enzymes with novel function. In order to include catalytic mechanism in protein design, a high-energy state (HES) rotamer (side chain representation) was constructed. In this rotamer, substrate atoms are in a HES. In addition, at least one amino acid side chain is positioned to interact favorably with substrate atoms in their HES and facilitate the reaction. Including an amino acid side chain in the HES rotamer automatically positions substrate relative to a protein scaffold and allows protein design algorithms to search for sequences capable of interacting favorably with the substrate. Because chemical similarity exists between the transition state and the high-energy state, optimizing the protein sequence to interact favorably with the HES rotamer should lead to transition state stabilization. In addition, the HES rotamer model focuses the subsequent computational active site design on a relevant phase space where an amino acid is capable of interacting in a catalytically active geometry with substrate. Using a HES rotamer model of the histidine mediated nucleophilic hydrolysis of p-nitrophenyl acetate, the catalytically inert 108 residue E. coli thioredoxin as a scaffold, and the ORBIT protein design software to compute sequences, an active site scan identified two promising active site designs. Experimentally, both candidate ?protozymes? demonstrated catalytic activity significantly above background. In addition, the rate enhancement of one of these ?protozymes? was the same order of magnitude as the first catalytic antibodies. Because polar groups are frequently buried at enzyme-substrate interfaces, improved modeling of buried polar interactions may benefit enzyme design. By studying native protein structures, rules have been

  6. Significance of Dynamic and Transient Analysis in the Design and Operation of Hybrid Energy Systems

    SciTech Connect

    Panwar, Mayank; Mohanpurkar, Manish; Hovsapian, Rob; Osorio, Julian D.

    2015-02-01

    Energy systems were historically designed and operated with a specific energy conversion objective, while managing loads and resources. In the recent years, the increased utilization of non-dispatchable renewable sources such as wind and solar has played a role in power quality and the reliability of power systems. In order to mitigate the risk associated with the non-dispatchable resources an integrated approach, such as Hybrid Energy Systems (HES), has to be taken, integrating the loads and resource management between the traditional thermal power plants and the non-dispatchable resources. As our electric energy becomes more diverse in its generation resources, the HES with its operational control system, its real-time view and its dynamic decisions making will become an essential part of the integrated energy systems and improve the overall grid reliability. The operational constraints of the energy sources on both the thermal power plants and the non-dispatchable resources in HES, plays a vital role in the planning and design stage. It is an established fact that the choice of energy source depends on the available natural resources and possible infrastructure. A critical component of decision-making depends on the complementary nature and controllability of the energy sources to supply the load demands with high reliability. Controllability of complex HES to achieve desired performance and flexibility is implemented via coordinated control systems while simultaneously generating electricity and other useful products such as useful heat or hydrogen. These systems are based on instrumentation, signal processing, control theory, and engineering system design. The entire HES along with the control systems are characterized by widely varying time constants. Hence, for a well-coordinated control and operation, we propose physics based modeling of the subsystems to assist in a dynamic and transient analysis. Dynamic and transient analysis in real and non-real time

  7. Immunostaining of phospho-histone H3 and Ki-67 improves reproducibility of recurrence risk assessment of gastrointestinal stromal tumors.

    PubMed

    Uguen, Arnaud; Conq, Gwenaël; Doucet, Laurent; Talagas, Matthieu; Costa, Sebastian; De Braekeleer, Marc; Marcorelles, Pascale

    2015-07-01

    Gastrointestinal stromal tumors (GISTs) are the most common non-epithelial tumors in the digestive tract. Beyond surgery, therapeutic management depends on risk of recurrence. Risk evaluation of GIST takes into account location and size of the tumor, whether or not the tumor was intact or ruptured and mitotic index. The mitotic index lacks in intra- and interobserver reproducibility. In this study, we evaluated on 61 GISTs, the reproducibility of mitotic counting using classical hematoxylin-eosin-saffron (HES) staining, and its correlation with the mitotic count obtained through immunohistochemical staining for phospho-histone H3 (PHH3) and the proliferation index based upon Ki-67 immunostaining. Mitotic counts by HES and PHH3 staining and Ki-67 proliferation index were evaluated twice by three independent observers taking into account interpretation times per tumor for each technique. HES-based and PHH3-based mitotic counts and Ki-67 proliferation index correlated well and presented good intra- and interobserver reproducibility. PHH3 staining resulted in a slight but statistically significant difference of about two more mitotic figures per 5 mm(2) than the HES-based count, which might have put some borderline tumors in a different risk category. Immunohistochemical staining for PHH3 and Ki-67 allowed more rapid interpretation than mitotic counts based upon HES staining, but only PHH3 staining allows counting of mitoses. Immunostaining using anti-PHH3 and anti-Ki-67 antibodies will eventually provide improved recurrence risk stratification of GIST and may become effective ancillary tools in deciding on optimal therapeutic management. PMID:25823616

  8. Long-term safety of mepolizumab for the treatment of hypereosinophilic syndromes

    PubMed Central

    Roufosse, Florence E; Kahn, Jean-Emmanuel; Gleich, Gerald J; Schwartz, Lawrence B; Singh, Anish D; Rosenwasser, Lanny J; Denburg, Judah A; Ring, Johannes; Rothenberg, Marc E; Sheikh, Javed; Haig, Ann E; Mallett, Stephen A; Templeton, Deborah N; Ortega, Hector G; Klion, Amy D

    2012-01-01

    Background Hypereosinophilic syndromes (HES) are chronic disorders that require long-term therapy to suppress eosinophilia and clinical manifestations. Corticosteroids are usually effective, yet many patients become corticosteroid-refractory or develop corticosteroid toxicity. Mepolizumab, a humanised monoclonal anti-interleukin-5 antibody, demonstrated corticosteroid-sparing effects in a double-blind, placebo-controlled study of FIP1L1/PDGFRA-negative, corticosteroid-responsive subjects with HES. Objective To evaluate long-term safety and efficacy of mepolizumab (750 mg) in HES. Methods MHE100901 is an open-label extension study. The primary endpoint was the frequency of adverse events (AEs). Optimal dosing frequency, corticosteroid-sparing effect of mepolizumab, and development of anti-mepolizumab antibodies were also explored. Results Seventy-eight subjects received 1–66 mepolizumab infusions each (including mepolizumab infusions received in the placebo-controlled trial). Mean exposure was 251 weeks (range 4–302). The most common dosing interval was 9–12 weeks. The incidence of AEs was 932 events per 100 subject-years in the first year, declining to 461 events per 100 subject-years after 48 months. Serious AEs, including one death, were reported by the investigator as possibly due to mepolizumab in three subjects. The median daily prednisone dose decreased from 20.0 to 0 mg in the first 24 weeks. The median average daily dose for all subjects over the course of the study was 1.8 mg. Sixty-two percent of subjects were prednisone-free without other HES medications for ≥12 consecutive weeks. No neutralizing antibodies were detected. Twenty-four subjects withdrew prior to study completion for death (n=4), lack of efficacy (n=6), or other reasons. Conclusion Mepolizumab was well tolerated and effective as a long-term corticosteroid-sparing agent in PDGFRA-negative HES. PMID:23040887

  9. Structural basis for ligand and substrate recognition by torovirus hemagglutinin esterases

    PubMed Central

    Langereis, Martijn A.; Zeng, Qinghong; Gerwig, Gerrit J.; Frey, Barbara; von Itzstein, Mark; Kamerling, Johannis P.; de Groot, Raoul J.; Huizinga, Eric G.

    2009-01-01

    Hemagglutinin esterases (HEs), closely related envelope glycoproteins in influenza C and corona- and toroviruses, mediate reversible attachment to O-acetylated sialic acids (Sias). They do so by acting both as lectins and as receptor-destroying enzymes, functions exerted by separate protein domains. HE divergence was accompanied by changes in quaternary structure and in receptor and substrate specificity. The selective forces underlying HE diversity and the molecular basis for Sia specificity are poorly understood. Here we present crystal structures of porcine and bovine torovirus HEs in complex with receptor analogs. Torovirus HEs form homodimers with sialate-O-acetylesterase domains almost identical to corresponding domains in orthomyxo- and coronavirus HEs, but with unique lectin sites. Structure-guided biochemical analysis of the esterase domains revealed that a functionally, but not structurally conserved arginine–Sia carboxylate interaction is critical for the binding and positioning of glycosidically bound Sias in the catalytic pocket. Although essential for efficient de-O-acetylation of Sias, this interaction is not required for catalysis nor does it affect substrate specificity. In fact, the distinct preference of the porcine torovirus enzyme for 9-mono- over 7,9-di-O-acetylated Sias can be explained from a single-residue difference with HEs of more promiscuous specificity. Apparently, esterase and lectin pockets coevolved; also the porcine torovirus HE receptor-binding site seems to have been designed to use 9-mono- and exclude di-O-acetylated Sias, possibly as an adaptation to replication in swine. Our findings shed light on HE evolution and provide fundamental insight into mechanisms of substrate binding, substrate recognition, and receptor selection in this important class of virion proteins. PMID:19721004

  10. Human Embryonic Stem Cells Form Functional Thyroid Follicles

    PubMed Central

    Latif, Rauf; Davies, Terry F.

    2015-01-01

    Objective: The molecular events that lead to human thyroid cell speciation remain incompletely characterized. It has been shown that overexpression of the regulatory transcription factors Pax8 and Nkx2-1 (ttf-1) directs murine embryonic stem (mES) cells to differentiate into thyroid follicular cells by initiating a transcriptional regulatory network. Such cells subsequently organized into three-dimensional follicular structures in the presence of extracellular matrix. In the current study, human embryonic stem (hES) cells were studied with the aim of recapitulating this scenario and producing functional human thyroid cell lines. Methods: Reporter gene tagged pEZ-lentiviral vectors were used to express human PAX8-eGFP and NKX2-1-mCherry in the H9 hES cell line followed by differentiation into thyroid cells directed by Activin A and thyrotropin (TSH). Results: Both transcription factors were expressed efficiently in hES cells expressing either PAX8, NKX2-1, or in combination in the hES cells, which had low endogenous expression of these transcription factors. Further differentiation of the double transfected cells showed the expression of thyroid-specific genes, including thyroglobulin (TG), thyroid peroxidase (TPO), the sodium/iodide symporter (NIS), and the TSH receptor (TSHR) as assessed by reverse transcription polymerase chain reaction and immunostaining. Most notably, the Activin/TSH-induced differentiation approach resulted in thyroid follicle formation and abundant TG protein expression within the follicular lumens. On stimulation with TSH, these hES-derived follicles were also capable of dose-dependent cAMP generation and radioiodine uptake, indicating functional thyroid epithelial cells. Conclusion: The induced expression of PAX8 and NKX2-1 in hES cells was followed by differentiation into thyroid epithelial cells and their commitment to form functional three-dimensional neo-follicular structures. The data provide proof of principal that hES cells can be

  11. Contribution of osmotic changes to disintegrative globulization of single cortical fibers isolated from rat lens.

    PubMed

    Wang, L F; Dhir, P; Bhatnagar, A; Srivastava, S K

    1997-08-01

    In this study the contribution of osmotic changes to disintegrative globulization of lens cortical fibers was examined. Single fiber cells were isolated by trypsinization of adult rat lens cortex, and morphological changes elicited by exposure to different external solutions were monitored optically. The survival of the fiber-shaped cells was analysed in accordance with the Weibull distribution. Changes in [Ca2+]i were measured using the fluorescent calcium-sensitive dye-Fluo-3. Exposure of isolated fiber cells to Ringer's solution (containing 2 mm Ca2+) led to an exponential increase in [Ca2+]i with a time constant of 10.2+/-0.8 min, and caused disintegrative globulization in 25+/-4 min (=Tg). The process of globulization as well as the rate of increase in [Ca2+]i was delayed by removing Cl- ions from the external media. Globulization was also delayed by adding 20% bovine serum albumin (Tg=107+/-3 min) or chloride channel inhibitors 5, nitro-2-(3-phenylpropylamino) benzoate (NPPB), dideoxyforskolin, niflumic acid, and tamoxifen. When the fiber cells were suspended in isotonic (280 mm sucrose) HEPES-sucrose (HS) or HEPES-EDTA-sucrose (HES) solution, no globulization was observed for an observation time of 120 min. However, exposure to hypotonic (180 mm) HES solution led to disintegration of fiber cells in 75+/-7 min. Disintegration of the fiber induced by hypotonic HES solution could be delayed by either 0. 05 mm leupeptin (Tg=97+/-6 min) or by pre-loading the fibers with BAPTA (Tg=100+/-4 min). Inhibition of membrane calcium transport by 0.5 mm La3+ had no effect on Tg in hypotonic HES. Addition of 2 mm Ca2+ to HES solution accelerated globulization, and Tg was 57+/-4, 69+/-5 and 102+/-6 min for hypo-, iso- and hyper- tonic solutions, respectively. Transient exposure to calcium also accelerated disintegrative globulization of fiber cells exposed subsequently to HES solution. These results suggest that in ionic media, part of the calcium influx in isolated fiber

  12. Home Energy Saver v.2.0

    Energy Science and Technology Software Center (ESTSC)

    2008-09-01

    A web-based residential energy calculator. Provides customized estimates of residential energy use, energy bills, and CO2 emissions, based on building description information provided by the user. Energy use is estimated by end-use and device, using engineering models. Space heating and cooling use is based on the DOE-2.1E building simulation model. Other end-uses (water heating, appliances, lighting, and miscellaneous equipment) are based on engineering models developed by LBNL. Users can estimate their household carbon footprint andmore » compare it to average vaules for their neighborhood and other regions, displayed using the Google Maps API. Energy bills can be calculated using either average energy price data or actual utility tariffs (including time-of-use) contained in the LBNL Tariff Analysis Project (TAP). HES includes a link to the TAP energy bill calculator web service. The HES software also includes extensive default input data for required user inputs.« less

  13. U.S. Hydropower Resource Assessment - California

    SciTech Connect

    A. M. Conner; B. N. Rinehart; J. E. Francfort

    1998-10-01

    The U.S. Department of Energy is developing an estimate of the underdeveloped hydropower potential in the United States. For this purpose, the Idaho National Engineering and Environmental Laboratory developed a computer model called Hydropower Evaluation Software (HES). HES measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of California.

  14. Inhibitory effect of CT domain of CCN3/NOV on proliferation and differentiation of osteogenic mesenchymal stem cells, Kusa-A1

    SciTech Connect

    Katsuki, Yuko; Sakamoto, Kei; Minamizato, Tokutaro; Makino, Hatsune; Umezawa, Akihiro; Ikeda, Masa-aki; Perbal, Bernard; Amagasa, Teruo; Yamaguchi, Akira; Katsube, Ken-ichi

    2008-04-11

    CCN3/NOV activates the Notch signal through the carboxyl terminal cysteine-rich (CT) domain. CCN3 transfection to Kusa-A1 inhibited osteogenic differentiation and cell proliferation, which is accompanied by upregulation of Hes/Hey, Notch downstream targets, and p21, a CDK inhibitor. Upregulation of Hes/Hey and p21 was abrogated by the deletion of CT domain. Anti-proliferative activity of CCN3 was also abrogated by CT domain deletion whereas anti-osteogenic activity was not completely abrogated. We found that CT domain-deleted CCN3 still possesses antagonistic effect on BMP-2. These results suggest that CCN3 employs Notch and BMP pathways in anti-osteogenic activity while it inhibits cell proliferation uniquely by Notch/p21 pathway.

  15. U.S. hydropower resource assessment for Nebraska

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-04-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Nebraska.

  16. U.S. hydropower resource assessment for Michigan

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-02-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering and Environmental Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Michigan.

  17. U.S. hydropower resource assessment for Nevada

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-10-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Nevada.

  18. U.S. hydropower resource assessment for Tennessee

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-07-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Tennessee.

  19. US hydropower resource assessment for Montana

    SciTech Connect

    Francfort, J.E.

    1993-12-01

    The Department of Energy is developing an estimate of the hydropower development potential in this country. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE menu-driven software application that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the state of Montana.

  20. U.S. hydropower resource assessment for North Carolina

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-10-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of North Carolina.

  1. U.S. hydropower resource assessment for West Virginia

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-02-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of West Virginia.

  2. Formation and properties of silicon/fluorite heterostructures. Final report, June 1, 1994--May 31, 1997

    SciTech Connect

    Olmstead, M.A.

    1997-10-01

    Our primary goal during the previous support periods was to determine the interrelationship among the structure of the CaF{sub 2}/Si interface, the growth kinetics, the structure and morphology of the overlying film, and the intrinsic dielectric properties of the interface itself. The experiments were designed (i) to determine the CaF{sub 2} overlayer structure and morphology as a function of kinetic conditions [Den93a, Den93b, Won93, Den95, Hes95], (ii) to develop and test models explaining the observed structures [Den95, Hes95], (iii) to investigate the stability of the interface structure [Les97] and (iv) to determine and interpret the photoelectron kinetic energy distributions as a function of the emitting atom location [Rot93, Rot94, Rot96]. As summarized below, we were successful in accomplishing these experiments and in largely fulfilling our original goal.

  3. Condenser performance test and back-pressure improvement: Final report

    SciTech Connect

    Piskorowski, J.; Beckett, G.; Bell, R.

    1988-04-01

    This document describes condenser performance test and analyses experiences. The testing was performed by Indianapolis Power and Light Company (IPL) on the Petersburg Unit 3 condenser. The initial testing revealed a performance deficiency. Modifications were made to the condenser, air in-leakage was reduced and the vacuum pumps were brought back to their original design capacity. Testing was reperformed after these activities and although a significant performance improvement was achieved deficiencies were still evident. Heat Exchanger Systems, Inc. (HES) was retained as consultants during this testing program. The Central Electricity Generating Board's (CEGB) Central Electricity Research Laboratory (CERL) acting as a subcontractor to HES were retained to perform an analysis of the Petersburg Unit 3 condenser using their EPOC computer code. The results of this analysis are also contained in this document. 3 refs., 48 figs., 3 tabs.

  4. US hydropower resource assessment for Kansas

    SciTech Connect

    Francfort, J.E.

    1993-12-01

    The Department of Energy is developing an estimate of the hydropower development potential in this country. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE menu-driven software application that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the state of Kansas.

  5. U.S. hydropower resource assessment for Alaska

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-11-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Alaska.

  6. Caveolin-1 regulates neural differentiation of rat bone mesenchymal stem cells into neurons by modulating Notch signaling.

    PubMed

    Wang, Shuyang; Kan, Quancheng; Sun, Yingpu; Han, Rui; Zhang, Guangyu; Peng, Tao; Jia, Yanjie

    2013-02-01

    Bone marrow mesenchymal stem cells (MSCs) are known to differentiate into neurons in vitro. However, the mechanism underlying MSC differentiation remains controversial. A recent analysis has shown that Notch signaling is involved in regulating the differentiation of MSCs. This study examines the potential mechanism of the differentiation of MSCs into neurons, and it considers the role of caveolin-1 in this process. We investigated neuron differentiation and Notch signaling by detecting the expression levels of microtubule-associated protein 2 (MAP-2), Neuron-specific Enolase (NSE), Notch-1, Notch intracellular domain (NICD) and hairy enhancer of split 5 (Hes5). We found that by down-regulating caveolin-1 during induction, MSCs were prone to neural differentiation and expressed high levels of neuronal markers. Meanwhile, the expression levels of Notch-1, NICD and Hes5 decreased. Our results indicate that down-regulation of caveolin-1 promotes the neuronal differentiation of MSCs by modulating the Notch signaling pathway. PMID:23031836

  7. Random mitotic activities across human embryonic stem cell colonies.

    SciTech Connect

    Jin, Q.; Duggan, R.; Dasa, S.; Li, F.; Chen, L.

    2010-08-01

    A systemic and quantitative study was performed to examine whether different levels of mitotic activities, assessed by the percentage of S-phase cells at any given time point, existed at different physical regions of human embryonic stem (hES) cell colonies at 2, 4, 6 days after cell passaging. Mitotically active cells were identified by the positive incorporation of 5-bromo-2-deoxyuridine (BrdU) within their newly synthesized DNA. Our data indicated that mitotically active cells were often distributed as clusters randomly across the colonies within the examined growth period, presumably resulting from local deposition of newly divided cells. This latter notion was further demonstrated by the confined growth of enhanced green florescence protein (EGFP) expressing cells amongst non-GFP expressing cells. Furthermore, the overall percentage of mitotically active cells remained constantly at about 50% throughout the 6-day culture period, indicating mitotic activities of hES cell cultures were time-independent under current growth conditions.

  8. U.S. hydropower resource assessment for New Mexico

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-03-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of New Mexico.

  9. U.S. hydropower resource assessment for Maryland

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-11-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Maryland.

  10. Home Energy Saver v.2.0

    SciTech Connect

    Mills, Evan; Brown, Richard; Pinckard, Maggie; Warner, Jeff

    2008-09-01

    A web-based residential energy calculator. Provides customized estimates of residential energy use, energy bills, and CO2 emissions, based on building description information provided by the user. Energy use is estimated by end-use and device, using engineering models. Space heating and cooling use is based on the DOE-2.1E building simulation model. Other end-uses (water heating, appliances, lighting, and miscellaneous equipment) are based on engineering models developed by LBNL. Users can estimate their household carbon footprint and compare it to average vaules for their neighborhood and other regions, displayed using the Google Maps API. Energy bills can be calculated using either average energy price data or actual utility tariffs (including time-of-use) contained in the LBNL Tariff Analysis Project (TAP). HES includes a link to the TAP energy bill calculator web service. The HES software also includes extensive default input data for required user inputs.

  11. Work-related injuries and occupational health and safety factors in smaller enterprises--a prospective study.

    PubMed

    Bull, N; Riise, T; Moen, B E

    2002-03-01

    The aim of this study was to determine whether any of the health, environmental and safety (HES) factors registered by visiting small mechanical enterprises in Norway at the start of the study could predict the risk of occupational injuries in subsequent years. Twelve HES factors, including injury awareness, programme for action, employee participation, training and use of personal safety devices, were registered. A questionnaire was completed by interviewing the employer and observing production. Two variables based on observation of the use of safety equipment were significantly correlated with occupational injuries. There is potential for prevention in smaller enterprises by increasing the use of personal protection devices and safety equipment on machines. Frequent inspection with feedback to the workers is probably the most effective means of attaining the desired result of reducing injuries. PMID:11967348

  12. U.S. hydropower resource assessment for Ohio

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-12-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Ohio.

  13. U.S. hydropower resource assessment for New York

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-08-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering and Environmental Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of New York.

  14. [Efficacy of plasma substitutes of different molecular weight in acute haemorrhagic shock in dogs (author's transl)].

    PubMed

    Klose, R; Hartung, H J; Ruffmann, R; Lutz, H

    1979-08-01

    Dogs were bled into haemorrhagic shock. They were then given isovolaemic infusions of dextran 60 and 40 and of hydroxyethyl starch with an average molecular weight of 200,000 and 40,000 respectively with a view of assessing the haemodynamic efficacy of these plasma substitutes. Solutions of high molecular weight hydroxyethyl starch (HES 400) were retained in the circulation for about the same length of time as was dextran 60. HES 40 (molecular weight 40,000) was retained for about 3-4 hours as measured by cardiac output. For normalizing a relative hypovolaemia, e.g. during anaesthesia or in some emergencies, colloidal plasma substitutes which will be retained for only a short time are entirely satisfactory. PMID:91161

  15. U.S. Hydropower Resource Assessment - Georgia

    SciTech Connect

    A. M. Conner; B. N. Rinehart; J. E. Francfort

    1998-10-01

    The U.S. Department of Energy is developing an estimate of the undeveloped hydropower potential in the United States. For this purpose, the Idaho National Engineering and Environmental Laboratory developed a computer model called Hydropower Evaluation Software (HES). HES measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Georgia.

  16. U.S. hydropower resource assessment for Alabama

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-02-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Alabama.

  17. US hydropower resource assessment for Texas

    SciTech Connect

    Francfort, J.E.

    1993-12-01

    The Department of Energy is developing an estimate of the hydropower development potential in this country. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE menu-driven software application that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the state of Texas.

  18. A flux-splitting method for hyperbolic-equation system of magnetized electron fluids in quasi-neutral plasmas

    NASA Astrophysics Data System (ADS)

    Kawashima, Rei; Komurasaki, Kimiya; Schönherr, Tony

    2016-04-01

    A flux-splitting method is proposed for the hyperbolic-equation system (HES) of magnetized electron fluids in quasi-neutral plasmas. The numerical fluxes are split into four categories, which are computed by using an upwind method which incorporates a flux-vector splitting (FVS) and advection upstream splitting method (AUSM). The method is applied to a test calculation condition of uniformly distributed and angled magnetic lines of force. All of the pseudo-time advancement terms converge monotonically and the conservation laws are strictly satisfied in the steady state. The calculation results are compared with those computed by using the elliptic-parabolic-equation system (EPES) approach using a magnetic-field-aligned mesh (MFAM). Both qualitative and quantitative comparisons yield good agreements of results, indicating that the HES approach with the flux-splitting method attains a high computational accuracy.

  19. The orientation-dependent expression of angiostatin-endostatin hybrid proteins and their characterization for the synergistic effects of antiangiogenesis.

    PubMed

    Paek, Sun-Yeol; Kim, Yong-Seok; Choi, Shin-Geon

    2010-10-01

    Two angiostatic fusion proteins (hAE and hEA) of human angiostatin (hAS) and endostatin (hES) proteins differed in tandem connection manner were constructed and evaluated for synergistic anti-angiogenic effects. The 65 kDa secreted fusion proteins from Pichia pastoris expression were verified by mass-spec analysis and western blotting assay. Luciferase reporter gene assay using VEGF promoter revealed that angiostatin-endostatin fusion protein (hAE) and its corresponding fusion gene delivery on Human Microvascular Endothelial Cells (HMEC-1) resulted in more potent synergistic anti-angiogenic effects than endostatin-angiostatin fusion protein (hEA). These facts suggest that the orientation of fusion genes between hAS and hES might be an important factor for developing therapeutic proteins. PMID:21030828

  20. U.S. hydropower resource assessment for Oregon

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-03-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering and Environmental Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Oregon.

  1. U.S. hydropower resource assessment for Pennsylvania

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-12-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Pennsylvania.

  2. Is hydroxyethyl starch necessary for sedimentation of bone marrow?

    PubMed

    Dijkstra-Tiekstra, Margriet J; Setroikromo, Airies C; Kraan, Marcha; Gkoumassi, Effimia; de Wildt-Eggen, Janny

    2015-02-01

    Hydroxyethyl starch (HES) is used to separate hematopoietic progenitor cells after bone marrow (BM) collection from red blood cells. The aims were to study alternatives for HAES-steril (200 kDa; not available anymore) and to optimize the sedimentation process. Using WBC-enriched product (10 × 10(9) WBC/L), instead of BM, sedimentation at 10% hematocrit using final 0.6 or 0.39% Voluven (130 kDa) or without HES appeared to be good alternatives for 0.6% HAES-steril. MNC recovery >80% and RBC depletion >90% was reached. Optimal sedimentation was reached using 110-140 mL volume. Centrifugation appeared not suitable for sedimentation. Additional testing with BM might be necessary to confirm these results. PMID:25544385

  3. Hawaii Energy Strategy program. Annual report 1993

    SciTech Connect

    Not Available

    1993-12-31

    This is the second annual report on the Hawaii Energy Strategy (HES) program which began on March 2, 1992, under a Cooperative Agreement (FCO3-92F19l68) with the United States Department of Energy (USDOE). The HES program is scheduled for completion by December 31, 1994. As outlined in the Statement of Joint Objectives. The purpose of the study is to develop an integrated State of Hawaii energy strategy, including an assessment of the State`s fossil fuel reserve requirements and the most effective way to meet those needs, the availability and practicality of increasing the use of native energy resources, potential alternative fossil energy technologies such as coal gasification and potential energy efficiency measures which could lead to demand reduction. This work contributes to the (US)DOE mission, will reduce the State`s vulnerability to energy supply disruptions and contributes to the public good.

  4. U.S. hydropower resource assessment for Illinois

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-01-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Illinois.

  5. U.S. hydropower resource assessment for Idaho

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-08-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering and Environmental Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Idaho.

  6. US hydropower resource assessment for Utah

    SciTech Connect

    Francfort, J.E.

    1993-12-01

    The Department of Energy is developing an estimate of the hydropower development potential in this country. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE menu-driven software application that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the state of Utah.

  7. U.S. hydropower resource assessment for Kentucky

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-07-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering and Environmental Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Kentucky.

  8. The roles of apoptotic pathways in the low recovery rate after cryopreservation of dissociated human embryonic stem cells

    PubMed Central

    Xu, Xia; Cowley, Sally; Flaim, Christopher J; James, William; Seymour, Leonard; Cui, Zhanfeng

    2010-01-01

    Human embryonic stem (hES) cells have enormous potential for clinical applications. However, one major challenge is to achieve high cell recovery rate after cryopreservation. Understanding how the conventional cryopreservation protocol fails to protect the cells is a prerequisite for developing efficient and successful cryopreservation methods for hES cell lines and banks. We investigated how the stimuli from cryopreservation result in apoptosis, which causes the low cell recovery rate after cryopreservation. The level of reactive oxygen species (ROS) is significantly increased, F-actin content and distribution is altered, and caspase-8 and caspase-9 are activated after cryopreservation. p53 is also activated and translocated into nucleus. During cryopreservation apoptosis is induced by activation of both caspase-8 through the extrinsic pathway and caspase-9 through the intrinsic pathway. However, exactly how the extrinsic pathway is activated is still unclear and deserves further investigation. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 PMID:20077485

  9. U.S. hydropower resource assessment for Arizona

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-10-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Arizona.

  10. US hydropower resource assessment for Wyoming

    SciTech Connect

    Francfort, J.E.

    1993-12-01

    The Department of Energy is developing an estimate of the hydropower development potential in this country. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE menu-driven software application that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the state of Wyoming.

  11. U.S. hydropower resource assessment for Florida

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1998-02-01

    The US Department of Energy is developing an estimate of the undeveloped hydropower potential in the US. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the US, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Florida.

  12. US hydropower resource assessment for Washington

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-07-01

    The U.S. Department of Energy is developing an estimate of the undeveloped hydropower potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. HES measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report describes the resource assessment results for the State of Washington.

  13. US hydropower resource assessment for Oklahoma

    SciTech Connect

    Francfort, J.E.

    1993-12-01

    The Department of Energy is developing an estimate of the hydropower development potential in this country. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose, The HES measures the potential hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a dBASE menu-driven software application that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the state of Oklahoma.

  14. Effects of Different Resuscitation Fluids on Pulmonary Expression of Aquaporin1 and Aquaporin5 in a Rat Model of Uncontrolled Hemorrhagic Shock and Infection

    PubMed Central

    Ge, Yali; Lin, Shunyan; Du, Jin

    2013-01-01

    Background To investigate the effects of fluids resuscitation on pulmonary expression of aquaporin1 and aquaporin5 in a rat model of uncontrolled hemorrhagic shock and infection. Methods Sixty Sprague-Dawley rats were randomly assigned to five groups, sham operation group (Group C) and four treated groups: no fluid resuscitation group (Group NF), groups resuscitated with Lactated Ringer's (LR),7.5% NaCl (HTS) and Hydroxyl ethyl starch (HES) respectively. Three-phased uncontrolled hemorrhagic shock and infection model was used. Phase I: Massive hemorrhage with a mean arterial pressure of 35–40 mmHg for 60 min, and followed by infection of lipopolysaccharide. Then some animals were resuscitated with solutions mentioned above, until 90 min. Phase II: At hemorrhagic shock 90 minutes, phase II of 60 minutes began with hemostasis and returning of all the initial shed blood. Phase III: Observation phase for 3.5 hours. After phase III, arterial blood gas analysis and the survival rates of the rats were recorded, Wet-to-dry lung weight ratio, BALF protein, pulmonary permeability index, and expressions of aquaporin1 and aquaporin5 were tested. Results The expressions of aquaporin1 and aquaporin5 were decreased in treatment groups comparing with sham operation group. Group HES and Group HTS decreased pulmonary vascular permeability and Wet-to-dry lung weight ratio, improved arterial blood gas analysis and survival rates, and attenuated the decreased pulmonary expression of aquaporin1 and aquaporin5 after the “two-hit”, comparing with groups NF and LR,but these beneficial effects were blunted in group HTS. Conclusion The expression of aquaporin1 and aquaporin5 may play important roles in formation of pulmonary edema. Resuscitation with HTS and HES, especially HES can reduce lung injury after hemorrhagic shock, partly by up-regulating the expressions of aquaporin1 and aquaporin5. PMID:23741323

  15. Comparison of Melatonin, Hypertonic Saline, and Hydroxyethyl Starch for Resuscitation of Secondary Intra-Abdominal Hypertension in an Animal Model.

    PubMed

    Chang, Mingtao; Tang, Hao; Liu, Dong; Li, Yang; Zhang, Lianyang

    2016-01-01

    A variety of agents may have a beneficial effect in reducing injury-induced intestinal edema of fluid, but studies confirming the efficacy and mechanisms of these agents in secondary intra-abdominal hypertension (IAH) are lacking. This study was to compare the effectiveness of melatonin, 7.5% hypertonic saline (HS), and hydroxyethyl starch 130/0.4 (HES) on the resuscitation of secondary IAH in a rat model. Female SD rats were divided into: sham group, shock group, lactated Ringer solution (LR) group, melatonin group, HS group, and HES group. Except for the sham group, all rats underwent a combination of inducing portal hypertension, hemorrhaging to a MAP of 40 mmHg for 2 hr, and using an abdominal restraint device. The collected blood was reinfused and the rats were treated with LR (30ml/h), melatonin (50 mg/kg) + LR, HS (6 ml/kg) + LR, and HES (30 ml/kg) + LR, respectively. The shock group received no fluids. LR was continuously infused for 6hr. The intestinal permeability, immunofluorescence of tight junction proteins, transmission electron microscopy, level of inflammatory mediators (TNF-a, IL-1β, IL-6) and of biochemical markers of oxidative stress (malondialdehyde, myeloperoxidase activity, and glutathione peroxidase) were assessed. Expressions of the protein kinase B (Akt) and of tight junction proteins were detected by Western blot. Compared with LR, HS, and HES, melatonin was associated with less inflammatory and oxidative injury, less intestinal permeability and injury, and lower incidence of secondary IAH in this model. The salutary effect of melatonin in this model was associated with the upregulation of intestinal Akt phosphorylation. PMID:27560478

  16. Derivation of human embryonic stem cell from spinal muscular atrophy patient.

    PubMed

    Xie, Pingyuan; Zhou, Hao; Zhou, Xiaoying; Zhao, Xiaomeng; Du, Juan; Lu, Guangxiu; Lin, Ge; Ouyang, Qi

    2016-03-01

    We established a human embryonic stem cell (hESC) line chHES-427 from the abnormal embryo carrying homozygous deletion of exon 7 of survival motor neuron gene (SMN). This cell line maintained a normal karyotype 46, XX during long-term culture. Further characteristic analysis suggested that the cells expressed the pluripotency-related markers and had the capacity to differentiate into the derivatives from the three germ layers in vitro. PMID:27345972

  17. The impact of NOTCH1, FBW7 and PTEN mutations on prognosis and downstream signaling in pediatric T- cell acute lymphoblastic leukemia: A report from the Children's Oncology Group

    PubMed Central

    Gedman, Amanda Larson; Chen, Qing; Desmoulin, Sita Kugel; Ge, Yubin; LaFiura, Katherine; Haska, Christina L.; Cherian, Christina; Devidas, Meenakshi; Linda, Stephen B.; Taub, Jeffrey W.; Matherly, Larry H.

    2009-01-01

    We explored the impact of mutations in the NOTCH1, FBW7 and PTEN genes on prognosis and downstream signaling in a well-defined cohort of 47 pediatric T-cell acute lymphoblastic leukemia (T-ALL) patients. In T-ALL lymphoblasts, we identified high frequency mutations in NOTCH1 (n=16), FBW7 (n=5) and PTEN (n=26). NOTCH1 mutations resulted in 1.3-3.3-fold increased transactivation of a HES1 reporter construct over wild-type NOTCH1; mutant FBW7 resulted in further augmentation of reporter gene activity. NOTCH1 and FBW7 mutations were accompanied by increased median transcripts for NOTCH1 target genes (HES1, DELTEX1, cMYC). However, none of these mutations were associated with treatment outcome. Elevated HES1, DELTEX1 and cMYC transcripts were associated with significant increases in transcript levels of several chemotherapy relevant genes, including MDR1, ABCC5, reduced folate carrier, asparagine synthetase, thiopurine methyltranserase, Bcl-2 and dihydrofolate reductase. PTEN transcripts positively correlated with HES1 and cMYC transcript levels. Our results suggest that (1) multiple factors should be considered with attempting to identify molecular-based prognostic factors for pediatric T-ALL, and (2) depending on the NOTCH1 signaling status, modifications in the types or dosing of standard chemotherapy drugs for T-ALL, or combinations of agents capable of targeting NOTCH1, AKT and/or mTOR with standard chemotherapy agents may be warranted. PMID:19340001

  18. Effects of goal-directed fluid therapy with different lactated Ringer's: hydroxyethyl starch ratios in hemorrhagic shock dogs.

    PubMed

    Tao, J P; Huang, Q Q; Huang, H Q; Yang, J J; Shi, M; Zhou, Y; Wan, L J; Zhou, C; Ou, Y J; Tong, Y Y; Yang, D G; Si, Y Y

    2015-01-01

    The effects of goal-directed fluid therapy, with lactated Ringer's (LR) and 6% hydroxyethyl starch (HES) solution, on hemorrhagic shock dogs are unknown. We aimed to determine the optimal LR: HES ratio for the resuscitation of hemorrhagic shock dogs. Hemorrhagic shock was induced in 40 ventilated dogs by drawing an estimated 60% blood volume. The animals were randomly divided into five groups (N = 8) according to the LR: HES ratio of the resuscitation fluid (3:1, 2:1, 1:1, 1:2, and 1:3), and were then resuscitated for 24 h to reach the stroke volume variation (SVV) and hemoglobin (Hb) goals by fluid infusion and autologous blood perfusion. The extravascular lung water index (EVLWI), pH, partial pressure of oxygen (PaO2), base excess (BE), sodium, chloride, Hb and creatinine clearance (Clearcrea) were checked after 24 h (R24). The EVLWI of the 3:1 group at R24 were higher than that of the 1:3 group and the baseline value (P < 0.05), whereas the PaO2 was lower (P < 0.05). In contrast to the 3:1 group at R24 and baseline, plasma chloride and sodium in the 1:3 and 1:2 groups increased; however, pH, BE, and Clearcrea decreased (P < 0.05). No significant differences were found in the 1:1 and 2:1 groups at R24 compared with baseline (P > 0.05). Resuscitation with LR and HES at 2:1 and 1:1 ratios are superior in maintaining the acid-base, electrolyte, and lung water balances as well as renal function in hemorrhagic shock dogs than at ratios of 3:l, 1:2, and1:3. PMID:26125873

  19. A SEARCH FOR UNRECOGNIZED CARBON-ENHANCED METAL-POOR STARS IN THE GALAXY

    SciTech Connect

    Placco, Vinicius M.; Rossi, Silvia; Kennedy, Catherine R.; Beers, Timothy C.; Lee, Young Sun; Christlieb, Norbert; Sivarani, Thirupathi; Reimers, Dieter; Wisotzki, Lutz

    2010-03-15

    We have developed a new procedure to search for carbon-enhanced metal-poor (CEMP) stars from the Hamburg/ESO (HES) prism-survey plates. This method employs an extended line index for the CH G band, which we demonstrate to have superior performance when compared to the narrower G-band index formerly employed to estimate G-band strengths for these spectra. Although CEMP stars have been found previously among candidate metal-poor stars selected from the HES, the selection on metallicity undersamples the population of intermediate-metallicity CEMP stars (-2.5 {<=} [Fe/H] {<=} -1.0); such stars are of importance for constraining the onset of the s-process in metal-deficient asymptotic giant branch stars (thought to be associated with the origin of carbon for roughly 80% of CEMP stars). The new candidates also include substantial numbers of warmer carbon-enhanced stars, which were missed in previous HES searches for carbon stars due to selection criteria that emphasized cooler stars. A first subsample, biased toward brighter stars (B< 15.5), has been extracted from the scanned HES plates. After visual inspection (to eliminate spectra compromised by plate defects, overlapping spectra, etc., and to carry out rough spectral classifications), a list of 669 previously unidentified candidate CEMP stars was compiled. Follow-up spectroscopy for a pilot sample of 132 candidates was obtained with the Goodman spectrograph on the SOAR 4.1 m telescope. Our results show that most of the observed stars lie in the targeted metallicity range, and possess prominent carbon absorption features at 4300 A. The success rate for the identification of new CEMP stars is 43% (13 out of 30) for [Fe/H] < -2.0. For stars with [Fe/H] < -2.5, the ratio increases to 80% (four out of five objects), including one star with [Fe/H] < -3.0.

  20. Evaluation of a minor eye conditions scheme delivered by community optometrists

    PubMed Central

    Konstantakopoulou, E; Edgar, D F; Harper, R A; Baker, H; Sutton, M; Janikoun, S; Larkin, G; Lawrenson, J G

    2016-01-01

    Background The establishment of minor eye conditions schemes (MECS) within community optometric practices provides a mechanism for the timely assessment of patients presenting with a range of acute eye conditions. This has the potential to reduce waiting times and avoid unnecessary referrals to hospital eye services (HES). Objective To evaluate the clinical effectiveness, impact on hospital attendances and patient satisfaction with a minor eye service provided by community optometrists. Methods Activity and outcome data were collected for 12 months in the Lambeth and Lewisham MECS. A patient satisfaction questionnaire was given to patients at the end of their MECS appointment. A retrospective difference-in-differences analysis of hospital activity compared changes in the volume of referrals by general practitioners (GPs) from a period before (April 2011–March 2013) to after (April 2013–March 2015) the introduction of the scheme in Lambeth and Lewisham relative to a neighbouring area (Southwark) where the scheme had not been commissioned. Appropriateness of case management was assessed by consensus using clinical members of the research team. Results A total of 2123 patients accessed the scheme. Approximately two-thirds of patients (67.5%) were referred by their GP. The commonest reasons for patients attending for a MECS assessment were ‘red eye’ (36.7% of patients), ‘painful white eye’ (11.1%) and ‘flashes and floaters’ (10.2%). A total of 64.1% of patients were managed in optometric practice and 18.9% were referred to the HES; of these, 89.2% had been appropriately referred. First attendances to HES referred by GPs reduced by 26.8% (95% CI −40.5% to −13.1%) in Lambeth and Lewisham compared to Southwark. Conclusions The Lambeth and Lewisham MECS demonstrates clinical effectiveness, reduction in hospital attendances and high patient satisfaction and represents a successful collaboration between commissioners, local HES units and primary

  1. Influence of colloid infusion on coagulation during off-pump coronary artery bypass grafting

    PubMed Central

    Muralidhar, K; Garg, Rajnish; Mohanty, SK; Banakal, Sanjay

    2010-01-01

    This study was conducted to determine the influence of colloid infusion on coagulation in patients undergoing off-pump coronary artery bypass grafting (OP-CABG). Thirty patients undergoing elective OP-CABG received medium molecular weight hydroxyethyl starch group I (MMW-HES 200/0.5), low molecular weight hydroxyethyl starch group II (LMW-HES 130/0.4) or gelatin group III (GEL) in a prospective randomized trial. Blood samples were assessed for hemoglobin (Hb), activated coagulation time (ACT), prothrombin time (PT), activated partial thromboplastin time (aPPT), platelet count, fibrinogen and von Willebrand factor (vWF) at specified intervals. Total volume of the colloid infused and postoperative chest-time drainage was also measured. There was a significant decrease in Hb, platelet count, fibrinogen levels in all these groups, which did not warrant blood transfusion. After the colloid infusion, vWF decreased significantly to 67% from baseline in group I as compared to 85 and 79% in group II and group III, respectively. vWF levels remained lower than the baseline value in the first 24 hours in group I, whereas this factor level increased above the baseline values in groups II and III, 6 hours postoperatively. Postoperative chest tube drainage in 24 hours was significantly higher in group I (856 ± 131 ml) as compared to group II (550 ± 124 ml) and group III (582 ± 159 ml). LMW-HES 130/0.4 was superior to MMW-HES 200/0.5 and gelatin in patients undergoing OP-CABG, in terms of better preservation of coagulation associated with enhanced volume effect. PMID:20661354

  2. Haemostasis monitored in stored red blood cells, plasma and platelet concentrates in the proportion of 4 :  4 :  1 diluted with crystalloids and colloids.

    PubMed

    Ågren, Anna; Edgren, Gustaf; Ambrosio, Daniela; Gryfelt, Gunilla; Östlund, Anders; Wikman, Agneta

    2016-04-01

    The aim of this in-vitro study was to evaluate haemostasis analysed with thromboelastometry and blood gas and blood count variables, in stored blood components and the effects after dilution with Ringer[Combining Acute Accent]s acetate, albumin and hydroxyethyl starch (HES). Aliquots from stored red blood cells, plasma and platelet concentrates were mixed in the proportion of 4 : 4 : 1 and analysed with rotational thromboelastometry (ROTEM), blood count [haemoglobin (Hb), haematocrit, platelet count] and blood gas (pH, calcium, sodium, potassium, glucose levels). The blood mix was thereafter diluted 20 and 33% with Ringer's acetate, albumin or HES. The stored blood component mix in a ratio of 4 : 4 : 1 had a low pH (7.11 ± 0.03, mean ± standard deviation), nonmeasurable calcium level, and high concentrations of sodium, potassium and glucose but ROTEM curves within normal range after recalcification. With Ringer's acetate dilution, the ROTEM variables changed almost linearly with increasing dilution volume. When albumin was used in the 33% dilution, the clot firmness of the fibrin clot (FibTEM) was further reduced, and with HES dilution, there was a pronounced impairment. The stored blood mix had a low pH and calcium level, both of which might have a significant influence on the coagulation process but normal ROTEM curves after recalcification. Dilution with Ringer's acetate and albumin resulted in moderate deterioration, while dilution with HES showed severely impaired haemostasis. PMID:26963027

  3. Comparison of Melatonin, Hypertonic Saline, and Hydroxyethyl Starch for Resuscitation of Secondary Intra-Abdominal Hypertension in an Animal Model

    PubMed Central

    Liu, Dong; Li, Yang; Zhang, Lianyang

    2016-01-01

    A variety of agents may have a beneficial effect in reducing injury-induced intestinal edema of fluid, but studies confirming the efficacy and mechanisms of these agents in secondary intra-abdominal hypertension (IAH) are lacking. This study was to compare the effectiveness of melatonin, 7.5% hypertonic saline (HS), and hydroxyethyl starch 130/0.4 (HES) on the resuscitation of secondary IAH in a rat model. Female SD rats were divided into: sham group, shock group, lactated Ringer solution (LR) group, melatonin group, HS group, and HES group. Except for the sham group, all rats underwent a combination of inducing portal hypertension, hemorrhaging to a MAP of 40 mmHg for 2 hr, and using an abdominal restraint device. The collected blood was reinfused and the rats were treated with LR (30ml/h), melatonin (50 mg/kg) + LR, HS (6 ml/kg) + LR, and HES (30 ml/kg) + LR, respectively. The shock group received no fluids. LR was continuously infused for 6hr. The intestinal permeability, immunofluorescence of tight junction proteins, transmission electron microscopy, level of inflammatory mediators (TNF-a, IL-1β, IL-6) and of biochemical markers of oxidative stress (malondialdehyde, myeloperoxidase activity, and glutathione peroxidase) were assessed. Expressions of the protein kinase B (Akt) and of tight junction proteins were detected by Western blot. Compared with LR, HS, and HES, melatonin was associated with less inflammatory and oxidative injury, less intestinal permeability and injury, and lower incidence of secondary IAH in this model. The salutary effect of melatonin in this model was associated with the upregulation of intestinal Akt phosphorylation. PMID:27560478

  4. Influence of laser induced hot electrons on the threshold for shock ignition of fusion reactions

    NASA Astrophysics Data System (ADS)

    Colaïtis, A.; Ribeyre, X.; Le Bel, E.; Duchateau, G.; Nicolaï, Ph.; Tikhonchuk, V.

    2016-07-01

    The effects of Hot Electrons (HEs) generated by the nonlinear Laser-Plasma Interaction (LPI) on the dynamics of Shock Ignition Inertial Confinement Fusion targets are investigated. The coupling between the laser beam, plasma dynamics and hot electron generation and propagation is described with a radiative hydrodynamics code using an inline model based on Paraxial Complex Geometrical Optics [Colaïtis et al., Phys. Rev. E 92, 041101 (2015)]. Two targets are considered: the pure-DT HiPER target and a CH-DT design with baseline spike powers of the order of 200-300 TW. In both cases, accounting for the LPI-generated HEs leads to non-igniting targets when using the baseline spike powers. While HEs are found to increase the ignitor shock pressure, they also preheat the bulk of the imploding shell, notably causing its expansion and contamination of the hotspot with the dense shell material before the time of shock convergence. The associated increase in hotspot mass (i) increases the ignitor shock pressure required to ignite the fusion reactions and (ii) significantly increases the power losses through Bremsstrahlung X-ray radiation, thus rapidly cooling the hotspot. These effects are less prominent for the CH-DT target where the plastic ablator shields the lower energy LPI-HE spectrum. Simulations using higher laser spike powers of 500 TW suggest that the CH-DT capsule marginally ignites, with an ignition window width significantly smaller than without LPI-HEs, and with three quarters of the baseline target yield. The latter effect arises from the relation between the shock launching time and the shell areal density, which becomes relevant in presence of a LPI-HE preheating.

  5. The Second Generation Hypernuclear Spectroscopy at JLab Hall C (E01-011 experiment)

    SciTech Connect

    Matsumura, Akihiko

    2008-11-01

    The second generation Λ hypernuclear spectroscopy by (e,e’K{sup +}) reaction has been carried out successfully at JLab in 2005. New configurations, HKS and Tilt method, significantly improved both energy resolution and statistics. Systematic error depend on tuning procedure was estimated by the blind analysis. Analysis is in the final stage. Third generation experiment (JLab E05-115) will be performed in the summer of 2009 w/ new e’ spectrometer (HES).

  6. Exploring unintended consequences of policy initiatives in mental health: the example of Child and Adolescent Mental Health Services (CAMHS) in England

    PubMed Central

    Foreman, David M

    2016-01-01

    Objectives The impact of policy and funding on Child and Adolescent Mental Health Service (CAMHS) activity and capacity, from 2003 to 2012, was assessed. The focus was on preschool children (aged 0–4 years), as current and 2003 policy initiatives stressed the importance of ‘early intervention’. Settings National service capacity from English CAMHS mapping was obtained from 2003 to 2008 inclusive. English Hospital Episode Statistics (HES) for English CAMHS was obtained from 2003 to 2012. The Child and Adolescent Faculty of the Royal College of Psychiatrists surveyed its members about comparative 0–4-year service activity and attitudes in 2012. Participants CAMHS services in England provided HES and CAMHS mapping data. The Child and Adolescent Faculty of the Royal College of Psychiatrists are child psychiatrists, including trainees. Outcome measures CAMHS mapping data provided national estimates of total numbers of CAMHS patients, whereas HES data counted appointments or episodes of inpatient care. The survey reported on Child Psychiatrists' informal estimates of service activity and attitudes towards children aged 0–4 years. Results The association between service capacity and service activity was moderated by an interaction between specified funding and age, the youngest children benefiting least from specified funding and suffering most when it was withdrawn (Pr=0.005). Policy review and significant differences between age-specific HES trends (Pr<0.001) suggested this reflected prioritisation of older children. Clinicians were unaware of this effect at local level, though it significantly influenced their attitudes to prioritising this group (Pr=0.02). Conclusions If the new policy initiative for CAMHS is to succeed, it will need to have time-limited priorities attached to sustained, specified funding, with planning for limits as well as expansion. Data collection for policy evaluation should include measures of capacity and activity. PMID:27496228

  7. Feeder-free and xeno-free culture of human pluripotent stem cells using UCBS matrix.

    PubMed

    Ding, Yan; Yang, Hua; Yu, Li; Xu, Chang Long; Zeng, Yi; Qiu, Ying; Li, Dong Sheng

    2015-10-01

    The ideal medium for human pluripotent stem cells (hPSCs) culture should be feeder-free, xeno-free, and completely defined. The present study aims to establish a new feeder-free and xeno-free system for culturing hPSCs. The system consists of the matrix, which was prepared from human umbilical cord blood serum (UCBS) and used to coat the culture plates, and the xeno-free medium, which was conventional serum-free hES medium supplemented with high concentrations of bFGF and Fibronectin. Compared with matrigel and mouse embryonic fibroblasts (MEFs), the UCBS matrix was proved to be equally suitable for the growth of hPSCs. After a series of experiments with different media and cytokins, the UCBS matrix was found worked the best with the basic medium (BM) supplemented with 20 ng/mL bFGF, 10 ug/mL fibronectin and Y-27632 for culture of hES cells. The hPSCs maintained normal karyotype, high proliferation rate and self-renewal ability after continuous culture more than 10 passages in this feeder-free and xeno-free system. Furthermore, a new human embryonic stem (hES) cell line was derived from discarded day 3 embryos in this newly developed culture system. In conclusion, this feeder-free and xeno-free system could not only be used to the culture hPSCs, but could also be used to derive new hES cell lines. PMID:25939663

  8. Quantifying the prevalence of frailty in English hospitals

    PubMed Central

    Soong, J; Poots, AJ; Scott, S; Donald, K; Woodcock, T; Lovett, D; Bell, D

    2015-01-01

    Objectives Population ageing has been associated with an increase in comorbid chronic disease, functional dependence, disability and associated higher health care costs. Frailty Syndromes have been proposed as a way to define this group within older persons. We explore whether frailty syndromes are a reliable methodology to quantify clinically significant frailty within hospital settings, and measure trends and geospatial variation using English secondary care data set Hospital Episode Statistics (HES). Setting National English Secondary Care Administrative Data HES. Participants All 50 540 141 patient spells for patients over 65 years admitted to acute provider hospitals in England (January 2005—March 2013) within HES. Primary and secondary outcome measures We explore the prevalence of Frailty Syndromes as coded by International Statistical Classification of Diseases, Injuries and Causes of Death (ICD-10) over time, and their geographic distribution across England. We examine national trends for admission spells, inpatient mortality and 30-day readmission. Results A rising trend of admission spells was noted from January 2005 to March 2013(daily average admissions for month rising from over 2000 to over 4000). The overall prevalence of coded frailty is increasing (64 559 spells in January 2005 to 150 085 spells by Jan 2013). The majority of patients had a single frailty syndrome coded (10.2% vs total burden of 13.9%). Cognitive impairment and falls (including significant fracture) are the most common frailty syndromes coded within HES. Geographic variation in frailty burden was in keeping with known distribution of prevalence of the English elderly population and location of National Health Service (NHS) acute provider sites. Overtime, in-hospital mortality has decreased (>65 years) whereas readmission rates have increased (esp.>85 years). Conclusions This study provides a novel methodology to reliably quantify clinically significant frailty

  9. Does delay in referral of proliferative diabetic retinopathy from the diabetic eye screening programme lead to visual loss?

    PubMed

    Negretti, G S; Amin, R; Webster, L; Egan, C A

    2016-06-01

    AimsTo ascertain the effect on visual acuity (VA) of a delay in Hospital Eye Service (HES) consultation for patients referred with proliferative diabetic retinopathy (PDR; R3) from the Diabetic Eye Screening Programme (DESP).MethodsAll patients referred to Moorfields Eye Hospital from DESP between April and December 2013 with a referral diagnosis of PDR in at least one eye were eligible. Screening programme VA was compared with VA at first HES appointment and final follow-up appointment. Reasons for any VA loss were noted.ResultsA total of 86 patients were included. Of these, 28 (33%) were seen in more than 4 weeks after their DESP referral. At first HES appointment, 39 (45%) patients were graded as having active PDR in at least one eye. Delay in referral did not significantly predict the likelihood of vision loss in all patients referred (χ(2), P=0.49) or in just those patients with a definitive HES diagnosis of active PDR (χ(2), P=1.00). In only 3 patients with active PDR was a delay in presentation thought to have led directly to VA loss.ConclusionsThere may be minimal short-term visual consequence in several weeks of delayed referral for many patients with a diagnosis of R3. However, the national guidance remains important. This is due to the occasional patient at very high risk of vision loss and the many gains for the patients in terms of time to properly assess medical and ocular conditions and counsel and consent them for treatment where necessary. PMID:27055673

  10. Impact of experimental haemodilution on platelet function, thrombin generation and clot firmness: effects of different coagulation factor concentrates

    PubMed Central

    Caballo, Carolina; Escolar, Gines; Diaz-Ricart, Maribel; Lopez-Vílchez, Irene; Lozano, Miguel; Cid, Joan; Pino, Marcos; Beltrán, Joan; Basora, Misericordia; Pereira, Arturo; Galan, Ana M.

    2013-01-01

    Background Haemodilution during resuscitation after massive haemorrhage may worsen the coagulopathy and perpetuate bleeding. Materials and methods Blood samples from healthy donors were diluted (30 and-60%) using crystalloids (saline, Ringer’s lactate, PlasmalyteTM) or colloids (6% hydroxyethylstarch [HES130/0.4], 5% human albumin, and gelatin). The effects of haemodilution on platelet adhesion (Impact R), thrombin generation (TG), and thromboelastometry (TEM) parameters were analysed as were the effects of fibrinogen, prothrombin complex concentrates (PCC), activated recombinant factor VII (FVIIa), and cryoprecipates on haemodilution. Results Platelet interactions was already significantly reduced at 30% haemodilution. Platelet reactivity was not improved by addition of any of the concentrates tested. A decrease in TG and marked alterations of TEM parameters were noted at 60% haemodilution. HES130/0.4 was the expander with the most deleterious action. TG was significantly enhanced by PCC whereas rFVIIa only caused a mild acceleration of TG initiation. Fibrinogen restored the alterations of TEM parameters caused by haemodilution including those caused by HES 130/0.4. Cryoprecipitates significantly improved the alterations caused by haemodilution on TG and TEM parameters; the effects on TG disappeared after ultracentrifugation of the cryoprecipitates. Discussion The haemostatic alterations caused by haemodilution are multifactorial and affect both blood cells and coagulation. In our in vitro approach, HES 130/0.4 had the most deleterious effect on haemostasis parameters. Coagulation factor concentrates did not improve platelet interactions in the Impact R, but did have favourable effects on coagulation parameters measured by TG and TEM. Fibrinogen notably improved TEM parameters without increasing thrombin generation, suggesting that this concentrate may help to preserve blood clotting abilities during haemodilution without enhancing the prothrombotic risk. PMID

  11. A case of hypereosinophilic syndrome presenting with intractable gastric ulcers

    PubMed Central

    Park, Tae Young; Choi, Chang Hwan; Yang, Suh Yoon; Oh, In Soo; Song, In-Do; Lee, Hyun Woong; Kim, Hyung Joon; Do, Jae Hyuk; Chang, Sae Kyung; Cho, Ah Ra; Cha, Young Joo

    2009-01-01

    We report a rare case of hypereosinophilic syndrome (HES) presenting with intractable gastric ulcers. A 71-year-old man was admitted with epigastric pain. Initial endoscopic findings revealed multiple, active gastric ulcers in the gastric antrum. He underwent Helicobacter pylori (H pylori) eradication therapy followed by proton pump inhibitor (PPI) therapy. However, follow-up endoscopy at 4, 6, 10 and 14 mo revealed persistent multiple gastric ulcers without significant improvement. The proportion of his eosinophil count increased to 43% (total count: 7903/mm3). Abdominal-pelvic and chest computed tomography scans showed multiple small nodules in the liver and both lungs. The endoscopic biopsy specimen taken from the gastric antrum revealed prominent eosinophilic infiltration, and the liver biopsy specimen also showed eosinophilic infiltration in the portal tract and sinusoid. A bone marrow biopsy disclosed eosinophilic hyperplasia as well as increased cellularity of 70%. The patient was finally diagnosed with HES involving the stomach, liver, lung, and bone marrow. When gastric ulcers do not improve despite H pylori eradication and prolonged PPI therapy, infiltrative gastric disorders such as HES should be considered. PMID:20027690

  12. [Interventions to solve overcrowding in hospital emergency services: a systematic review].

    PubMed

    Bittencourt, Roberto José; Hortale, Virginia Alonso

    2009-07-01

    This review discusses interventions aimed at solving the problem of overcrowding in hospital emergency services (HES), characteristic of low organizational effectiveness. In free-access and restricted-access databases, the target descriptors were 'overcrowding; emergency; medicine; first aid'. The survey identified 66 citations of interventions, grouped in 47 related interventions. The majority of the studies used observational designs that evaluated the results before and after interventions. Of the 47 related interventions, 34 had results for organizational performance. Four were the most frequently cited: implementation of an observation unit for patients already hospitalized and awaiting diagnosis or clinical stabilization; designation of a dedicated nursing station for the patient's admission, discharge, or transfer; establishment of protocols with indicators of operational saturation and implementation of an emergency care unit. Analyzing 21 interventions that were intended to solve overcrowding in HES and with positive results for length-of-stay, 15 were related to patient stream improvement, interfering actively and positively in patient stream. Interventions that increased access barriers or that merely improved the HES structure were ineffective. PMID:19578565

  13. Identifying developmental toxicity pathways for a subset of ToxCast chemicals using human embryonic stem cells and metabolomics.

    PubMed

    Kleinstreuer, N C; Smith, A M; West, P R; Conard, K R; Fontaine, B R; Weir-Hauptman, A M; Palmer, J A; Knudsen, T B; Dix, D J; Donley, E L R; Cezar, G G

    2011-11-15

    Metabolomics analysis was performed on the supernatant of human embryonic stem (hES) cell cultures exposed to a blinded subset of 11 chemicals selected from the chemical library of EPA's ToxCast™ chemical screening and prioritization research project. Metabolites from hES cultures were evaluated for known and novel signatures that may be indicative of developmental toxicity. Significant fold changes in endogenous metabolites were detected for 83 putatively annotated mass features in response to the subset of ToxCast chemicals. The annotations were mapped to specific human metabolic pathways. This revealed strong effects on pathways for nicotinate and nicotinamide metabolism, pantothenate and CoA biosynthesis, glutathione metabolism, and arginine and proline metabolism pathways. Predictivity for adverse outcomes in mammalian prenatal developmental toxicity studies used ToxRefDB and other sources of information, including Stemina Biomarker Discovery's predictive DevTox® model trained on 23 pharmaceutical agents of known developmental toxicity and differing potency. The model initially predicted developmental toxicity from the blinded ToxCast compounds in concordance with animal data with 73% accuracy. Retraining the model with data from the unblinded test compounds at one concentration level increased the predictive accuracy for the remaining concentrations to 83%. These preliminary results on a 11-chemical subset of the ToxCast chemical library indicate that metabolomics analysis of the hES secretome provides information valuable for predictive modeling and mechanistic understanding of mammalian developmental toxicity. PMID:21925528

  14. Nitric Oxide Receptor Soluble Guanylyl Cyclase Undergoes Splicing Regulation in Differentiating Human Embryonic Cells

    PubMed Central

    Sharin, Vladislav G.; Mujoo, Kalpana; Kots, Alexander Y.; Martin, Emil; Murad, Ferid

    2011-01-01

    Nitric oxide (NO), an important mediator molecule in mammalian physiology, initiates a number of signaling mechanisms by activating the enzyme soluble guanylyl cyclase (sGC). Recently, a new role for NO/cyclic guanosine monophosphate signaling in embryonic development and cell differentiation has emerged. The changes in expression of NO synthase isoforms and various sGC subunits has been demonstrated during human and mouse embryonic stem (ES) cells differentiation. Previously, our laboratory demonstrated that nascent α1 sGC transcript undergoes alternative splicing and that expression of α1 sGC splice forms directly affects sGC activity. Expression of sGC splice variants in the process of human ES (hES) cells differentiation has not been investigated. In this report, we demonstrate that α1 sGC undergoes alternative splicing during random hES differentiation for the first time. Our results indicate that C-α1 sGC splice form is expressed at high levels in differentiating cells and its intracellular distribution varies from canonical α1 sGC subunit. Together, our data suggest that alternative splicing of sGC subunits is associated with differentiation of hES cells. PMID:20964618

  15. Hesperidin increases intestinal β,β-carotene 15-15' mono-oxygenase 1 (BCMO1) activity in Mongolian gerbils (Meriones unguiculatus) fed with β-carotene-free diet.

    PubMed

    Poulaert, Marie; Gunata, Ziya; During, Alexandrine; Reboul, Emmanuelle; Laurent, Caroline; Gaillet, Sylvie; Dhuique-Mayer, Claudie

    2014-09-15

    β,β-Carotene 15-15' mono-oxygenase 1 (BCMO1) is a key enzyme in vitamin A (VitA) metabolism in mammals. Dietary compounds, such as carotenoids and polyphenols, were reported to influence BCMO1 activity. The aim of this study was to evaluate the effect of hesperidin (Hes), on the VitA bioefficacy of β-carotene (Bc) from orange-fleshed sweet potato, using Mongolian gerbils, focussing on BCMO1 activity. Gerbils (n=50) depleted in VitA were divided into five groups fed with basal diet containing 3% white- or orange-fleshed sweet potatoes supplemented or not with Hes. Liver BCMO1 activity was low, with no significant differences between groups. Interestingly, intestinal mucosal BCMO1 activity was significantly higher in the gerbils fed without Bc or VitA than those fed with a VitA/Bc-supplemented diet. Finally, our results show that, under a low VitA status, Hes dramatically stimulated intestinal BCMO1 activity, an effect that could possibly be related to its action as an agonist of PPARγ. PMID:24767085

  16. SOX1 links the function of neural patterning and Notch signalling in the ventral spinal cord during the neuron-glial fate switch

    SciTech Connect

    Genethliou, Nicholas; Panayiotou, Elena; Panayi, Helen; Orford, Michael; Mean, Richard; Lapathitis, George; Gill, Herman; Raoof, Sahir; Gasperi, Rita De; Elder, Gregory; Kessaris, Nicoletta; Richardson, William D.; Malas, Stavros

    2009-12-25

    During neural development the transition from neurogenesis to gliogenesis, known as the neuron-glial ({Nu}/G) fate switch, requires the coordinated function of patterning factors, pro-glial factors and Notch signalling. How this process is coordinated in the embryonic spinal cord is poorly understood. Here, we demonstrate that during the N/G fate switch in the ventral spinal cord (vSC) SOX1 links the function of neural patterning and Notch signalling. We show that, SOX1 expression in the vSC is regulated by PAX6, NKX2.2 and Notch signalling in a domain-specific manner. We further show that SOX1 regulates the expression of Hes1 and that loss of Sox1 leads to enhanced production of oligodendrocyte precursors from the pMN. Finally, we show that Notch signalling functions upstream of SOX1 during this fate switch and is independently required for the acquisition of the glial fate perse by regulating Nuclear Factor I A expression in a PAX6/SOX1/HES1/HES5-independent manner. These data integrate functional roles of neural patterning factors, Notch signalling and SOX1 during gliogenesis.

  17. The Eosinophil in Health and Disease: from Bench to Bedside and Back.

    PubMed

    Liao, Wei; Long, Hai; Chang, Christopher Chia-Chi; Lu, Qianjin

    2016-04-01

    Historically, eosinophils have been considered as end-stage cells involved in host protection against parasitic infection and in the mechanisms of hypersensitivity. However, later studies have shown that this multifunctional cell is also capable of producing immunoregulatory cytokines and soluble mediators and is involved in tissue homeostasis and modulation of innate and adaptive immune responses. In this review, we summarize the biology of eosinophils, including the function and molecular mechanisms of their granule proteins, cell surface markers, mediators, and pathways, and present comprehensive reviews of research updates on the genetics and epigenetics of eosinophils. We describe recent advances in the development of epigenetics of eosinophil-related diseases, especially in asthma. Likewise, recent studies have provided us with a more complete appreciation of how eosinophils contribute to the pathogenesis of various diseases, including hypereosinophilic syndrome (HES). Over the past decades, the definition and criteria of HES have been evolving with the progress of our understanding of the disease and some aspects of this disease still remain controversial. We also review recent updates on the genetic and molecular mechanisms of HES, which have spurred dramatic developments in the clinical strategies of diagnosis and treatment for this heterogeneous group of diseases. The conclusion from this review is that the biology of eosinophils provides significant insights as to their roles in health and disease and, furthermore, demonstrates that a better understanding of eosinophil will accelerate the development of new therapeutic strategies for patients. PMID:26410377

  18. STB-HO, a novel mica fine particle, inhibits the teratoma-forming ability of human embryonic stem cells after in vivo transplantation.

    PubMed

    Choi, Soon Won; Shin, Tae-Hoon; Uddin, Md Hafiz; Shin, Ji-Hee; Kang, Tae-Wook; Lee, Byung-Chul; Kim, Hyung-Sik; Seo, Yoojin; Shams, Sulaiman; Jung, Yeon-Kwon; Kang, Kyung-Sun

    2016-01-19

    Although pluripotent stem cell (PSC) therapy has advantages for clinical applications because of the self-renewal and multi-lineage differentiation abilities of PSCs, it also has disadvantages in terms of the potential for PSCs to undergo malignant transformation or unexpected differentiation. The prevention of teratoma formation is the largest hurdle of all. Despite intensive studies that have investigated ways to block teratomas, such methods have yet to be further developed for clinical use. Here, a new approach has focused on exerting anti-tumorigenic effects using a novel mica fine particle (MFP) designated STB-HO. Treatment with STB-HO regulated pluripotency- and apoptosis-related genes in differentiating human embryonic stem (hES) cells, while there is no effects in undifferentiated hES cells. In particular, STB-HO blocked the anti-apoptotic gene BIRC5 and activated p53, p21 and the pro-apoptotic proteins Bim, Puma and p-Bad during early spontaneous differentiation. Moreover, STB-HO-pretreated differentiating hES cells did not give rise to teratomas following in vivo stem cell transplantation. Our in vitro and in vivo results suggest a method for teratoma prevention in the context of PSC-derived cell transplantation. This novel MFP could break through the limitations of PSC therapy. PMID:26646796

  19. Combinatorial Development of Biomaterials for Clonal Growth of Human Pluripotent Stem Cells

    PubMed Central

    Mei, Ying; Saha, Krishanu; Bogatyrev, Said R.; Yang, Jing; Hook, Andrew L.; Kalcioglu, Z. Ilke; Cho, Seung-Woo; Mitalipova, Maisam; Pyzocha, Neena; Rojas, Fredrick; Van Vliet, Krystyn J.; Davies, Martyn C.; Alexander, Morgan R.; Langer, Robert; Jaenisch, Rudolf; Anderson, Daniel G.

    2010-01-01

    Both human embryonic stem (hES) cells and induced pluripotent stem (hiPS) cells can self-renew indefinitely in culture, however current methods to clonally grow them are inefficient and poorly-defined for genetic manipulation and therapeutic purposes. Here we develop the first chemically-defined, xeno-free, feeder-free synthetic substrates to support robust self-renewal of fully-dissociated hES and hiPS cells. Materials properties including wettability, surface topography, surface chemistry and indentation elastic modulus of all polymeric substrates were quantified using high-throughput methods to develop structure/function relationships between materials properties and biological performance. These analyses show that optimal hES cell substrates are generated from monomers with high acrylate content, have a moderate wettability, and employ integrin αvβ3 and αvβ5 engagement with adsorbed vitronectin to promote colony formation. The structure/function methodology employed herein provides a general framework for the combinatorial development of synthetic substrates for stem cell culture. PMID:20729850

  20. Solar gamma-ray and neutron registration capabilities of the GRIS instrument onboard the International Space Station

    NASA Astrophysics Data System (ADS)

    Trofimov, Yury; Kochemasov, Alexey; Yurov, Vitaly; Glyanenko, Alexander; Kotov, Yury; Lupar, Evgeny; Faradzhaev, Rodion

    2016-07-01

    GRIS (Gamma and Roentgen radiation of the Sun) is a prospective hard X-ray and gamma-ray spectrometer of solar flares with the energy range from 50 keV to 200 MeV. It is also designed for registration of high energy neutron fluxes (>30 MeV). The apparatus will be mounted on an oriented platform outside the Russian Orbital Segment of the International Space Station. The instrument includes two detector heads: a low energy spectrometer (LES) based on a fast scintillator with relatively high energy resolution 3.5-4.5% at 662 keV (LaBr _{3}(Ce) or CeBr _{3}) and size of ø7.62×7.62 cm, and a high energy spectrometer (HES) based on ø12×15 cm CsI(Tl) scintillator. Thanks to n/γ discrimination capability of CsI(Tl) crystals, the HES spectrometer is also intended for neutron registration. To estimate GRIS instrument registration capabilities, simulation of the HES neutron and gamma registration channels response to background radiation and to solar flares of different magnitude and spectral compositions was performed. Expected spectral and n/γ discrimination performances based on measurements with detectors prototypes are represented.

  1. Hawaii Energy Strategy Project 2: Fossil Energy Review. Task IV. Scenario development and analysis

    SciTech Connect

    Yamaguchi, N.D.; Breazeale, K.

    1993-12-01

    The Hawaii Energy Strategy (HES) Program is a seven-project effort led by the State of Hawaii Department of Business, Economic Development & Tourism (DBEDT) to investigate a wide spectrum of Hawaii energy issues. The East-West Center`s Program on Resources: Energy and Minerals, has been assigned HES Project 2, Fossil Energy Review, which focuses on fossil energy use in Hawaii and the greater regional and global markets. HES Project 2 has four parts: Task I (World and Regional Fossil Energy Dynamics) covers petroleum, natural gas, and coal in global and regional contexts, along with a discussion of energy and the environment. Task II (Fossil Energy in Hawaii) focuses more closely on fossil energy use in Hawaii: current utilization and trends, the structure of imports, possible future sources of supply, fuel substitutability, and energy security. Task III`s emphasis is Greenfield Options; that is, fossil energy sources not yet used in Hawaii. This task is divided into two sections: first, an in-depth {open_quotes}Assessment of Coal Technology Options and Implications for the State of Hawaii,{close_quotes} along with a spreadsheet analysis model, which was subcontracted to the Environmental Assessment and Information Sciences Division of Argonne National Laboratory; and second, a chapter on liquefied natural gas (LNG) in the Asia-Pacific market and the issues surrounding possible introduction of LNG into the Hawaii market.

  2. Electric utility applications of hydrogen energy storage systems

    SciTech Connect

    Swaminathan, S.; Sen, R.K.

    1997-10-15

    This report examines the capital cost associated with various energy storage systems that have been installed for electric utility application. The storage systems considered in this study are Battery Energy Storage (BES), Superconducting Magnetic Energy Storage (SMES) and Flywheel Energy Storage (FES). The report also projects the cost reductions that may be anticipated as these technologies come down the learning curve. This data will serve as a base-line for comparing the cost-effectiveness of hydrogen energy storage (HES) systems in the electric utility sector. Since pumped hydro or compressed air energy storage (CAES) is not particularly suitable for distributed storage, they are not considered in this report. There are no comparable HES systems in existence in the electric utility sector. However, there are numerous studies that have assessed the current and projected cost of hydrogen energy storage system. This report uses such data to compare the cost of HES systems with that of other storage systems in order to draw some conclusions as to the applications and the cost-effectiveness of hydrogen as a electricity storage alternative.

  3. Disrupting NOTCH Slows Diffuse Intrinsic Pontine Glioma Growth, Enhances Radiation Sensitivity, and Shows Combinatorial Efficacy With Bromodomain Inhibition.

    PubMed

    Taylor, Isabella C; Hütt-Cabezas, Marianne; Brandt, William D; Kambhampati, Madhuri; Nazarian, Javad; Chang, Howard T; Warren, Katherine E; Eberhart, Charles G; Raabe, Eric H

    2015-08-01

    NOTCH regulates stem cells during normal development and stemlike cells in cancer, but the roles of NOTCH in the lethal pediatric brain tumor diffuse intrinsic pontine glioma (DIPG) remain unknown. Because DIPGs express stem cell factors such as SOX2 and MYCN, we hypothesized that NOTCH activity would be critical for DIPG growth. We determined that primary DIPGs expressed high levels of NOTCH receptors, ligands, and downstream effectors. Treatment of the DIPG cell lines JHH-DIPG1 and SF7761 with the γ-secretase inhibitor MRK003 suppressed the level of the NOTCH effectors HES1, HES4, and HES5; inhibited DIPG growth by 75%; and caused a 3-fold induction of apoptosis. Short hairpin RNAs targeting the canonical NOTCH pathway caused similar effects. Pretreatment of DIPG cells with MRK003 suppressed clonogenic growth by more than 90% and enhanced the efficacy of radiation therapy. The high level of MYCN in DIPG led us to test sequential therapy with the bromodomain inhibitor JQ1 and MRK003, and we found that JQ1 and MRK003 inhibited DIPG growth and induced apoptosis. Together, these results suggest that dual targeting of NOTCH and MYCN in DIPG may be an effective therapeutic strategy in DIPG and that adding a γ-secretase inhibitor during radiation therapy may be efficacious initially or during reirradiation. PMID:26115193

  4. Effects of Acute Bleeding Followed by Hydroxyethyl Starch 130/0.4 or a Crystalloid on Propofol Concentrations, Cerebral Oxygenation, and Electroencephalographic and Haemodynamic Variables in Pigs

    PubMed Central

    Venâncio, Carlos; Souza, Almir P.; Ferreira, Luísa Maria; Branco, Paula Sério; de Pinho, Paula Guedes; Amorim, Pedro; Ferreira, David A.

    2014-01-01

    Bleeding changes the haemodynamics, compromising organ perfusion. In this study, the effects of bleeding followed by replacement with hydroxyethyl starch 130/0.4 (HES) or lactated Ringer's (LR) on cerebral oxygenation and electroencephalogram-derived parameters were investigated. Twelve young pigs under propofol-remifentanil anaesthesia were bled 30 mL/kg and, after a 20-minute waiting period, volume replacement was performed with HES (GHES; N = 6) or LR (GRL; N = 6). Bleeding caused a decrease of more than 50% in mean arterial pressure (P < 0.01) and a decrease in cerebral oximetry (P = 0.039), bispectral index, and electroencephalogram total power (P = 0.04 and P < 0.01, resp.), while propofol plasma concentrations increased (P < 0.01). Both solutions restored the haemodynamics and cerebral oxygenation similarly and were accompanied by an increase in electroencephalogram total power. No differences between groups were found. However, one hour after the end of the volume replacement, the cardiac output (P = 0.03) and the cerebral oxygenation (P = 0.008) decreased in the GLR and were significantly lower than in GHES (P = 0.02). Volume replacement with HES 130/0.4 was capable of maintaining the cardiac output and cerebral oxygenation during a longer period than LR and caused a decrease in the propofol plasma concentrations. PMID:24971192

  5. STB-HO, a novel mica fine particle, inhibits the teratoma-forming ability of human embryonic stem cells after in vivo transplantation

    PubMed Central

    Uddin, Md. Hafiz; Shin, Ji-Hee; Kang, Tae-Wook; Lee, Byung-Chul; Kim, Hyung-Sik; Seo, Yoojin; Shams, Sulaiman; Jung, Yeon-Kwon; Kang, Kyung-Sun

    2016-01-01

    Although pluripotent stem cell (PSC) therapy has advantages for clinical applications because of the self-renewal and multi-lineage differentiation abilities of PSCs, it also has disadvantages in terms of the potential for PSCs to undergo malignant transformation or unexpected differentiation. The prevention of teratoma formation is the largest hurdle of all. Despite intensive studies that have investigated ways to block teratomas, such methods have yet to be further developed for clinical use. Here, a new approach has focused on exerting anti-tumorigenic effects using a novel mica fine particle (MFP) designated STB-HO. Treatment with STB-HO regulated pluripotency- and apoptosis-related genes in differentiating human embryonic stem (hES) cells, while there is no effects in undifferentiated hES cells. In particular, STB-HO blocked the anti-apoptotic gene BIRC5 and activated p53, p21 and the pro-apoptotic proteins Bim, Puma and p-Bad during early spontaneous differentiation. Moreover, STB-HO-pretreated differentiating hES cells did not give rise to teratomas following in vivo stem cell transplantation. Our in vitro and in vivo results suggest a method for teratoma prevention in the context of PSC-derived cell transplantation. This novel MFP could break through the limitations of PSC therapy. PMID:26646796

  6. A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cells.

    PubMed

    Wang, Dachun; Haviland, David L; Burns, Alan R; Zsigmond, Eva; Wetsel, Rick A

    2007-03-13

    Alveolar epithelial type II (ATII) cells are small, cuboidal cells that constitute approximately 60% of the pulmonary alveolar epithelium. These cells are crucial for repair of the injured alveolus by differentiating into alveolar epithelial type I cells. ATII cells derived from human ES (hES) cells are a promising source of cells that could be used therapeutically to treat distal lung diseases. We have developed a reliable transfection and culture procedure, which facilitates, via genetic selection, the differentiation of hES cells into an essentially pure (>99%) population of ATII cells (hES-ATII). Purity, as well as biological features and morphological characteristics of normal ATII cells, was demonstrated for the hES-ATII cells, including lamellar body formation, expression of surfactant proteins A, B, and C, alpha-1-antitrypsin, and the cystic fibrosis transmembrane conductance receptor, as well as the synthesis and secretion of complement proteins C3 and C5. Collectively, these data document the successful generation of a pure population of ATII cells derived from hES cells, providing a practical source of ATII cells to explore in disease models their potential in the regeneration and repair of the injured alveolus and in the therapeutic treatment of genetic diseases affecting the lung. PMID:17360544

  7. The Bellamy challenge: it's about time.

    PubMed

    Dobson, Geoffrey P; Letson, H L; Tadaki, D

    2014-03-01

    In 1984, Col. Ronald Bellamy launched a worldwide challenge to develop a new resuscitation fluid to aid survival after catastrophic blood loss on the battlefield. In 1996, after careful compromise among need, cube weight and efficacy, the US military and later coalition forces adopted 6% hetastarch (HES) fluids for early resuscitation. In the intervening years, evidence has amassed indicating that the HES fluids may not be safe, and in June 2013 the US Food and Drug Administration issued a warning that HES solutions should not be used to treat patients with hypovolaemia or the critically ill. We review the unique challenges of early battlefield resuscitation, why the 'Bellamy challenge' remains open and discuss a number of forward-looking strategies that may help to solve the problem. The first two pillars of resuscitation that we believe have not been adequately addressed are rescuing and stabilising the heart (and brain) and the vascular system. The 'ideal' resuscitation fluid needs to nurture the heart and body slowly back to health, and not 'shock' it a second time with unnatural colloids or large volumes of unphysiological saline-based solutions. PMID:24142962

  8. The retinoic acid receptor agonist Am80 increases hippocampal ADAM10 in aged SAMP8 mice.

    PubMed

    Kitaoka, Kazuyoshi; Shimizu, Noriyuki; Ono, Koji; Chikahisa, Sachiko; Nakagomi, Madoka; Shudo, Koichi; Ishimura, Kazunori; Séi, Hiroyoshi; Yoshizaki, Kazuo

    2013-09-01

    The retinoic acid (RA, a vitamin A metabolite) receptor (RAR) is a transcription factor. Vitamin A/RA administration improves the Alzheimer's disease (AD)- and age-related attenuation of memory/learning in mouse models. Recently, a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) was identified as a key molecule in RA-mediated anti-AD mechanisms. We investigated the effect of chronic administration of the RAR agonist Am80 (tamibarotene) on ADAM10 expression in senescence-accelerated mice (SAMP8). Moreover, we estimated changes in the expression of the amyloid precursor protein (APP), amyloid beta (Aβ), and hairy/enhancer of split (Hes), which are mediated by ADAM10. Spatial working memory and the levels of a hippocampal proliferation marker (Ki67) were also assessed in these mice. ADAM10 mRNA and protein expression was significantly reduced in the hippocampus of 13-month-old SAMP8 mice; their expression improved significantly after Am80 administration. Further, after Am80 administration, the expression levels of Hes5 and Ki67 were restored and the deterioration of working memory was suppressed, whereas APP and Aβ levels remained unchanged. Our results suggest that Am80 administration effectively improves dementia by activating the hippocampal ADAM10-Notch-Hes5 proliferative pathway. PMID:23624141

  9. EXAMINING A HOME ENVIRONMENTAL STRATEGY TO REDUCE AVAILABILITY OF LEGAL PRODUCTS THAT CAN BE MISUSED BY YOUTH

    PubMed Central

    Collins, David A.; Johnson, Knowlton W.; Shamblen, Stephen R.

    2013-01-01

    This article presents results from a study of a home environmental strategy (HES) designed to reduce availability of harmful legal products (HLPs) in the home that can be used by youth to get high. HLPs include inhalants, prescription and non-prescription drugs, and household products that can be ingested to get high. Availability is one of the most consistent predictors of substance use among youth. Parents of 5th to 7th-grade students in four Alaskan communities participated in telephone interviews as part of a larger study of a multi-component Community Prevention Model that included a HES. The strategy was designed to encourage parents to reduce availability of HLPs by removing them from the home, and by locking up and monitoring the supplies of HLPs in the home. Data from 402 parents at Wave 1 and 371 parents at Wave 2 were analyzed using Hierarchical Non-Linear Modeling (HNLM). Results show there was a significant decrease in HLPs in the home from Wave 1 to Wave 2, mostly inhalants, prescription and non-prescription drugs. Parents also reported a significant increase in locking up prescription and non-prescription drugs in the home. Parents’ direct exposure to the HES was marginally associated with the change over time in HLP availability in the home. Indirect exposure through others and media was not associated with this change. Study lessons learned and conclusions are highlighted. PMID:22943304

  10. In Vivo Evaluation of the Ameliorating Effects of Small-Volume Resuscitation with Four Different Fluids on Endotoxemia-Induced Kidney Injury.

    PubMed

    Wang, Yan-ling; Chen, Jing-hui; Zhu, Qiong-fang; Yu, Gao-feng; Luo, Chen-fang; Luo, Gang-jian; Li, Shang-rong; Hei, Zi-qing

    2015-01-01

    Acute kidney injury associated with renal hypoperfusion is a frequent and severe complication during sepsis. Fluid resuscitation is the main therapy. However, heart failure is usually lethal for those patients receiving large volumes of fluids. We compared the effects of small-volume resuscitation using four different treatment regimens, involving saline, hypertonic saline (HTS), hydroxyethyl starch (HES), or hypertonic saline hydroxyethyl starch (HSH), on the kidneys of rats treated with lipopolysaccharide (LPS) to induce endotoxemia. LPS injection caused reduced and progressively deteriorated systemic (arterial blood pressure) and renal hemodynamics (renal blood flow and renal vascular resistance index) over time. This deterioration was accompanied by marked renal functional and pathological injury, as well as an oxidative and inflammatory response, manifesting as increased levels of tumor necrosis factor-α, nitric oxide, and malondialdehyde and decreased activity of superoxide dismutase. Small-volume perfusion with saline failed to improve renal and systemic circulation. However, small-volume perfusion with HES and HSH greatly improved the above parameters, while HTS only transiently improved systemic and renal hemodynamics with obvious renal injury. Therefore, single small-volume resuscitation with HES and HSH could be valid therapeutic approaches to ameliorate kidney injury induced by endotoxemia, while HTS transiently delays injury and saline shows no protective effects. PMID:26273142

  11. In Vivo Evaluation of the Ameliorating Effects of Small-Volume Resuscitation with Four Different Fluids on Endotoxemia-Induced Kidney Injury

    PubMed Central

    Wang, Yan-ling; Chen, Jing-hui; Zhu, Qiong-fang; Yu, Gao-feng; Luo, Chen-fang; Luo, Gang-jian; Li, Shang-rong; Hei, Zi-qing

    2015-01-01

    Acute kidney injury associated with renal hypoperfusion is a frequent and severe complication during sepsis. Fluid resuscitation is the main therapy. However, heart failure is usually lethal for those patients receiving large volumes of fluids. We compared the effects of small-volume resuscitation using four different treatment regimens, involving saline, hypertonic saline (HTS), hydroxyethyl starch (HES), or hypertonic saline hydroxyethyl starch (HSH), on the kidneys of rats treated with lipopolysaccharide (LPS) to induce endotoxemia. LPS injection caused reduced and progressively deteriorated systemic (arterial blood pressure) and renal hemodynamics (renal blood flow and renal vascular resistance index) over time. This deterioration was accompanied by marked renal functional and pathological injury, as well as an oxidative and inflammatory response, manifesting as increased levels of tumor necrosis factor-α, nitric oxide, and malondialdehyde and decreased activity of superoxide dismutase. Small-volume perfusion with saline failed to improve renal and systemic circulation. However, small-volume perfusion with HES and HSH greatly improved the above parameters, while HTS only transiently improved systemic and renal hemodynamics with obvious renal injury. Therefore, single small-volume resuscitation with HES and HSH could be valid therapeutic approaches to ameliorate kidney injury induced by endotoxemia, while HTS transiently delays injury and saline shows no protective effects. PMID:26273142

  12. Notch regulation of bone development and remodeling and related skeletal disorders.

    PubMed

    Zanotti, Stefano; Canalis, Ernesto

    2012-02-01

    Notch signaling mediates cell-to-cell interactions that are critical for embryonic development and tissue renewal. In the canonical signaling pathway, the Notch receptor is cleaved following ligand binding, resulting in the release and nuclear translocation of the Notch intracellular domain (NICD). NICD induces gene expression by forming a ternary complex with the DNA binding protein CBF1/Rbp-Jk, Suppressor of Hairless, Lag1, and Mastermind-Like (Maml). Hairy Enhancer of Split (Hes) and Hes related with YRPW motif (Hey) are classic Notch targets. Notch canonical signaling plays a central role in skeletal development and bone remodeling by suppressing the differentiation of skeletal cells. The skeletal phenotype of mice misexpressing Hes1 phenocopies partially the effects of Notch misexpression, suggesting that Hey proteins mediate most of the skeletal effects of Notch. Dysregulation of Notch signaling is associated with diseases affecting human skeletal development, such as Alagille syndrome, brachydactyly and spondylocostal dysostosis. Somatic mutations in Notch receptors and ligands are found in tumors of the skeletal system. Overexpression of NOTCH1 is associated with osteosarcoma, and overexpression of NOTCH3 or JAGGED1 in breast cancer cells favors the formation of osteolytic bone metastasis. Activating mutations in NOTCH2 cause Hajdu-Cheney syndrome, which is characterized by skeletal defects and fractures, and JAG1 polymorphisms, are associated with variations in bone mineral density. In conclusion, Notch is a regulator of skeletal development and bone remodeling, and abnormal Notch signaling is associated with developmental and postnatal skeletal disorders. PMID:22002679

  13. Cognitive influence of a 5-h ENERGY® shot: Are effects perceived or real?

    PubMed

    Buckenmeyer, Philip J; Bauer, Jeffery A; Hokanson, James F; Hendrick, Joy L

    2015-12-01

    With the gain in popularity and use of 'functional energy drinks' (FEDs), manufacturers of these beverages have been making greater claims as to their benefits on a number of performance factors including mental alertness, energy, and physical performance. Few experimental studies have been conducted on FEDs, and no study to date has examined their effects over time. This study looked at the effects of consumption of a 5-h ENERGY® shot (5-HES) on various cognitive functions across five hours on 24 college-aged students using a double-blind, cross-over, placebo-based design. Participants completed a series of five computer-based tests before ingesting the beverage (either 5-HES or placebo) and then completed the tests for each of the next five hours (morning to midday). One week later, they repeated the process with the other beverage. While 90% of participants subjectively thought that the 5-HES was effective at one-hour post-ingestion, no evidence was found to support an enhanced effect on recognition, reaction time, short-term and working memory, or attention capacity. In conclusion, the 5-h Energy Shot® did not significantly improve short- or long-term cognitive function for selected computer-based tasks despite a high level of perception that it was working effectively compared to a placebo with college-aged participants. PMID:26394126

  14. Distribution and mobility of heavy elements in floodplain agricultural soils along the Ibar River (Southern Serbia and Northern Kosovo). Chemometric investigation of pollutant sources and ecological risk assessment.

    PubMed

    Barać, Nemanja; Škrivanj, Sandra; Bukumirić, Zoran; Živojinović, Dragana; Manojlović, Dragan; Barać, Milan; Petrović, Rada; Ćorac, Aleksandar

    2016-05-01

    This work investigates the influence of a high-magnitude flood event on heavy elements (HEs) pollution and mobility in the agricultural soils along Ibar River in Southern Serbia and Northern Kosovo. The study area was one of the most important Pb/Zn industrial regions in Europe. Soil samples (n = 50) collected before and after the floods in May 2014 were subjected to the sequential extraction procedure proposed by the Community Bureau of Reference (BCR). The results indicated that the floods significantly increased not only the pseudo total concentrations of HEs in the soil but also their mobile and potentially bioavailable amounts. Moreover, higher concentrations (both pseudo total and potentially bioavailable) were found in the agricultural soils closer to the industrial hotspots. Principal component analysis and hierarchical cluster analysis successfully grouped the analyzed elements according to their anthropogenic or natural origin. The floods significantly increased the potential ecological risk of HEs associated with Pb/Zn industrial activities in the study area. The potential ecological risk of Cd after the floods was highest and should be of special concern. PMID:26822217

  15. Disrupting NOTCH Slows Diffuse Intrinsic Pontine Glioma Growth, Enhances Radiation Sensitivity, and Shows Combinatorial Efficacy with Bromodomain Inhibition

    PubMed Central

    Taylor, Isabella C.; Hütt-Cabezas, Marianne; Brandt, William D.; Kambhampati, Madhuri; Nazarian, Javad; Chang, Howard T.; Warren, Katherine E.; Eberhart, Charles G.; Raabe, Eric H.

    2015-01-01

    NOTCH regulates stem cells during normal development and stem-like cells in cancer but the roles of NOTCH in the lethal pediatric brain tumor diffuse intrinsic pontine glioma (DIPG) remain unknown. Because DIPGs express stem cell factors such as SOX2 and MYCN, we hypothesized that NOTCH activity would be critical for DIPG growth. We determined that primary DIPGs expressed high levels of NOTCH receptors, ligands, and downstream effectors. Treatment of the DIPG cell lines JHH-DIPG1 and SF7761 with the γ-secretase inhibitor MRK003 suppressed the level of the NOTCH effectors HES1, HES4, HES5, inhibited DIPG growth by 75%, and caused a 3-fold induction of apoptosis. Short hairpin RNAs targeting the canonical NOTCH pathway caused similar effects. Pre-treatment of DIPG cells with MRK003 suppressed clonogenic growth by more than 90% and enhanced the efficacy of radiation therapy. The high level of MYCN in DIPG led us to test sequential therapy with the bromodomain inhibitor JQ1 and MRK003, and we found that JQ1 and MRK003 inhibited DIPG growth and induced apoptosis. Together, these results suggest that dual targeting of NOTCH and MYCN in DIPG may be an effective therapeutic strategy in DIPG and that adding a γ-secretase inhibitor during radiation therapy may be efficacious initially or during re-irradiation. PMID:26115193

  16. Efficient generation of retinal progenitor cells from human embryonic stem cells

    PubMed Central

    Lamba, Deepak A.; Karl, Mike O.; Ware, Carol B.; Reh, Thomas A.

    2006-01-01

    The retina is subject to degenerative conditions, leading to blindness. Although retinal regeneration is robust in lower vertebrates, regeneration does not occur in the adult mammalian retina. Thus, we have developed efficient methods for deriving retinal neurons from human embryonic stem (hES) cells. Under appropriate culture conditions, up to 80% of the H1 line can be directed to the retinal progenitor fate, and express a gene expression profile similar to progenitors derived from human fetal retina. The hES cell-derived progenitors differentiate primarily into inner retinal neurons (ganglion and amacrine cells), with functional glutamate receptors. Upon coculture with retinas derived from a mouse model of retinal degeneration, the hES cell derived retinal progenitors integrate with the degenerated mouse retina and increase in their expression of photoreceptor-specific markers. These results demonstrate that human ES cells can be selectively directed to a neural retinal cell fate and thus may be useful in the treatment of retinal degenerations. PMID:16908856

  17. Effect of sugars on headgroup mobility in freeze-dried dipalmitoylphosphatidylcholine bilayers: solid-state 31P NMR and FTIR studies.

    PubMed

    Tsvetkova, N M; Phillips, B L; Crowe, L M; Crowe, J H; Risbud, S H

    1998-12-01

    The effect of the carbohydrates trehalose, glucose, and hydroxyethyl starch (HES) on the motional properties of the phosphate headgroup of freeze-dried dipalmitoylphosphatidylcholine (DPPC) liposomes was studied by means of 31P NMR, Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The results show that trehalose, which is a strong glass former (Tg = 115 degreesC), elevates the onset of the lipid headgroup rotations and preserves some rotational mobility of the phosphate headgroups after cooling from the liquid-crystalline state. Glucose (Tg = 30 degreesC), a very effective depressant of the phase transition temperature of freeze-dried DPPC, markedly elevates the initiation of the temperature of headgroup rotations. On the other hand, the monosaccharide does not preserve the headgroup disordering when cooled from the liquid-crystalline state. These effects are consistent with formation of hydrogen bonds between the OH groups of the sugar and the polar headgroups of DPPC. They show, however, that hydrogen bonding is not sufficient for preservation of the dynamic properties of freeze-dried DPPC. HES, although a very good glass former (Tg > 110 degreesC), does not depress the phase transition temperature and affects only slightly the rotational properties of freeze-dried DPPC. This lack of effect of HES is associated with the absence of direct interactions with the lipid phosphates, as evidenced by the FTIR results. These data show that vitrification of the additive is not sufficient to affect the dynamic properties of dried DPPC. PMID:9826615

  18. Peroral Endoscopic Myotomy Can Improve Esophageal Motility in Patients with Achalasia from a Large Sample Self-Control Research (66 Patients)

    PubMed Central

    Yao, Shuangzhe; Linghu, Enqiang

    2015-01-01

    Background Peroral endoscopic myotomy (POEM) as a new approach to achalasia attracts broad attention. The primary objective of this study was to evaluate the results with esophageal motility after POEM through the first large sample clinical research. Patients and Methods We have a self-control research with all patients (205 in total) who underwent POEM from 2010 to 2014 at our Digestive Endoscopic Center, 66 patients of which underwent high resolution manometry (HRM) before and after POEM in our motility laboratory. Follow-ups last for 5.6 months on average. Outcome variables analyzed included upper esophageal sphincter pressure (UESP), upper esophageal sphincter residual pressure (UESRP), lower esophageal sphincter pressure (LESP), lower esophageal sphincter residual pressure (LESRP) and esophageal body peristalsis. We have a statistical analysis to illustrate how POEM impacts on the change of esophageal motility. Results The symptoms related to dysphagia were relieved in 95% of patients in recent term after POEM. While HRM showed a statistically significant reduction of URSRP, LESP and LESRP (P<0.01), however, peristalsis was not consistently affected. There were 11 patients who had undergone other prior endoscopic treatment (endoscopic dilation or botulinum toxin injection) and 55 patients had not. The statistical difference (P>0.05) did not occur for these two groups on LESP and LESRP reduction. Conclusions POEM clearly relieved the symptoms related to dysphagia by lowering the pressure of upper esophageal sphincter (UES) and lower esophageal sphincter (LES),and other endoscopic treatment before POEM did not affect the improvement of LES pressure. These results are concluded from our short-term follow-up study, while the long-term efficacy remains to be further illustrated. Trial Registration Chinese Clinical Trial Register ChiCTR-TRC-12002204) PMID:25993648

  19. The vascular delta-like ligand-4 (DLL4)-Notch4 signaling correlates with angiogenesis in primary glioblastoma: an immunohistochemical study.

    PubMed

    Zhang, Jin-Feng; Chen, Yao; Qiu, Xian-Xin; Tang, Wen-Long; Zhang, Jian-Dong; Huang, Jian-Huang; Lin, Guo-Shi; Wang, Xing-Fu; Lin, Zhi-Xiong

    2016-03-01

    Delta-like ligand-4 (DLL4)-Notch signaling is known to play a pivotal role in the regulation of tumor angiogenesis. We had previously found that DLL4 was overexpressed, while Notch1 receptor, which binds to DLL4 during angiogenesis, was absent in the majority of human primary glioblastomas. Thus, DLL4-Notch signaling pathway in the regulation of tumor angiogenesis in primary glioblastoma remains unknown. Tumor tissues from 70 patients with primary glioblastoma were analyzed by immunohistochemistry for expression of components of DLL4-Notch signaling, vascular endothelial growth factor (VEGF), and microvessel density (MVD). Immunohistochemistry results showed that the positive staining of DLL4 and Notch4 was primarily distributed in tumor vascular endothelial cells but rarely detected in tumor cells. However, VEGF, hairy/enhancer of split-1 (HES1; a target gene of Notch signaling), and Notch1-3 expression was seen in both tumor vascular endothelial cells and tumor cells. Univariate analysis showed that the expression levels of VEGF and DLL4, HES1, and Notch4 in tumor endothelial cells were significantly associated with MVD in primary glioblastoma (P < 0.001). Binary logistic regression analysis showed that high expression levels of DLL4, HES1, and Notch4 in tumor endothelial cells were associated with a decrease of MVD in primary glioblastoma, while MVD increased with elevated VEGF expression in contrast. In addition, DLL4, Notch4, and HES1 expression were positively correlated in tumor vascular endothelial cells (P < 0.05). We conclude that the vascular DLL4-Notch4 signaling and VEGF signaling complementing each other plays an important role in the progression of tumor angiogenesis in primary glioblastoma. Graphical abstract A, positive staining of DLL4 in human kidney; B, positive staining of VEGF in human breast cancer; C, positive staining of CD34 in human lung cancer; D, positive staining of HES1 in human breast cancer; E-H, positive staining of Notch1

  20. Bacteria Regulate Intestinal Epithelial Cell Differentiation Factors Both In Vitro and In Vivo

    PubMed Central

    Becker, Svetlana; Oelschlaeger, Tobias A.; Wullaert, Andy; Pasparakis, Manolis; Wehkamp, Jan; Stange, Eduard F.; Gersemann, Michael

    2013-01-01

    Background The human colon harbours a plethora of bacteria known to broadly impact on mucosal metabolism and function and thought to be involved in inflammatory bowel disease pathogenesis and colon cancer development. In this report, we investigated the effect of colonic bacteria on epithelial cell differentiation factors in vitro and in vivo. As key transcription factors we focused on Hes1, known to direct towards an absorptive cell fate, Hath1 and KLF4, which govern goblet cell. Methods Expression of the transcription factors Hes1, Hath1 and KLF4, the mucins Muc1 and Muc2 and the defensin HBD2 were measured by real-time PCR in LS174T cells following incubation with several heat-inactivated E. coli strains, including the probiotic E. coli Nissle 1917+/− flagellin, Lactobacilli and Bifidobacteria. For protein detection Western blot experiments and chamber-slide immunostaining were performed. Finally, mRNA and protein expression of these factors was evaluated in the colon of germfree vs. specific pathogen free vs. conventionalized mice and colonic goblet cells were counted. Results Expression of Hes1 and Hath1, and to a minor degree also of KLF4, was reduced by E. coli K-12 and E. coli Nissle 1917. In contrast, Muc1 and HBD2 expression were significantly enhanced, independent of the Notch signalling pathway. Probiotic E. coli Nissle 1917 regulated Hes1, Hath1, Muc1 and HBD2 through flagellin. In vivo experiments confirmed the observed in vitro effects of bacteria by a diminished colonic expression of Hath1 and KLF4 in specific pathogen free and conventionalized mice as compared to germ free mice whereas the number of goblet cells was unchanged in these mice. Conclusions Intestinal bacteria influence the intestinal epithelial differentiation factors Hes1, Hath1 and KLF4, as well as Muc1 and HBD2, in vitro and in vivo. The induction of Muc1 and HBD2 seems to be triggered directly by bacteria and not by Notch. PMID:23418447

  1. Cardiovascular surveys: manual of operations.

    PubMed

    Primatesta, Paola; Allender, Steven; Ciccarelli, Paola; Doring, Angela; Graff-Iversen, Sidsel; Holub, Jiri; Panico, Salvatore; Trichopoulou, Antonia; Verschuren, W M Monique

    2007-12-01

    Cardiovascular disease (CVD) is the leading cause of death and hospitalization in both men and women in nearly all countries of Europe. The most frequent forms of CVD are those of an atherosclerotic origin, mainly ischaemic heart disease, stroke and heart failure. The magnitude of the problem contrasts with the usual paucity and poor quality of data available on incidence and prevalence of CVD, except for few rigorous but limited studies. The objectives of the health interview and health examination surveys (HIS/HES) are to evaluate the frequency and the distribution of the disease, to evaluate trends and treatment effectiveness, to estimate risk factors distribution and prevalence of high risk conditions and to monitor prevention programmes. According to the EUROCISS project (EUROpean Cardiovascular Surveillance Set) recommendations, surveys are aimed at describing the prevalence of the following CVD conditions: myocardial infarction, heart failure, angina pectoris, peripheral arterial disease, stroke, and ischaemic heart disease.HIS and HES were developed to supplement information collected from routine databases and population-based registers to implement consistent public health policies. HIS can be repeated periodically in a new sample of the population, or can follow up over time the population recruited at baseline. Procedures and methods to collect information from participants include self-administered questionnaires, direct interviewer-administered questions and telephone interviews. A minimum set of questions to be administered every year, along with a longer, more detailed module to be administered periodically are recommended to evaluate CVD prevalence. The addition of HES provides more detailed and objective information that can be used to improve estimates regarding prevalence of both risk factors and disease status. The selection of more specialized CVD-specific tests will depend on the objective the survey is designed to achieve, the assumed

  2. Validation of a survey instrument to assess home environments for physical activity and healthy eating in overweight children

    PubMed Central

    Gattshall, Michelle L; Shoup, Jo Ann; Marshall, Julie A; Crane, Lori A; Estabrooks, Paul A

    2008-01-01

    Background Few measures exist to measure the overall home environment for its ability to support physical activity (PA) and healthy eating in overweight children. The purpose of this study was to develop and test the reliability and validity of such a measure. Methods The Home Environment Survey (HES) was developed to reflect availability, accessibility, parental role modelling, and parental policies related to PA resources, fruits and vegetables (F&V), and sugar sweetened drinks and snacks (SS). Parents of overweight children (n = 219) completed the HES and concurrent behavioural assessments. Children completed the Block Kids survey and wore an accelerometer for one week. A subset of parents (n = 156) completed the HES a second time to determine test-retest reliability. Finally, 41 parent dyads living in the same home (n = 41) completed the survey to determine inter-rater reliability. Initial psychometric analyses were completed to trim items from the measure based on lack of variability in responses, moderate or higher item to scale correlation, or contribution to strong internal consistency. Inter-rater and test-retest reliability were completed using intraclass correlation coefficients. Validity was assessed using Pearson correlations between the HES scores and child and parent nutrition and PA. Results Eight items were removed and acceptable internal consistency was documented for all scales (α = .66–84) with the exception of the F&V accessibility. The F&V accessibility was reduced to a single item because the other two items did not meet reliability standards. Test-retest reliability was high (r > .75) for all scales. Inter-rater reliability varied across scales (r = .22–.89). PA accessibility, parent role modelling, and parental policies were all related significantly to child (r = .14–.21) and parent (r = .15–.31) PA. Similarly, availability of F&V and SS, parental role modelling, and parental policies were related to child (r = .14–36) and

  3. TSG attenuates LPC-induced endothelial cells inflammatory damage through notch signaling inhibition.

    PubMed

    Zhao, Jing; Liang, Yuan; Song, Fan; Xu, Shouzhu; Nian, Lun; Zhou, Xuanxuan; Wang, Siwang

    2016-01-01

    Lysophosphatidylcholine (LPC) induces inflammation in endothelial cells (ECs) but the mechanism is not fully understood. The Notch signaling pathway is involved in chronic EC inflammation, but its functions in LPC-induced endothelial inflammatory damage and 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside's (TSG) protective effect during LPC-induced inflammatory damage in human umbilical vein endothelial cells (HUVECs) is largely unknown. We report that Notch signaling activation contributed to LPC-induced injury in HUVECs, and that TSG protected HUVECs from LPC-induced injury by antagonizing Notch signaling activation by LPC. γ-secretase inhibitor (DAPT), a specific inhibitor of the Notch signaling pathway, and Notch1 siRNA were used to inhibit Notch activity. HUVECs were exposed to LPC in the presence or absence of TSG, DAPT, and Notch1 siRNA. LPC treatment of HUVECs resulted in reduced cell viability, and Notch1 and Hes1 upregulation. Either silencing of Notch1 by siRNA or pharmacological inhibition of Notch signaling by DAPT prevented the loss of cell viability, and induction of apoptosis, and enhanced expression Notch1, Hes1 and MCP-1 by LPC in HUVECs. Similarly, TSG reduced LPC stimulation of Notch1, Hes1, and MCP-1 expression, prevented the release of IL-6 and CRP and rescued HUVECs from LPC-induced cell damage. Our data indicate that the Notch signaling pathway is a crucial mediator of endothelial inflammatory damage and that TSG protects against endothelial inflammatory damage by inhibiting the Notch signaling pathway. Our findings suggest that targeting Notch signaling by natural products such as TSG is a promising strategy for the prevention and treatment of chronic inflammation associated diseases, including atherosclerosis. © 2015 IUBMB Life, 68(1):37-50, 2016. PMID:26662286

  4. Literature review of the lifetime of DOE materials: Aging of plastic bonded explosives and the explosives and polymers contained therein

    SciTech Connect

    Burgess, C.E.; Woodyard, J.D.; Rainwater, K.A.; Lightfoot, J.M.; Richardson, B.R.

    1998-09-01

    There are concerns about the lifetime of the nation`s stockpile of high explosives (HEs) and their components. The DOE`s Core Surveillance and Enhanced Surveillance programs specifically target degradation of HE, binders, and plastic-bonded explosives (PBXs) for determination of component lifetimes and handling procedures. The principal goal of this project is to identify the decomposition mechanisms of HEs, plasticizers, and plastic polymer binders resulting from exposure to ionizing radiation, heat, and humidity. The primary HEs of concern are 1,3,5-triamino-2,4,6-trinitrobenzene (TATB) and 1,3,5,7-tetranitro-1,3,5,7-tetraazocyclooctane (HMX). Hexahydro-1,3,5-triazine (RDX) is closely related to these two compounds and is also included in the literature review. Both Kel-F 800 and Estane are polymers of interest. A stabilizer, Irganox 1010, and an energetic plasticizer that is a blend of acetaldehyde 2,2-dinitropropyl acetal, are also of interest, but the focus of this report will be on the explosives and polymers. This presents a literature review that provides background on the synthesis, degradation, and techniques to analyze TATB, HMX, RDX, Kel-F 800, Estane, and the PBXs of these compounds. As there are many factors that can influence degradation of materials, the degradation discussion will be divided into sections based on each factor and how it might affect the degradation mechanism. The factors reviewed that influence the degradation of these materials are exposure to heat, UV- and {gamma}-irradiation, and the chemistry of these compounds. The report presents a recently compiled accounting of the available literature. 80 refs., 7 figs.

  5. Lhx2 Is an Essential Factor for Retinal Gliogenesis and Notch Signaling.

    PubMed

    de Melo, Jimmy; Zibetti, Cristina; Clark, Brian S; Hwang, Woochang; Miranda-Angulo, Ana L; Qian, Jiang; Blackshaw, Seth

    2016-02-24

    Müller glia (MG) are the only glial cell type produced by the neuroepithelial progenitor cells that generate the vertebrate retina. MG are required to maintain retinal homeostasis and support the survival of retinal neurons. Furthermore, in certain vertebrate classes, MG function as adult stem cells, mediating retinal regeneration in response to injury. However, the mechanisms that regulate MG development are poorly understood because there is considerable overlap in gene expression between retinal progenitor cells and differentiated MG. We show that the LIM homeodomain transcription factor Lhx2 is required for the development of MG in the mouse retina. Temporally controlled knock-out studies reveal a requirement for Lhx2 during all stages of MG development, ranging from the proliferation of gliocompetent retinal progenitors, activation of Müller-specific gene expression, and terminal differentiation of MG morphological features. We show that Lhx2 regulates gliogenesis in part by regulating directly the expression of Notch pathway genes including Notch1, Dll1, and Dll3 and gliogenic transcription factors such as Hes1, Hes5, Sox8, and Rax. Conditional knock-out of Lhx2 resulted in a rapid downregulation of Notch pathway genes and loss of Notch signaling. We further demonstrate that Müller gliogenesis induced by misexpression of the potently gliogenic Notch pathway transcriptional effector Hes5 requires Lhx2 expression. These results indicate that Lhx2 not only directly regulates expression of Notch signaling pathway components, but also acts together with the gliogenic Notch pathway to drive MG specification and differentiation. PMID:26911688

  6. Precise cis-regulatory control of spatial and temporal expression of the alx-1 gene in the skeletogenic lineage of s. purpuratus.

    PubMed

    Damle, Sagar; Davidson, Eric H

    2011-09-15

    Deployment of the gene-regulatory network (GRN) responsible for skeletogenesis in the embryo of the sea urchin Strongylocentrotus purpuratus is restricted to the large micromere lineage by a double negative regulatory gate. The gate consists of a GRN subcircuit composed of the pmar1 and hesC genes, which encode repressors and are wired in tandem, plus a set of target regulatory genes under hesC control. The skeletogenic cell state is specified initially by micromere-specific expression of these regulatory genes, viz. alx1, ets1, tbrain and tel, plus the gene encoding the Notch ligand Delta. Here we use a recently developed high throughput methodology for experimental cis-regulatory analysis to elucidate the genomic regulatory system controlling alx1 expression in time and embryonic space. The results entirely confirm the double negative gate control system at the cis-regulatory level, including definition of the functional HesC target sites, and add the crucial new information that the drivers of alx1 expression are initially Ets1, and then Alx1 itself plus Ets1. Cis-regulatory analysis demonstrates that these inputs quantitatively account for the magnitude of alx1 expression. Furthermore, the Alx1 gene product not only performs an auto-regulatory role, promoting a fast rise in alx1 expression, but also, when at high levels, it behaves as an auto-repressor. A synthetic experiment indicates that this behavior is probably due to dimerization. In summary, the results we report provide the sequence level basis for control of alx1 spatial expression by the double negative gate GRN architecture, and explain the rising, then falling temporal expression profile of the alx1 gene in terms of its auto-regulatory genetic wiring. PMID:21723273

  7. Identifying developmental toxicity pathways for a subset of ToxCast chemicals using human embryonic stem cells and metabolomics

    SciTech Connect

    Kleinstreuer, N.C.; Smith, A.M.; West, P.R.; Conard, K.R.; Fontaine, B.R.; Weir-Hauptman, A.M.; Palmer, J.A.; Knudsen, T.B.; Dix, D.J.; Donley, E.L.R.; Cezar, G.G.

    2011-11-15

    Metabolomics analysis was performed on the supernatant of human embryonic stem (hES) cell cultures exposed to a blinded subset of 11 chemicals selected from the chemical library of EPA's ToxCast Trade-Mark-Sign chemical screening and prioritization research project. Metabolites from hES cultures were evaluated for known and novel signatures that may be indicative of developmental toxicity. Significant fold changes in endogenous metabolites were detected for 83 putatively annotated mass features in response to the subset of ToxCast chemicals. The annotations were mapped to specific human metabolic pathways. This revealed strong effects on pathways for nicotinate and nicotinamide metabolism, pantothenate and CoA biosynthesis, glutathione metabolism, and arginine and proline metabolism pathways. Predictivity for adverse outcomes in mammalian prenatal developmental toxicity studies used ToxRefDB and other sources of information, including Stemina Biomarker Discovery's predictive DevTox Registered-Sign model trained on 23 pharmaceutical agents of known developmental toxicity and differing potency. The model initially predicted developmental toxicity from the blinded ToxCast compounds in concordance with animal data with 73% accuracy. Retraining the model with data from the unblinded test compounds at one concentration level increased the predictive accuracy for the remaining concentrations to 83%. These preliminary results on a 11-chemical subset of the ToxCast chemical library indicate that metabolomics analysis of the hES secretome provides information valuable for predictive modeling and mechanistic understanding of mammalian developmental toxicity. -- Highlights: Black-Right-Pointing-Pointer We tested 11 environmental compounds in a hESC metabolomics platform. Black-Right-Pointing-Pointer Significant changes in secreted small molecule metabolites were observed. Black-Right-Pointing-Pointer Perturbed mass features map to pathways critical for normal development and

  8. Measurement of Blood Volume in Adult Rhesus Macaques (Macaca mulatta)

    PubMed Central

    Hobbs, Theodore R; Blue, Steven W; Park, Byung S; Greisel, Jennifer J; Conn, P Michael; Pau, Francis K-Y

    2015-01-01

    Most biomedical facilities that use rhesus macaques (Macaca mulatta) limit the amount of blood that may be collected for experimental purposes. These limits typically are expressed as a percentage of blood volume (BV), estimated by using a fixed ratio of blood (mL) per body weight (kg). BV estimation ratios vary widely among facilities and typically do not factor in variables known to influence BV in humans: sex, age, and body condition. We used indicator dilution methodology to determine the BV of 20 adult rhesus macaques (10 male, 10 female) that varied widely in body condition. We measured body composition by using dual-energy X-ray absorptiometry, weight, crown-to-rump length, and body condition score. Two indicators, FITC-labeled hydroxyethyl starch (FITC–HES) and radioiodinated rhesus serum albumin (125I-RhSA), were injected simultaneously, followed by serial blood collection. Plasma volume at time 0 was determined by linear regression. BV was calculated from the plasma volume and Hct. We found that BV calculated by using FITC–HES was consistently lower than BV calculated by using 125I-RhSA. Sex and age did not significantly affect BV. Percentage body fat was significantly associated with BV. Subjects categorized as having ‘optimal’ body condition score had 18% body fat and 62.1 mL/kg BV (by FITC–HES; 74.5 mL/kg by 125I-RhSA). Each 1% increase in body fat corresponded to approximately 1 mL/kg decrease in BV. Body condition score correlated with the body fat percentage (R2 = 0.7469). We provide an equation for calculating BV from weight and body condition score. PMID:26632777

  9. Characterization of HIFU ablation using DNA fragmentation labeling as apoptosis stain

    NASA Astrophysics Data System (ADS)

    Anquez, Jeremie; Corréas, Jean-Michel; Pau, Bernard; Lacoste, François; Yon, Sylvain

    2012-11-01

    The goal of this work was to compare modalities to precisely quantify the extent of thermally induced lesions: gross pathology vs. histopathology vs. devascularization. Liver areas of 14 rabbits were targeted with HIFU and RF ablations in an acute study. Contrast enhanced computorized tomography (CE-CT) scan images were acquired two hours after HIFU and RF treatment to obtain the devascularized volumes of the livers. The animals were then euthanized and deep frozen. The livers were sliced and each slice was photographed and stacked yielding a volume of gross pathology. The volume VGP of the HIFU lesions were derived. The area AGP of the lesions were computed on a particular slice. The lesions were segmented as hypo intense (devascularized) regions on CE-CT images and their volumes VC were computed. The ratios VC/VGP were computed for all the HIFU lesions on all the 14 subjects with a mean value of 1.2. Histology was performed on the livers using Hematoxyline Eosine Staining (HES) and DNA Fragmentation labeling (TUNEL® technology) which characterizes apoptosis. Apoptotic regions of area AT were segmented on the images stained by TUNEL®. No necrosis was identified on the HES data. While TUNEL® did not mark the cores of the RF lesions as apoptotic, the periphery of HIFU and RF lesions was always recognized with TUNEL® as apoptotic. The ratio AGP/AT was computed. The mean value was 0.95 and 0.25 for HIFU and RF lesions respectively. These findings show that the devascularized territory seen on CE-CT scan coincide with the coagulated territories seen with gross pathology. Those actually correspond to cells in apoptosis. It is confirmed that HES stain does not show necrosis 2 hours after thermal ablation. TUNEL® technology for DNA fragmentation labeling appears as a useful marker for thermally induced acute lesions in the liver.

  10. A feeder-free culture using autogeneic conditioned medium for undifferentiated growth of human embryonic stem cells: Comparative expression profiles of mRNAs, microRNAs and proteins among different feeders and conditioned media

    PubMed Central

    2010-01-01

    Background Human embryonic stem (hES) cell lines were derived from the inner cell mass of human blastocysts, and were cultured on mouse embryonic fibroblast (MEF) feeder to maintain undifferentiated growth, extensive renewal capacity, and pluripotency. The hES-T3 cell line with normal female karyotype was previously used to differentiate into autogeneic fibroblast-like cells (T3HDF) as feeder to support the undifferentiated growth of hES-T3 cells (T3/HDF) for 14 passages. Results A feeder-free culture on Matrigel in hES medium conditioned by the autogeneic feeder cells (T3HDF) was established to maintain the undifferentiated growth of hES-T3 cells (T3/CMHDF) for 8 passages in this investigation. The gene expression profiles of mRNAs, microRNAs and proteins between the undifferentiated T3/HDF and T3/CMHDF cells were shown to be very similar, and their expression profiles were also found to be similar to those of T3/MEF and T3/CMMEF cells grown on MEF feeder and feeder-free Matrigel in MEF-conditioned medium, respectively. The undifferentiated state of T3/HDF and T3/CMHDF as well as T3/MEF andT3/CMMEF cells was evidenced by the very high expression levels of "stemness" genes and low expression levels of differentiation markers of ectoderm, mesoderm and endoderm in addition to the strong staining of OCT4 and NANOG. Conclusion The T3HDF feeder and T3HDF-conditioned medium were able to support the undifferentiated growth of hES cells, and they would be useful for drug development and toxicity testing in addition to the reduced risks of xenogeneic pathogens when used for medical applications such as cell therapies. PMID:20937144

  11. Precise cis-regulatory control of spatial and temporal expression of the alx-1 gene in the skeletogenic lineage of s. purpuratus

    PubMed Central

    Damle, Sagar; Davidson, Eric H.

    2011-01-01

    Deployment of the gene regulatory network (GRN) responsible for skeletogenesis in the embryo of the sea urchin Strongylocentrotus purpuratus is restricted to the large micromere lineage by a double negative regulatory gate. The gate consists of a GRN subcircuit composed of the pmar1 and hesC genes, which encode repressors and are wired in tandem, plus a set of target regulatory genes under hesC control. The skeletogenic cell state is specified initially by micromere-specific expression of these regulatory genes, viz. alx1, ets1, tbrain and tel, plus the gene encoding the Notch ligand Delta. Here we use a recently developed high throughput methodology for experimental cis-regulatory analysis to elucidate the genomic regulatory system controlling alx1 expression in time and embryonic space. The results entirely confirm the double negative gate control system at the cis-regulatory level, including definition of the functional HesC target sites, and add the crucial new information that the drivers of alx1 expression are initially Ets1, and then Alx1 itself plus Ets1. Cis-regulatory analysis demonstrates that these inputs quantitatively account for the magnitude of alx1 expression. Furthermore, the Alx1 gene product not only performs an auto-regulatory role, promoting a fast rise in alx1 expression, but also, when at high levels, it behaves as an autorepressor. A synthetic experiment indicates that this behavior is probably due to dimerization. In summary, the results we report provide the sequence level basis for control of alx1 spatial expression by the double negative gate GRN architecture, and explain the rising, then falling temporal expression profile of the alx1 gene in terms of its auto-regulatory genetic wiring. PMID:21723273

  12. Antitumor activity of S116836, a novel tyrosine kinase inhibitor, against imatinib-resistant FIP1L1-PDGFRα-expressing cells

    PubMed Central

    Ding, Ke; Zhang, Zhang; Wang, Deping; Pan, Jingxuan

    2014-01-01

    The FIP1-like-1-platelet-derived growth factor receptor alpha (FIP1L1-PDGFRα) fusion oncogene is the driver factor in a subset of patients with hypereosinophilic syndrome (HES)/chronic eosinophilic leukemia (CEL). Most FIP1L1-PDGFRα-positive patients respond well to the tyrosine kinase inhibitor (TKI) imatinib. Resistance to imatinib in HES/CEL has been described mainly due to the T674I mutation in FIP1L1-PDGFRα, which is homologous to the imatinib-resistant T315I mutation in BCR-ABL. Development of novel TKIs is imperative to overcome resistance to imatinib. We synthesized S116836, a novel TKI. In this study, we evaluated the antitumor activity of S116836 in FIP1L1-PDGFRα-expressing cells. The results showed that S116836 potently inhibited PDGFRα and its downstream signaling molecules such as STAT3, AKT, and Erk1/2. S116836 effectively inhibited the growth of the WT and T674I FIP1L1-PDGFRα-expressing neoplastic cells in vitro and in nude mouse xenografts. Moreover, S116836 induced intrinsic pathway of apoptosis as well as the death receptor pathway, coincided with up-regulation of the proapoptotic BH3-only protein Bim-EL through the Erk1/2 pathway. In conclusion, S116836 is active against WT and T674I FIP1L1-PDGFRα-expressing cells, and may be a prospective agent for the treatment of HES/CEL. PMID:25431951

  13. Coronavirus receptor switch explained from the stereochemistry of protein-carbohydrate interactions and a single mutation.

    PubMed

    Bakkers, Mark J G; Zeng, Qinghong; Feitsma, Louris J; Hulswit, Ruben J G; Li, Zeshi; Westerbeke, Aniek; van Kuppeveld, Frank J M; Boons, Geert-Jan; Langereis, Martijn A; Huizinga, Eric G; de Groot, Raoul J

    2016-05-31

    Hemagglutinin-esterases (HEs) are bimodular envelope proteins of orthomyxoviruses, toroviruses, and coronaviruses with a carbohydrate-binding "lectin" domain appended to a receptor-destroying sialate-O-acetylesterase ("esterase"). In concert, these domains facilitate dynamic virion attachment to cell-surface sialoglycans. Most HEs (type I) target 9-O-acetylated sialic acids (9-O-Ac-Sias), but one group of coronaviruses switched to using 4-O-Ac-Sias instead (type II). This specificity shift required quasisynchronous adaptations in the Sia-binding sites of both lectin and esterase domains. Previously, a partially disordered crystal structure of a type II HE revealed how the shift in lectin ligand specificity was achieved. How the switch in esterase substrate specificity was realized remained unresolved, however. Here, we present a complete structure of a type II HE with a receptor analog in the catalytic site and identify the mutations underlying the 9-O- to 4-O-Ac-Sia substrate switch. We show that (i) common principles pertaining to the stereochemistry of protein-carbohydrate interactions were at the core of the transition in lectin ligand and esterase substrate specificity; (ii) in consequence, the switch in O-Ac-Sia specificity could be readily accomplished via convergent intramolecular coevolution with only modest architectural changes in lectin and esterase domains; and (iii) a single, inconspicuous Ala-to-Ser substitution in the catalytic site was key to the emergence of the type II HEs. Our findings provide fundamental insights into how proteins "see" sugars and how this affects protein and virus evolution. PMID:27185912

  14. Diagnostic challenges in the work up of hypereosinophilia: pitfalls in bone marrow core biopsy interpretation.

    PubMed

    Schwaab, Juliana; Jawhar, Mohamad; Naumann, Nicole; Schmitt-Graeff, Annette; Fabarius, Alice; Horny, Hans-Peter; Cross, Nicholas C P; Hofmann, Wolf-Karsten; Reiter, Andreas; Metzgeroth, Georgia

    2016-03-01

    The FIP1L1-PDGFRA (FP) fusion gene is identified in a substantial proportion of patients with eosinophilia-associated myeloproliferative neoplasms (MPN-eo) who subsequently achieve rapid and durable remissions on imatinib. In the initial diagnostic work-up of hypereosinophilia (HE), histologic and immunohistochemical evaluation of a bone marrow (BM) core biopsy is considered essential for the differentiation between reactive hypereosinophilia (HER), MPN-eo and hypereosinophilic syndrome (HES). We therefore retrospectively analysed the initial reports of BM core biopsies from 116 patients who were subsequently identified as FP positive (FP+, n = 56) or FP negative/corticosteroid-responsive HER or HES (n = 60). Compared to HER or HES, detection of FP was more frequently associated with increased numbers of blasts (11/56 vs. 2/60, p = 0.007) and mast cells (23/33 vs. 7/23, p = 0.006; with expression of CD25 [11/18 vs. 2/13, p = 0.025]), and/or fibrosis (25/35 vs. 1/23, p < 0.0001). In FP+ patients, HE was correctly associated with an underlying clonal haematologic disorder in only 36/56 (64 %) of cases, but final BM diagnoses included a variety of diagnoses such as MPN-eo (n = 15), acute myeloid leukaemia (n = 8), systemic mastocytosis (n = 6), chronic myeloid leukaemia (n = 5) or unclassified MPN (n = 2). We conclude that the final evaluation of BM core biopsies in the diagnostic work-up of HE should include comprehensive morphologic (stains for myeloid blast cells, mast cells and fibres) and genetic analyses before a final diagnosis is established. PMID:26797429

  15. Id4, a new candidate gene for senile osteoporosis, acts as a molecular switch promoting osteoblast differentiation.

    PubMed

    Tokuzawa, Yoshimi; Yagi, Ken; Yamashita, Yzumi; Nakachi, Yutaka; Nikaido, Itoshi; Bono, Hidemasa; Ninomiya, Yuichi; Kanesaki-Yatsuka, Yukiko; Akita, Masumi; Motegi, Hiromi; Wakana, Shigeharu; Noda, Tetsuo; Sablitzky, Fred; Arai, Shigeki; Kurokawa, Riki; Fukuda, Toru; Katagiri, Takenobu; Schönbach, Christian; Suda, Tatsuo; Mizuno, Yosuke; Okazaki, Yasushi

    2010-07-01

    Excessive accumulation of bone marrow adipocytes observed in senile osteoporosis or age-related osteopenia is caused by the unbalanced differentiation of MSCs into bone marrow adipocytes or osteoblasts. Several transcription factors are known to regulate the balance between adipocyte and osteoblast differentiation. However, the molecular mechanisms that regulate the balance between adipocyte and osteoblast differentiation in the bone marrow have yet to be elucidated. To identify candidate genes associated with senile osteoporosis, we performed genome-wide expression analyses of differentiating osteoblasts and adipocytes. Among transcription factors that were enriched in the early phase of differentiation, Id4 was identified as a key molecule affecting the differentiation of both cell types. Experiments using bone marrow-derived stromal cell line ST2 and Id4-deficient mice showed that lack of Id4 drastically reduces osteoblast differentiation and drives differentiation toward adipocytes. On the other hand knockdown of Id4 in adipogenic-induced ST2 cells increased the expression of Ppargamma2, a master regulator of adipocyte differentiation. Similar results were observed in bone marrow cells of femur and tibia of Id4-deficient mice. However the effect of Id4 on Ppargamma2 and adipocyte differentiation is unlikely to be of direct nature. The mechanism of Id4 promoting osteoblast differentiation is associated with the Id4-mediated release of Hes1 from Hes1-Hey2 complexes. Hes1 increases the stability and transcriptional activity of Runx2, a key molecule of osteoblast differentiation, which results in an enhanced osteoblast-specific gene expression. The new role of Id4 in promoting osteoblast differentiation renders it a target for preventing the onset of senile osteoporosis. PMID:20628571

  16. Thermodynamic and structural insights into CSL-DNA complexes

    SciTech Connect

    Friedmann, David R.; Kovall, Rhett A.

    2010-10-28

    The Notch pathway is an intercellular signaling mechanism that plays important roles in cell fates decisions throughout the developing and adult organism. Extracellular complexation of Notch receptors with ligands ultimately results in changes in gene expression, which is regulated by the nuclear effector of the pathway, CSL (C-promoter binding factor 1 (CBF-1), suppressor of hairless (Su(H)), lin-12 and glp-1 (Lag-1)). CSL is a DNA binding protein that is involved in both repression and activation of transcription from genes that are responsive to Notch signaling. One well-characterized Notch target gene is hairy and enhancer of split-1 (HES-1), which is regulated by a promoter element consisting of two CSL binding sites oriented in a head-to-head arrangement. Although previous studies have identified in vivo and consensus binding sites for CSL, and crystal structures of these complexes have been determined, to date, a quantitative description of the energetics that underlie CSL-DNA binding is unknown. Here, we provide a thermodynamic and structural analysis of the interaction between CSL and the two individual sites that comprise the HES-1 promoter element. Our comprehensive studies that analyze binding as a function of temperature, salt, and pH reveal moderate, but distinct, differences in the affinities of CSL for the two HES-1 binding sites. Similarly, our structural results indicate that overall CSL binds both DNA sites in a similar manner; however, minor changes are observed in both the conformation of CSL and DNA. Taken together, our results provide a quantitative and biophysical basis for understanding how CSL interacts with DNA sites in vivo.

  17. Development and application of a UPLC-MS/MS method for the pharmacokinetic study of 10-hydroxy camptothecin and hydroxyethyl starch conjugate in rats.

    PubMed

    Li, Guofei; Cai, Cuifang; Ren, Tianyang; Tang, Xing

    2014-01-01

    With the purpose to carry out the pharmacokinetic studies of 10-hydroxy camptothecin (10-HCPT) and hydroxyethyl starch (10-HCPT-HES) conjugate, an ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated. The analytes, 10-HCPT and the internal standard, Diphenhydramine hydrochloride were extracted with ethyl acetate-isopropanol (95:5, v/v) and separated on an ACQUITY UPLC™ BEH C18 column using a mobile phase composed of acetonitrile and water (containing 0.1% formic acid) with a linear gradient program. With positive ion electrospray ionization (ESI), the analytes were monitored on a triple quadrupole mass spectrometer in the multiple reaction monitoring (MRM) mode. Linear calibration curves were obtained over the concentration ranges of 0.5-2500ng/mL. The intra- and inter-day precisions were less than 9.8% and 10.8%, respectively. The accuracy was within 12.1%. The mean recoveries of 10-HCPT at three concentrations of 2.5, 100, 2000ng/mL were higher than 87.2%. Commercial 10-HCPT injection and 10-HCPT-HES conjugate were administered intravenously at an equal dose of 10-HCPT at 0.5mg/kg. The biological half-life of conjugate was increased significantly from 10min to 3.15h and the bioavailability was 40 times higher than 10-HCPT injection. Consequently, the proposed UPLC-ESI-MS/MS method was proved to be sensitive, specific and reliable to analyze 10-HCPT in biological samples; 10-HCPT and HES conjugate is a promising strategy for delivery of 10-HCPT with prolonged half time and improved bioavailability. PMID:24140449

  18. Notch/Rbpjκ signaling regulates progenitor maintenance and differentiation of hypothalamic arcuate neurons

    PubMed Central

    Aujla, Paven K.; Naratadam, George T.; Xu, Liwen; Raetzman, Lori T.

    2013-01-01

    The hypothalamic arcuate nucleus (Arc), containing pro-opoiomelanocortin (POMC), neuropeptide Y (NPY) and growth hormone releasing hormone (GHRH) neurons, regulates feeding, energy balance and body size. Dysregulation of this homeostatic mediator underlies diseases ranging from growth failure to obesity. Despite considerable investigation regarding the function of Arc neurons, mechanisms governing their development remain unclear. Notch signaling factors such as Hes1 and Mash1 are present in hypothalamic progenitors that give rise to Arc neurons. However, how Notch signaling controls these progenitor populations is unknown. To elucidate the role of Notch signaling in Arc development, we analyzed conditional loss-of-function mice lacking a necessary Notch co-factor, Rbpjκ, in Nkx2.1-cre-expressing cells (Rbpjκ cKO), as well as mice with expression of the constitutively active Notch1 intracellular domain (NICD) in Nkx2.1-cre-expressing cells (NICD Tg). We found that loss of Rbpjκ results in absence of Hes1 but not of Hes5 within the primordial Arc at E13.5. Additionally, Mash1 expression is increased, coincident with increased proliferation and accumulation of Arc neurons at E13.5. At E18.5, Rbpjκ cKO mice have few progenitors and show increased numbers of differentiated Pomc, NPY and Ghrh neurons. By contrast, NICD Tg mice have increased hypothalamic progenitors, show an absence of differentiated Arc neurons and aberrant glial differentiation at E18.5. Subsequently, both Rbpjκ cKO and NICD Tg mice have changes in growth and body size during postnatal development. Taken together, our results demonstrate that Notch/Rbpjκ signaling regulates the generation and differentiation of Arc neurons, which contribute to homeostatic regulation of body size. PMID:23884446

  19. Hydroxyethyl Starch Could Save a Patient With Acute Aluminum Phosphide Poisoning.

    PubMed

    Marashi, Sayed Mahdi; Nasri Nasrabadi, Zeynab; Jafarzadeh, Mostafa; Mohammadi, Sogand

    2016-07-01

    A 40-year-old male patient with suicidal ingestion of one tablet of aluminium phosphide was referred to the department of toxicology emergency of Baharloo Hospital, Tehran, Iran. The garlic odor was smelled from the patient and abdominal pain and continuous vomiting as well as agitation and heartburn were the first signs and symptoms. Systolic and diastolic blood pressures at the arrival time were 95 and 67 mmHg, respectively. Gastric lavage with potassium permanganate (1:10,000), and 2 vials of sodium bicarbonate through a nasogastric tube was started for the patient and the management was continued with free intravenous infusion of 1 liter of NaCl 0.9% serum plus NaHCO3, hydrocortisone acetate (200 mg), calcium gluconate (1 g) and magnesium sulfate (1 g). Regarding the large intravenous fluid therapy and vasoconstrictor administering (norepinephrine started by 5 µg/min and continued till 15 µg/min), there were no signs of response and the systolic blood pressure was 49 mmHg. At this time, hydroxyethyl starch (HES) (6% hetastarch 600/0.75 in 0.9% sodium chloride) with a dose of 600 cc in 6 hours was started for the patient. At the end of therapy with HES, the patient was stable with systolic and diastolic blood pressure of 110 and 77 mmHg, respectively. He was discharged on the 6th day after the psychological consultation, with normal clinical and paraclinical examinations. This is the first report of using HES in the management of AlP poisoning and its benefit to survive the patient. PMID:27424021

  20. Heligmosomoides polygyrus elicits a dominant nonprotective antibody response directed against restricted glycan and peptide epitopes.

    PubMed

    Hewitson, James P; Filbey, Kara J; Grainger, John R; Dowle, Adam A; Pearson, Mark; Murray, Janice; Harcus, Yvonne; Maizels, Rick M

    2011-11-01

    Heligmosomoides polygyrus is a widely used gastrointestinal helminth model of long-term chronic infection in mice, which has not been well-characterized at the antigenic level. We now identify the major targets of the murine primary Ab response as a subset of the secreted products in H. polygyrus excretory-secretory (HES) Ag. An immunodominant epitope is an O-linked glycan (named glycan A) carried on three highly expressed HES glycoproteins (venom allergen Ancylostoma-secreted protein-like [VAL]-1, -2, and -5), which stimulates only IgM Abs, is exposed on the adult worm surface, and is poorly represented in somatic parasite extracts. A second carbohydrate epitope (glycan B), present on both a non-protein high molecular mass component and a 65-kDa molecule, is widely distributed in adult somatic tissues. Whereas the high molecular mass component and 65-kDa molecules bear phosphorylcholine, the glycan B epitope itself is not phosphorylcholine. Class-switched IgG1 Abs are found to glycan B, but the dominant primary IgG1 response is to the polypeptides of VAL proteins, including also VAL-3 and VAL-4. Secondary Ab responses include the same specificities while also recognizing VAL-7. Although vaccination with HES conferred complete protection against challenge H. polygyrus infection, mAbs raised against each of the glycan epitopes and against VAL-1, VAL-2, and VAL-4 proteins were unable to do so, even though these specificities (with the exception of VAL-2) are also secreted by tissue-phase L4 larvae. The primary immune response in susceptible mice is, therefore, dominated by nonprotective Abs against a small subset of antigenic epitopes, raising the possibility that these act as decoy specificities that generate ineffective humoral immunity. PMID:21964031

  1. Heligmosomoides polygyrus elicits a dominant non-protective antibody response directed against restricted glycan and peptide epitopes

    PubMed Central

    Hewitson, James P; Filbey, Kara J; Grainger, John R; Dowle, Adam A; Pearson, Mark; Murray, Janice; Harcus, Yvonne; Maizels, Rick M

    2014-01-01

    Summary Heligmosomoides polygyrus is a widely-used gastrointestinal helminth model of long-term chronic infection in mice, which has not been well-characterized at the antigenic level. We now identify the major targets of the murine primary antibody response as a subset of the secreted products in H. polygyrus Excretory-Secretory (HES) antigen. An immunodominant epitope is an O-linked glycan (named Glycan A) carried on 3 highly-expressed HES glycoproteins (VAL-1, -2 and -5), which stimulates only IgM antibodies, is exposed on the adult worm surface, and is poorly represented in somatic parasite extracts. A second carbohydrate epitope (Glycan B), present on both a non-protein high molecular weight component (HM) and a 65-kDa molecule, is widely distributed in adult somatic tissues; while HM-65 molecules bear phosphorylcholine (PC), the Glycan B epitope itself is not PC. Class-switched IgG1 antibodies are found to Glycan B, but the dominant primary IgG1 response is to the polypeptides of VAL proteins, including also VAL-3 and VAL-4. Secondary antibody responses are similar while recognizing in addition VAL-7. Whilst vaccination with HES conferred complete protection against challenge H. polygyrus infection, monoclonal antibodies raised against each of the glycan epitopes, and against VAL-1, -2 and -4 proteins were unable to do so, even though these specificities (with the exception of VAL-2) are also secreted by tissue-phase L4 larvae. The primary immune response in susceptible mice is, therefore, dominated by nonprotective antibodies against a small subset of antigenic epitopes, raising the possibility that these act as decoy specificities that generate ineffective humoral immunity. PMID:21964031

  2. Measurement of Blood Volume in Adult Rhesus Macaques (Macaca mulatta).

    PubMed

    Hobbs, Theodore R; Blue, Steven W; Park, Byung S; Greisel, Jennifer J; Conn, P Michael; Pau, Francis K-Y

    2015-11-01

    Most biomedical facilities that use rhesus macaques (Macaca mulatta) limit the amount of blood that may be collected for experimental purposes. These limits typically are expressed as a percentage of blood volume (BV), estimated by using a fixed ratio of blood (mL) per body weight (kg). BV estimation ratios vary widely among facilities and typically do not factor in variables known to influence BV in humans: sex, age, and body condition. We used indicator dilution methodology to determine the BV of 20 adult rhesus macaques (10 male, 10 female) that varied widely in body condition. We measured body composition by using dual-energy X-ray absorptiometry, weight, crown-to-rump length, and body condition score. Two indicators, FITC-labeled hydroxyethyl starch (FITC-HES) and radioiodinated rhesus serum albumin ((125)I-RhSA), were injected simultaneously, followed by serial blood collection. Plasma volume at time 0 was determined by linear regression. BV was calculated from the plasma volume and Hct. We found that BV calculated by using FITC-HES was consistently lower than BV calculated by using (125)I-RhSA. Sex and age did not significantly affect BV. Percentage body fat was significantly associated with BV. Subjects categorized as having 'optimal' body condition score had 18% body fat and 62.1 mL/kg BV (by FITC-HES; 74.5 mL/kg by (125)I-RhSA). Each 1% increase in body fat corresponded to approximately 1 mL/kg decrease in BV. Body condition score correlated with the body fat percentage (R(2) = 0.7469). We provide an equation for calculating BV from weight and body condition score. PMID:26632777

  3. Enhanced cell growth and tumorigenicity of rat glioma cells by stable expression of human CD133 through multiple molecular actions.

    PubMed

    Fang, Kuan-Min; Lin, Tzu-Chien; Chan, Ti-Chun; Ma, Shi-Zhang; Tzou, Bo-Cheng; Chang, Wen-Ruei; Liu, Jun-Jen; Chiou, Shih-Hwa; Yang, Chung-Shi; Tzeng, Shun-Fen

    2013-09-01

    CD133 (Prominin-1/AC133) is generally treated as a cell surface marker found on multipotent stem cells and tumor stem-like cells, and its biological function remains debated. Genetically modified rat glioma cell lines were generated by lentiviral gene delivery of human CD133 into rat C6 glioma cells (hCD133(+) -C6) or by infection of C6 cells with control lentivirus (mock-C6). Stable hCD133 expression promoted the self-renewal ability of C6-formed spheres with an increase in the expression of the stemness markers, Bmi-1 and SOX2. Akt phosphorylation, Notch-1 activation, and Notch-1 target gene expression (Hes-1, Hey1 and Hey2) were increased in hCD133(+) -C6 when compared to mock-C6. The inhibition of Akt phosphorylation, Notch-1 activation, and Hes-1 in hCD133(+) -C6 cells effectively suppressed their clonogenic ability, indicating that these factors are involved in expanding the growth of hCD133(+) -C6. An elevated expression of GTPase-activating protein 27 (Arhgap27) was detected in hCD133(+) -C6. A decline in the invasion of hCD133(+) -C6 by knockdown of Arhgap27 expression indicated the critical role of Arhgap27 in promoting cell migration of hCD133(+) -C6. In vivo study further showed that hCD133(+) -C6 formed aggressive tumors in vivo compared to mock-C6. Exposure of hCD133(+) -C6 to arsenic trioxide not only reduced Akt phosphorylation, Notch-1 activation and Hes-1 expression in vitro, but also inhibited their tumorigenicity in vivo. The results show that C6 glioma cells with stable hCD133 expression enhanced their stemness properties with increased Notch-1/Hes-1 signaling, Akt activation, and Arhgap27 action, which contribute to increased cell proliferation and migration of hCD133(+) -C6 in vitro, as well as progressive tumor formation in vivo. PMID:23832679

  4. Endothelial Cells Stimulate Self-Renewal and Expand Neurogenesis of Neural Stem Cells

    NASA Astrophysics Data System (ADS)

    Shen, Qin; Goderie, Susan K.; Jin, Li; Karanth, Nithin; Sun, Yu; Abramova, Natalia; Vincent, Peter; Pumiglia, Kevin; Temple, Sally

    2004-05-01

    Neural stem cells are reported to lie in a vascular niche, but there is no direct evidence for a functional relationship between the stem cells and blood vessel component cells. We show that endothelial cells but not vascular smooth muscle cells release soluble factors that stimulate the self-renewal of neural stem cells, inhibit their differentiation, and enhance their neuron production. Both embryonic and adult neural stem cells respond, allowing extensive production of both projection neuron and interneuron types in vitro. Endothelial coculture stimulates neuroepithelial cell contact, activating Notch and Hes1 to promote self-renewal. These findings identify endothelial cells as a critical component of the neural stem cell niche.

  5. Modelling Ultradian Oscillations and Segmentation

    NASA Astrophysics Data System (ADS)

    Jensen, Mogens

    2008-03-01

    We model ultradian oscillations in four different eucaryotic systems: Hes1, p53-mdm2, NF-kB and Wnt-Notch. In each of the systems we identify the feed-back loops for the genetic regulations. Oscillations are possible when time delays are present, either by directly introducing a delay, by many steps in the loops or by saturated degradation. The oscillations are important for apoptosis and control of inflammation. The Wnt-Notch system is essential in embryo segmentation and we introduce a model in which the Wnt oscillates by itself but drives the Notch cycle out of phase with the Wnt cycle, in good agreement with experimental observations.

  6. From C-Enhanced, Metal Poor Stars to AGB Nucleosynthesis

    NASA Astrophysics Data System (ADS)

    Lucatello, S.; Gratton, R.; Beers, T.; Carretta, E.

    The largest to date surveys for metal poor stars (i.e. HK survey Beers et al. 1992 and HES Christlieb et al. 2001) find that as many as ~25% of stars with [Fe/H]≤ -2.5 dex are have [C/Fe]>1 dex (CEMP stars). High resolution studies have revealed that the C-enhancements is accompanied by different abundance patterns, s and/or r-process enrichment, but there are also cases with no ncapture elements overabundance,and with or without extraordinary α elements enhancements. The mechanisms that originate the range of phenomena observed are far from being fully understood.

  7. MicroRNA and gene expression patterns in the differentiation of human embryonic stem cells

    PubMed Central

    Ren, Jiaqiang; Jin, Ping; Wang, Ena; Marincola, Francesco M; Stroncek, David F

    2009-01-01

    Background The unique features of human embryonic stem (hES) cells make them the best candidate resource for both cell replacement therapy and development research. However, the molecular mechanisms responsible for the simultaneous maintenance of their self-renewal properties and undifferentiated state remain unclear. Non-coding microRNAs (miRNA) which regulate mRNA cleavage and inhibit encoded protein translation exhibit temporal or tissue-specific expression patterns and they play an important role in development timing. Results In this study, we analyzed miRNA and gene expression profiles among samples from 3 hES cell lines (H9, I6 and BG01v), differentiated embryoid bodies (EB) derived from H9 cells at different time points, and 5 adult cell types including Human Microvascular Endothelial Cells (HMVEC), Human Umbilical Vein Endothelial Cells (HUVEC), Umbilical Artery Smooth Muscle Cells (UASMC), Normal Human Astrocytes (NHA), and Lung Fibroblasts (LFB). This analysis rendered 104 miRNAs and 776 genes differentially expressed among the three cell types. Selected differentially expressed miRNAs and genes were further validated and confirmed by quantitative real-time-PCR (qRT-PCR). Especially, members of the miR-302 cluster on chromosome 4 and miR-520 cluster on chromosome 19 were highly expressed in undifferentiated hES cells. MiRNAs in these two clusters displayed similar expression levels. The members of these two clusters share a consensus 7-mer seed sequence and their targeted genes had overlapping functions. Among the targeted genes, genes with chromatin structure modification function are enriched suggesting a role in the maintenance of chromatin structure. We also found that the expression level of members of the two clusters, miR-520b and miR-302c, were negatively correlated with their targeted genes based on gene expression analysis Conclusion We identified the expression patterns of miRNAs and gene transcripts in the undifferentiation of human embryonic

  8. The Equation of State of PBX 9502

    NASA Astrophysics Data System (ADS)

    Aslam, Tariq

    Reactive flow modeling of high explosives (HEs) requires accurate equation of states (EOS) for both the HE's reactants and products. The Wescott-Stewart-Davis ``wide-ranging EOS'' model will be examined. A procedure for calibrating both the reactants and products for this EOS will be presented. Several thermodynamic pathways will be explored for the plastic bonded HE PBX 9502. These include: isothermal compression, isentropic compression, single and multiple shock compression, isobaric thermal expansion, adiabatic expansion of the products and the overdriven detonation state. Data from several different experimental techniques are employed to constrain model parameters. Validation tests of the model EOS will also be presented.

  9. Lhx2 Is an Essential Factor for Retinal Gliogenesis and Notch Signaling

    PubMed Central

    de Melo, Jimmy; Zibetti, Cristina; Clark, Brian S.; Hwang, Woochang; Miranda-Angulo, Ana L.; Qian, Jiang

    2016-01-01

    Müller glia (MG) are the only glial cell type produced by the neuroepithelial progenitor cells that generate the vertebrate retina. MG are required to maintain retinal homeostasis and support the survival of retinal neurons. Furthermore, in certain vertebrate classes, MG function as adult stem cells, mediating retinal regeneration in response to injury. However, the mechanisms that regulate MG development are poorly understood because there is considerable overlap in gene expression between retinal progenitor cells and differentiated MG. We show that the LIM homeodomain transcription factor Lhx2 is required for the development of MG in the mouse retina. Temporally controlled knock-out studies reveal a requirement for Lhx2 during all stages of MG development, ranging from the proliferation of gliocompetent retinal progenitors, activation of Müller-specific gene expression, and terminal differentiation of MG morphological features. We show that Lhx2 regulates gliogenesis in part by regulating directly the expression of Notch pathway genes including Notch1, Dll1, and Dll3 and gliogenic transcription factors such as Hes1, Hes5, Sox8, and Rax. Conditional knock-out of Lhx2 resulted in a rapid downregulation of Notch pathway genes and loss of Notch signaling. We further demonstrate that Müller gliogenesis induced by misexpression of the potently gliogenic Notch pathway transcriptional effector Hes5 requires Lhx2 expression. These results indicate that Lhx2 not only directly regulates expression of Notch signaling pathway components, but also acts together with the gliogenic Notch pathway to drive MG specification and differentiation. SIGNIFICANCE STATEMENT Müller glia (MG) are radial glial cells located in the vertebrate retina that are essential for the function and survival of retinal neurons. We found the LIM homeodomain transcription factor Lhx2 to be expressed in both retinal progenitor cells and MG. Using conditional knock-outs, we show that Lhx2 is required

  10. VizieR Online Data Catalog: A low-luminosity type-1 QSO sample. III. (Tremou+, 2015)

    NASA Astrophysics Data System (ADS)

    Tremou, E.; Garcia-Marin, M.; Zuther, J.; Eckart, A.; Valencia-Schneider, M.; Vitale, M.; Shan, C.

    2015-11-01

    The optical spectroscopic analysis of the LLQSO sample is based on two sets of observations. The first data set is from the HES (Reimers et al., 1996A&AS..115..235R, Cat. J/A+AS/115/235), in which follow-up observations were carried out to confirm the type-1 character of the survey candidates spectroscopically. In addition to this, we have used data from the 6 Degree Field Galaxy survey (6dFGS), an optical spectroscopy public database (Jones et al., 2004MNRAS.355..747J, 2009MNRAS.399..683J, Cat. VII/259). (3 data files).

  11. Preliminary studies on the heat exchanger option for S-CO{sub 2} power conversion cycle coupled to water cooled SMR

    SciTech Connect

    Ahn, Y.; Lee, J.; Lee, J. I.

    2012-07-01

    For more than a half century, the steam Rankine cycle had been the major power conversion cycle for a nuclear power plant. However, as the interest on the next generation reactors grows, a variety of alternative power conversion systems have been studied. Among them, the S-CO{sub 2} cycle (Supercritical carbon dioxide Brayton cycle) is considered as a promising candidate due to several benefits such as 1) Relatively high thermal efficiency at relatively low turbine inlet temperature, 2) High efficiency with simple lay-out 3) Compactness of turbo-machineries. 4) Compactness of total cycle combined with PCHE (Printed Circuit Heat Exchanger). According to the conventional classification of heat exchangers (HE), there are three kind of HE, 1) Tubular HEs, 2) Plate-type HEs, 3) Extended surface HEs. So far, the researcher has mostly assumed PCHE type HE for the S-CO{sub 2} cycle due to its compactness with reasonably low pressure drop. However, PCHE is currently one of the most expensive components in the cycle, which can have a negative effect on the economics of the cycle. Therefore, an alternative for the HE should be seriously investigated. By comparing the operating condition (pressure and temperature) there are three kind of HE in the S-CO{sub 2} cycle, 1) IHX (Intermediate Heat exchanger) 2) Recuperator and 3) Pre-cooler. In each heat exchanger, hot side and cold side coolants are different, i.e. reactor coolant to S-CO{sub 2} (IHX), S-CO{sub 2} to S-CO{sub 2}(Recuperator), S-CO{sub 2} to water (Pre-cooler). By considering all the attributes mentioned above, all existing types of heat exchangers are compared to find a possible alternative to PCHE. The comparing factors are 1) Size(volume), 2) Cost. Plate fin type HEs are considered to be the most competitive heat exchanger regarding the size and the cost after some improvements on the design limit are made. (authors)

  12. Silencing of core transcription factors in human EC cells highlights the importance of autocrine FGF signaling for self-renewal

    PubMed Central

    Greber, Boris; Lehrach, Hans; Adjaye, James

    2007-01-01

    Background Despite their distinct origins, human embryonic stem (hES) and embryonic carcinoma (hEC) cells share a number of similarities such as surface antigen expression, growth characteristics, the ability to either self-renew or differentiate, and control of the undifferentiated state by the same core transcription factors. To obtain further insights into the regulation of self-renewal, we have silenced hES/hEC cell-specific genes in NCCIT hEC cells and analysed the downstream effects by means of microarrays. Results RNAi-mediated silencing of OCT4 and SOX2 induced differentiation with mesodermal characteristics. Markers of trophoblast induction were only transiently up-regulated in the OCT4 knock-down. Independent knock-downs of NANOG produced a proliferation rather than a differentiation phenotype, which may be due to high NANOG expression levels in the cell line used. Published ChIP-chip data from hES cells were used to identify putative direct targets. RNAi-mediated differentiation was accompanied by direct down-regulation of known hES/hEC cell markers. This included all three core transcription factors in the case of the OCT4 and SOX2 knock-downs, confirming previous findings of reciprocal activation in ES cells. Furthermore, large numbers of histone genes as well as epigenetic regulators were differentially expressed, pointing at chromatin remodeling as an additional regulatory level in the differentiation process. Moreover, loss of self-renewal was accompanied by the down-regulation of genes involved in FGF signaling. FGF receptor inhibition for short and prolonged periods of time revealed that the ERK/MAPK cascade is activated by endogenously expressed fibroblast growth factors and that FGF signaling is cruicial for maintaining the undifferentiated state of hEC cells, like in hES cells. Conclusion Control of self-renewal appears to be very similar in hEC and hES cells. This is supported by large numbers of common transcription factor targets and the

  13. Detection of occult carcinomatous diffusion in lymph nodes from head and neck squamous cell carcinoma using real-time RT–PCR detection of cytokeratin 19 mRNA

    PubMed Central

    Tao, L; Lefèvre, M; Ricci, S; Saintigny, P; Callard, P; Périé, S; Lacave, R; Bernaudin, J-F; Lacau St Guily, J

    2006-01-01

    The aim of the present study was to evaluate the occult lymph node carcinomatous diffusion in head and neck squamous cell carcinoma (HNSCC). A total of 1328 lymph nodes from 31 patients treated between 2004 and 2005 were prospectively evaluated by routine haematoxylin–eosin–safran (HES) staining, immunohistochemistry (IHC) and real-time Taqman reverse–transcriptase polymerase chain reaction (real-time RT–PCR) assay. Amplification of cytokeratin 19 (CK19) mRNA transcripts using real-time RT–PCR was used to quantify cervical micrometastatic burden. The cervical lymph node metastatic rates determined by routine HES staining and real-time RT–PCR assay were 16.3 and 36.0%, respectively (P<0.0001). A potential change in the nodal status was observed in 13 (42.0%) of the 31 patients and an atypical pattern of lymphatic spread was identified in four patients (12.9%). Moreover, CK19 mRNA expression values in histologically positive lymph nodes were significantly higher than those observed in histologically negative lymph nodes (P<0.0001). These results indicate that real-time RT–PCR assay for the detection of CK19 mRNA is a sensitive and reliable method for the detection of carcinomatous cells in lymph nodes. This type of method could be used to reassess lymph node status according to occult lymphatic spread in patients with HNSCC. PMID:16622440

  14. Characterization of sodium hyaluronate blends using frit inlet asymmetrical flow field-flow fractionation and multiangle light scattering.

    PubMed

    Ali, Muhammad; Hwang, Euijin; Cho, Il-Hwan; Moon, Myeong Hee

    2012-01-01

    We characterized ultrahigh molecular weight sodium hyaluronate (NaHA) and blended pharmaceutical products containing NaHA using flow field-flow fractionation and multiangle light scattering-differential refractive index (FlFFF-MALS-DRI). NaHA is a water-soluble polysaccharide with a range of molecular weights (MW; 10(5)~10(8) Da) that is found in body fluids and tissues. NaHA is also used commercially in pharmaceutical and cosmetic applications. We used a frit inlet asymmetrical FlFFF channel to separate aqueous polymers according to their hydrodynamic size, and we used on-line measurements of light scattering to obtain the MW distribution (MWD) as well as structural information about NaHA in aqueous solution. In this study, we investigated NaHA and anti-adhesive blend mixtures of NaHA (a commercial NaHA blend mixture containing sodium carboxymethyl cellulose and a new blend with hydroxyethyl starch (HES)) to determine the molecular weight distribution MWD of NaHA and the blend mixtures and to obtain structural information about these compounds in aqueous solution. We also examined the characteristics of NaHA-HES-polylactic-co-glycolic acid film products exposed to gamma radiation for sterilization purposes. PMID:22101460

  15. Structural and mechanistic insights into cooperative assembly of dimeric Notch transcription complexes

    SciTech Connect

    Arnett, Kelly L.; Hass, Matthew; McArthur, Debbie G.; Ilagan, Ma Xenia G.; Aster, Jon C.; Kopan, Raphael; Blacklow, Stephen C.

    2010-11-12

    Ligand-induced proteolysis of Notch produces an intracellular effector domain that transduces essential signals by regulating the transcription of target genes. This function relies on the formation of transcriptional activation complexes that include intracellular Notch, a Mastermind co-activator and the transcription factor CSL bound to cognate DNA. These complexes form higher-order assemblies on paired, head-to-head CSL recognition sites. Here we report the X-ray structure of a dimeric human Notch1 transcription complex loaded on the paired site from the human HES1 promoter. The small interface between the Notch ankyrin domains could accommodate DNA bending and untwisting to allow a range of spacer lengths between the two sites. Cooperative dimerization occurred on the human and mouse Hes5 promoters at a sequence that diverged from the CSL-binding consensus at one of the sites. These studies reveal how promoter organizational features control cooperativity and, thus, the responsiveness of different promoters to Notch signaling.

  16. Cloning and Expression of Recombinant Human Endostatin in Periplasm of Escherichia coli Expression System

    PubMed Central

    Mohajeri, Abbas; Pilehvar-Soltanahmadi, Yones; Pourhassan-Moghaddam, Mohammad; Abdolalizadeh, Jalal; Karimi, Pouran; Zarghami, Nosratollah

    2016-01-01

    Purpose: Recombinant human endostatin (rhEs) is an angiogenesis inhibitor which is used as a specific drug in the treatment of non-small-cell lung cancer. In the current research, we developed an efficient method for expressing soluble form of the rhEs protein in the periplasmic space of Escherichia coli via fusing with pelB signal peptide. Methods: The human endostatin (hEs) gene was amplified using synthetic (hEs) gene as a template; then, cloned and expressed under T7 lac promoter. IPTG was used as an inducer for rhEs expression. Next, the osmotic shock was used to extraction of protein from the periplasmic space. The presence of rhEs in the periplasmic space was approved by SDS-PAGE and Western blotting. Results: The results show the applicability of pelB fusion protein system usage for secreting rhEs in the periplasm of E. coli in the laboratory scale. The rhEs represents approximately 35 % (0.83mg/l) of the total cell protein. Conclusion: The present study apparently is the first report of codon-optimized rhEs expression as a fusion with pelB signal peptide. The results presented the successful secretion of soluble rhEs to the periplasmic space. PMID:27478780

  17. Coupled hydrodynamic model for laser-plasma interaction and hot electron generation.

    PubMed

    Colaïtis, A; Duchateau, G; Ribeyre, X; Maheut, Y; Boutoux, G; Antonelli, L; Nicolaï, Ph; Batani, D; Tikhonchuk, V

    2015-10-01

    We present a formulation of the model of laser-plasma interaction (LPI) at hydrodynamical scales that couples the plasma dynamics with linear and nonlinear LPI processes, including the creation and propagation of high-energy electrons excited by parametric instabilities and collective effects. This formulation accounts for laser beam refraction and diffraction, energy absorption due to collisional and resonant processes, and hot electron generation due to the stimulated Raman scattering, two-plasmon decay, and resonant absorption processes. Hot electron (HE) transport and absorption are described within the multigroup angular scattering approximation, adapted for transversally Gaussian electron beams. This multiscale inline LPI-HE model is used to interpret several shock ignition experiments, highlighting the importance of target preheating by HEs and the shortcomings of standard geometrical optics when modeling the propagation and absorption of intense laser pulses. It is found that HEs from parametric instabilities significantly increase the shock pressure and velocity in the target, while decreasing its strength and the overall ablation pressure. PMID:26565161

  18. Coupled hydrodynamic model for laser-plasma interaction and hot electron generation

    NASA Astrophysics Data System (ADS)

    Colaïtis, A.; Duchateau, G.; Ribeyre, X.; Maheut, Y.; Boutoux, G.; Antonelli, L.; Nicolaï, Ph.; Batani, D.; Tikhonchuk, V.

    2015-10-01

    We present a formulation of the model of laser-plasma interaction (LPI) at hydrodynamical scales that couples the plasma dynamics with linear and nonlinear LPI processes, including the creation and propagation of high-energy electrons excited by parametric instabilities and collective effects. This formulation accounts for laser beam refraction and diffraction, energy absorption due to collisional and resonant processes, and hot electron generation due to the stimulated Raman scattering, two-plasmon decay, and resonant absorption processes. Hot electron (HE) transport and absorption are described within the multigroup angular scattering approximation, adapted for transversally Gaussian electron beams. This multiscale inline LPI-HE model is used to interpret several shock ignition experiments, highlighting the importance of target preheating by HEs and the shortcomings of standard geometrical optics when modeling the propagation and absorption of intense laser pulses. It is found that HEs from parametric instabilities significantly increase the shock pressure and velocity in the target, while decreasing its strength and the overall ablation pressure.

  19. Hypereosinophilic syndrome: cardiac diagnosis and management.

    PubMed

    Mankad, Rekha; Bonnichsen, Crystal; Mankad, Sunil

    2016-01-01

    Hypereosinophilic syndrome (HES) is a heterogeneous group of conditions that is defined at its core by hypereosinophilia (HE) (blood eosinophil count of >1.5×10(9)/L) and organ damage directly attributable to the HE. Cardiac dysfunction occurs frequently in all forms of HES and is a major cause of morbidity and mortality. Once a significantly elevated eosinophil count is identified, it must be confirmed on repeat testing and the aetiology for the HE must be rigorously sought out with a focus on identifying whether organ dysfunction is occurring. Echocardiography is routinely performed to assess for cardiac involvement, looking for evidence of left ventricular and/or right ventricular apical obliteration or thrombi or a restrictive cardiomyopathy. Cardiac magnetic resonance imaging and CT are often useful adjuncts to establish the diagnosis but endomyocardial biopsy remains the gold standard. To decrease the degree of eosinophilia, treatment can include corticosteroids and/or imatinib based on the aetiology. Anticoagulation, standard heart failure therapy for a restrictive cardiomyopathy and finally cardiac transplantation may be indicated in the treatment algorithm. PMID:26567231

  20. Harvesting of granulocytes using a hydroxyethyl starch solution.

    PubMed

    Sussman, L N; Colli, W; Pichetshote, C

    1975-01-01

    The need for sophisticated ocmponent therapy has resulted in improved techniques for obtaining concentrates of platelets and granulocytes. The use of single donors as a source for these products is advisable to avoid multiple sensitizations. Obtaining concentrated granulocytes represents a problem because of the difficulty in separating granulocytes from red blood cells by differential centrifugation or sedimentation since the specific gravities are similar. Hydroxyethyl starch (HES) makes the separation more effective. A solution made of 250 ml of 6 per cent HES, 250 ml of distilled water, and 15 g of sodium citrate in 30 ml distilled water provided a satisfactory anticoagulant solution for this purpose. The granulocytes collected averaged 49 per cent of the total available in the processed blood; the platelets averages 82 per cent. A satisfactory yield could thus be obtained from a single donor, and this could be repeated several times in a month. The value of ABO, Rh, and HL-A compatibility between donor and recipient probably increases the viability and safety of this procedure. The Haemonetic No. 30 Cell Separator provided an easy and rapid method for this procedure. PMID:53918

  1. Atractylenolide I-mediated Notch pathway inhibition attenuates gastric cancer stem cell traits

    SciTech Connect

    Ma, Li; Mao, Rurong; Shen, Ke; Zheng, Yuanhong; Li, Yueqi; Liu, Jianwen; Ni, Lei

    2014-07-18

    Highlights: • This paper supports the anti-tumor effects of AT-I on gastric cancer in vitro. • AT-I attenuates gastric cancer stem cell traits. • It is the systematic study regarding AT-I suppression of Notch pathway in GC and GCSLCs. - Abstract: Atractylenolide I (AT-I), one of the main naturally occurring compounds of Rhizoma Atractylodis Macrocephalae, has remarkable anti-cancer effects on various cancers. However, its effects on the treatment of gastric cancer remain unclear. Via multiple cellular and molecular approaches, we demonstrated that AT-I could potently inhibit cancer cell proliferation and induce apoptosis through inactivating Notch pathway. AT-I treatment led to the reduction of expressions of Notch1, Jagged1, and its downstream Hes1/ Hey1. Our results showed that AT-I inhibited the self-renewal capacity of gastric stem-like cells (GCSLCs) by suppression of their sphere formation capacity and cell viability. AT-I attenuated gastric cancer stem cell (GCSC) traits partly through inactivating Notch1, leading to reducing the expressions of its downstream target Hes1, Hey1 and CD44 in vitro. Collectively, our results suggest that AT-I might develop as a potential therapeutic drug for the treatment of gastric cancer.

  2. Quetiapine Ameliorates Schizophrenia-Like Behaviors and Protects Myelin Integrity in Cuprizone Intoxicated Mice: The Involvement of Notch Signaling Pathway

    PubMed Central

    Wang, Hua-ning; Liu, Gao-hua; Zhang, Rui-guo; Xue, Fen; Wu, Di; Chen, Yun-chun; Peng, Ye

    2016-01-01

    Background: White matter disturbances and myelin impairment are key features of schizophrenia. The antipsychotic drug quetiapine can promote the maturation of oligodendrocytes, but the molecular mechanisms remain largely unknown. Methods: The schizophrenia-like behaviors, degrees of demyelination, and levels of Notch signaling molecules in forebrains of adult male C57BL/6 mice were examined after fed with cuprizone (0.2% wt/wt) in the presence or absence of 10mg/kg/d quetiapine for 6 weeks. These parameters were also observed after the transcranial injection of Notch signaling inhibitor MW167 (1mM) daily during the last week of the treatment period. Results: Quetiapine ameliorated the schizophrenia-like behaviors and decreased expression of myelin basic protein and inhibition of Notch signaling molecules, such as Notch1, Hes1, and Hes5, in the forebrain that induced by cuprizone. These beneficial effects of quetiapine were abolished by MW167. Conclusions: The antipsychotic and myelin protective effects of quetiapine are mediated by Notch signaling in a mouse model of cuprizone-induced demyelination associated with schizophrenia-like behaviors. The Notch pathway might therefore be a novel target for the development of antipsychotic drugs. PMID:26232790

  3. Complete mitochondrial genome of the medicinal fungus Ophiocordyceps sinensis

    PubMed Central

    Li, Yi; Hu, Xiao-Di; Yang, Rui-Heng; Hsiang, Tom; Wang, Ke; Liang, De-Quan; Liang, Fan; Cao, De-Ming; Zhou, Fan; Wen, Ge; Yao, Yi-Jian

    2015-01-01

    As part of a genome sequencing project for Ophiocordyceps sinensis, strain 1229, a complete mitochondrial (mt) genome was assembled as a single circular dsDNA of 157,510 bp, one of the largest reported for fungi. Conserved genes including the large and small rRNA subunits, 27 tRNA and 15 protein-coding genes, were identified. In addition, 58 non-conserved open reading frames (ncORFs) in the intergenic and intronic regions were also identified. Transcription analyses using RNA-Seq validated the expression of most conserved genes and ncORFs. Fifty-two introns (groups I and II) were found within conserved genes, accounting for 68.5% of the genome. Thirty-two homing endonucleases (HEs) with motif patterns LAGLIDADG (21) and GIY-YIG (11) were identified in group I introns. The ncORFs found in group II introns mostly encoded reverse transcriptases (RTs). As in other hypocrealean fungi, gene contents and order were found to be conserved in the mt genome of O. sinensis, but the genome size was enlarged by longer intergenic regions and numerous introns. Intergenic and intronic regions were composed of abundant repetitive sequences usually associated with mobile elements. It is likely that intronic ncORFs, which encode RTs and HEs, may have contributed to the enlarged mt genome of O. sinensis. PMID:26370521

  4. Pollination Mode and Mating System Explain Patterns in Genetic Differentiation in Neotropical Plants.

    PubMed

    Ballesteros-Mejia, Liliana; Lima, Natácia E; Lima-Ribeiro, Matheus S; Collevatti, Rosane G

    2016-01-01

    We studied genetic diversity and differentiation patterns in Neotropical plants to address effects of life history traits (LHT) and ecological attributes based on an exhaustive literature survey. We used generalized linear mixed models (GLMMs) to test the effects as fixed and random factors of growth form, pollination and dispersal modes, mating and breeding systems, geographical range and habitat on patterns of genetic diversity (HS, HeS, π and h), inbreeding coefficient (FIS), allelic richness (AR) and differentiation among populations (FST) for both nuclear and chloroplast genomes. In addition, we used phylogenetic generalized least squares (pGLS) to account for phylogenetic independence on predictor variables and verify the robustness of the results from significant GLMMs. In general, GLMM revealed more significant relationships among LHTs and genetic patterns than pGLS. After accounting for phylogenetic independence (i.e., using pGLS), FST for nuclear microsatellites was significantly related to pollination mode, mating system and habitat. Plants specifically with outcrossing mating system had lower FST. Moreover, AR was significantly related to pollination mode and geographical range and HeS for nuclear dominant markers was significantly related to habitat. Our findings showed that different results might be retrieved when phylogenetic non-independence is taken into account and that LHTs and ecological attributes affect substantially the genetic pattern in Neotropical plants, hence may drive key evolutionary processes in plants. PMID:27472384

  5. Corilagin suppresses cholangiocarcinoma progression through Notch signaling pathway in vitro and in vivo

    PubMed Central

    GU, YUE; XIAO, LINFENG; MING, YANLIN; ZHENG, ZHIZHONG; LI, WENGANG

    2016-01-01

    Corilagin is a natural plant polyphenol tannic acid with antitumor, anti-inflammatory, and anti-oxidative properties. However, the mechanisms of its actions are largely unknown. Our group reported that corilagin could induce cell inhibition in human breast cancer cell line MCF-7 and human liver hepatocellular carcinoma cell lines HepG2. We report here that corilagin inhibits cholangiocarcinoma (CCA) development through regulating Notch signaling pathway. We found that, in vitro, corilagin inhibited CCA cell proliferation, migration and invasion, promoted CCA cell apoptosis, and inhibited Notch1 and Notch signaling pathway protein expression. Co-immunoprecipitation was used to establish Notch intracellular domain (NICD) interaction with MAML1 and P300 in CCA. Importantly, corilagin reduced Hes1 mRNA level through inhibiting Hes1 promoter activity. In nude mice, corilagin inhibited CCA growth and repressed the expression of Notch1 and mTOR. These results indicate that corilagin may control CCA cell growth by downregulating the expression of Notch1. Therefore, our findings suggest that corilagin may have the potential to become a new therapeutic drug for human CCA. PMID:26935808

  6. Strong thermal nonequilibrium in hypersonic CO and CH4 probed by CRDS.

    PubMed

    Louviot, M; Suas-David, N; Boudon, V; Georges, R; Rey, M; Kassi, S

    2015-06-01

    A new experimental setup coupling a High Enthalpy Source (HES) reaching 2000 K to a cw-cavity ring-down spectrometer has been developed to investigate rotationally cold hot bands of polyatomic molecules in the [1.5, 1.7] μm region. The rotational and vibrational molecular degrees of freedom are strongly decoupled in the hypersonic expansion produced by the HES and probed by cavity ring-down spectroscopy. Carbon monoxide has been used as a first test molecule to validate the experimental approach. Its expansion in argon led to rotational and vibrational temperatures of 6.7 ± 0.8 K and 2006 ± 476 K, respectively. The tetradecad polyad of methane (1.67 μm) was investigated under similar conditions leading to rotational and vibrational temperatures of 13 ± 5 K and 750 ± 100 K, respectively. The rotationally cold structure of the spectra reveals many hot bands involving highly excited vibrational states of methane. PMID:26049494

  7. The transcriptome of utricle hair cell regeneration in the avian inner ear.

    PubMed

    Ku, Yuan-Chieh; Renaud, Nicole A; Veile, Rose A; Helms, Cynthia; Voelker, Courtney C J; Warchol, Mark E; Lovett, Michael

    2014-03-01

    Sensory hair cell loss is the major cause of hearing and balance disorders. Mammals are incapable of sustained hair cell regeneration, but lower vertebrates can regenerate these mechano-electrical transducers. We present the first comprehensive transcriptome (by mRNA-Seq) of hair cell regeneration in the chick utricle. We provide pathway and pattern annotations and correlate these with the phenotypic events that occur during regeneration. These patterns are surprisingly synchronous and highly punctuated. We show how these patterns are a new resource for identifying components of the hair cell transcriptome and identify 494 new putative hair-cell-specific genes and validate three of these (of three tested) by immunohistochemical staining. We describe many surprising new components and dynamic expression patterns, particularly within NOTCH signaling. For example, we show that HES7 is specifically expressed during utricle hair cell regeneration and closely parallels the expression of HES5. Likewise, the expression of ATOH1 is closely correlated with HEYL and the HLH inhibitory transcription factors ID1, ID2, and ID4. We investigate the correlation between fibroblast growth factor signaling and supporting cell proliferation and show that FGF20 inhibits supporting cell proliferation. We also present an analysis of 212 differentially expressed transcription factor genes in the regenerative time course that fall into nine distinct gene expression patterns, many of which correlate with phenotypic events during regeneration and represent attractive candidates for future analysis and manipulation of the regenerative program in sensory epithelia and other vertebrate neuroepithelia. PMID:24599453

  8. [Cryopreservation of vascular mixed cell for tissue engineering in cardiovascular surgery].

    PubMed

    Hibino, N; Shin'oka, T; Matsumura, G; Watanabe, M; Toyama, S; Imai, Y

    2001-06-01

    Tissue engineering (TE) is a new discipline that offers the potential to create replacement structures from autologous cells and biodegradable polymer scaffold. Various vascular and valvular grafts have been tried to create with this TE approach. In clinical use of this technique, harvested and cultured cells have to keep viability until implantation as tissue engineered tissue. But few research for cryopreservation of vascular mixed cells has been performed. So, we investigated the proper method for cryopreservation of vascular mixed cells harvested from femoral artery and vein of dogs. Cells were cultured and divide into three groups, A: cryopreserving in 5% dimethylsulfoxide (DMSO), hydroxyethyl starch (HES), and fetal bovine serum (FBS) with -80 degrees C freezer; B: cryopreserving in 10% DMSO and FBS with programmed freezer; C: control (continuous culture in media). After rapid thawing at 40 degrees C, group A showed higher viability than group B with flow cytometry. The results means that vascular mixed cells can be successfully cryopreserved in the DMSO/HES mixture simply and inexpensively, without rate controlled freezing. PMID:11424498

  9. Pulmonary embolism and mortality following total ankle replacement: a data linkage study using the NJR data set

    PubMed Central

    Zaidi, Razi; MacGregor, Alexander; Cro, Suzie; Goldberg, Andy

    2016-01-01

    Objective To determine the mortality rate following total ankle replacement (TAR) and incidence of 90 day pulmonary embolism (PE) along with the associated risk factors. Design Data-linkage study of the UK National Joint Registry (NJR) data and Hospital Episodes Statistics (HES) database. Linkage was performed in a deterministic fashion. HES episodes 90 days after the index procedure were analysed for PE. Mortality data were obtained pertaining to all the index procedures from the NJR for analysis. Participants All primary and revision ankle replacement patients captured on the NJR between February 2008 and February 2013. Results The 90-day mortality following TAR was 0.13% (95% CI 0.03 to 0.52) and 1-year mortality was 0.72% (95% CI 0.40 to 1.30); no deaths were as a result of PE. The incidence of PE within 90 days following primary TAR was 0.51% (95% CI 0.23 to 1.13). There was only one PE following revision surgery. Patients with an Royal College of Surgeons Charlson score greater than zero were at 13 times greater risk of PE (p=0.003). Conclusions There is low incidence of PE following TAR, but multiple comorbidities are a leading risk factor for its occurrence. PMID:27329444

  10. Effect of Mesenchymal Stem Cells and a Novel Curcumin Derivative on Notch1 Signaling in Hepatoma Cell Line

    PubMed Central

    Abdel Aziz, Mohamed Talaat; Khaled, Hussien Mostafa; El Hindawi, Ali; Roshdy, Nagwa Kamal; Rashed, Laila A.; Hassouna, Amira A.; Taha, Fatma; Ali, Walaa Ibrahim

    2013-01-01

    This study was conducted to evaluate the effect of mesenchymal stem cells (MSCs) and a novel curcumin derivative (NCD) on HepG2 cells (hepatoma cell line) and to investigate their effect on Notch1 signaling pathway target genes. HepG2 cells were divided into HepG2 control group, HepG2 cells treated with MSC conditioned medium (MSCs CM), HepG2 cells treated with a NCD, HepG2 cells treated with MSCs CM and NCD, and HepG2 cells treated with MSCs CM (CM of MSCs pretreated with a NCD). Expression of Notch1, Hes1, VEGF, and cyclin D1 was assessed by real-time, reverse transcription-polymerase chain reaction (RT-PCR) in HepG2 cells. In addition, HepG2 proliferation assay was performed in all groups. Notch1 and its target genes (Hes1 and cyclin D1) were downregulated in all treated groups with more suppressive effect in the groups treated with both MSCs and NCD. Also, treated HepG2 cells showed significant decrease in cell proliferation rate. These data suggest that modulation of Notch1 signaling pathway by MSCs and/or NCD can be considered as a therapeutic target in HCC. PMID:24024180

  11. Plasma substitutes therapy in pediatrics.

    PubMed

    Pietrini, Domenico; De Luca, Daniele; Tosi, Federica; Luca, Ersilia; Cavaliere, Franco; Conti, Giorgio; Piastra, Marco

    2012-06-01

    Hypovolemia is the most common cause of circulatory failure in children and may lead to critical tissue perfusion and eventually multiple-organ failure. Administration of fluids to maintain or restore intravascular volume represents a common intervention after hemorrhagic shock occurring during surgical procedures or in patients with trauma. Notwithstanding, there is uncertainty whether the type of fluid may significantly influence the outcome, especially in pediatrics. Both human albumin and crystalloids are usually administered: the advantages of crystalloids include low cost, lack of effect on coagulation, no risk of anaphylactic reaction or transmission of infectious agents. However, large amount of crystalloid infusion has been correlated with pulmonary oedema, bilateral pleural effusions, intestinal intussusception, excessive bowel edema, impairing closure of surgical wounds and peripheral edema. Moreover, intravascular volume expansion obtained by crystalloids is known to be significantly shorter and less efficacious than colloids. Among synthetic colloids, gelatins have been used for many years in children, also in early infancy, to treat intravascular fluid deficits. Hydroxyethylstarch (HES) preparations have been introduced recently, becoming very popular for vascular loading both in adults and children. However, the number of pediatric studies aimed at evaluating HES efficacy and tolerance is limited. Given the ongoing controversies on the use of colloids in childhood, this review will focus on the pharmacodynamics of synthetic and non synthetic colloids for the treatment of critical blood loss in pediatrics. PMID:22512388

  12. Impact of the National Institute for Health and Care Excellence (NICE) guidance on medical technology uptake: analysis of the uptake of spinal cord stimulation in England 2008–2012

    PubMed Central

    Vyawahare, Bharati; Hallas, Natalie; Brookes, Morag; Taylor, Rod S; Eldabe, Sam

    2014-01-01

    Background The National Institute for Health and Care Excellence (NICE) Technology Appraisal Guidance on spinal cord stimulation (SCS) was published in 2008 and updated in 2012 with no change. This guidance recommends SCS as a cost-effective treatment for patients with neuropathic pain. Objective To assess the impact of NICE guidance by comparing SCS uptake in England pre-NICE (2008–2009) and post-NICE (2009–2012) guidance. We also compared the English SCS uptake rate with that of Belgium, the Netherlands, France and Germany. Design SCS implant data for England was obtained from the Hospital Episode Statistics (HES) database and compared with other European countries where comparable data were available. Results The HES data showed small increases in SCS implantation and replacement/revision procedures, and a large increase in SCS trials between 2008 and 2012. The increase in the total number of SCS procedures per million of population in England is driven primarily by revision/replacements and increased trial activity. Marked variability in SCS uptake at both health regions and primary care trust level was observed. Conclusions Despite the positive NICE recommendation for the routine use of SCS, we found no evidence of a significant impact on SCS uptake in England. Rates of SCS implantation in England are lower than many other European countries. PMID:24398364

  13. Endocrine Cell Clustering During Human Pancreas Development

    PubMed Central

    Jeon, Jongmin; Correa-Medina, Mayrin; Ricordi, Camillo; Edlund, Helena; Diez, Juan A.

    2009-01-01

    The development of efficient, reproducible protocols for directed in vitro differentiation of human embryonic stem (hES) cells into insulin-producing β cells will benefit greatly from increased knowledge regarding the spatiotemporal expression profile of key instructive factors involved in human endocrine cell generation. Human fetal pancreases 7 to 21 weeks of gestational age, were collected following consent immediately after pregnancy termination and processed for immunostaining, in situ hybridization, and real-time RT-PCR expression analyses. Islet-like structures appear from approximately week 12 and, unlike the mixed architecture observed in adult islets, fetal islets are initially formed predominantly by aggregated insulin- or glucagon-expressing cells. The period studied (7–22 weeks) coincides with a decrease in the proliferation and an increase in the differentiation of the progenitor cells, the initiation of NGN3 expression, and the appearance of differentiated endocrine cells. The present study provides a detailed characterization of islet formation and expression profiles of key intrinsic and extrinsic factors during human pancreas development. This information is beneficial for the development of efficient protocols that will allow guided in vitro differentiation of hES cells into insulin-producing cells. (J Histochem Cytochem 57:811–824, 2009) PMID:19365093

  14. Sox17 regulates organ lineage segregation of ventral foregut progenitor cells

    PubMed Central

    Spence, Jason R.; Lange, Alex W.; Lin, Suh-Chin J.; Kaestner, Klaus H.; Lowy, Andrew M.; Kim, Injune; Whitsett, Jeffrey A.; Wells, James M.

    2009-01-01

    SUMMARY The ventral pancreas, biliary system and liver arise from the posterior ventral foregut, but the cell-intrinsic pathway by which these organ lineages are separated is not known. Here we show that the extrahepatobiliary system shares a common origin with the ventral pancreas and not the liver, as previously thought. These pancreatobiliary progenitor cells coexpress the transcription factors Pdx1 and Sox17 at e8.5 and their segregation into a Pdx1+ ventral pancreas and a Sox17+ biliary primordium is Sox17-dependant. Deletion of Sox17 at e8.5 results in the loss of biliary structures and ectopic pancreatic tissue in the liver bud and common duct, while Sox17 overexpression suppresses pancreas development and promotes ectopic biliary-like tissue throughout the Pdx1+ domain. Restricting Sox17+ biliary progenitor cells to the ventral region of the gut requires the notch effector Hes1. Our results highlight the role of Sox17 and Hes1 in patterning and morphogenetic segregation of ventral foregut lineages. PMID:19619492

  15. Notch Signaling and the Skeleton.

    PubMed

    Zanotti, Stefano; Canalis, Ernesto

    2016-06-01

    Notch 1 to 4 receptors are important determinants of cell fate and function, and Notch signaling plays an important role in skeletal development and bone remodeling. After direct interactions with ligands of the Jagged and Delta-like families, a series of cleavages release the Notch intracellular domain (NICD), which translocates to the nucleus where it induces transcription of Notch target genes. Classic gene targets of Notch are hairy and enhancer of split (Hes) and Hes-related with YRPW motif (Hey). In cells of the osteoblastic lineage, Notch activation inhibits cell differentiation and causes cancellous bone osteopenia because of impaired bone formation. In osteocytes, Notch1 has distinct effects that result in an inhibition of bone resorption secondary to an induction of osteoprotegerin and suppression of sclerostin with a consequent enhancement of Wnt signaling. Notch1 inhibits, whereas Notch2 enhances, osteoclastogenesis and bone resorption. Congenital disorders of loss- and gain-of-Notch function present with severe clinical manifestations, often affecting the skeleton. Enhanced Notch signaling is associated with osteosarcoma, and Notch can influence the invasive potential of carcinoma of the breast and prostate. Notch signaling can be controlled by the use of inhibitors of Notch activation, small peptides that interfere with the formation of a transcriptional complex, or antibodies to the extracellular domain of specific Notch receptors or to Notch ligands. In conclusion, Notch plays a critical role in skeletal development and homeostasis, and serious skeletal disorders can be attributed to alterations in Notch signaling. PMID:27074349

  16. Mechanism of flocculate formation of highly concentrated phospholipid vesicles suspended in a series of water-soluble biopolymers.

    PubMed

    Sakai, Hiromi; Sato, Atsushi; Takeoka, Shinji; Tsuchida, Eishun

    2009-08-10

    Polyethylene glycol-modified vesicles (liposomes) encapsulating hemoglobin (HbV) are artificial oxygen carriers that have been developed as a transfusion alternative. The HbV suspension in an albumin solution is nearly Newtonian; other biopolymers, hydroxyethyl starch (HES), dextran (DEX), and modified fluid gelatin, induce flocculation of HbVs through depletion interaction and render the suspensions as non-Newtonian. The flocculation level increased with hydrodynamic radius (R(h)) or radius of gyration (R(g)) of series of HES or DEX with different molecular weights at a constant polymer concentration (4 wt %). However, the flocculation level differed markedly among the polymers. A crowding index (C(i)) representing the crowding level of a polymer solution is defined as (excluded volume of one polymer) x (molar concentration) x Avogadro's number, using R(h) or R(g). All polymers' flocculation level increases when C(i) approaches 1: when the theoretical total excluded volumes approach the entire solution volume, the excluded HbV particles are forced to flocculate. PMID:19572642

  17. LIF-dependent primitive neural stem cells derived from mouse ES cells represent a reversible stage of neural commitment.

    PubMed

    Tsang, Wan-Hong; Wang, Bin; Wong, Wing Ki; Shi, Shuo; Chen, Xiao; He, Xiangjun; Gu, Shen; Hu, Jiabiao; Wang, Chengdong; Liu, Pi-Chu; Lu, Gang; Chen, Xiongfong; Zhao, Hui; Poon, Wai-Sang; Chan, Wai-Yee; Feng, Bo

    2013-11-01

    Primitive neural stem cells (NSCs) define an early stage of neural induction, thus provide a model to understand the mechanism that controls initial neural commitment. In this study, we investigated primitive NSCs derived from mouse embryonic stem cells (ESCs). By genome-wide transcriptional profiling, we revealed their unique signature and depicted the molecular changes underlying critical cell fate transitions during early neural induction at a global level. Together with qRT-PCR analysis, our data illustrated that primitive NSCs retained expression of key pluripotency genes Oct4 and Nanog, while exhibiting repression of other pluripotency-related genes Zscan4, Foxp1 and Dusp9 and up-regulation of neural markers Sox1 and Hes1. The early differentiation feature in primitive NSCs was also supported by their intermediate characters on cell cycle profiles. Moreover, re-plating primitive NSCs back to ESC culture condition could reverse them back to ESC stage, as shown by reversible regulation of marker genes, cell cycle profile changes and enhanced embryoid body formation. In addition, our microarray analysis also identified genes differentially expressed in primitive NSCs, and loss-of-function analysis demonstrated that Hes1 and Ccdc141 play important function at this stage, opening up an opportunity to further understand the regulation of early neural commitment. PMID:23973799

  18. Extracorporal-circulation-induced leukocyte/endothelial cell interaction is inhibited by dextran.

    PubMed

    Kamler, Markus; Pizanis, Nikolaus; Hagl, Siegfried; Gebhard, Martha Maria; Jakob, Heinz

    2004-01-01

    Clinical complications of Extracorporeal Circulation (ECC) are the consequence of a systemic activation of cellular and humoral factors resulting in a dysregulation of the microcirculation. Previously we developed an animal model to study effects of ECC on the microcirculation in vivo. To prevent the systemic inflammatory reaction (SIRS) seen after ECC, colloids were used as priming solution. Intravital fluorescence microscopy was used on the hamster skinfold chamber model. ECC was introduced and the ECC-tube system was flushed with Ringer solution (control), with Dextran 60 (group I) or HES 10% (group II). ECC for 30 minutes resulted in an increase in rolling and adherent leukocytes in postcapillary venules (Roller: 11 +/- 3% to 38% +/- 20% 4 h after ECC, p < 0.05, Sticker: 19 +/- 16 cells/mm2 to 215 +/- 145 cells/mm2 4 h after ECC, p < 0.05; n = 7). Use of Dextran prevented L/E cell interaction (10 +/- 5%; 63 +/- 40 cells/mm2 at 4 h), whereas HES affected only adherent white cells. L/E cell interaction in the microcirculation is an indicator of the systemic activation induced by ECC. Dextran 60 prevented L/E cell interaction without side effects and may be preferable for ECC in regard to inhibition of SIRS. PMID:15310949

  19. Predicting human developmental toxicity of pharmaceuticals using human embryonic stem cells and metabolomics

    SciTech Connect

    West, Paul R.; Weir, April M.; Smith, Alan M.; Donley, Elizabeth L.R.; Cezar, Gabriela G.

    2010-08-15

    Teratogens, substances that may cause fetal abnormalities during development, are responsible for a significant number of birth defects. Animal models used to predict teratogenicity often do not faithfully correlate to human response. Here, we seek to develop a more predictive developmental toxicity model based on an in vitro method that utilizes both human embryonic stem (hES) cells and metabolomics to discover biomarkers of developmental toxicity. We developed a method where hES cells were dosed with several drugs of known teratogenicity then LC-MS analysis was performed to measure changes in abundance levels of small molecules in response to drug dosing. Statistical analysis was employed to select for specific mass features that can provide a prediction of the developmental toxicity of a substance. These molecules can serve as biomarkers of developmental toxicity, leading to better prediction of teratogenicity. In particular, our work shows a correlation between teratogenicity and changes of greater than 10% in the ratio of arginine to asymmetric dimethylarginine levels. In addition, this study resulted in the establishment of a predictive model based on the most informative mass features. This model was subsequently tested for its predictive accuracy in two blinded studies using eight drugs of known teratogenicity, where it correctly predicted the teratogenicity for seven of the eight drugs. Thus, our initial data shows that this platform is a robust alternative to animal and other in vitro models for the prediction of the developmental toxicity of chemicals that may also provide invaluable information about the underlying biochemical pathways.

  20. In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease.

    PubMed

    Aubert, Martine; Boyle, Nicole M; Stone, Daniel; Stensland, Laurence; Huang, Meei-Li; Magaret, Amalia S; Galetto, Roman; Rawlings, David J; Scharenberg, Andrew M; Jerome, Keith R

    2014-01-01

    Following acute infection, herpes simplex virus (HSV) establishes latency in sensory neurons, from which it can reactivate and cause recurrent disease. Available antiviral therapies do not affect latent viral genomes; therefore, they do not prevent reactivation following therapy cessation. One possible curative approach involves the introduction of DNA double strand breaks in latent HSV genomes by rare-cutting endonucleases, leading to mutagenesis of essential viral genes. We tested this approach in an in vitro HSV latency model using the engineered homing endonuclease (HE) HSV1m5, which recognizes a sequence in the HSV-1 gene UL19, encoding the virion protein VP5. Coexpression of the 3'-exonuclease Trex2 with HEs increased HE-mediated mutagenesis frequencies up to sixfold. Following HSV1m5/Trex2 delivery with adeno-associated viral (AAV) vectors, the target site was mutated in latent HSV genomes with no detectable cell toxicity. Importantly, HSV production by latently infected cells after reactivation was decreased after HSV1m5/Trex2 exposure. Exposure to histone deacetylase inhibitors prior to HSV1m5/Trex2 treatment increased mutagenesis frequencies of latent HSV genomes another two- to fivefold, suggesting that chromatin modification may be a useful adjunct to gene-targeting approaches. These results support the continuing development of HEs and other nucleases (ZFNs, TALENs, CRISPRs) for cure of chronic viral infections.Molecular Therapy-Nucleic Acids (2014) 3, e1; doi:10.1038/mtna.2013.75; published online 4 February 2014. PMID:24496438

  1. Proteomic analysis of secretory products from the model gastrointestinal nematode Heligmosomoides polygyrus reveals dominance of Venom Allergen-Like (VAL) proteins

    PubMed Central

    Hewitson, James P.; Harcus, Yvonne; Murray, Janice; van Agtmaal, Maaike; Filbey, Kara J.; Grainger, John R.; Bridgett, Stephen; Blaxter, Mark L.; Ashton, Peter D.; Ashford, David; Curwen, Rachel S.; Wilson, R. Alan; Dowle, Adam A.; Maizels, Rick M.

    2016-01-01

    The intestinal helminth parasite, Heligmosomoides polygyrus offers a tractable experimental model for human hookworm infections such as Ancylostoma duodenale and veterinary parasites such as Haemonchus contortus. Parasite excretory-secretory (ES) products represent the major focus for immunological and biochemical analyses, and contain immunomodulatory molecules responsible for nematode immune evasion. In a proteomic analysis of adult H. polygyrus secretions (termed HES) matched to an extensive transcriptomic dataset, we identified 374 HES proteins by LC-MS/MS, which were distinct from those in somatic extract HEx, comprising 446 identified proteins, confirming selective export of ES proteins. The predominant secreted protein families were proteases (astacins and other metalloproteases, aspartic, cysteine and serine-type proteases), lysozymes, apyrases and acetylcholinesterases. The most abundant products were members of the highly divergent venom allergen-like (VAL) family, related to Ancylostoma secreted protein (ASP); 25 homologues were identified, with VAL-1 and -2 also shown to be associated with the parasite surface. The dominance of VAL proteins is similar to profiles reported for Ancylostoma and Haemonchus ES products. Overall, this study shows that the secretions of H. polygyrus closely parallel those of clinically important GI nematodes, confirming the value of this parasite as a model of helminth infection. PMID:21722761

  2. Human health risks from TNT, RDX, and HMX in environmental media and consideration of the US Regulatory Environment

    SciTech Connect

    Daniels, J.I.; Knezovich, J.P.

    1994-12-01

    Although the most economical method for disposing of unwanted energetic high explosives [HEs; e.g., 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-triazine (RDX, also known as Cyclonite), and octahydro-1,3,5,7-tetrazocine (HMX, also known as Octogen)] involves open burning and open or underground detonation [OB/O(U)D]; federal, state, and even local government agencies in the United States (U.S.) are implementing stricter environmental regulations that eventually may prevent such activities. These stricter regulations will promote alternative technologies that are designed to be environmentally benign. However, past HE-waste disposal practices at manufacturing and fabrication facilities in the U.S. have included uncontrolled OB/O(U)D, as well as direct surface discharge of HE-contaminated waste water, resulting in contaminated environmental media (e.g., ground water, soil, and perhaps even edible vegetation) near residential areas. Using TNT, RDX, and HMX as examples, this paper describes how risk-based standards for HEs can be derived that account for potential multimedia exposures (associated with contaminated air, water, food, and soil) by individuals near a contaminated site, and used to (1) protect public health and safety; (2)prevent limited resources from being dedicated to unnecessary cleanup activities; and (3) identify the most cost-effective, practical, and environmentally benign technologies suitable for integrating with the handling of the large quantity of high explosives scheduled for demilitarization.

  3. The aryl hydrocarbon receptor (AHR) transcription factor regulates megakaryocytic polyploidization

    PubMed Central

    Lindsey, Stephan; T. Papoutsakis, Eleftherios

    2012-01-01

    Summary We propose that the aryl hydrocarbon receptor (AHR) is a novel transcriptional regulator of megakaryopoietic polyploidization. Functional evidence was obtained that AHR impacts in vivo megakaryocytic differentiation and maturation; compared to wild-type mice, AHR-null mice had lower platelet counts, fewer numbers of newly synthesized platelets, increased bleeding times and lower-ploidy megakaryocytes (Mks). AHR mRNA increased 3·6-fold during ex vivo megakaryocytic differentiation, but reduced or remained constant during parallel isogenic granulocytic or erythroid differentiation. We interrogated the role of AHR in megakaryopoiesis using a validated Mk model of megakaryopoiesis, the human megakaryoblastic leukaemia CHRF cell line. Upon CHRF Mk differentiation, AHR mRNA and protein levels increased, AHR protein shifted from the cytoplasm to the nucleus and AHR binding to its consensus DNA binding sequence increased. Protein and mRNA levels of the AHR transcriptional target HES1 also increased. Mk differentiation of CHRF cells where AHR or HES1 was knocked-down using RNAi resulted in lower ploidy distributions and cells that were incapable of reaching ploidy classes ≥16n. AHR knockdown also resulted in increased DNA synthesis of lower ploidy cells, without impacting apoptosis. Together, these data support a role for AHR in Mk polyploidization and in vivo platelet function, and warrant further detailed investigations. PMID:21226706

  4. Dissecting neural differentiation regulatory networks through epigenetic footprinting

    PubMed Central

    Yaffe, Yakey; Donaghey, Julie; Pop, Ramona; Mallard, William; Issner, Robbyn; Gifford, Casey A.; Goren, Alon; Xing, Jeff; Gu, Hongcang; Cachiarelli, Davide; Tsankov, Alexander; Epstein, Chuck; Rinn, John R.; Mikkelsen, Tarjei S.; Kohlbacher, Oliver; Gnirke, Andreas; Bernstein, Bradley E.

    2014-01-01

    Human pluripotent stem cell derived models that accurately recapitulate neural development in vitro and allow for the generation of specific neuronal subtypes are of major interest to the stem cell and biomedical community. Notch signaling, particularly through the Notch effector HES5, is a major pathway critical for the onset and maintenance of neural progenitor cells (NPCs) in the embryonic and adult nervous system1-3. This can be exploited to isolate distinct populations of human embryonic stem (ES) cell derived NPCs4. Here, we report the transcriptional and epigenomic analysis of six consecutive stages derived from a HES5-GFP reporter ES cell line5 differentiated along the neural trajectory aimed at modeling key cell fate decisions including specification, expansion and patterning during the ontogeny of cortical neural stem and progenitor cells. In order to dissect the regulatory mechanisms that orchestrate the stage-specific differentiation process, we developed a computational framework to infer key regulators of each cell state transition based on the progressive remodeling of the epigenetic landscape and then validated these through a pooled shRNA screen. We were also able to refine our previous observations on epigenetic priming at transcription factor binding sites and show here that they are mediated by combinations of core and stage- specific factors. Taken together, we demonstrate the utility of our system and outline a general framework, not limited to the context of the neural lineage, to dissect regulatory circuits of differentiation. PMID:25533951

  5. Regulation of somatic cell reprogramming through inducible mir-302 expression.

    PubMed

    Lin, Shi-Lung; Chang, Donald C; Lin, Chun-Hung; Ying, Shao-Yao; Leu, Davey; Wu, David T S

    2011-02-01

    Global demethylation is required for early zygote development to establish stem cell pluripotency, yet our findings reiterate this epigenetic reprogramming event in somatic cells through ectopic introduction of mir-302 function. Here, we report that induced mir-302 expression beyond 1.3-fold of the concentration in human embryonic stem (hES) H1 and H9 cells led to reprogramming of human hair follicle cells (hHFCs) to induced pluripotent stem (iPS) cells. This reprogramming mechanism functioned through mir-302-targeted co-suppression of four epigenetic regulators, AOF2 (also known as KDM1 or LSD1), AOF1, MECP1-p66 and MECP2. Silencing AOF2 also caused DNMT1 deficiency and further enhanced global demethylation during somatic cell reprogramming (SCR) of hHFCs. Re-supplementing AOF2 in iPS cells disrupted such global demethylation and induced cell differentiation. Given that both hES and iPS cells highly express mir-302, our findings suggest a novel link between zygotic reprogramming and SCR, providing a regulatory mechanism responsible for global demethylation in both events. As the mechanism of conventional iPS cell induction methods remains largely unknown, understanding this microRNA (miRNA)-mediated SCR mechanism may shed light on the improvements of iPS cell generation. PMID:20870751

  6. Corilagin suppresses cholangiocarcinoma progression through Notch signaling pathway in vitro and in vivo.

    PubMed

    Gu, Yue; Xiao, Linfeng; Ming, Yanlin; Zheng, Zhizhong; Li, Wengang

    2016-05-01

    Corilagin is a natural plant polyphenol tannic acid with antitumor, anti-inflammatory, and anti-oxidative properties. However, the mechanisms of its actions are largely unknown. Our group reported that corilagin could induce cell inhibition in human breast cancer cell line MCF-7 and human liver hepatocellular carcinoma cell lines HepG2. We report here that corilagin inhibits cholangiocarcinoma (CCA) development through regulating Notch signaling pathway. We found that, in vitro, corilagin inhibited CCA cell proliferation, migration and invasion, promoted CCA cell apoptosis, and inhibited Notch1 and Notch signaling pathway protein expression. Co-immunoprecipitation was used to establish Notch intracellular domain (NICD) interaction with MAML1 and P300 in CCA. Importantly, corilagin reduced Hes1 mRNA level through inhibiting Hes1 promoter activity. In nude mice, corilagin inhibited CCA growth and repressed the expression of Notch1 and mTOR. These results indicate that corilagin may control CCA cell growth by downregulating the expression of Notch1. Therefore, our findings suggest that corilagin may have the potential to become a new therapeutic drug for human CCA. PMID:26935808

  7. Folic Acid Supplementation Stimulates Notch Signaling and Cell Proliferation in Embryonic Neural Stem Cells

    PubMed Central

    Liu, Huan; Huang, Guo-wei; Zhang, Xu-mei; Ren, Da-lin; X. Wilson, John

    2010-01-01

    The present study investigated the effect of folic acid supplementation on the Notch signaling pathway and cell proliferation in rat embryonic neural stem cells (NSCs). The NSCs were isolated from E14–16 rat brain and grown as neurospheres in serum-free suspension culture. Individual cultures were assigned to one of 3 treatment groups that differed according to the concentration of folic acid in the medium: Control (baseline folic acid concentration of 4 mg/l), low folic acid supplementation (4 mg/l above baseline, Folate-L) and high folic acid supplementation (40 mg/l above baseline, Folate-H). NSCs were identified by their expression of immunoreactive nestin and proliferating cells by incorporation of 5'bromo-2'deoxyuridine. Cell proliferation was also assessed by methyl thiazolyl tetrazolium assay. Notch signaling was analyzed by real-time PCR and western blot analyses of the expression of Notch1 and hairy and enhancer of split 5 (Hes5). Supplementation of NSCs with folic acid increased the mRNA and protein expression levels of Notch1 and Hes5. Folic acid supplementation also stimulated NSC proliferation dose-dependently. Embryonic NSCs respond to folic acid supplementation with increased Notch signaling and cell proliferation. This mechanism may mediate the effects of folic acid supplementation on neurogenesis in the embryonic nervous system. PMID:20838574

  8. Investigating Students' Ideas About Buoyancy and the Influence of Haptic Feedback

    NASA Astrophysics Data System (ADS)

    Minogue, James; Borland, David

    2016-04-01

    While haptics (simulated touch) represents a potential breakthrough technology for science teaching and learning, there is relatively little research into its differential impact in the context of teaching and learning. This paper describes the testing of a haptically enhanced simulation (HES) for learning about buoyancy. Despite a lifetime of everyday experiences, a scientifically sound explanation of buoyancy remains difficult to construct for many. It requires the integration of domain-specific knowledge regarding density, fluid, force, gravity, mass, weight, and buoyancy. Prior studies suggest that novices often focus on only one dimension of the sinking and floating phenomenon. Our HES was designed to promote the integration of the subconcepts of density and buoyant forces and stresses the relationship between the object itself and the surrounding fluid. The study employed a randomized pretest-posttest control group research design and a suite of measures including an open-ended prompt and objective content questions to provide insights into the influence of haptic feedback on undergraduate students' thinking about buoyancy. A convenience sample (n = 40) was drawn from a university's population of undergraduate elementary education majors. Two groups were formed from haptic feedback (n = 22) and no haptic feedback (n = 18). Through content analysis, discernible differences were seen in the posttest explanations sinking and floating across treatment groups. Learners that experienced the haptic feedback made more frequent use of "haptically grounded" terms (e.g., mass, gravity, buoyant force, pushing), leading us to begin to build a local theory of language-mediated haptic cognition.

  9. Notch Signaling and Atoh1 Expression During Hair Cell Regeneration in the Mouse Utricle

    PubMed Central

    Wang, Guo-Peng; Chatterjee, Ishani; Batts, Shelley A.; Wong, Hiu Tung; Gong, Tzy-Wen; Gong, Shu-Sheng; Raphael, Yehoash

    2010-01-01

    The mammalian vestibular epithelium has a limited capacity for spontaneous hair cell regeneration. The mechanism underlying the regeneration is not well understood. Because the Notch signaling pathway mediates the formation of the sensory epithelial mosaic patterning during ear development, it may also play a role in hair cell regeneration in the mature mammalian vestibular epithelium after a lesion. To investigate the process of spontaneous regeneration in the vestibular epithelium vis-à-vis changes in Notch signaling, we induced a unilateral lesion by infusing streptomycin into the mouse posterior semicircular canal, and examined Notch signaling molecules and their mRNA expression levels by immunohistochemistry and quantitative real-time polymerase chain reaction (qRTPCR), respectively. We detected Jagged1 in supporting cells in both normal and lesioned utricles. Atoh1, a marker for early developing hair cells, was absent in the intact mature tissue, but re-appeared after the lesion. Many cells were either positive for both Atoh1 and myosin VIIa, or for one of them. qRTPCR data showed a post trauma decrease of Hes5 and an increase in Atoh1. Atoh1 up-regulation may either be a result of Hes5 down-regulation or mediated by another signaling pathway. PMID:20433915

  10. Inhibition of Notch Signaling Ameliorates Acute Kidney Failure and Downregulates Platelet-Derived Growth Factor Receptor β in the Mouse Model.

    PubMed

    Kramer, Jan; Schwanbeck, Ralf; Pagel, Horst; Cakiroglu, Figen; Rohwedel, Jürgen; Just, Ursula

    2016-01-01

    Ischemic acute kidney injury (AKI) is associated with high morbidity and frequent complications. Repeated episodes of AKI may lead to end-stage renal failure. The pathobiology of regeneration in AKI is not well understood and there is no effective clinical therapy that improves regeneration. The Notch signaling pathway plays an essential role in kidney development and has been implicated in tissue repair in the adult kidney. Here, we found that kidneys after experimental AKI in mice showed increased expression of Notch receptors, specifically Notch1-3, of the Notch ligands Jagged-1 (Jag1), Jag2 and Delta-like-4 (Dll4) and of the Notch target genes Hes1, Hey2, HeyL, Sox9 and platelet-derived growth factor receptor β (Pdgfrb). Treatment of ischemic mice with the x03B3;-secretase inhibitor DBZ blocked Notch signaling and specifically downregulated the expression of Notch3 and the Notch target genes Hes1, Hey2, HeyL and Pdgfrb. After DBZ treatment, the mice developed less interstitial edema and displayed altered interstitial inflammation patterns. Furthermore, serum urea and creatinine levels were significantly decreased from 6 h onwards when compared to control mice treated with DMSO only. Our data are consistent with an amelioration of the severity of kidney injury by blocking Notch activation following AKI, and suggest an involvement of Notch-regulated Pdgfrb in AKI pathogenesis. PMID:26939110

  11. Chronic rapamycin treatment on the nutrient utilization and metabolism of juvenile turbot (Psetta maxima).

    PubMed

    Wang, Qingchao; He, Gen; Mai, Kangsen; Xu, Wei; Zhou, Huihui; Wang, Xuan; Mei, Lin

    2016-01-01

    High dietary protein inclusion is necessary in fish feeds and also represents a major cost in the aquaculture industry, which demands improved dietary conversion into body proteins in fish. In mammals, the target of rapamycin (TOR) is a key nutritionally responsive molecule governing postprandial anabolism. However, its physiological significance in teleosts has not been fully examined. In the present study, we examined the nutritional physiology of turbot after chronic rapamycin inhibition. Our results showed that a 6-week inhibition of TOR using dietary rapamycin inclusion (30 mg/kg diet) reduced growth performance and feed utilization. The rapamycin treatment inhibited TOR signaling and reduced expression of key enzymes in glycolysis, lipogenesis, cholesterol biosynthesis, while increasing the expression of enzymes involved in gluconeogenesis. Furthermore, rapamycin treatment increased intestinal goblet cell number in turbot, while the expressions of Notch and Hes1 were down regulated. It was possible that stimulated goblet cell differentiation by rapamycin was mediated through Notch-Hes1 pathway. Therefore, our results demonstrate the important role of TOR signaling in fish nutritional physiology. PMID:27305975

  12. Progress and prospects of engineered sequence-specific DNA modulating technologies for the management of liver diseases

    PubMed Central

    Nicholson, Samantha A; Moyo, Buhle; Arbuthnot, Patrick B

    2015-01-01

    Liver diseases are one of the leading causes of mortality in the world. The hepatic illnesses, which include inherited metabolic disorders, hemophilias and viral hepatitides, are complex and currently difficult to treat. The maturation of gene therapy has heralded new avenues for developing effective intervention for these diseases. DNA modification using gene therapy is now possible and available technology may be exploited to achieve long term therapeutic benefit. The ability to edit DNA sequences specifically is of paramount importance to advance gene therapy for application to liver diseases. Recent development of technologies that allow for this has resulted in rapid advancement of gene therapy to treat several chronic illnesses. Improvements in application of derivatives of zinc finger proteins (ZFPs), transcription activator-like effectors (TALEs), homing endonucleases (HEs) and clustered regularly interspaced palindromic repeats (CRISPR) and CRISPR associated (Cas) systems have been particularly important. These sequence-specific technologies may be used to modify genes permanently and also to alter gene transcription for therapeutic purposes. This review describes progress in development of ZFPs, TALEs, HEs and CRISPR/Cas for application to treating liver diseases. PMID:25937863

  13. Human papillomavirus 16E6 and NFX1-123 potentiate notch signaling and differentiation without activating cellular arrest

    SciTech Connect

    Vliet-Gregg, Portia A.; Hamilton, Jennifer R.; Katzenellenbogen, Rachel A.

    2015-04-15

    High-risk human papillomavirus (HR HPV) oncoproteins bind host cell proteins to dysregulate and uncouple apoptosis, senescence, differentiation, and growth. These pathways are important for both the viral life cycle and cancer development. HR HPV16 E6 (16E6) interacts with the cellular protein NFX1-123, and they collaboratively increase the growth and differentiation master regulator, Notch1. In 16E6 expressing keratinocytes (16E6 HFKs), the Notch canonical pathway genes Hes1 and Hes5 were increased with overexpression of NFX1-123, and their expression was directly linked to the activation or blockade of the Notch1 receptor. Keratinocyte differentiation genes Keratin 1 and Keratin 10 were also increased, but in contrast their upregulation was only indirectly associated with Notch1 receptor stimulation and was fully unlinked to growth arrest, increased p21{sup Waf1/CIP1}, or decreased proliferative factor Ki67. This leads to a model of 16E6, NFX1-123, and Notch1 differently regulating canonical and differentiation pathways and entirely uncoupling cellular arrest from increased differentiation. - Highlights: • 16E6 and NFX1-123 increased the Notch canonical pathway through Notch1. • 16E6 and NFX1-123 increased the differentiation pathway indirectly through Notch1. • 16E6 and NFX1-123 increased differentiation gene expression without growth arrest. • Increased NFX1-123 with 16E6 may create an ideal cellular phenotype for HPV.

  14. The Coumarin Derivative Osthole Stimulates Adult Neural Stem Cells, Promotes Neurogenesis in the Hippocampus, and Ameliorates Cognitive Impairment in APP/PS1 Transgenic Mice.

    PubMed

    Kong, Liang; Hu, Yu; Yao, Yingjia; Jiao, Yanan; Li, Shaoheng; Yang, Jingxian

    2015-01-01

    It is believed that neuronal death caused by abnormal deposition of amyloid-beta peptide is the major cause of the cognitive decline in Alzheimer's disease. Adult neurogenesis plays a key role in the rescue of impaired neurons and amelioration of cognitive impairment. In the present study, we demonstrated that osthole, a natural coumarin derivative, was capable of promoting neuronal stem cell (NSC) survival and inducing NSC proliferation in vitro. In osthole-treated APP/PS1 transgenic mice, a significant improvement in learning and memory function was seen, which was associated with a significant increase in the number of new neurons (Ki67(+)/NF-M(+)) and a decrease in apoptotic cells in the hippocampal region of the brain. These observations suggested that osthole promoted NSC proliferation, supported neurogenesis, and thus efficiently rescued impaired neurons in the hippocampus and ameliorated cognitive impairment. We also found that osthole treatment activated the Notch pathway and upregulated the expression of self-renewal genes Notch 1 and Hes 1 mRNA in NSCs. However, when Notch activity was blocked by the γ-secretase inhibitor DAPT, the augmentation of Notch 1 and Hes 1 protein was ameliorated, and the proliferation-inducing effect of osthole was abolished, suggesting that the effects of osthole are at least in part mediated by activation of the Notch pathway. PMID:26328484

  15. Minimal change glomerulopathy in a cat.

    PubMed

    Backlund, Brianna; Cianciolo, Rachel E; Cook, Audrey K; Clubb, Fred J; Lees, George E

    2011-04-01

    A 6-year-old domestic shorthair male castrated cat was evaluated for sudden onset of vomiting and anorexia. A diagnosis of hypereosinophilic syndrome (HES) was made, and the cat was treated with imatinib mesylate. The cat had an initial clinical improvement with the normalization of the peripheral eosinophil count. After approximately 8 weeks of treatment, lethargy and anorexia recurred despite the normal eosinophil count and a significant proteinuric nephropathy was identified. Treatment with imatinib was discontinued. Ultrasound guided renal biopsies exhibited histologic, ultrastructural, and immunostaining changes indicative of a minimal change glomerulopathy (MCG) which has not previously been reported in the literature in a cat. The proteinuria and HES initially improved while the cat was treated with more traditional medications; however, both the problems persisted for 30 months that the cat was followed subsequently. Previous studies demonstrating the safety and efficacy of imatinib in cats do not report any glomerular injury or significant adverse drug reactions, and the exact cause of this cat's proteinuric nephropathy is uncertain. Nonetheless, the possibility of an adverse drug reaction causing proteinuria should be considered when initiating treatment with imatinib in a cat. PMID:21414552

  16. Pollination Mode and Mating System Explain Patterns in Genetic Differentiation in Neotropical Plants

    PubMed Central

    Ballesteros-Mejia, Liliana; Lima, Natácia E.; Lima-Ribeiro, Matheus S.

    2016-01-01

    We studied genetic diversity and differentiation patterns in Neotropical plants to address effects of life history traits (LHT) and ecological attributes based on an exhaustive literature survey. We used generalized linear mixed models (GLMMs) to test the effects as fixed and random factors of growth form, pollination and dispersal modes, mating and breeding systems, geographical range and habitat on patterns of genetic diversity (HS, HeS, π and h), inbreeding coefficient (FIS), allelic richness (AR) and differentiation among populations (FST) for both nuclear and chloroplast genomes. In addition, we used phylogenetic generalized least squares (pGLS) to account for phylogenetic independence on predictor variables and verify the robustness of the results from significant GLMMs. In general, GLMM revealed more significant relationships among LHTs and genetic patterns than pGLS. After accounting for phylogenetic independence (i.e., using pGLS), FST for nuclear microsatellites was significantly related to pollination mode, mating system and habitat. Plants specifically with outcrossing mating system had lower FST. Moreover, AR was significantly related to pollination mode and geographical range and HeS for nuclear dominant markers was significantly related to habitat. Our findings showed that different results might be retrieved when phylogenetic non-independence is taken into account and that LHTs and ecological attributes affect substantially the genetic pattern in Neotropical plants, hence may drive key evolutionary processes in plants. PMID:27472384

  17. Disease modeling and cell based therapy with iPSC: future therapeutic option with fast and safe application

    PubMed Central

    2014-01-01

    Induced pluripotent stem cell (iPSC) technology has shown us great hope to treat various human diseases which have been known as untreatable and further endows personalized medicine for future therapy without ethical issues and immunological rejection which embryonic stem cell (hES) treatment has faced. It has been agreed that iPSCs knowledge can be harnessed from disease modeling which mimics human pathological development rather than trials utilizing conventional rodent and cell lines. Now, we can routinely generate iPSC from patient specific cell sources, such as skin fibroblast, hair follicle cells, patient blood samples and even urine containing small amount of epithelial cells. iPSC has both similarity and dissimilarity to hES. iPSC is similar enough to regenerate tissue and even full organism as ES does, however what we want for therapeutic advantage is limited to regenerated tissue and lineage specific differentiation. Depending on the lineage and type of cells, both tissue memory containing (DNA rearrangement/epigenetics) and non-containing iPSC can be generated. This makes iPSC even better choice to perform disease modeling as well as cell based therapy. Tissue memory containing iPSC from mature leukocytes would be beneficial for curing cancer and infectious disease. In this review, the benefit of iPSC for translational approaches will be presented. PMID:24724061

  18. RBP-J-interacting and tubulin-associated protein induces apoptosis and cell cycle arrest in human hepatocellular carcinoma by activating the p53–Fbxw7 pathway

    SciTech Connect

    Wang, Haihe; Yang, Zhanchun; Liu, Chunbo; Huang, Shishun; Wang, Hongzhi; Chen, Yingli; Chen, Guofu

    2014-11-07

    Highlights: • RITA overexpression increased protein expression of p53 and Fbxw7 and downregulated the expression of cyclin D1, cyclin E, CDK2, Hes-1 and NF-κB p65. • RITA can significantly inhibit the in vitro growth of SMMC7721 and HepG2 cells. • RITA exerts tumor-suppressive effects in hepatocarcinogenesis through induction of G0/G1 cell cycle arrest and apoptosis and suggest a therapeutic application of RITA in HCC. - Abstract: Aberrant Notch signaling is observed in human hepatocellular carcinoma (HCC) and has been associated with the modulation of cell growth. However, the role of Notch signaling in HCC and its underlying mechanism remain elusive. RBP-J-interacting and tubulin-associated (RITA) mediates the nuclear export of RBP-J to tubulin fibers and downregulates Notch-mediated transcription. In this study, we found that RITA overexpression increased protein expression of p53 and Fbxw7 and downregulated the expression of cyclin D1, cyclin E, CDK2, Hes-1 and NF-κB p65. These changes led to growth inhibition and induced G0/G1 cell cycle arrest and apoptosis in SMMC7721 and HepG2 cells. Our findings indicate that RITA exerts tumor-suppressive effects in hepatocarcinogenesis through induction of G0/G1 cell cycle arrest and apoptosis and suggest a therapeutic application of RITA in HCC.

  19. Distribution of genetic diversity among disjunct populations of the rare forest understory herb, Trillium reliquum.

    PubMed

    Gonzales, E; Hamrick, J L

    2005-10-01

    We assessed genetic diversity and its distribution in the rare southeastern US forest understory species, Trillium reliquum. In all, 21 loci were polymorphic (PS=95.5%) and the mean number of alleles per polymorphic locus was 3.05. However, genetic diversity was relatively low (Hes=0.120) considering the level of polymorphism observed for this outcrossing species. A relatively high portion of the genetic diversity (29.7%) was distributed among populations. There was no relationship between population size and genetic diversity, and we did not detect significant inbreeding. These results are best explained by the apparent self-incompatibility of this species, its longevity and clonal reproduction. To address questions regarding the history of T. reliquum's rarity, we compared results for T. reliquum with that of its more common and partially sympatric congener, T. cuneatum. Despite shared life history traits and history of land use, we observed significant genetic differences between the two species. Although T. cuneatum contains slightly lower polymorphism (Ps=85%), we detected significantly higher genetic diversity (Hes=0.217); most of its genetic diversity is contained within its populations (GST=0.092). Our results suggest that not only is there little gene flow among extant T. reliquum populations, but that rarity and population isolation in this species is of ancient origins, rather than due to more recent anthropogenic fragmentation following European colonization. The Chattahoochee River was identified as a major barrier to gene exchange. PMID:16094302

  20. Methotrexate supports in vivo selection of human embryonic stem cell derived-hematopoietic cells expressing dihydrofolate reductase

    PubMed Central

    Gori, Jennifer L; McIvor, R Scott

    2010-01-01

    Human embryonic stem cells (hES Cs) are an attractive alternative cell source for hematopoietic gene therapy applications as the cells are easily modified with lentiviral or other vectors and can be subsequently induced to differentiate into hematopoietic progenitor cells. However, demonstration of the full hematopoietic potential of hESC-derived progeny is challenging due to low marrow engraftment and the difficulty of detecting cells in the peripheral blood of human/mouse xenografts. Methotrexate (MTX) chemotherapy coupled with expression of a drug resistant dihydrofolate reductase such as Tyr22 (Tyr22DHFR) has the potential to selectively increase engraftment of gene-modified human hematopoietic cells in mice, which would allow for better phenotypic characterization of hESC-derived cells in vivo. We showed that hES Cs transduced with Tyr22DHFR-GFP encoding lentivirus vectors differentiate into MTX resistant (MTXr) hemato-endothelial cells. MTX treatment of immunodeficient mice infused with Tyr22DHFR hESC-derived hemato-endothelial cells increased the long-term engraftment of human cells in the bone marrow of MTX-treated mice. In contrast to previous studies, these results indicate that MTX administration has the potential to support in vivo selection that is maintained after cessation of treatment. The MTX/Tyr22DHFR system may therefore be useful for enrichment of gene-modified cell populations in human stem cell and gene therapy applications. PMID:21468213

  1. Regulation of dendritic-cell differentiation by bone marrow stroma via different Notch ligands

    PubMed Central

    Cheng, Pingyan; Nefedova, Yulia; Corzo, Cesar A.; Gabrilovich, Dmitry I.

    2007-01-01

    Notch is a major factor mediating interaction between hematopoietic progenitor cells (HPCs) and bone marrow stroma (BMS). However its contribution to dendritic cell (DC) differentiation is controversial. We found that main Notch ligands Delta-1 and Jagged-1 had the opposite effect on DC differentiation. Delta-1 promoted generation of fully differentiated DCs, whereas Jagged-1 stimulated accumulation of DC precursors but prevented their transition to terminally differentiated DCs. BMS expressed a substantially higher level of Jagged-1 than Delta-1. Just the opposite expression pattern was observed in spleen stroma (SS). The BMS effect on DC differentiation was similar to that of Jagged-1, whereas the effect of SS was similar to the effect of Delta-1. Down-regulation of Jagged-1 in BMS substantially increased DC differentiation. Experiments in vivo with adoptive transfer of DC precursors further supported the different roles of BMS and SS in DC development. Jagged-1 and Delta-1 equally activated CBF-1/RBPJκ transcription factor, which is a major Notch target. However, they produced a different pattern of activation of Notch target gene Hes1. Overexpression of Hes1 resulted in increased DC differentiation from HPCs. Thus, this study not only revealed the different role of Notch ligands in DC differentiation but also may provide a new insight into regulation of DC differentiation by BMS. PMID:16973960

  2. BAC transgenesis in human embryonic stem cells as a novel tool to define the human neural lineage.

    PubMed

    Placantonakis, Dimitris G; Tomishima, Mark J; Lafaille, Fabien; Desbordes, Sabrina C; Jia, Fan; Socci, Nicholas D; Viale, Agnes; Lee, Hyojin; Harrison, Neil; Tabar, Viviane; Studer, Lorenz

    2009-03-01

    Human embryonic stem cells (hESCs) have enormous potential for applications in basic biology and regenerative medicine. However, harnessing the potential of hESCs toward generating homogeneous populations of specialized cells remains challenging. Here we describe a novel technology for the genetic identification of defined hESC-derived neural cell types using bacterial artificial chromosome (BAC) transgenesis. We generated hESC lines stably expressing Hes5::GFP, Dll1::GFP, and HB9::GFP BACs that yield green fluorescent protein (GFP)(+) neural stem cells, neuroblasts, and motor neurons, respectively. Faithful reporter expression was confirmed by cell fate analysis and appropriate transgene regulation. Prospective isolation of HB9::GFP(+) cells yielded purified human motor neurons with proper marker expression and electrophysiological activity. Global mRNA and microRNA analyses of Hes5::GFP(+) and HB9::GFP(+) populations revealed highly specific expression signatures, suggesting that BAC transgenesis will be a powerful tool for establishing expression libraries that define the human neural lineage and for accessing defined cell types in applications of human disease. PMID:19074416

  3. Notch signaling in skeletal health and disease.

    PubMed

    Zanotti, Stefano; Canalis, Ernesto

    2013-06-01

    Notch receptors are single-pass transmembrane proteins that determine cell fate. Upon Notch ligand interactions, proteolytic cleavages release the Notch intracellular domain, which translocates to the nucleus to regulate the transcription of target genes, including Hairy enhancer of split (Hes) and Hes related to YRPW motif (Hey). Notch is critical for skeletal development and activity of skeletal cells, and dysregulation of Notch signaling is associated with human diseases affecting the skeleton. Inherited or sporadic mutations in components of the Notch signaling pathway are associated with spondylocostal dysostosis, spondylothoracic dysostosis and recessive brachydactyly, diseases characterized by skeletal patterning defects. Inactivating mutations of the Notch ligand JAG1 or of NOTCH2 are associated with Alagille syndrome, and activating mutations in NOTCH2 are associated with Hajdu-Cheney syndrome (HCS). Individuals affected by HCS exhibit osteolysis in distal phalanges and osteoporosis. NOTCH is activated in selected tumors, such as osteosarcoma, and in breast cancer cells that form osteolytic bone metastases. In conclusion, Notch regulates skeletal development and bone remodeling, and gain- or loss-of-function mutations of Notch signaling result in important skeletal diseases. PMID:23554451

  4. Allozyme diversity in the endangered pitcher plant Sarracenia rubra ssp. Alabamensis (Sarraceniaceae) and its close relative S. rubra ssp. rubra.

    PubMed

    Godt, M; Hamrick, J

    1998-06-01

    Genetic variability in the federally endangered pitcher plant Sarracenia rubra ssp. alabamensis was assessed in eight Alabama populations using starch gel electrophoresis. Ten populations of the more widespread Sarracenia rubra ssp. rubra were sampled in the southeastern United States for comparison. Fifteen allozyme loci representing 13 enzyme systems were scored for each species. In contrast to S. oreophila and S. jonesii, two previously analyzed endangered pitcher plants, genetic diversity was high for both S. rubra subspecies. Within ssp. alabamensis the percentage polymorphic loci (Ps) was 80.0, the mean number of alleles per polymorphic locus was (APs) = 2.58, and expected heterozygosity (Hes) was 0.209. Genetic diversity was slightly lower for ssp. rubra (Ps = 73.3, APs = 2.91, and Hes = 0.177). The proportion of total genetic diversity found among populations was fairly low for both species (GST = 0.09 for ssp. alabamensis and 0.14 for ssp. rubra). Little genetic divergence has occurred between the two subspecies as indicated by the lack of diagnostic alleles, the proportion of total genetic diversity between taxa (GST = 0.09), and the genetic identity estimate (I = 0.90). The relatively high genetic diversity found for ssp. alabamensis indicates that the maintenance of its evolutionary potential is possible if population sizes are maintained or increased. Low levels of genetic diversity found within small Georgia ssp. rubra populations indicate that genetic erosion may increase extinction risks for these populations. PMID:21684964

  5. Gene and MicroRNA Transcriptional Signatures of Angiotensin II in Endothelial Cells

    PubMed Central

    Mehta, Jawahar L.; Mercanti, Federico; Stone, Annjannette; Wang, Xianwei; Ding, Zufeng; Romeo, Francesco

    2015-01-01

    Abstract: Growth of atherosclerotic plaque requires neovascularization (angiogenesis). To elucidate the involvement of angiotensin II (Ang II) in angiogenesis, we performed gene microarray and microRNA (miRNA) polymerase chain reaction array analyses on human coronary artery endothelial cells exposed to moderate concentration of Ang II for 2 and 12 hours. At 12, but not 2, hours, cultures treated with Ang II exhibited shifts in transcriptional activity involving 267 genes (>1.5-fold difference; P < 0.05). Resulting transcriptome was most significantly enriched for genes associated with blood vessel development, angiogenesis, and regulation of proliferation. Majority of upregulated genes implicated in angiogenesis shared a commonality of being either regulators (HES1, IL-18, and CXCR4) or targets (ADM, ANPEP, HES1, KIT, NOTCH4, PGF, and SOX18) of STAT3. In line with these findings, STAT3 inhibition attenuated Ang II–dependent stimulation of tube formation in Matrigel assay. Expression analysis of miRNAs transcripts revealed that the pattern of differential expression for miRNAs was largely consistent with proangiogenic response with a prominent theme of upregulation of miRs targeting PTEN (miR-19b-3p, miR-21-5p, 23b-3p, and 24-3p), many of which are directly or indirectly STAT3 dependent. We conclude that STAT3 signaling may be an intrinsic part of Ang II–mediated proangiogenic response in human endothelial cells. PMID:24853489

  6. Epidermal growth factor promotes proliferation of dermal papilla cells via Notch signaling pathway.

    PubMed

    Zhang, Haihua; Nan, Weixiao; Wang, Shiyong; Zhang, Tietao; Si, Huazhe; Wang, Datao; Yang, Fuhe; Li, Guangyu

    2016-08-01

    The effect of epidermal growth factor (EGF) on the development and growth of hair follicle is controversial. In the present study, 2-20 ng/ml EGF promoted the growth of mink hair follicles in vitro, whereas 200 ng/ml EGF inhibited follicle growth. Further, dermal papilla (DP) cells, a group of mesenchymal cells that govern hair follicle development and growth, were isolated and cultured in vitro. Treatment with or forced expression of EGF accelerated proliferation and induced G1/S transition in DP cells. Moreover, EGF upregulated the expression of DP mesenchymal genes, such as alkaline phosphatase (ALP) and insulin-like growth factor (IGF-1), as well as the Notch pathway molecules including Notch1, Jagged1, Hes1 and Hes5. In addition, inhibition of Notch signaling pathway by DAPT significantly reduced the basal and EGF-enhanced proliferation rate, and also suppressed cell cycle progression. We also show that the expression of several follicle-regulatory genes, such as Survivin and Msx2, were upregulated by EGF, and was inhibited by DAPT. In summary, our study demonstrates that the concentration of EGF is critical for the switch between hair follicle growth and inhibition, and EGF promotes DP cell proliferation via Notch signaling pathway. PMID:27109378

  7. Numerical simulation of long-duration blast wave evolution in confined facilities

    NASA Astrophysics Data System (ADS)

    Togashi, F.; Baum, J. D.; Mestreau, E.; Löhner, R.; Sunshine, D.

    2010-10-01

    The objective of this research effort was to investigate the quasi-steady flow field produced by explosives in confined facilities. In this effort we modeled tests in which a high explosive (HE) cylindrical charge was hung in the center of a room and detonated. The HEs used for the tests were C-4 and AFX 757. While C-4 is just slightly under-oxidized and is typically modeled as an ideal explosive, AFX 757 includes a significant percentage of aluminum particles, so long-time afterburning and energy release must be considered. The Lawrence Livermore National Laboratory (LLNL)-produced thermo-chemical equilibrium algorithm, “Cheetah”, was used to estimate the remaining burnable detonation products. From these remaining species, the afterburning energy was computed and added to the flow field. Computations of the detonation and afterburn of two HEs in the confined multi-room facility were performed. The results demonstrate excellent agreement with available experimental data in terms of blast wave time of arrival, peak shock amplitude, reverberation, and total impulse (and hence, total energy release, via either the detonation or afterburn processes.

  8. Notch and the Skeleton▿

    PubMed Central

    Zanotti, Stefano; Canalis, Ernesto

    2010-01-01

    Notch receptors are transmembrane receptors that regulate cell fate decisions. There are four Notch receptors in mammals. Upon binding to members of the Delta and Jagged family of transmembrane proteins, Notch is cleaved and the Notch intracellular domain (NICD) is released. NICD then translocates to the nucleus, where it associates with the CBF-1, Suppressor of Hairless, and Lag-2 (CSL) and Mastermind-Like (MAML) proteins. This complex activates the transcription of Notch target genes, such as Hairy Enhancer of Split (Hes) and Hes-related with YRPF motif (Hey). Notch signaling is critical for the regulation of mesenchymal stem cell differentiation. Misexpression of Notch in skeletal tissue indicates a role as an inhibitor of skeletal development and postnatal bone formation. Overexpression of Notch inhibits endochondral bone formation and osteoblastic differentiation, causing severe osteopenia. Conditional inactivation of Notch in the skeleton causes an increase in cancellous bone volume and enhanced osteoblastic differentiation. Notch ligands are expressed in the hematopoietic stem cell niche and are critical for the regulation of hematopoietic stem cell self-renewal. Dysregulation of Notch signaling is the underlying cause of diseases affecting the skeletal tissue, including Alagille syndrome, spondylocostal dysostosis, and possibly, osteosarcoma. PMID:19995916

  9. Full-field OCT for fast diagnostic of head and neck cancer

    NASA Astrophysics Data System (ADS)

    De Leeuw, Frederic; Casiraghi, Odile; Ben Lakhdar, Aïcha; Abbaci, Muriel; Laplace-Builhé, Corinne

    2015-02-01

    Full-Field OCT (FFOCT) produces optical slices of tissue using white light interferometry providing in-depth 2D images, with an isotropic resolution around 1 micrometer. These optical biopsy images are similar to those obtained with established histological procedures, but without tissue preparation and within few minutes. This technology could be useful when diagnosing a lesion or at the time of its surgical management. Here we evaluate the clinical value of FFOCT imaging in the management of patients with Head and Neck cancers by assessing the accuracy of the diagnosis done on FFOCT images from resected specimen. FFOCT images from Head and Neck samples were first compared to the gold standard (HES-conventional histology). An image atlas dedicated to the training of pathologists was built and diagnosis criteria were identified. Then, we performed a morphological correlative study: both healthy and cancerous samples from patients who undergo Head and Neck surgery of oral cavity, pharynx, and larynx were imaged. Images were interpreted in a random way by two pathologists and the FFOCT based diagnostics were compared with HES (gold standard) of the same samples. Here we present preliminary results showing that FFOCT provides a quick assessment of tissue architecture at microscopic level that could guide surgeons for tumor margin delineation during intraoperative procedure.

  10. Chronic rapamycin treatment on the nutrient utilization and metabolism of juvenile turbot (Psetta maxima)

    PubMed Central

    Wang, Qingchao; He, Gen; Mai, Kangsen; Xu, Wei; Zhou, Huihui; Wang, Xuan; Mei, Lin

    2016-01-01

    High dietary protein inclusion is necessary in fish feeds and also represents a major cost in the aquaculture industry, which demands improved dietary conversion into body proteins in fish. In mammals, the target of rapamycin (TOR) is a key nutritionally responsive molecule governing postprandial anabolism. However, its physiological significance in teleosts has not been fully examined. In the present study, we examined the nutritional physiology of turbot after chronic rapamycin inhibition. Our results showed that a 6-week inhibition of TOR using dietary rapamycin inclusion (30 mg/kg diet) reduced growth performance and feed utilization. The rapamycin treatment inhibited TOR signaling and reduced expression of key enzymes in glycolysis, lipogenesis, cholesterol biosynthesis, while increasing the expression of enzymes involved in gluconeogenesis. Furthermore, rapamycin treatment increased intestinal goblet cell number in turbot, while the expressions of Notch and Hes1 were down regulated. It was possible that stimulated goblet cell differentiation by rapamycin was mediated through Notch-Hes1 pathway. Therefore, our results demonstrate the important role of TOR signaling in fish nutritional physiology. PMID:27305975

  11. In vitro Inactivation of Latent HSV by Targeted Mutagenesis Using an HSV-specific Homing Endonuclease

    PubMed Central

    Aubert, Martine; Boyle, Nicole M; Stone, Daniel; Stensland, Laurence; Huang, Meei-Li; Magaret, Amalia S; Galetto, Roman; Rawlings, David J; Scharenberg, Andrew M; Jerome, Keith R

    2014-01-01

    Following acute infection, herpes simplex virus (HSV) establishes latency in sensory neurons, from which it can reactivate and cause recurrent disease. Available antiviral therapies do not affect latent viral genomes; therefore, they do not prevent reactivation following therapy cessation. One possible curative approach involves the introduction of DNA double strand breaks in latent HSV genomes by rare-cutting endonucleases, leading to mutagenesis of essential viral genes. We tested this approach in an in vitro HSV latency model using the engineered homing endonuclease (HE) HSV1m5, which recognizes a sequence in the HSV-1 gene UL19, encoding the virion protein VP5. Coexpression of the 3′-exonuclease Trex2 with HEs increased HE-mediated mutagenesis frequencies up to sixfold. Following HSV1m5/Trex2 delivery with adeno-associated viral (AAV) vectors, the target site was mutated in latent HSV genomes with no detectable cell toxicity. Importantly, HSV production by latently infected cells after reactivation was decreased after HSV1m5/Trex2 exposure. Exposure to histone deacetylase inhibitors prior to HSV1m5/Trex2 treatment increased mutagenesis frequencies of latent HSV genomes another two- to fivefold, suggesting that chromatin modification may be a useful adjunct to gene-targeting approaches. These results support the continuing development of HEs and other nucleases (ZFNs, TALENs, CRISPRs) for cure of chronic viral infections. PMID:24496438

  12. Evidence for increasing severity of community-onset boils and abscesses in UK General Practice.

    PubMed

    Shallcross, L J; Hayward, A C; Johnson, A M; Petersen, I

    2015-08-01

    In England, hospital admissions for severe staphylococcal boils and abscesses trebled between 1989 and 2004. We investigated this trend using routine data from primary and secondary care. We used The Health Improvement Network (THIN), a large primary-care database and national data on hospital admissions from Hospital Episode Statistics (HES). Time trends in the incidence of primary-care consultations for boils and abscesses were estimated for 1995-2010. HES data were used to calculate age-standardized hospital admission rates for boils, abscesses and cellulitis. The incidence of boil or abscess was 450 [95% confidence interval (CI) 447-452] per 100 000 person-years and increased slightly over the study period (incidence rate ratio 1·005, 95% CI 1·004-1·007). The rate of repeat consultation for a boil or abscess increased from 66 (95% CI 59-73) per 100 000 person-years in 1995 to peak at 97 (95% CI 94-101) per 100 000 person-years in 2006, remaining stable thereafter. Hospital admissions for abscesses, carbuncles, furuncles and cellulitis almost doubled, from 123 admissions per 100 000 in 1998/1999 to 236 admissions per 100 000 in 2010/2011. Rising hospitalization and recurrence rates set against a background of stable community incidence suggests increased disease severity. Patients may be experiencing more severe and recurrent staphylococcal skin disease with limited treatment options. PMID:25530161

  13. Expression and reconstitution of the bioluminescent Ca(2+) reporter aequorin in human embryonic stem cells, and exploration of the presence of functional IP3 and ryanodine receptors during the early stages of their differentiation into cardiomyocytes.

    PubMed

    Chan, Harvey Y S; Cheung, Man Chun; Gao, Yi; Miller, Andrew L; Webb, Sarah E

    2016-08-01

    In order to develop a novel method of visualizing possible Ca(2+) signaling during the early differentiation of hESCs into cardiomyocytes and avoid some of the inherent problems associated with using fluorescent reporters, we expressed the bioluminescent Ca(2+) reporter, apo-aequorin, in HES2 cells and then reconstituted active holo-aequorin by incubation with f-coelenterazine. The temporal nature of the Ca(2+) signals generated by the holo-f-aequorin-expressing HES2 cells during the earliest stages of differentiation into cardiomyocytes was then investigated. Our data show that no endogenous Ca(2+) transients (generated by release from intracellular stores) were detected in 1-12-day-old cardiospheres but transients were generated in cardiospheres following stimulation with KCl or CaCl2, indicating that holo-f-aequorin was functional in these cells. Furthermore, following the addition of exogenous ATP, an inositol trisphosphate receptor (IP3R) agonist, small Ca(2+) transients were generated from day 1 onward. That ATP was inducing Ca(2+) release from functional IP3Rs was demonstrated by treatment with 2-APB, a known IP3R antagonist. In contrast, following treatment with caffeine, a ryanodine receptor (RyR) agonist, a minimal Ca(2+) response was observed at day 8 of differentiation only. Thus, our data indicate that unlike RyRs, IP3Rs are present and continually functional at these early stages of cardiomyocyte differentiation. PMID:27430888

  14. Canonical Wnt signaling transiently stimulates proliferation and enhances neurogenesis in neonatal neural progenitor cultures

    SciTech Connect

    Hirsch, Cordula; Campano, Louise M.; Woehrle, Simon; Hecht, Andreas . E-mail: andreas.hecht@mol-med.uni-freiburg.de

    2007-02-01

    Canonical Wnt signaling triggers the formation of heterodimeric transcription factor complexes consisting of {beta}-catenin and T cell factors, and thereby controls the execution of specific genetic programs. During the expansion and neurogenic phases of embryonic neural development canonical Wnt signaling initially controls proliferation of neural progenitor cells, and later neuronal differentiation. Whether Wnt growth factors affect neural progenitor cells postnatally is not known. Therefore, we have analyzed the impact of Wnt signaling on neural progenitors isolated from cerebral cortices of newborn mice. Expression profiling of pathway components revealed that these cells are fully equipped to respond to Wnt signals. However, Wnt pathway activation affected only a subset of neonatal progenitors and elicited a limited increase in proliferation and neuronal differentiation in distinct subsets of cells. Moreover, Wnt pathway activation only transiently stimulated S-phase entry but did not support long-term proliferation of progenitor cultures. The dampened nature of the Wnt response correlates with the predominant expression of inhibitory pathway components and the rapid actuation of negative feedback mechanisms. Interestingly, in differentiating cell cultures activation of canonical Wnt signaling reduced Hes1 and Hes5 expression suggesting that during postnatal neural development, Wnt/{beta}-catenin signaling enhances neurogenesis from progenitor cells by interfering with Notch pathway activity.

  15. A novel 3q29 deletion associated with autism, intellectual disability, psychiatric disorders, and obesity.

    PubMed

    Biamino, Elisa; Di Gregorio, Eleonora; Belligni, Elga Fabia; Keller, Roberto; Riberi, Evelise; Gandione, Marina; Calcia, Alessandro; Mancini, Cecilia; Giorgio, Elisa; Cavalieri, Simona; Pappi, Patrizia; Talarico, Flavia; Fea, Antonio M; De Rubeis, Silvia; Cirillo Silengo, Margherita; Ferrero, Giovanni Battista; Brusco, Alfredo

    2016-03-01

    Copy number variation (CNV) has been associated with a variety of neuropsychiatric disorders, including intellectual disability/developmental delay (ID/DD), autism spectrum disorder (ASD), and schizophrenia (SCZ). Often, individuals carrying the same pathogenic CNV display high clinical variability. By array-CGH analysis, we identified a novel familial 3q29 deletion (1.36 Mb), centromeric to the 3q29 deletion region, which manifests with variable expressivity. The deletion was identified in a 3-year-old girl diagnosed with ID/DD and autism and segregated in six family members, all affected by severe psychiatric disorders including schizophrenia, major depression, anxiety disorder, and personality disorder. All individuals carrying the deletion were overweight or obese, and anomalies compatible with optic atrophy were observed in three out of four cases examined. Amongst the 10 genes encompassed by the deletion, the haploinsufficiency of Optic Atrophy 1 (OPA1), associated with autosomal dominant optic atrophy, is likely responsible for the ophthalmological anomalies. We hypothesize that the haploinsufficiency of ATPase type 13A4 (ATP13A4) and/or Hairy/Enhancer of Split Drosophila homolog 1 (HES1) contribute to the neuropsychiatric phenotype, while HES1 deletion might underlie the overweight/obesity. In conclusion, we propose a novel contiguous gene syndrome due to a proximal 3q29 deletion variably associated with autism, ID/DD, psychiatric traits and overweight/obesity. PMID:26620927

  16. Nuclear Hybrid Energy Systems: Imperatives, Prospects, and Challenges

    SciTech Connect

    Steven E. Aumeier

    2010-10-01

    As global population reaches an expected 8 billion people by 2030, primary energy consumption is expected to increase by almost 40% from approximately 520 exajoules consumed today to almost 740 exajoules. Much of this increase is expected to come from non-Organization for Economic Cooperation and Development (OECD) nations, and Asia specifically. In these economies, energy used for transportation is expected to grow substantially, as is industrial, commercial and to a lesser degree residential energy use, creating considerable pressure on global and local energy markets. The magnitude and timing of growth in energy consumption likely will create a global imperative to deploy energy production technologies that balance the three pillars of energy security: • economic stability – related to the affordability of energy products, stability and predictability in their price, and the efficient and effective deployment of global capital resources in their development; • environmental sustainability – related to minimizing the negative impacts of energy production to air, land, and water systems and advancing the long-term viability of using a particular resource in a way that does not limit future generations ability to prosper; • resource security – related to the ability to access energy resources and products where and when necessary, in an affordable and predictable manner. One approach to meeting these objectives is hybrid energy systems (HES). Broadly described, HES are energy product production plants that take two or more energy resource inputs (typically includes both carbon and non-carbon based sources) and produce two or more energy products (e.g. electricity, liquid transportation fuels, industrial chemicals) in an integrated plant. Nuclear energy integration into HES offers intriguing potential, particularly if smaller (<300 MWe) reactors are available. Although the concept of using nuclear energy in a variety of non-electrical process

  17. Modelling the hydrological behaviour of a coffee agroforestry basin in Costa Rica

    NASA Astrophysics Data System (ADS)

    Gómez-Delgado, F.; Roupsard, O.; Moussa, R.; Le Maire, G.; Taugourdeau, S.; Bonnefond, J. M.; Pérez, A.; van Oijen, M.; Vaast, P.; Rapidel, B.; Voltz, M.; Imbach, P.; Harmand, J. M.

    2010-05-01

    The profitability of hydropower in Costa Rica is affected by soil erosion and sedimentation in dam reservoirs, which are in turn influenced by land use, infiltration and aquifer interactions with surface water. In order to foster the provision and payment of Hydrological Environmental Services (HES), a quantitative assessment of the impact of specific land uses on the functioning of drainage-basins is required. The present paper aims to study the water balance partitioning in a volcanic coffee agroforestry micro-basin (1 km2, steep slopes) in Costa Rica, as a first step towards evaluating sediment or contaminant loads. The main hydrological processes were monitored during one year, using flume, eddy-covariance flux tower, soil water profiles and piezometers. A new Hydro-SVAT lumped model is proposed, that balances SVAT (Soil Vegetation Atmosphere Transfer) and basin-reservoir routines. The purpose of such a coupling was to achieve a trade-off between the expected performance of ecophysiological and hydrological models, which are often employed separately and at different spatial scales, either the plot or the basin. The calibration of the model to perform streamflow yielded a NS coefficient equal to 0.80, while the validation of the water balance partitioning was consistent with the independent measurements of actual evapotranspiration (R2=0.79, energy balance closed independently), soil water content (R2=0.49) and water table level (R2=0.90). An uncertainty analysis showed that the streamflow modelling was precise for nearly every time step, while a sensitivity analysis revealed which parameters mostly affected model precision, depending on the season. It was observed that 64% of the incident rainfall R flowed out of the basin as streamflow, 25% as evapotranspiration and the remaining 11% was attributed to deep percolation. The model indicated an interception loss equal to 4% of R, a surface runoff of 5% and an infiltration component of 91%. The modelled

  18. A novel reporter of notch signalling indicates regulated and random notch activation during vertebrate neurogenesis

    PubMed Central

    2011-01-01

    Background Building the complex vertebrate nervous system involves the regulated production of neurons and glia while maintaining a progenitor cell population. Neurogenesis starts asynchronously in different regions of the embryo and occurs over a long period of time, allowing progenitor cells to be exposed to multiple extrinsic signals that regulate the production of different cell types. Notch-mediated cell-cell signalling is one of the mechanisms that maintain the progenitor pool, however, little is known about how the timing of Notch activation is related to the cell cycle and the distinct modes of cell division that generate neurons. An essential tool with which to investigate the role of Notch signalling on cell by cell basis is the development a faithful reporter of Notch activity. Results Here we present a novel reporter for Notch activity based on the promoter of the well characterised Notch target chick Hes5-1, coupled with multiple elements that confer instability, including a destabilized nuclear Venus fluorescent protein and the 3' untranslated region (UTR) of Hes5-1. We demonstrate that this reporter faithfully recapitulates the endogenous expression of Hes5-1 and that it robustly responds to Notch activation in the chick neural tube. Analysis of the patterns of Notch activity revealed by this reporter indicates that although Notch is most frequently activated prior to mitosis it can be activated at any time within the cell cycle. Notch active progenitors undergoing mitosis generate two daughters that both continue to experience Notch signalling. However, cells lacking Notch activity before and during mitosis generate daughters with dissimilar Notch activity profiles. Conclusions A novel Notch reporter with multiple destabilisation elements provides a faithful read-out of endogenous Notch activity on a cell-by-cell basis, as neural progenitors progress through the cell cycle in the chick neural tube. Notch activity patterns in this cell population

  19. Opposing role of Notch1 and Notch2 in a Kras(G12D)-driven murine non-small cell lung cancer model.

    PubMed

    Baumgart, A; Mazur, P K; Anton, M; Rudelius, M; Schwamborn, K; Feuchtinger, A; Behnke, K; Walch, A; Braren, R; Peschel, C; Duyster, J; Siveke, J T; Dechow, T

    2015-01-29

    Lung cancer is the leading cause of cancer-related deaths worldwide. Recently, we have shown that Notch1 inhibition resulted in substantial cell death of non-small cell lung cancer (NSCLC) cells in vitro. New compounds targeting Notch signal transduction have been developed and are now being tested in clinical trials. However, the tumorigenic role of individual Notch receptors in vivo remains largely unclear. Using a Kras(G12D)-driven endogenous NSCLC mouse model, we analyzed the effect of conditional Notch1 and Notch2 receptor deletion on NSCLC tumorigenesis. Notch1 deficiency led to a reduced early tumor formation and lower activity of MAPK compared with the controls. Unexpectedly, Notch2 deletion resulted in a dramatically increased carcinogenesis and increased MAPK activity. These mice died significantly earlier due to rapidly growing tumor burden. We found that Notch1 regulates Ras/MAPK pathway via HES1-induced repression of the DUSP1 promoter encoding a phosphatase specifically suppressing pERK1/2. Interestingly, Notch1 but not Notch2 ablation leads to decreased HES1 and DUSP1 expression. However, Notch2-depleted tumors showed an appreciable increase in β-catenin expression, a known activator of HES1 and important lung cancer oncogene. Characteristically for β-catenin upregulation, we found that the majority of Notch2-deficient tumors revealed an undifferentiated phenotype as determined by their morphology, E-Cadherin and TTF1 expression levels. In addition, these carcinomas showed aggressive growth patterns with bronchus invasion and obstruction. Together, we show that Notch2 mediates differentiation and has tumor suppressor functions during lung carcinogenesis, whereas Notch1 promotes tumor initiation and progression. These data are further supported by immunohistochemical analysis of human NSCLC samples showing loss or downregulation of Notch2 compared with normal lung tissue. In conclusion, this is the first study characterizing the in vivo functions of

  20. SEARCHES FOR METAL-POOR STARS FROM THE HAMBURG/ESO SURVEY USING THE CH G BAND

    SciTech Connect

    Placco, Vinicius M.; Rossi, Silvia; Kennedy, Catherine R.; Beers, Timothy C.; Lee, Young Sun; Christlieb, Norbert; Sivarani, Thirupathi; Reimers, Dieter; Wisotzki, Lutz

    2011-12-15

    We describe a new method to search for metal-poor candidates from the Hamburg/ESO objective-prism survey (HES) based on identifying stars with apparently strong CH G-band strengths for their colors. The hypothesis we exploit is that large overabundances of carbon are common among metal-poor stars, as has been found by numerous studies over the past two decades. The selection was made by considering two line indices in the 4300 A region, applied directly to the low-resolution prism spectra. This work also extends a previously published method by adding bright sources to the sample. The spectra of these stars suffer from saturation effects, compromising the index calculations and leading to an undersampling of the brighter candidates. A simple numerical procedure, based on available photometry, was developed to correct the line indices and overcome this limitation. Visual inspection and classification of the spectra from the HES plates yielded a list of 5288 new metal-poor (and by selection, carbon-rich) candidates, which are presently being used as targets for medium-resolution spectroscopic follow-up. Estimates of the stellar atmospheric parameters, as well as carbon abundances, are now available for 117 of the first candidates, based on follow-up medium-resolution spectra obtained with the SOAR 4.1 m and Gemini 8 m telescopes. We demonstrate that our new method improves the metal-poor star fractions found by our pilot study by up to a factor of three in the same magnitude range, as compared with our pilot study based on only one CH G-band index. Our selection scheme obtained roughly a 40% success rate for identification of stars with [Fe/H] <-1.0; the primary contaminant is late-type stars with near-solar abundances and, often, emission line cores that filled in the Ca II K line on the prism spectrum. Because the selection is based on carbon, we greatly increase the numbers of known carbon-enhanced metal-poor stars from the HES with intermediate metallicities -2

  1. At Least Three Transporters Likely Mediate Threonine Uptake Needed for Mouse Embryonic Stem Cell Proliferation

    PubMed Central

    Formisano, Tara M.; Van Winkle, Lon J.

    2016-01-01

    Stem cells are at the forefront of current regenerative and biomedical research. Thus, there exists an imperative and urgent need to understand the mechanisms that drive stem cell function in order to exploit their use as a therapeutic tool. Amino acids are potent inducers of signaling cascades that drive stem cell proliferation and differentiation. With a focus on mouse embryonic stem (mES) cells, Threonine (Thr) is the only amino acid required in culture media for mES cell proliferation. Current research associates this need for Thr with threonine dehydrogenase (TDH), which catabolizes Thr to glycine and acetyl-CoA in mES cells. This theory depends, in part, on the ability of 3- hydroxynorvaline (3-HNV) to inhibit both TDH and mES cell proliferation. However, the concentration of 3-HNV needed to inhibit mES cell proliferation is more than an order of magnitude less than its apparent Ki for TDH inhibition. Additionally, 3-HNV inhibits human embryonic stem (hES) cell proliferation, but hES cells do not express a functional tdh gene. Such findings indicate another mechanism for Thr stimulated mES and hES cell proliferation. Since amino acid transporters may be inducers of signaling cascades, we characterized the Thr transport systems in mES cells. We found that there is a Na+-dependent and a Na+-independent component of substrate-saturable transport, with the Na+-dependent component predominating. We also found that of 20 amino acids tested, the amino acids that were the strongest inhibitors of the Na+-dependent component of radiolabeled Thr transport were Ser, Cys, 4-OH-Pro, Asn, Met, and non-radiolabeled Thr itself. Such findings are consistent with characteristics of the ASC transport system, suggesting that this ASC system is responsible for the majority of Thr transport in mES cells. We confirmed expression of mRNA encoding the ASC system transporters, ASCT1 and ASCT2, in mES cells using RT-PCR. In conclusion, mES cells likely express at least three

  2. The choice of the intravenous fluid influences the tolerance of acute normovolemic anemia in anesthetized domestic pigs

    PubMed Central

    2012-01-01

    Introduction The correction of hypovolemia with acellular fluids results in acute normovolemic anemia. Whether the choice of the infusion fluid has an impact on the maintenance of oxygen (O2) supply during acute normovolemic anemia has not been investigated so far. Methods Thirty-six anesthetized and mechanically ventilated pigs were hemodiluted to their physiological limit of anemia tolerance, reflected by the individual critical hemoglobin concentration (Hbcrit). Hbcrit was defined as the Hb-concentration corresponding with the onset of supply-dependency of total body O2-consumption (VO2). The hemodilution protocol was randomly performed with either tetrastarch (6% HES 130/0.4, TS-group, n = 9), gelatin (3.5% urea-crosslinked polygeline, GEL-group, n = 9), hetastarch (6% HES 450/0.7, HS-group, n = 9) or Ringer's solution (RS-group, n = 9). The primary endpoint was the dimension of Hbcrit, secondary endpoints were parameters of central hemodynamics, O2 transport and tissue oxygenation. Results In each animal, normovolemia was maintained throughout the protocol. Hbcrit was met at 3.7 ± 0.6 g/dl (RS), 3.0 ± 0.6 g/dl (HS P < 0.05 vs. RS), 2.7 ± 0.6 g/dl (GEL, P < 0.05 vs. RS) and 2.1 ± 0.4 g/dl (TS, P < 0.05 vs. GEL, HS and RS). Hemodilution with RS resulted in a significant increase of extravascular lung water index (EVLWI) and a decrease of arterial oxygen partial pressure (paO2), and O2 extraction ratio was increased, when animals of the TS-, GEL- and HS-groups met their individual Hbcrit. Conclusions The choice of the intravenous fluid has an impact on the tolerance of acute normovolemic anemia induced by acellular volume replacement. Third-generation tetrastarch preparations (e.g., HES 130/0.4) appear most advantageous regarding maintenance of tissue oxygenation during progressive anemia. The underlying mechanism includes a lower degree of extravasation and favourable effects on microcirculatory function. PMID:22546374

  3. The Notch pathway mediates the angiotensin II-induced synthesis of extracellular matrix components in podocytes.

    PubMed

    Yao, Min; Wang, Xiaomei; Wang, Xiaomeng; Zhang, Tao; Chi, Yanqing; Gao, Feng

    2015-07-01

    The Notch pathway is known to contribute to the development of glomerular disease. Angiotensin II (Ang II), an important member of the renin-angiotensin system, stimulates the accumulation of extracellular matrix components in glomerular disease; however, the exact mechanisms involved remain to be elucidated. In the present study, we aimed to investigate the effects of the Notch pathway on the synthesis of extracellular matrix components in Ang II-stimulated podocytes. Mouse podocytes were stimulated with Ang II (10-6 mol/l). The activation of the Notch pathway was inhibited by a vector carrying short hairpin RNA (shRNA) targeting Notch1 (sh-Notch1) or by γ-secretase inhibitor (GSI). The protein levels of Notch1, Notch intracellular domain 1 (NICD1), hairy and enhancer of split-1 (Hes1), matrix metalloproteinase (MMP)-2, MMP-9, transforming growth factor-β1 (TGF-β1), type IV collagen and laminin were determined by western blot analysis. The Notch1, Hes1, MMP-2, MMP-9, TGF-β1, type IV collagen and laminin mRNA levels were detected by RT-PCR. The MMP-2 and MMP-9 activity was measured using a cell active fluorescence assay kit. The levels of TGF-β1, type IV collagen and laminin were determined in the culture medium of the podocytes by enzyme-linked immunosorbent assay (ELISA). Our results revealed that Ang II upregulated Notch1, NICD1, Hes1, TGF-β1, type IV collagen and laminin expression and downregulated MMP-2 and MMP-9 expression in the cultured podocytes. The inhibition of the Notch pathway by sh-Notch1 or GSI increased MMP-2 and MMP-9 expression, decreased the TGF-β1 level and suppressed type IV collagen and laminin expression. The inhibition of the Notch pathway by sh-Notch1 or GSI also increased MMP-2 and MMP-9 activity, and decreased TGF-β1 levels, type IV collagen levels and laminin secretion. These findings indicate that the Notch pathway potentially mediates the Ang II-induced synthesis of extracellular matrix components in podocytes through the

  4. Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers: objective comparison between cellular and acellular types.

    PubMed

    Sakai, H; Yuasa, M; Onuma, H; Takeoka, S; Tsuchida, E

    2000-01-01

    A series of hemoglobin (Hb)-based O(2) carriers, acellular and cellular types, were synthesized and their physicochemical characteristics were compared. The acellular type includes intramolecularly cross-linked Hb (XLHb), polyoxyethylene (POE)-conjugated pyridoxalated Hb (POE-PLP-Hb), hydroxyethylstarch-conjugated Hb (HES-XLHb), and glutaraldehyde-polymerized XLHb (Poly-XLHb). The cellular type is Hb-vesicles (HbV) of which the surface is modified with POE (POE-HbV). Their particle diameters are 7 +/- 2, 22 +/- 2, 47 +/- 17, 68 +/- 24, and 224 +/- 76 nm, respectively, thus all the materials penetrate across membrane filters with 0.4 microm pore size, though only the POE-HbV cannot penetrate across the filter with 0.2 microm pore size. These characteristics of permeability are important to consider an optimal particle size in microcirculation in vivo. POE-PLP-Hb ([Hb] = 5 g/dL) showed viscosity of 6.1 cP at 332 s(-1) and colloid osmotic pressure (COP) of 70.2 Torr, which are beyond the physiological conditions (human blood, viscosity = 3-4 cP, COP = ca. 25 Torr). XLHb and Poly-XLHb showed viscosities of 1.0 and 1.5 cp, respectively, which are significantly lower than that of blood. COP of POE-HbV is regulated to 20 Torr in 5% human serum albumin (HSA). HES-XLHb and POE-HbV/HSA showed comparable viscosity with human blood. Microscopic observation of human red blood cells (RBC) after mixing blood with POE-PLP-Hb or HES-XLHb disclosed aggregates of RBC, a kind of sludge, indicating a strong interaction with RBC, which is anticipated to modify peripheral blood flow in vivo. On the other hand, XLHb and POE-HbV showed no rouleaux or aggregates of RBC. The acellular Hbs (P(50) = 14-32 Torr) have their specific O(2) affinities determined by their structures, while that of the cellular POE-HbV is regulated by coencapsulating an appropriate amount of an allosteric effector (e.g., P(50) = 18, 32 Torr). These differences in physicochemical characteristics between the acellular

  5. Interference and complementarity for two-photon hybrid entangled states

    SciTech Connect

    Nogueira, W. A. T.; Santibanez, M.; Delgado, A.; Saavedra, C.; Neves, L.; Lima, G.; Padua, S.

    2010-10-15

    In this work we generate two-photon hybrid entangled states (HESs), where the polarization of one photon is entangled with the transverse spatial degree of freedom of the second photon. The photon pair is created by parametric down-conversion in a polarization-entangled state. A birefringent double-slit couples the polarization and spatial degrees of freedom of these photons, and finally, suitable spatial and polarization projections generate the HES. We investigate some interesting aspects of the two-photon hybrid interference and present this study in the context of the complementarity relation that exists between the visibility of the one-photon and that of the two-photon interference patterns.

  6. US Hydropower Resource Assessment for Massachusetts

    SciTech Connect

    Francfort, J.E.; Rinehart, B.N.

    1995-07-01

    The Department of Energy is developing an estimate of the undeveloped hydropower potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The software measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven software program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the Commonwealth of Massachusetts.

  7. Comparation of enhanced green fluorescent protein gene transfected and wild-type porcine neural stem cells.

    PubMed

    Zheng, Yue-Mao; An, Zhi-Xing; Zhao, Xiao-E; Quan, Fu-Sheng; Zhao, Hui-Ying; Zhang, Ya-Rong; Liu, Jun; He, Xiao-Ying; He, Xiao-Ning

    2010-02-01

    The aim of this study was to transfect and express the enhanced green fluorescence protein (EGFP) gene into porcine neural stem cells (NSCs) to determine whether EGFP can be used as a marker to monitor NSCs. NSCs were isolated from embryonic day 30 fetal pig brain and transfected with EGFP gene using lipofection. Transfected and wild-type NSCs were induced to differentiate into cells of neuronal and myogenic lineages. Markers of passage three NSCs and their differentiated cells were tested by reverse transcription polymerase chain reaction. The results showed that EGFP could be expressed in NSCs and the differentiated cells. NSCs expressed Nestin, NogoA, DCX, Hes1, Oct4, CD-90 and Sox2. NSCs could differentiated into astrocyte (GFAP(+)), oligodendrocyte (GalC(+)), neuron (NF(+), NSE(+) and MAP2(+)) and myocyte (myf-6(+) and myoD(+)). We concluded that EGFP can be used as a marker in monitoring NSCs. PMID:19580981

  8. Thermophysical properties of a monolayer tissue with respect to freeze-drying.

    PubMed

    Ferrando, M; Glasmacher, B; Rau, G

    2002-01-01

    Conservation of tissue structures by means of freeze-drying is still limited as the complex mechanisms taking place at molecular/cellular level are not fully understood. The successful application of hydroxyethyl starch (HES) in combination with maltose, sucrose, and trehalose as stabilizers of lipid bilayers/membranes in red blood cells suggests an extended use of this mixture of cryoprotectants. The effectiveness of such cryoprotectant solutions has been linked to changes in the thermophysical properties of cellular structures. This work deals, in a first step, with the thermophysical properties of a model monolayer tissue--onion epidermis--in binary aqueous solutions of dissacharides. First and second order phase transitions, i.e. melting, crystallisation, and glass transition, are characterised by means of Modulated Differential Scanning Calorimetry (MDSC). PMID:12451873

  9. Coherent structures in the Es layer and neutral middle atmosphere

    NASA Astrophysics Data System (ADS)

    Mošna, Zbyšek; Knížová, Petra Koucká; Potužníková, Kateřina

    2015-12-01

    The present paper shows results from the summer campaign performed during geomagnetically quiet period from June 1 to August 31, 2009. Within time-series of stratospheric and mesospheric temperatures at pressure levels 10-0.1 hPa, mesospheric winds measured in Collm, Germany, and the sporadic E-layer parameters foEs and hEs measured at the Pruhonice station we detected specific coherent wave-bursts in planetary wave domain. Permanent wave-like activity is observed in all analyzed data sets. However, the number of wave-like structures persistent in large range of height from the stratosphere to lower ionosphere is limited. The only coherent modes that are detected on consequent levels of the atmosphere are those corresponding to eigenmodes of planetary waves.

  10. Notch Lineages and Activity in Intestinal Stem Cells Determined by a New Set of Knock-In Mice

    PubMed Central

    Fre, Silvia; Hannezo, Edouard; Sale, Sanja; Huyghe, Mathilde; Lafkas, Daniel; Kissel, Holger; Louvi, Angeliki; Greve, Jeffrey; Louvard, Daniel; Artavanis-Tsakonas, Spyros

    2011-01-01

    The conserved role of Notch signaling in controlling intestinal cell fate specification and homeostasis has been extensively studied. Nevertheless, the precise identity of the cells in which Notch signaling is active and the role of different Notch receptor paralogues in the intestine remain ambiguous, due to the lack of reliable tools to investigate Notch expression and function in vivo. We generated a new series of transgenic mice that allowed us, by lineage analysis, to formally prove that Notch1 and Notch2 are specifically expressed in crypt stem cells. In addition, a novel Notch reporter mouse, Hes1-EmGFPSAT, demonstrated exclusive Notch activity in crypt stem cells and absorptive progenitors. This roster of knock-in and reporter mice represents a valuable resource to functionally explore the Notch pathway in vivo in virtually all tissues. PMID:21991352

  11. Interferon-γ regulates cellular metabolism and mRNA translation to potentiate macrophage activation.

    PubMed

    Su, Xiaodi; Yu, Yingpu; Zhong, Yi; Giannopoulou, Eugenia G; Hu, Xiaoyu; Liu, Hui; Cross, Justin R; Rätsch, Gunnar; Rice, Charles M; Ivashkiv, Lionel B

    2015-08-01

    Interferon-γ (IFN-γ) primes macrophages for enhanced microbial killing and inflammatory activation by Toll-like receptors (TLRs), but little is known about the regulation of cell metabolism or mRNA translation during this priming. We found that IFN-γ regulated the metabolism and mRNA translation of human macrophages by targeting the kinases mTORC1 and MNK, both of which converge on the selective regulator of translation initiation eIF4E. Physiological downregulation of mTORC1 by IFN-γ was associated with autophagy and translational suppression of repressors of inflammation such as HES1. Genome-wide ribosome profiling in TLR2-stimulated macrophages showed that IFN-γ selectively modulated the macrophage translatome to promote inflammation, further reprogram metabolic pathways and modulate protein synthesis. These results show that IFN-γ-mediated metabolic reprogramming and translational regulation are key components of classical inflammatory macrophage activation. PMID:26147685

  12. Functional genomics, proteomics, and regulatory DNA analysis in isogenic settings using zinc finger nuclease-driven transgenesis into a safe harbor locus in the human genome

    PubMed Central

    DeKelver, Russell C.; Choi, Vivian M.; Moehle, Erica A.; Paschon, David E.; Hockemeyer, Dirk; Meijsing, Sebastiaan H.; Sancak, Yasemin; Cui, Xiaoxia; Steine, Eveline J.; Miller, Jeffrey C.; Tam, Phillip; Bartsevich, Victor V.; Meng, Xiangdong; Rupniewski, Igor; Gopalan, Sunita M.; Sun, Helena C.; Pitz, Kathleen J.; Rock, Jeremy M.; Zhang, Lei; Davis, Gregory D.; Rebar, Edward J.; Cheeseman, Iain M.; Yamamoto, Keith R.; Sabatini, David M.; Jaenisch, Rudolf; Gregory, Philip D.; Urnov, Fyodor D.

    2010-01-01

    Isogenic settings are routine in model organisms, yet remain elusive for genetic experiments on human cells. We describe the use of designed zinc finger nucleases (ZFNs) for efficient transgenesis without drug selection into the PPP1R12C gene, a “safe harbor” locus known as AAVS1. ZFNs enable targeted transgenesis at a frequency of up to 15% following transient transfection of both transformed and primary human cells, including fibroblasts and hES cells. When added to this locus, transgenes such as expression cassettes for shRNAs, small-molecule-responsive cDNA expression cassettes, and reporter constructs, exhibit consistent expression and sustained function over 50 cell generations. By avoiding random integration and drug selection, this method allows bona fide isogenic settings for high-throughput functional genomics, proteomics, and regulatory DNA analysis in essentially any transformed human cell type and in primary cells. PMID:20508142

  13. U.S. hydropower resource assessment for Connecticut

    SciTech Connect

    Francfort, J.E.; Rinehart, B.N.

    1995-07-01

    The Department of Energy is developing an estimate of the undeveloped hydro-power potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The software measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven software program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the State of Connecticut.

  14. Structure of the human paralemmin gene (PALM), mapping to human chromosome 19p13.3 and mouse chromosome 10, and exclusion of coding mutations in grizzled, mocha, jittery, and hesitant mice.

    PubMed

    Burwinkel, B; Miglierini, G; Jenne, D E; Gilbert, D J; Copeland, N G; Jenkins, N A; Ring, H Z; Francke, U; Kilimann, M W

    1998-05-01

    Paralemmin is a newly identified protein that is associated with the plasma membrane and with intracellular membranes through a lipid anchor. It is abundant in brain, is expressed at intermediate levels in the kidney and in endocrine cells, and occurs at low levels in many other tissues. As it is a candidate for genetic disorders that affect membrane functions, we have determined the structure of the human paralemmin gene, PALM, showing that it is organized into nine exons. Moreover, we have performed chromosomal assignments of the human and mouse paralemmin genes, localizing them to regions of homology at human 19p13.3 and the central mouse chromosome 10. Finally, mutation analysis using RNA from mice homozygous for the mutant genes grizzled (gr), mocha (mh), mocha 2J (mh2J), jittery (ji) and hesitant (ji(hes)), which map to this area, excluded mutations in their Palm coding sequences. PMID:9615234

  15. Oscillations and temporal signalling in cells.

    PubMed

    Tiana, G; Krishna, S; Pigolotti, S; Jensen, M H; Sneppen, K

    2007-06-01

    The development of new techniques to quantitatively measure gene expression in cells has shed light on a number of systems that display oscillations in protein concentration. Here we review the different mechanisms which can produce oscillations in gene expression or protein concentration using a framework of simple mathematical models. We focus on three eukaryotic genetic regulatory networks which show 'ultradian' oscillations, with a time period of the order of hours, and involve, respectively, proteins important for development (Hes1), apoptosis (p53) and immune response (NF-kappaB). We argue that underlying all three is a common design consisting of a negative feedback loop with time delay which is responsible for the oscillatory behaviour. PMID:17664651

  16. Human Embryonic Stem Cell-Derived Neural Precursors Develop Into Neurons and Integrate Into the Host Brain

    PubMed Central

    Guillaume, Daniel J.; Johnson, M. Austin; Li, Xue-Jun; Zhang, Su-Chun

    2009-01-01

    Whether and how in-vitro-produced human neural precursors mature and integrate into the brain are crucial to the utility of human embryonic stem (hES) cells in treating neurological disorders. After transplantation into the ventricles of neonatal immune-deficient mice, hES-cell-derived neural precursors stopped expressing the cell division marker Ki67, except in neurogenic areas, and differentiated into neurons and then glia in a temporal course intrinsic to that of human cells regardless of location. The human cells located in the gray matter became neurons in the olfactory bulb and striatum, whereas those in the white matter produced exclusively glia. Importantly, the grafted human cells formed synapses. Thus, the in-vitro-produced human neural precursors follow their intrinsic temporal program to produce neurons and glia and, in response to environmental signals, generate cells appropriate to their target regions and integrate into the brain. PMID:16941479

  17. Chemical/hydrogen energy systems

    NASA Astrophysics Data System (ADS)

    1987-06-01

    This report describes activities conducted during 1986 within the Chemical/Hydrogen Energy Systems (C/HES) Program, for which Bookhaven National Laboratory provides technical and management support to the U.S. Department of Energy. Research and exploratory efforts under this program have been directed towards developing a base technology that will apply to hydrogen production, storage, and transport. Major areas of interest include: (1) High Temperature Water Vapor Electrolysis for Hydrogen Generation (Westinghouse); (2) Evaluation of Materials for Medium Temperature Water Vapor Electrolysis; (3) Cryoadsorption of Hydrogen on Activated Carbon (Syracuse). Contributions in 1986 made by private sector contractors, university, researchers, and BNL in-house technical staff are summarized in the report. Also included is a summary of related International Energy Agency (IEA) cooperative efforts as well as plans and major activities scheduled for 1987.

  18. Reprogramming homing endonuclease specificity through computational design and directed evolution.

    PubMed

    Thyme, Summer B; Boissel, Sandrine J S; Arshiya Quadri, S; Nolan, Tony; Baker, Dean A; Park, Rachel U; Kusak, Lara; Ashworth, Justin; Baker, David

    2014-02-01

    Homing endonucleases (HEs) can be used to induce targeted genome modification to reduce the fitness of pathogen vectors such as the malaria-transmitting Anopheles gambiae and to correct deleterious mutations in genetic diseases. We describe the creation of an extensive set of HE variants with novel DNA cleavage specificities using an integrated experimental and computational approach. Using computational modeling and an improved selection strategy, which optimizes specificity in addition to activity, we engineered an endonuclease to cleave in a gene associated with Anopheles sterility and another to cleave near a mutation that causes pyruvate kinase deficiency. In the course of this work we observed unanticipated context-dependence between bases which will need to be mechanistically understood for reprogramming of specificity to succeed more generally. PMID:24270794

  19. Ethics and synthetic gametes.

    PubMed

    Testa, Giuseppe; Harris, John

    2005-04-01

    The recent in vitro derivation of gamete-like cells from mouse embryonic stem (mES) cells is a major breakthrough and lays down several challenges, both for the further scientific investigation and for the bioethical and biolegal discourse. We refer here to these cells as gamete-like (sperm-like or oocyte-like, respectively), because at present there is still no evidence that these cells behave fully like bona fide sperm or oocytes, lacking the fundamental proof, i.e. combination with a normally derived gamete of the opposite sex to yield a normal individual. However, the results published so far do show that these cells share some defining features of gametes. We discuss these results in the light of the bioethical and legal questions that are likely to arise would the same process become possible with human embryonic stem (hES) cells. PMID:15943023

  20. High resolution spectroscopic study of Be10Lambda;

    NASA Astrophysics Data System (ADS)

    Gogami, T.; Chen, C.; Kawama, D.; Achenbach, P.; Ahmidouch, A.; Albayrak, I.; Androic, D.; Asaturyan, A.; Asaturyan, R.; Ates, O.; Baturin, P.; Badui, R.; Boeglin, W.; Bono, J.; Brash, E.; Carter, P.; Chiba, A.; Christy, E.; Danagoulian, S.; de Leo, R.; Doi, D.; Elaasar, M.; Ent, R.; Fujii, Y.; Fujita, M.; Furic, M.; Gabrielyan, M.; Gan, L.; Garibaldi, F.; Gaskell, D.; Gasparian, A.; Han, Y.; Hashimoto, O.; Horn, T.; Hu, B.; Hungerford, Ed. V.; Jones, M.; Kanda, H.; Kaneta, M.; Kato, S.; Kawai, M.; Khanal, H.; Kohl, M.; Liyanage, A.; Luo, W.; Maeda, K.; Margaryan, A.; Markowitz, P.; Maruta, T.; Matsumura, A.; Maxwell, V.; Mkrtchyan, A.; Mkrtchyan, H.; Nagao, S.; Nakamura, S. N.; Narayan, A.; Neville, C.; Niculescu, G.; Niculescu, M. I.; Nunez, A.; Nuruzzaman, Okayasu, Y.; Petkovic, T.; Pochodzalla, J.; Qiu, X.; Reinhold, J.; Rodriguez, V. M.; Samanta, C.; Sawatzky, B.; Seva, T.; Shichijo, A.; Tadevosyan, V.; Tang, L.; Taniya, N.; Tsukada, K.; Veilleux, M.; Vulcan, W.; Wesselmann, F. R.; Wood, S. A.; Yamamoto, T.; Ya, L.; Ye, Z.; Yokota, K.; Yuan, L.; Zhamkochyan, S.; Zhu, L.; Hksjlab E05-115 Collaboration

    2016-03-01

    Spectroscopy of a Be10Lambda; hypernucleus was carried out at JLab Hall C using the (e ,e'K+) reaction. A new magnetic spectrometer system (SPL+HES+HKS), specifically designed for high resolution hypernuclear spectroscopy, was used to obtain an energy spectrum with a resolution of ˜0.78 MeV (FWHM). The well-calibrated spectrometer system of the present experiment using p (e ,e'K+)Λ ,Σ0 reactions allowed us to determine the energy levels; and the binding energy of the ground-state peak (mixture of 1- and 2- states) was found to be BΛ=8.55 ±0.07 (stat . ) ±0.11 (sys . ) MeV. The result indicates that the ground-state energy is shallower than that of an emulsion study by about 0.5 MeV which provides valuable experimental information on the charge symmetry breaking effect in the Λ N interaction.

  1. Distinguishing short and long Fermi gamma-ray bursts

    NASA Astrophysics Data System (ADS)

    Tarnopolski, M.

    2015-11-01

    Two classes of gamma-ray bursts (GRBs), short and long, have been determined without any doubts, and are usually ascribed to different progenitors, yet these classes overlap for a variety of descriptive parameters. A subsample of 46 long and 22 short Fermi GRBs with estimated Hurst Exponents (HEs), complemented by minimum variability time-scales (MVTS) and durations (T90) is used to perform a supervised machine learning and Monte Carlo simulation using a support vector machine algorithm. It is found that while T90 itself performs very well in distinguishing short and long GRBs, the overall success ratio is higher when the training set is complemented by MVTS and HE. These results may allow us to introduce a new (non-linear) parameter that might provide less ambiguous classification of GRBs.

  2. Notch in the intestine: regulation of homeostasis and pathogenesis.

    PubMed

    Noah, Taeko K; Shroyer, Noah F

    2013-01-01

    The small and large intestines are tubular organs composed of several tissue types. The columnar epithelium that lines the inner surface of the intestines distinguishes the digestive physiology of each region of the intestine and consists of several distinct cell types that are rapidly and continually renewed by intestinal stem cells that reside near the base of the crypts of Lieberkühn. Notch signaling controls the fate of intestinal stem cells by regulating the expression of Hes genes and by repressing Atoh1. Alternate models of Notch pathway control of cell fate determination are presented. Roles for Notch signaling in development of the intestine, including mesenchymal and neural cells, are discussed. The oncogenic activities of Notch in colorectal cancer, as well as the tumor suppressive activities of Atoh1, are reviewed. Therapeutic targeting of the Notch pathway in colorectal cancers is discussed, along with potential caveats. PMID:23190077

  3. India gas well blowout capped and killed in 17 days

    SciTech Connect

    1995-06-01

    On January 8, 1995, the Pasarlapudi 19 gas well being drilled by India`s ONGC (oil and Natural Gas Corp.) near Amalapuram, India, 295 miles east of the state capital, Hyderabad, blew out while the operator was retrieving a stuck fish in deviated hole. On February 26, ONGC awarded a well control contract to International Well Control (IWC), Houston. On March 15, IWC and ONGC`s Crisis Management Team (CMT) completed extinguishing the fire, capping the well and killing the blowout, which was described by the experienced team as one of the two or three biggest they had ever seen. The article describes how the fire was extinguished and the well was capped, procedures heavily dependent on successful application of an abrasive fluid cutter supplied by Halliburton Energy Services (HES).

  4. Notch1 Signaling Is Activated in Cells Expressing Embryonic Stem Cell Proteins in Human Primary Nasopharyngeal Carcinoma

    PubMed Central

    Zhang, Yue; Peng, Jianhua; Zhang, Huxiang; Zhu, Yi; Wan, Li; Chen, Jianfu; Chen, Xiaoyun; Lin, Renyu; Li, He; Mao, XiaoOu; Jin, Kunlin

    2010-01-01

    Objective: To explore the expression of Notch1 signaling pathway in nasopharyngeal carcinoma (NPC). Methods: We performed immunocytochemistry on surgically resected NPC using antibodies against embryonic stem (ES) cell proteins and against Notch1 signaling components. Results: We found that ES cell protein markers SOX2 and OCT4 were expressed in a subpopulation of cells for all three subtypes of NPC but barely in the normal control. Double immunostaining shows that SOX2- and OCT4-positive cells coexpressed proliferative markers, suggesting that human NPC may contain cancer stem–like cells. In addition, we found that Notch1 signaling was activated in NPC. Confocal images show that the Notch1 signaling activated form and Hes1, a downstream target of Notch1 signaling, was predominantly found in SOX2- and OCT4-positive cells. Conclusion: Our findings suggest that the Notch1 signaling pathway might be a regulator of cancer stem–like cells in NPC. PMID:20211102

  5. The relationship between leadership styles and empathy among student nurses.

    PubMed

    Gunther, Mary; Evans, Ginger; Mefford, Linda; Coe, Thomas R

    2007-01-01

    Much of the nursing literature on leadership describes the qualities of existing nursing leaders, while emphasizing the need for leadership development in student nurses for both managerial and clinical practice. However, there is a lack of research literature on the characteristics of current students. Conducted by the University of Tennessee College of Nursing Empathy Research Group, this pilot study explores the relationship between leadership styles and empathy (cognitive and affective) levels. This correlational descriptive study involved self-report using 3 instruments. Hogan Empathy Scale (HES) and Emotional Empathy Tendency Scale (EETS) measured cognitive and affective empathy levels. The Multifactoral Leadership Questionnaire (MLQ-5x) was used to determine leadership style. Data analysis yielded evidence of a weak positive correlation between the predominant transformational leadership style and empathy levels in both junior and senior students. This correlation has implications for both nurse educators and future employers. PMID:17678685

  6. U.S. hydropower resource assessment for Virginia

    SciTech Connect

    Conner, A.M.; Francfort, J.E.

    1997-12-01

    In June 1989, the US Department of Energy initiated the development of a National Energy Strategy to identify the energy resources available to support the expanding demand for energy in the US. Public hearings conducted as part of the strategy development process indicated that undeveloped hydropower resources were not well defined. As a result, the Department of Energy established an interagency Hydropower Resource Assessment Team to ascertain the undeveloped hydropower potential. In connection with these efforts by the Department of Energy, the Idaho National Engineering Laboratory designed the Hydropower Evaluation Software (HES), which has been used to perform a resource assessment of the undeveloped conventional hydropower potential in over 30 states. This report presents the results of the hydropower resource assessment for the State of Virginia. Undeveloped pumped storage hydropower potential is not included.

  7. Determination of sound velocities of "overcompressed" detonation in HMX-based explosive

    NASA Astrophysics Data System (ADS)

    Zhernokletov, Mikhail V.; Kovalev, Alexey E.; Bel'sky, Vladimir M.; Bogdanov, Evgeny N.

    2015-09-01

    The authors present results of determination of sound velocities in explosion products (EP) of HMX-based explosive overcompressed up to the pressures of 50-85 GPa by overtaking unloading method. The radiowave and optical methods are used to record the time when a front of overcompressed detonation wave in investigated sample of high explosive (HE) is overtaken by expansion wave, which propagates from the back surface of impactor with sound velocity. The data on sound velocities, which were independently obtained by two different methods, were in agreement. The methods with use of radiointerferometer and indicator liquid are rather effective for determination of sound velocities in overcompresed EP and for investigation of parameters at the Jouget point of various HEs, which are required for calibration of their equations of state (EOS).

  8. Id2 IS REQUIRED FOR SPECIFICATION OF DOPAMINERGIC NEURONS DURING ADULT OLFACTORY NEUROGENESIS

    PubMed Central

    Havrda, Matthew C.; Harris, Brent T.; Mantani, Akio; Ward, Nora M.; Paolella, Brenton R.; Cuzon, Verginia C.; Yeh, Hermes H.; Israel, Mark A.

    2009-01-01

    Understanding the biology of adult neural stem cells has important implications for nervous system development and may contribute to our understanding of neurodegenerative disorders and their treatment. We have characterized the process of olfactory neurogenesis in adult mice lacking Inhibitor of DNA Binding 2 (Id2). We found a diminished olfactory bulb containing reduced numbers of granular and periglomerular neurons with a distinct paucity of dopaminergic periglomerular neurons. While no deficiency of the stem cell compartment was detectable, migrating neuroblasts in Id2−/− mutant mice prematurely undergo astroglial differentiation within a disorganized rostral migratory stream. Further, when evaluated in vitro loss of Id2 results in decreased proliferation of neural progenitors and decreased expression of the Hes1 and Mash1 transcription factors, known mediators of neuronal differentiation. These data support a novel role for sustained Id2 expression in migrating neural progenitors mediating olfactory dopaminergic neuronal differentiation in adult animals. PMID:19109490

  9. Hey bHLH transcription factors.

    PubMed

    Weber, David; Wiese, Cornelia; Gessler, Manfred

    2014-01-01

    Hey bHLH transcription factors are direct targets of canonical Notch signaling. The three mammalian Hey proteins are closely related to Hes proteins and they primarily repress target genes by either directly binding to core promoters or by inhibiting other transcriptional activators. Individual candidate gene approaches and systematic screens identified a number of Hey target genes, which often encode other transcription factors involved in various developmental processes. Here, we review data on interaction partners and target genes and conclude with a model for Hey target gene regulation. Furthermore, we discuss how expression of Hey proteins affects processes like cell fate decisions and differentiation, e.g., in cardiovascular, skeletal, and neural development or oncogenesis and how this relates to the observed developmental defects and phenotypes observed in various knockout mice. PMID:25248480

  10. How I treat hypereosinophilic syndromes

    PubMed Central

    2015-01-01

    Hypereosinophilic syndromes (HESs) are a group of rare disorders characterized by peripheral blood eosinophilia of 1.5 × 109/L or higher and evidence of end organ manifestations attributable to the eosinophilia and not otherwise explained in the clinical setting. HESs are pleomorphic in clinical presentation and can be idiopathic or associated with a variety of underlying conditions, including allergic, rheumatologic, infectious, and neoplastic disorders. Moreover, the etiology of the eosinophilia in HESs can be primary (myeloid), secondary (lymphocyte-driven), or unknown. Although corticosteroids remain the first-line therapy for most forms of HESs, the availability of an increasing number of novel therapeutic agents, including tyrosine kinase inhibitors and monoclonal antibodies, has necessarily altered the approach to treatment of HESs. This review presents an updated treatment-based approach to the classification of patients with presumed HES and discusses the roles of conventional and novel agents in the management of these patients. PMID:25964669

  11. Downregulation of expression of mater genes SOX9, FOXA2, and GATA4 in pancreatic cancer cells stimulated with TGFβ1 epithelial-mesenchymal transition.

    PubMed

    Kondratyeva, L G; Sveshnikova, A A; Grankina, E V; Chernov, I P; Kopantseva, M R; Kopantzev, E P; Sverdlov, E D

    2016-07-01

    We show characteristic morphological changes corresponding to epithelial-mesenchymal transition (EMT) program fulfillment in PANC1 cell line stimulated with TGFβ1. Our results support downregulation of E-cadherin protein. We show 5- and 28-fold increase in SNAI1 and SNAI2 expression levels and 25- and 15-fold decrease in CDH1 and KRT8 expression levels, respectively, which confirms the EMT-program fulfillment. We demonstrate downregulation of expression of pancreatic master genes SOX9, FOXA2, and GATA4 (2-, 5-, and 4-fold, respectively) and absence of significant changes in HES1, NR5A2, and GATA6 expression levels in the cells stimulated with TGFβ1. Our results indicate the absence of induction of expression of PTF1A, PDX1, HNF1b, NEUROG3, RPBJL, NKX6.1, and ONECUT1 genes, which are inactive in PANC1 cell line after the EMT stimulated by TGFβ1. PMID:27599506

  12. Genome editing and the next generation of antiviral therapy.

    PubMed

    Stone, Daniel; Niyonzima, Nixon; Jerome, Keith R

    2016-09-01

    Engineered endonucleases such as homing endonucleases (HEs), zinc finger nucleases (ZFNs), Tal-effector nucleases (TALENS) and the RNA-guided engineered nucleases (RGENs or CRISPR/Cas9) can target specific DNA sequences for cleavage, and are proving to be valuable tools for gene editing. Recently engineered endonucleases have shown great promise as therapeutics for the treatment of genetic disease and infectious pathogens. In this review, we discuss recent efforts to use the HE, ZFN, TALEN and CRISPR/Cas9 gene-editing platforms as antiviral therapeutics. We also discuss the obstacles facing gene-editing antiviral therapeutics as they are tested in animal models of disease and transition towards human application. PMID:27272125

  13. Organization of monitoring the state of structures at the Sayano-Shushenskoe Hydroelectric Station

    SciTech Connect

    Bryzgalov, V.I.; Stafievskii, V.A.

    1994-06-01

    Engineering art when reference is to the reliability of an engineering structure should not depend on the political situation in the country. For example, in stagnant years it was decided to replace the type of dam at the Krasnoyarsk hydroelectric station (HES). Despite the insistence of the design organization for a so-called buttress dam, the engineering corps of like-minded persons - the construction engineers and customer headed by the chief of the construction project - categorically objected to the proposed design: in the construction industry the technology was not ready for openwork construction under harsh climatic conditions, and, what really matters, millions of people living downstream of the dam should be safeguarded against disasters. As a result a gravity dam was adopted.

  14. Cool Carbon Stars in the Galactic Halo

    NASA Astrophysics Data System (ADS)

    Gigoyan, K. S.

    2016-06-01

    In this paper we report current status of search and study for Faint High Latitude Carbon Stars (FHLCs). Data for more than 1800 spectroscopically confirmed FHLCs are known, which are found thanks to objective prism surveys and photometric selections. More than half of the detected objects belongs to group of dwarf Carbon (dC) stars. Many-sided investigations based on modern astrophysical databases are necessary to study the space distribution of different groups of the FHLC stars and their possible origin in the Halo of our Galaxy. We report about the selection of FHLCs by the spectroscopic surveys: First Byurakan Survey (FBS), Hamburg/ESO Survey (HES), LAMOST Pilot Survey and SDSS, as well as by photometric selection: APM Survey for Cool Carbon Stars in the Galactic Halo, SDSS and 2MASS JHK colours.

  15. Hawaii Energy Strategy program. [First Annual Report

    SciTech Connect

    Not Available

    1992-01-01

    The Hawaii Energy Strategy (HES) program began on March 2, 1992, under United States Department of Energy Cooperative Agreement DE-FC03-92F19168, and is scheduled for completion by December 31, 1994. As outlined in the Statement of Joint Objectives: The purpose of the study is to develop an integrated State of Hawaii energy strategy, including an assessment of the State's fossil fuel reserve requirements and the most effective way to meet those needs, the availability and practicality of increasing the use of native energy resources, potential alternative fossil energy technologies such as coal gasification and potential energy efficiency measures which could lead to demand reduction. This work contributes to the DOE mission, will reduce the State's vulnerability to energy supply disruptions and contributes to the public good.

  16. Hawaii Energy Strategy program. Annual report, 1992

    SciTech Connect

    Not Available

    1992-12-31

    The Hawaii Energy Strategy (HES) program began on March 2, 1992, under United States Department of Energy Cooperative Agreement DE-FC03-92F19168, and is scheduled for completion by December 31, 1994. As outlined in the Statement of Joint Objectives: The purpose of the study is to develop an integrated State of Hawaii energy strategy, including an assessment of the State`s fossil fuel reserve requirements and the most effective way to meet those needs, the availability and practicality of increasing the use of native energy resources, potential alternative fossil energy technologies such as coal gasification and potential energy efficiency measures which could lead to demand reduction. This work contributes to the DOE mission, will reduce the State`s vulnerability to energy supply disruptions and contributes to the public good.

  17. NOTCH1 inhibition enhances the efficacy of conventional chemotherapeutic agents by targeting head neck cancer stem cell

    PubMed Central

    Zhao, Zhi-Li; Zhang, Lu; Huang, Cong-Fa; Ma, Si-Rui; Bu, Lin-Lin; Liu, Jian-Feng; Yu, Guang-Tao; Liu, Bing; Gutkind, J. Silvio; Kulkarni, Ashok B.; Zhang, Wen-Feng; Sun, Zhi-Jun

    2016-01-01

    Cancer stem cells (CSCs) are considered responsible for tumor initiation and chemoresistance. This study was aimed to investigate the possibility of targeting head neck squamous cell carcinoma (HNSCC) by NOTCH1 pathway inhibition and explore the synergistic effect of combining NOTCH inhibition with conventional chemotherapy. NOTCH1/HES1 elevation was found in human HNSCC, especially in tissue post chemotherapy and lymph node metastasis, which is correlated with CSCs markers. NOTCH1 inhibitor DAPT (GSI-IX) significantly reduces CSCs population and tumor self-renewal ability in vitro and in vivo. Flow cytometry analysis showed that NOTCH1 inhibition reduces CSCs frequency either alone or in combination with chemotherapeutic agents, namely, cisplatin, docetaxel, and 5-fluorouracil. The combined strategy of NOTCH1 blockade and chemotherapy synergistically attenuated chemotherapy-enriched CSC population, promising a potential therapeutic exploitation in future clinical trial. PMID:27108536

  18. Long-Term Quiescent Fibroblast Cells Transit into Senescence

    PubMed Central

    Marthandan, Shiva; Priebe, Steffen; Hemmerich, Peter; Klement, Karolin; Diekmann, Stephan

    2014-01-01

    Cellular senescence is described to be a consequence of telomere erosion during the replicative life span of primary human cells. Quiescence should therefore not contribute to cellular aging but rather extend lifespan. Here we tested this hypothesis and demonstrate that cultured long-term quiescent human fibroblasts transit into senescence due to similar cellular mechanisms with similar dynamics and with a similar maximum life span as proliferating controls, even under physiological oxygen conditions. Both, long-term quiescent and senescent fibroblasts almost completely fail to undergo apoptosis. The transition of long-term quiescent fibroblasts into senescence is also independent of HES1 which protects short-term quiescent cells from becoming senescent. Most significantly, DNA damage accumulates during senescence as well as during long-term quiescence at physiological oxygen levels. We suggest that telomere-independent, potentially maintenance driven gradual induction of cellular senescence during quiescence is a counterbalance to tumor development. PMID:25531649

  19. Physical growth of St. Lawrence Island Eskimos: body size, proportion, and composition.

    PubMed

    Johnston, F E; Laughlin, W S; Harper, A B; Ensroth, A E

    1982-08-01

    Growth patterns of body size, proportion, and composition were analyzed in 57 male and 56 female Eskimos from St. Lawrence Island in the Bering Sea, ranging in age from 1.23 through 19.82 years. Age-groups means for whites and blacks of the U.S. Health Examination Survey served as reference data. Relative to HES data, the Eskimo sample were shorter with lower values for leg length, while there were no differences from the reference values for sitting height. The Eskimos also had higher values of Quetelet's Index, the sitting height/height ratio, and the upper arm muscle circumference, while there were no differences in body weight or triceps skinfold thickness. Differences from the reference data were more pronounced in males than in females. The growth patterns for size and body proportion are in conformity with known relationships between morphology and climate. PMID:7124933

  20. U.S. hydropower resource assessment for Maine

    SciTech Connect

    Francfort, J.E.; Rinehart, B.N.

    1995-07-01

    The Department of Energy is developing an estimate of the undeveloped hydro-power potential in the United States. The Hydropower Evaluation Software (HES) is a computer model that was developed by the Idaho National Engineering Laboratory for this purpose. The software measures the undeveloped hydropower resources available in the United States, using uniform criteria for measurement. The software was developed and tested using hydropower information and data provided by the Southwestern Power Administration. It is a menu-driven software program that allows the personal computer user to assign environmental attributes to potential hydropower sites, calculate development suitability factors for each site based on the environmental attributes present, and generate reports based on these suitability factors. This report details the resource assessment results for the State of Maine.

  1. Chemical Equilibrium Detonation

    NASA Astrophysics Data System (ADS)

    Bastea, Sorin; Fried, Laurence E.

    Energetic materials are unique for having a strong exothermic reactivity, which has made them desirable for both military and commercial applications. The fundamental principles outlined in this chapter pertain to the study of detonation in both gas-phase and condensed-phase energetic materials, but our main focus will be on the condensed ones, particularly on high explosives (HEs). They share many properties with other classes of condensed energetic compounds such as propellants and pyrotechnics, but a detailed understanding of detonation is especially important for numerous HE applications. The usage and study of HE materials goes back more than a century, but many questions remain to be answered, e.g., on their reaction pathways at high pressures and temperatures, chemical properties, etc.

  2. Honokiol inhibits melanoma stem cells by targeting notch signaling.

    PubMed

    Kaushik, Gaurav; Venugopal, Anand; Ramamoorthy, Prabhu; Standing, David; Subramaniam, Dharmalingam; Umar, Shahid; Jensen, Roy A; Anant, Shrikant; Mammen, Joshua M V

    2015-12-01

    Melanoma is an aggressive disease with limited therapeutic options. Here, we determined the effects of honokiol (HNK), a biphenolic natural compound on melanoma cells and stemness. HNK significantly inhibited melanoma cell proliferation, viability, clonogenicity and induced autophagy. In addition, HNK significantly inhibited melanosphere formation in a dose dependent manner. Western blot analyses also demonstrated reduction in stem cell markers CD271, CD166, Jarid1b, and ABCB5. We next examined the effect of HNK on Notch signaling, a pathway involved in stem cell self-renewal. Four different Notch receptors exist in cells, which when cleaved by a series of enzymatic reactions catalyzed by Tumor Necrosis Factor-α-Converting Enzyme (TACE) and γ-secretase protein complex, results in the release of the Notch intracellular domain (NICD), which then translocates to the nucleus and induces target gene expression. Western blot analyses demonstrated that in HNK treated cells there is a significant reduction in the expression of cleaved Notch-2. In addition, there was a reduction in the expression of downstream target proteins, Hes-1 and cyclin D1. Moreover, HNK treatment suppressed the expression of TACE and γ-secretase complex proteins in melanoma cells. To confirm that suppression of Notch-2 activation is critical for HNK activity, we overexpressed NICD1, NICD2, and performed HNK treatment. NICD2, but not NICD1, partially restored the expression of Hes-1 and cyclin D1, and increased melanosphere formation. Taken together, these data suggest that HNK is a potent inhibitor of melanoma cells, in part, through the targeting of melanoma stem cells by suppressing Notch-2 signaling. PMID:25491779

  3. Fluxgate magnetorelaxometry: a new approach to study the release properties of hydrogel cylinders and microspheres.

    PubMed

    Wöhl-Bruhn, S; Heim, E; Schwoerer, A; Bertz, A; Harling, S; Menzel, H; Schilling, M; Ludwig, F; Bunjes, H

    2012-10-15

    Hydrogels are under investigation as long term delivery systems for biomacromolecules as active pharmaceutical ingredients. The release behavior of hydrogels can be tailored during the fabrication process. This study investigates the applicability of fluxgate magnetorelaxometry (MRX) as a tool to characterize the release properties of such long term drug delivery depots. MRX is based on the use of superparamagnetic core-shell nanoparticles as model substances. The feasibility of using superparamagnetic nanoparticles to study the degradation of and the associated release from hydrogel cylinders and hydrogel microspheres was a major point of interest. Gels prepared from two types of photo crosslinkable polymers based on modified hydroxyethylstarch, specifically hydroxyethyl starch-hydroxyethyl methacrylate (HES-HEMA) and hydroxyethyl starch-polyethylene glycol methacrylate (HES-P(EG)(6)MA), were analyzed. MRX analysis of the incorporated nanoparticles allowed to evaluate the influence of different crosslinking conditions during hydrogel production as well as to follow the increase in nanoparticle mobility as a result of hydrogel degradation during release studies. Conventional release studies with fluorescent markers (half-change method) were performed for comparison. MRX with superparamagnetic nanoparticles as model substances is a promising method to analyze pharmaceutically relevant processes such as the degradation of hydrogel drug carrier systems. In contrast to conventional release experiments MRX allows measurements in closed vials (reducing loss of sample and sampling errors), in opaque media and at low magnetic nanoparticle concentrations. Magnetic markers possess a better long-term stability than fluorescent ones and are thus also promising for the use in in vivo studies. PMID:22820133

  4. The matrix metalloproteinase inhibitor marimastat promotes neural progenitor cell differentiation into neurons by gelatinase-independent TIMP-2-dependent mechanisms.

    PubMed

    Sinno, Maddalena; Biagioni, Stefano; Ajmone-Cat, Maria Antonietta; Pafumi, Irene; Caramanica, Pasquale; Medda, Virginia; Tonti, Gaetana; Minghetti, Luisa; Mannello, Ferdinando; Cacci, Emanuele

    2013-02-01

    Metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs), produced in the brain by cells of non-neural and neural origin, including neural progenitors (NPs), are emerging as regulators of nervous system development and adult brain functions. In the present study, we explored whether MMP-2, MMP-9, and TIMP-2, abundantly produced in the brain, modulate NP developmental properties. We found that treatment of NPs, isolated from the murine fetal cerebral cortex or adult subventricular zone, with the clinically tested broad-spectrum MMP inhibitor Marimastat profoundly affected the NP differentiation fate. Marimastat treatment allowed for an enrichment of our cultures in neuronal cells, inducing NPs to generate higher percentage of neurons and a lower percentage of astrocytes, possibly affecting NP commitment. Consistently with its proneurogenic effect, Marimastat early downregulated the expression of Notch target genes, such as Hes1 and Hes5. MMP-2 and MMP-9 profiling on proliferating and differentiating NPs revealed that MMP-9 was not expressed under these conditions, whereas MMP-2 increased in the medium as pro-MMP-2 (72 kDa) during differentiation; its active form (62 kDa) was not detectable by gel zymography. MMP-2 silencing or administration of recombinant active MMP-2 demonstrated that MMP-2 does not affect NP neuronal differentiation, nor it is involved in the Marimastat proneurogenic effect. We also found that TIMP-2 is expressed in NPs and increases during late differentiation, mainly as a consequence of astrocyte generation. Endogenous TIMP-2 did not modulate NP neurogenic potential; however, the proneurogenic action of Marimastat was mediated by TIMP-2, as demonstrated by silencing experiments. In conclusion, our data exclude a major involvement of MMP-2 and MMP-9 in the regulation of basal NP differentiation, but highlight the ability of TIMP-2 to act as key effector of the proneurogenic response to an inducing stimulus such as Marimastat. PMID

  5. Characterization of Notch Signaling During Osteogenic Differentiation in Human Osteosarcoma Cell Line MG63.

    PubMed

    Ongaro, Alessia; Pellati, Agnese; Bagheri, Leila; Rizzo, Paola; Caliceti, Cristiana; Massari, Leo; De Mattei, Monica

    2016-12-01

    Osteogenic differentiation is a multi-step process controlled by a complex molecular framework. Notch is an evolutionarily conserved intercellular signaling pathway playing a prominent role in cell fate and differentiation, although the mechanisms by which this pathway regulates osteogenesis remain controversial. This study aimed to investigate, in vitro, the involvement of Notch pathway during all the developmental stages of osteogenic differentiation in human osteosarcoma cell line MG63. Cells were cultured in basal condition (control) and in osteoinductive medium (OM). Notch inhibitors were also added in OM to block Notch pathway. During osteogenic differentiation, early (alkaline phosphatase activity and collagen type I) and late osteogenic markers (osteocalcin levels and matrix mineralization), as well as the gene expression of the main osteogenic transcription factors (Runx2, Osterix, and Dlx5) increased. Time dependent changes in the expression of specific Notch receptors were identified in OM versus control with a significant reduction in the expression of Notch1 and Notch3 receptors in the early phase of differentiation, and an increase of Notch2 and Notch4 receptors in the late phase. Among Notch nuclear target genes, Hey1 expression was significantly higher in OM than control, while Hes5 expression decreased. Osteogenic markers were reduced and Hey1 was significantly inhibited by Notch inhibitors, suggesting a role for Notch through the canonical pathway. In conclusion, Notch pathway might be involved with a dual role in osteogenesis of MG63, through the activation of Notch2, Notch4, and Hey1, inducing osteoblast differentiation and the depression of Notch1, Notch3, and Hes5, maintaining an undifferentiated status. J. Cell. Physiol. 231: 2652-2663, 2016. © 2016 Wiley Periodicals, Inc. PMID:26946465

  6. A standardized technique for efficient platelet and leukocyte collection using the Model 30 Blood Processor.

    PubMed

    Aisner, J; Schiffer, C A; Wolff, J H; Wiernik, P H

    1976-01-01

    The Model 30 Blood Processor is a safe and simple means of harvesting blood cell components. Presently cell collection depends on a visual assessment by the operator of the indistinct boundaries of cell fractions. To determine when each cell component could best be harvested, serial samples were taken from the output port at fixed intervals anf the results of counts and differentials were graphed and tabulated. Studies in normal donors were done using acid-citrate-dextrose (ACD), 2 per cent sodium citrate in 6 per cent hydroxyethyl starch (HES), or heparin as anticoagulants. There was considerable overlap between the latter part of the platelet band, the leukocyte band and the rising hematocrit with all three anticoagulants. Normally functional lymphocytes could be harvested efficiently (approximately 80%) using ACD or heparin. Platelets could be harvested from ACD very efficiently (approximately 90%). Granulocytes could not be harvested from ACD (less than 10%) since they were dispersed in the red blood cell (RBC) layer. Using HES, granulocytes could be harvested efficiently (approximately 70%) by extending collection into the RBC layer. Based on these data, a standard technique for cell collection has been devised. The flow rate is slowed to 20 ml/min and collection is carried 30 ml (90 seconds at a rate of 20 ml/min) for platelets. The RBC loss is approximately 6 to 8 and 2 to 3 ml/pass respectively. These studies indicate that the Model 30 is a highly efficient apparatus for blood cell separation, but the volume of blood processed is limited by the intermittent blood flow. PMID:62425

  7. Principle of Spacetime and Black Hole Equivalence

    NASA Astrophysics Data System (ADS)

    Zhang, Tianxi

    2016-06-01

    Modelling the universe without relying on a set of hypothetical entities (HEs) to explain observations and overcome problems and difficulties is essential to developing a physical cosmology. The well-known big bang cosmology, widely accepted as the standard model, stands on two fundamentals, which are Einstein’s general relativity (GR) that describes the effect of matter on spacetime and the cosmological principle (CP) of spacetime isotropy and homogeneity. The field equation of GR along with the Friedmann-Lemaitre-Robertson-Walker (FLRW) metric of spacetime derived from CP generates the Friedmann equation (FE) that governs the development and dynamics of the universe. The big bang theory has made impressive successes in explaining the universe, but still has problems and solutions of them rely on an increasing number of HEs such as inflation, dark matter, dark energy, and so on. Recently, the author has developed a new cosmological model called black hole universe, which, instead of making many those hypotheses, only includes a new single postulate (or a new principle) to the cosmology - Principle of Spacetime and Black Hole Equivalence (SBHEP) - to explain all the existing observations of the universe and overcome all the existing problems in conventional cosmologies. This study thoroughly demonstrates how this newly developed black hole universe model, which therefore stands on the three fundamentals (GR, CP, and SBHEP), can fully explain the universe as well as easily conquer the difficulties according to the well-developed physics, thus, neither needing any other hypotheses nor existing any unsolved difficulties. This work was supported by NSF/REU (Grant #: PHY-1263253) at Alabama A & M University.

  8. Notch3 and Hey-1 as Prognostic Biomarkers in Pancreatic Adenocarcinoma

    PubMed Central

    Mann, Christopher D.; Bastianpillai, Christopher; Neal, Christopher P.; Masood, Muhammad M.; Jones, Donald J. L.; Teichert, Friederike; Singh, Rajinder; Karpova, Elena; Berry, David P.; Manson, Margaret M.

    2012-01-01

    In order to achieve a better outcome for pancreatic cancer patients, reliable biomarkers are required which allow for improved diagnosis. These may emanate from a more detailed molecular understanding of the aggressive nature of this disease. Having previously reported that Notch3 activation appeared to be associated with more aggressive disease, we have now examined components of this pathway (Notch1, Notch3, Notch4, HES-1, HEY-1) in more detail in resectable (n = 42) and non-resectable (n = 50) tumours compared to uninvolved pancreas. All three Notch family members were significantly elevated in tumour tissue, compared to uninvolved pancreas, with expression maintained within matched lymph node metastases. Furthermore, significantly higher nuclear expression of Notch1, -3 and -4, HES-1, and HEY-1 (all p≤0.001) was noted in locally advanced and metastatic tumours compared to resectable cancers. In survival analyses, nuclear Notch3 and HEY-1 expression were significantly associated with reduced overall and disease-free survival following tumour resection with curative intent, with nuclear HEY-1 maintaining independent prognostic significance for both outcomes on multivariate analysis. These data further support a central role for Notch signalling in pancreatic cancer and suggest that nuclear expression of Notch3 and its target gene, HEY-1, merit validation in biomarker panels for diagnosis, prognosis and treatment efficacy. A peptide fragment of Notch3 was detected in plasma from patients with inoperable pancreatic cancer, but due to wide inter-individual variation, mean levels were not significantly different compared to age-matched controls. PMID:23226563

  9. Defects in subventricular zone pigmented epithelium-derived factor niche signaling in the senescence-accelerated mouse prone-8.

    PubMed

    Castro-Garcia, Paola; Díaz-Moreno, María; Gil-Gas, Carmen; Fernández-Gómez, Francisco J; Honrubia-Gómez, Paloma; Álvarez-Simón, Carmen Belén; Sánchez-Sánchez, Francisco; Cano, Juan Carlos Castillo; Almeida, Francisco; Blanco, Vicente; Jordán, Joaquín; Mira, Helena; Ramírez-Castillejo, Carmen

    2015-04-01

    We studied potential changes in the subventricular zone (SVZ) stem cell niche of the senescence-accelerated mouse prone-8 (SAM-P8) aging model. Bromodeoxyuridine (BrdU) assays with longtime survival revealed a lower number of label-retaining stem cells in the SAM-P8 SVZ compared with the SAM-Resistant 1 (SAM-R1) control strain. We also found that in SAM-P8 niche signaling is attenuated and the stem cell pool is less responsive to the self-renewal niche factor pigmented epithelium-derived factor (PEDF). Protein analysis demonstrated stable amounts of the PEDF ligand in the SAM-P8 SVZ niche; however, SAM-P8 stem cells present a significant expression decrease of patatin-like phospholipase domain containing 2, a receptor for PEDF (PNPLA2-PEDF) receptor, but not of laminin receptor (LR), a receptor for PEDF (LR-PEDF) receptor. We observed changes in self-renewal related genes (hairy and enhancer of split 1 (Hes1), hairy and enhancer of split 1 (Hes5), Sox2] and report that although these genes are down-regulated in SAM-P8, differentiation genes (Pax6) are up-regulated and neurogenesis is increased. Finally, sheltering mammalian telomere complexes might be also involved given a down-regulation of telomeric repeat binding factor 1 (Terf1) expression was observed in SAM-P8 at young age periods. Differences between these 2 models, SAM-P8 and SAM-R1 controls, have been previously detected at more advanced ages. We now describe alterations in the PEDF signaling pathway and stem cell self-renewal at a very young age, which could be involved in the premature senescence observed in the SAM-P8 model. PMID:25636741

  10. Biodegradation of high explosives on granular activated carbon [GAC]: Enhanced desorption of high explosives from GAC -- Batch studies

    SciTech Connect

    Morley, M.C.; Speitel, G.E. Jr.

    1999-03-01

    Adsorption to GAC is an effective method for removing high explosives (HE) compounds from water, but no permanent treatment is achieved. Bioregeneration, which treats adsorbed contaminants by desorption and biodegradation, is being developed as a method for reducing GAC usage rates and permanently degrading RDX and HMX. Because desorption is often the limiting mass transfer mechanism in bioregeneration systems, several methods for increasing the rate and extent of desorption of RDX and HMX are being studied. These include use of cosolvents (methanol and ethanol), surfactants (both anionic and nonionic), and {beta}- and {gamma}-cyclodextrins. Batch experiments to characterize the desorption of these HEs from GAC have been completed using Northwestern LB-830, the GAC being used at Pantex. Over a total of 11 days of desorption, about 3% of the adsorbed RDX was desorbed from the GAC using buffered water as the desorption fluid. In comparison, about 96% of the RDX was extracted from the GAC by acetonitrile over the same desorption period. Ethanol and methanol were both effective in desorbing RDX and HMX; higher alcohol concentrations were able to desorb more HE from the GAC. Surfactants varied widely in their abilities to enhance desorption of HEs. The most effective surfactant that was studied was sodium dodecyl sulfate (SDS), which desorbed 56.4% of the adsorbed RDX at a concentration of 500 mg SDS/L. The cyclodextrins that were used were marginally more effective than water. Continuous-flow column tests are underway for further testing the most promising of these methods. These results will be compared to column experiments that have been completed under baseline conditions (using buffered water as the desorption fluid). Results of this research will support modeling and design of further desorption and bioregeneration experiments.

  11. Single-cell mRNA profiling identifies progenitor subclasses in neurospheres.

    PubMed

    Narayanan, Gunaseelan; Poonepalli, Anuradha; Chen, Jinmiao; Sankaran, Shvetha; Hariharan, Srivats; Yu, Yuan Hong; Robson, Paul; Yang, Henry; Ahmed, Sohail

    2012-12-10

    Neurospheres are widely used to propagate and investigate neural stem cells (NSCs) and neural progenitors (NPs). However, the exact cell types present within neurospheres are still unknown. To identify cell types, we used single-cell mRNA profiling of 48 genes in 187 neurosphere cells. Using a clustering algorithm, we identified 3 discrete cell populations within neurospheres. One cell population [cluster unsorted (US) 1] expresses high Bmi1 and Hes5 and low Myc and Klf12. Cluster US2 shows intermediate expression of most of the genes analyzed. Cluster US3 expresses low Bmi1 and Hes5 and high Myc and Klf12. The mRNA profiles of these 3 cell populations correlate with a developmental timeline of early, intermediate, and late NPs, as seen in vivo from the mouse brain. We enriched the cell population for neurosphere-forming cells (NFCs) using morphological criteria of forward scatter (FSC) and side scatter (SSC). FSC/SSC(high) cells generated 2.29-fold more neurospheres than FSC/SSC(low) cells at clonal density. FSC/SSC(high) cells were enriched for NSCs and Lewis-X(+ve) cells, possessed higher phosphacan levels, and were of a larger cell size. Clustering of both FSC/SSC(high) and FSC/SSC(low) cells identified an NFC cluster. Significantly, the mRNA profile of the NFC cluster drew close resemblance to that of early NPs. Taken together, data suggest that the neurosphere culture system can be used to model central nervous system development, and that early NPs are the cell population that gives rise to neurospheres. In future work, it may be possible to further dissect the NFCs and reveal the molecular signature for NSCs. PMID:22834539

  12. Energy supplementation and productivity of Guatemalan sugar-cane cutters: a longitudinal approach.

    PubMed

    Immink, M D; Blake, C C; Viteri, F E; Flores, R; Torún, B

    1986-06-01

    A long-term energy supplementation program was carried out to determine its effect on the productivity of agricultural workers in Guatemala. The program provided, free of charge, a low-energy (24 Kcal) and a high-energy (350 Kcal) bottled, orange-flavored soft drink to two groups of long-term resident sugar-cane cutters who worked on the same plantation, located in the Pacific Coast. Previous to, and periodically thereafter during implementation of the program, data relative to energy intake and anthropometry were collected. Through data obtained from payroll lists, a longitudinal series of average productivity (tons of sugar cane cut and loaded per day) covering 48 weeks of pre-supplementation, 90 weeks of supplementation and 21 weeks post-supplementation, was constructed. Control of the supplement consumption was daily observed. Random assignment of workers to the high-energy supplement (HES) and the low-energy (LES) groups, was not possible. Prior to supplementation both groups presented the same characteristics in terms of age, energy intake level, weight, height, tricipital adiposity and daily productivity. Little variation was found throughout the time the supplement was consumed by the HES Group. Energy intake of workers increased significantly in absolute terms in relation to the LES Group, except towards the end of the 28 months' supplementation period. Energy balance was maintained by workers throughout the study period. A time series of the difference in mean productivity of the two supplement groups (Yt) was modeled using the ARIMA techniques. No auto-regressive term was present in the Yt series. The ARIMA (0,0,1) model was fitted and expanded with different intervention components. None of the estimated parameters of the intervention components were statistically significant. It was therefore concluded that no abrupt, or gradual and sustained energy supplementation effect on productivity was present. PMID:3632204

  13. Involvement of Numb-mediated HIF-1α inhibition in anti-proliferative effect of PNA-antimiR-182 in trastuzumab-sensitive and -resistant SKBR3 cells.

    PubMed

    Sajadimajd, Soraya; Yazdanparast, Razieh; Akram, Sadeghirizi

    2016-04-01

    Trastuzumab is a humanized monoclonal antibody against the human epidermal growth factor receptor 2 (HER2) that is overexpressed in about 25 % of breast cancer patients. However, primary and/or acquired resistance to trastuzumab develops in most affected persons. In this study, we explored the functional role of miR-182 inhibition with aiming the sensitization of SKBR3 cells to trastuzumab. Cell viability, apoptosis, colony formation, and migration capacities of SKBR3(S) (sensitive) and SKBR3(R) (resistant) cells were assessed to determine the anti-proliferative effects of PNA-antimiR-182. In addition, the expression levels of miR-182, mRNA of FOXO1, and Bim as well as the protein levels of HER2 and Notch1 signaling factors were evaluated by stem-loop RT-qPCR, RT-qPCR, and Western blot, respectively. The results indicated that miR-182 might play a causal role in the mechanism of trastuzumab. In line with that, PNA-antimiR-182 inhibited synergistically the viability of both the sensitive and resistant cell groups. Furthermore, the inhibitory effect of PNA-anitmiR-182 on migration in SKBR3 cells was more than the induction of apoptosis. In addition, PNA-antimiR-182 reduced the levels of NICD, Hes1, HIF-1α, and p-Akt in both cell groups, while it augmented the intracellular content of FOXO1 and Numb suppressor proteins. In other words, PNA-antimiR-182-mediated upregulation of Numb was associated with downregulation of HIF-1α and Hes1. Consequently, downregulation of miR-182 might find therapeutical value for overcoming trastuzumab resistance. Graphical Abstract The crosstalk between HER2 and Notch1 signaling pathway is mediated by miR-182. PMID:26563369

  14. Height and critical frequency variations of the sporadic-E layer at midlatitudes

    NASA Astrophysics Data System (ADS)

    Šauli, P.; Bourdillon, A.

    2008-12-01

    The present study concerns variations in height and critical frequency of sporadic E layer over a wide period range of hours to several days, covering tidal and planetary oscillation domain. Besides periodicities in the tidal and planetary range that are known to occur within time series of critical frequencies (foEs) of sporadic-E layer [Pancheva, D., Haldoupis, C., Meek, C.E., Manson, A.H., Mitchell, N.J., 2003. Evidence of a role for modulated atmospheric tides in the dependence of sporadic E layers on planetary waves. Journal of Geophysical Research 108, Art. No. 1176; Haldoupis, C., Pancheva, D., Michell, N.J., 2004. A study of tidal and planetary wave periodicities present in midlatitude sporadic E layers. Journal of Geophysical Research 109, Art. No. A02302] among others, we evidence the existence of the 4-day planetary wave well developed in the height of sporadic E time serie (hEs). Moreover it is shown that the central-period of the diurnal tidal component of hEs is not exactly 24 h but it varies between 22 and 26 h at the planetary wave period. At a first glance, this is surprising since the origin of the diurnal tide is a forced oscillation with a 24 h period due to the Earth rotation and the periodic heating of the atmosphere by the Sun. Our interpretation is based on the perturbation of the height of the Es layer imposed by the planetary wave. In this mechanism the Es layer is moved up and down by the planetary wave producing a Doppler effect and resulting in a shift of the central-period around 24 h. With this interpretation, the excursion of the central-period is related to the vertical velocity perturbation of the Es layer due to the planetary wave. For a central period varying between 22 and 26 h the peturbation velocities are 0.026 and , respectively.

  15. The mobilization of hazardous elements after a tropical storm event in a polluted estuary.

    PubMed

    Rodriguez-Iruretagoiena, Azibar; de Vallejuelo, Silvia Fdez-Ortiz; de Diego, Alberto; de Leão, Felipe B; de Medeiros, Diego; Oliveira, Marcos L S; Tafarel, Silvio R; Arana, Gorka; Madariaga, Juan Manuel; Silva, Luis F O

    2016-09-15

    The Tubarão River (Santa Catarina, Brazil) is affected by hazardous elements (HEs) pollution from abandoned coal mines, agricultural activities, urban discharges, industrial and leisure zones, etc. In order to study the distribution and sources of HEs contamination in a polluted estuary after a tropical storm, waters and surface sediments were collected from 15 sampling sites along the Tubarão River. The concentration of 24 elements (Ag, Al, As, Ba, Cd, Co, Cr, Cu, Fe, Hg, Mg, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Ti, Tl, V, W, and Zn) were measured by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and the mineralogical composition of the sediments by Raman spectroscopy and X-ray diffraction (XRD). The metal concentrations in water and sediment showed wide spatial variation due to the variability in water discharges and anthropogenic inputs after a storm. In general, higher metal concentration in water and lower in sediments were found upstream (closer to coal mining). Downstream sampling sites and the Oratorio River sampling site (one of the eight tributaries of the estuary) showed the highest values in sediment samples. Normalized and Weighed Average Concentrations (NWAC) were calculated, which allow us to identify, in a very simple way, the sampling sites of higher concern (hotspots of contamination) in the studied area. NWAC suggested that the strong rainfall events could affect to the metal distribution in sediments. The results of this study were compared with a previous study in the same area during dry season by Principal Component Analysis (PCA), showing changes in environmental pollution of the sediment after a strong storm event. PMID:27232719

  16. Quality measures for total ankle replacement, 30-day readmission and reoperation rates within 1 year of surgery: a data linkage study using the NJR data set

    PubMed Central

    Zaidi, Razi; Macgregor, Alexander J; Goldberg, Andy

    2016-01-01

    Objective To report on the rate of 30-day readmission and the rate of additional or revision surgery within 12 months following total ankle replacement (TAR). Design A data-linkage study of the UK National Joint Registry (NJR) data and Hospital Episodes Statistics (HES) database. These two databases were linked in a deterministic fashion. HES episodes 12 months following the index procedure were isolated and analysed. Logistic regression was used to model predictors of reoperation and revision for primary ankle replacement. Participants All patients who underwent primary and revision ankle replacements according to the NJR between February 2008 and February 2013. Results The rate of 30-day readmission following primary and revision ankle replacement was 2.2% and 1.3%, respectively. In the 12 months following primary and revision ankle replacements, the revision rate (where implants needed to be removed) was 1.2% with increased odds in those orthopaedic units preforming <20 ankle replacements per year and patients with a preoperative fixed equinus deformity. The reoperation other than revision (where implants were not removed) in the 12 months following primary and revision TARs was 6.6% and 9.3%, respectively. Rheumatoid arthritis, cemented prosthesis and high ASA grade significantly increased the odds of reoperation. Conclusions TAR has a 30-day readmission rate of 2.2%, which is similar to that of knee replacement but lower than that of total hip replacement. 6.6% of patients undergoing primary TAR require a reoperation within 12 months of the index procedure. Early revision rates are significantly higher in low-volume centres. PMID:27217286

  17. Neurotrophins Acting Via TRKB Receptors Activate the JAGGED1-NOTCH2 Cell-Cell Communication Pathway to Facilitate Early Ovarian Development

    PubMed Central

    Dorfman, Mauricio D.; Kerr, Bredford; Garcia-Rudaz, Cecilia; Paredes, Alfonso H.; Dissen, Gregory A.

    2011-01-01

    Tropomyosin-related kinase (TRK) receptor B (TRKB) mediates the supportive actions of neurotrophin 4/5 and brain-derived neurotrophic factor on early ovarian follicle development. Absence of TRKB receptors reduces granulosa cell (GC) proliferation and delays follicle growth. In the present study, we offer mechanistic insights into this phenomenon. DNA array and quantitative PCR analysis of ovaries from TrkB-null mice revealed that by the end of the first week of postnatal life, Jagged1, Hes1, and Hey2 mRNA abundance is reduced in the absence of TRKB receptors. Although Jagged1 encodes a NOTCH receptor ligand, Hes1 and Hey2 are downstream targets of the JAGGED1-NOTCH2 signaling system. Jagged1 is predominantly expressed in oocytes, and the abundance of JAGGED1 is decreased in TrkB−/− oocytes. Lack of TRKB receptors also resulted in reduced expression of c-Myc, a NOTCH target gene that promotes entry into the cell cycle, but did not alter the expression of genes encoding core regulators of cell-cycle progression. Selective restoration of JAGGED1 synthesis in oocytes of TrkB−/− ovaries via lentiviral-mediated transfer of the Jagged1 gene under the control of the growth differentiation factor 9 (Gdf9) promoter rescued c-Myc expression, GC proliferation, and follicle growth. These results suggest that neurotrophins acting via TRKB receptors facilitate early follicle growth by supporting a JAGGED1-NOTCH2 oocyte-to-GC communication pathway, which promotes GC proliferation via a c-MYC-dependent mechanism. PMID:22028443

  18. [In vivo rheologic studies of plasma substitutes].

    PubMed

    Dewachter, P; Laxenaire, M C; Donner, M; Kurtz, M; Stoltz, J F

    1992-01-01

    The aim of this study was to compare in 60 ASA1 patients, the rheological effects of a 500 ml plasma substitute infusion at induction of general anaesthesia. The 60 patients were allocated into 6 groups of 10. Each group received either albumin 4%, or dextran 40 3.5%, or dextran 60 6%, or hydroxyethylstarch (HES) 200 6%, or modified fluid gelatin or Ringer lactate. The infusion extended over 30 minutes. In blood samples obtained before infusion, immediately after the end, three and 24 hours after the end of infusion, osmotic pressure, oncotic pressure, proteins and fibrinogen concentration were measured. Following rheological parameters were also assessed: plasma viscosity, blood viscosity at two shear rates (0.5 and 128 s-1), erythrocyte aggregation by primary and final aggregation times as well as total and partial dissociation thresholds. The determinations were carried out at haematocrit corrected to 40%. At intergroup analysis of the different substitutes compared to albumin 4%, with the exception of Ringer lactate, there was no significant modification of osmotic and oncotic pressures or fibrinogen concentrations. Only gelatin and dextran 60 modified the rheological parameters. The intragroup comparison did not demonstrate significant variations of osmotic and oncotic pressures. Fibrinogen concentrations remained unchanged up to the 24th hours, where they increased as a reaction to surgery. Similar changes of rheological parameters occurred for Ringer lactate, albumin 4% and dextran 40: decrease of plasma viscosity (< 10%) and blood viscosity (< 20% at shear rate of 0.5 s-1), increase of primary aggregation time (30-50%) with decrease of total dissociation threshold (10-20%). These changes ended 24 hours after infusion. Dextran 60 and gelatin elicited a modification of blood rheology until the 24th hour after the end of infusion. Such modifications did not occur with HES. It is concluded that when a rheological effect is required albumin 4% or dextran 40 3

  19. Effects of fibrinogen concentrate after shock/resuscitation – A comparison between in vivo microvascular clot formation and thromboelastometry

    PubMed Central

    Martini, Judith; Cabrales, Pedro; Fries, Dietmar; Intaglietta, Marcos; Tsai, Amy G.

    2014-01-01

    Objective Dilutional coagulopathy after resuscitation with crystalloids/colloids clinically often appears as diffuse microvascular bleeding. Administration of fibrinogen reduces bleeding and increases maximum clot firmness (MCF), measured by thromboelastometry. Study objective was to implement a model where microvascular bleeding can be directly assessed by visualizing clot formation in microvessels, and correlations can be made to thromboelastometry. Design Randomized animal study. Setting University research laboratory. Subjects Male Syrian Golden hamsters. Interventions Microvessels of Syrian Golden hamsters fitted with a dorsal window chamber were studied using videomicroscopy. After 50% hemorrhage followed by 1 hr of hypovolemia resuscitation with 35% of blood volume using a high molecular weight (MW) HES solution (Hextend®, Hospira, MW 670 kD) occurred. Animals were then treated with 250 mg/kg fibrinogen iv (Laboratoire français du Fractionnement et des Biotechnologies (LFB), Paris, France) or an equal volume of saline before venular vessel wall injuries were made by directed laser irradiation and the ability of microthrombus formation was assessed. Measurements and main results Thromboelastometric measurements of MCF were performed at the beginning and at the end of the experiment. Resuscitation with HES and sham treatment significantly decreased FIBTEM MCF from 32 ± 9 at baseline vs. 13 ± 5 mm after sham treatment (p < 0.001). Infusion of fibrinogen concentrate significantly increased MCF, restoring baseline levels (baseline 32 ± 9 mm; after fibrinogen administration 29 ± 2 mm). In vivo microthrombus formation in laser injured vessels significantly increased in fibrinogen treated animals compared with sham (77% vs. 18%). Conclusions Fibrinogen treatment leads to increased clot firmness in dilutional coagulopathy as measured with thromboelastometry. At the microvascular level this increased clot strength, corresponds to an increased incidence of

  20. Hawaii demand-side management resource assessment. Final report: DSM opportunity report

    SciTech Connect

    1995-08-01

    The Hawaii Demand-Side Management Resource Assessment was the fourth of seven projects in the Hawaii Energy Strategy (HES) program. HES was designed by the Department of Business, Economic Development, and Tourism (DBEDT) to produce an integrated energy strategy for the State of Hawaii. The purpose of Project 4 was to develop a comprehensive assessment of Hawaii`s demand-side management (DSM) resources. To meet this objective, the project was divided into two phases. The first phase included development of a DSM technology database and the identification of Hawaii commercial building characteristics through on-site audits. These Phase 1 products were then used in Phase 2 to identify expected energy impacts from DSM measures in typical residential and commercial buildings in Hawaii. The building energy simulation model DOE-2.1E was utilized to identify the DSM energy impacts. More detailed information on the typical buildings and the DOE-2.1E modeling effort is available in Reference Volume 1, ``Building Prototype Analysis``. In addition to the DOE-2.1E analysis, estimates of residential and commercial sector gas and electric DSM potential for the four counties of Honolulu, Hawaii, Maui, and Kauai through 2014 were forecasted by the new DBEDT DSM Assessment Model. Results from DBEDTs energy forecasting model, ENERGY 2020, were linked with results from DOE-2.1E building energy simulation runs and estimates of DSM measure impacts, costs, lifetime, and anticipated market penetration rates in the DBEDT DSM Model. Through its algorithms, estimates of DSM potential for each forecast year were developed. Using the load shape information from the DOE-2.1E simulation runs, estimates of electric peak demand impacts were developed. 10 figs., 55 tabs.

  1. Regulation of Nitrogenase Gene Expression by Transcript Stability in the Cyanobacterium Anabaena variabilis

    PubMed Central

    Pratte, Brenda S.

    2014-01-01

    The nitrogenase gene cluster in cyanobacteria has been thought to comprise multiple operons; however, in Anabaena variabilis, the promoter for the first gene in the cluster, nifB1, appeared to be the primary promoter for the entire nif cluster. The structural genes nifHDK1 were the most abundant transcripts; however, their abundance was not controlled by an independent nifH1 promoter, but rather, by RNA processing, which produced a very stable nifH1 transcript and a moderately stable nifD1 transcript. There was also no separate promoter for nifEN1. In addition to the nifB1 promoter, there were weak promoters inside the nifU1 gene and inside the nifE1 gene, and both promoters were heterocyst specific. In an xisA mutant, which effectively separated promoters upstream of an 11-kb excision element in nifD1 from the downstream genes, the internal nifE1 promoter was functional. Transcription of the nif1 genes downstream of the 11-kb element, including the most distant genes, hesAB1 and fdxH1, was reduced in the xisA mutant, indicating that the nifB1 promoter contributed to their expression. However, with the exception of nifK1 and nifE1, which had no expression, the downstream genes showed low to moderate levels of transcription in the xisA mutant. The hesA1 gene also had a promoter, but the fdxH gene had a processing site just upstream of the gene. The processing of transcripts at sites upstream of nifH1 and fdxH1 correlated with increased stability of these transcripts, resulting in greater amounts than transcripts that were not close to processing sites. PMID:25092030

  2. Influence of in vitro and in vivo oxygen modulation on β cell differentiation from human embryonic stem cells.

    PubMed

    Cechin, Sirlene; Alvarez-Cubela, Silvia; Giraldo, Jaime A; Molano, Ruth D; Villate, Susana; Ricordi, Camillo; Pileggi, Antonello; Inverardi, Luca; Fraker, Christopher A; Domínguez-Bendala, Juan

    2014-03-01

    The possibility of using human embryonic stem (hES) cell-derived β cells as an alternative to cadaveric islets for the treatment of type 1 diabetes is now widely acknowledged. However, current differentiation methods consistently fail to generate meaningful numbers of mature, functional β cells. In order to address this issue, we set out to explore the role of oxygen modulation in the maturation of pancreatic progenitor (PP) cells differentiated from hES cells. We have previously determined that oxygenation is a powerful driver of murine PP differentiation along the endocrine lineage of the pancreas. We hypothesized that targeting physiological oxygen partial pressure (pO2) levels seen in mature islets would help the differentiation of PP cells along the β-cell lineage. This hypothesis was tested both in vivo (by exposing PP-transplanted immunodeficient mice to a daily hyperbaric oxygen regimen) and in vitro (by allowing PP cells to mature in a perfluorocarbon-based culture device designed to carefully adjust pO2 to a desired range). Our results show that oxygen modulation does indeed contribute to enhanced maturation of PP cells, as evidenced by improved engraftment, segregation of α and β cells, body weight maintenance, and rate of diabetes reversal in vivo, and by elevated expression of pancreatic endocrine makers, β-cell differentiation yield, and insulin production in vitro. Our studies confirm the importance of oxygen modulation as a key variable to consider in the design of β-cell differentiation protocols and open the door to future strategies for the transplantation of fully mature β cells. PMID:24375542

  3. A Minimal Nitrogen Fixation Gene Cluster from Paenibacillus sp. WLY78 Enables Expression of Active Nitrogenase in Escherichia coli

    PubMed Central

    Zhao, Dehua; Liu, Xiaomeng; Zhang, Bo; Xie, Jianbo; Hong, Yuanyuan; Li, Pengfei; Chen, Sanfeng; Dixon, Ray; Li, Jilun

    2013-01-01

    Most biological nitrogen fixation is catalyzed by molybdenum-dependent nitrogenase, an enzyme complex comprising two component proteins that contains three different metalloclusters. Diazotrophs contain a common core of nitrogen fixation nif genes that encode the structural subunits of the enzyme and components required to synthesize the metalloclusters. However, the complement of nif genes required to enable diazotrophic growth varies significantly amongst nitrogen fixing bacteria and archaea. In this study, we identified a minimal nif gene cluster consisting of nine nif genes in the genome of Paenibacillus sp. WLY78, a gram-positive, facultative anaerobe isolated from the rhizosphere of bamboo. We demonstrate that the nif genes in this organism are organized as an operon comprising nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV and that the nif cluster is under the control of a σ70 (σA)-dependent promoter located upstream of nifB. To investigate genetic requirements for diazotrophy, we transferred the Paenibacillus nif cluster to Escherichia coli. The minimal nif gene cluster enables synthesis of catalytically active nitrogenase in this host, when expressed either from the native nifB promoter or from the T7 promoter. Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. Whereas nif transcription in Paenibacillus is regulated in response to nitrogen availability and by the external oxygen concentration, transcription from the nifB promoter is constitutive in E. coli, indicating that negative regulation of nif transcription is bypassed in the heterologous host. This study demonstrates the potential for engineering nitrogen fixation in a non-nitrogen fixing organism with a minimum set of nine nif genes. PMID:24146630

  4. MicroRNA-206 overexpression promotes apoptosis, induces cell cycle arrest and inhibits the migration of human hepatocellular carcinoma HepG2 cells

    PubMed Central

    LIU, WEIWEI; XU, CHUANMING; WAN, HUIFANG; LIU, CHUNJU; WEN, CAN; LU, HONGFEI; WAN, FUSHENG

    2014-01-01

    MicroRNA-206 (miR-206) is known to regulate cell proliferation and migration and is involved in various types of cancer. However, the role of miR-206 in human hepatocellular carcinoma (HHC) has not been previously reported. In the present study, the expression of Notch3 in HCC and adjacent non-neoplastic tissue was immunohistochemically assessed on formalin-fixed, paraffin-embedded sections. miR-206 mimics were transiently transfected into HepG2 cells using Lipofectamine™ 2000. Subsequently, we evaluated the role of miR-206 in cell proliferation, apoptosis, cell cycle arrest and migration by MTS assay, Hoechst 33342 staining, Annexin V-FITC/PI assay, flow cytometry and wound healing assay. Using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis, we detected the expression of Notch3, Bax, Bcl-2, Hes1, p57 and matrix metalloproteinase (MMP)-9 at the mRNA and protein level, respectively. In addition, we measured the expression of miR-206 at the mRNA level and that of caspase-3 at the protein level. After miR-206 was upregulated in HepG2 cells, Notch3, Hes1, Bcl-2 and MMP-9 were downregulated both at the mRNA and protein level, whereas p57 and Bax were upregulated. Cleaved caspase-3 protein expression was also markedly increased. Cell proliferation was significantly attenuated and apoptosis was markedly increased. Furthermore, miR-206 overexpression induced cell cycle arrest and inhibited the migration of HepG2 cells. Taken together, our results uggest that miR-206 is a potential regulator of apoptosis, the cell cycle and migration in HepG2 cells and that it has the potential for use in the targeted therapy of HCC and is a novel tumor suppressor. PMID:24919811

  5. Effects of economic status and education level on the height and weight of community adolescents in Nepal.

    PubMed

    Joshi, Nira; Rikimaru, Toru; Pandey, Sharada

    2005-08-01

    There is scarce information on the relative importance of socio-economic factors in determining the adolescent anthropometric measurements. The objective of this study was to examine the effects of economic status, education level, and food consumption on the height and weight of community adolescents in Nepal. The study was done in the communities of the Kathmandu Valley area in Nepal. All together 426 unmarried adolescent girls aged 14-19 y were selected. The adolescents were interviewed regarding socioeconomic background (education, occupation and property possessions) and frequency of foods consumption. Height and weight were determined and BMI was calculated. Z-scores of height-for-age and weight-for-age were calculated based on the WHO/NCHS standard to avoid bias by age. The adolescents participating in the survey were categorized into three groups using the various indicators of economic status: Low Economic Status (LES) group, Middle Economic Status (MES) group and High Economic Status (HES) group. The Z-scores of height and weight were significantly lower in the LES group than in the MES and HES groups (p<0.05). The Z-score of height was significantly increased with education level even under the condition of controlling economic level (p<0.05). Since the frequency of milk consumption was significantly related not only with height (p<0.05), but also with economic (chi2=31.6, df=4, p<0.001) and education levels (chi2=22.4, df=6, p<0.01), the increased height in the groups of the better economic status or the better education level was interpreted to be due to the outcome of the higher frequency of milk consumption. This study indicated that education was a more important factor affecting the height of the adolescents via improved food habits even under adverse economic conditions. PMID:16261994

  6. A person-time analysis of hospital activity among cancer survivors in England

    PubMed Central

    Maddams, J; Utley, M; Møller, H

    2011-01-01

    Background: There are around 2 million cancer survivors in the UK. This study describes the inpatient and day case hospital activity among the population of cancer survivors in England. This is one measure of the burden of cancer on the individual and the health service. Methods: The national cancer registry data set for England (1990–2006) is linked to the NHS Hospital Episode Statistics (HES) database. Cohorts of survivors were defined as those people recorded in the cancer registry data with a diagnosis of breast, colorectal, lung or prostate cancer before 2007. The person-time of prevalence in 2006 for each cohort of survivors was calculated according to the cancer type, sex, age and time since diagnosis. The corresponding HES episodes of care in 2006 were used to calculate the person-time of admitted hospital care for each cohort of survivors. The average proportion of time spent in hospital by survivors in each cohort was calculated as the summed person-time of hospital activity divided by the summed person-time of prevalence. The analysis was conducted separately for cancer-related episodes and non-cancer-related episodes. Results: Lung cancer survivors had the highest intensity of cancer-related hospital activity. For all cancers, cancer-related hospital activity was highest in the first year following diagnosis. Breast and prostate cancer survivors had peaks of cancer-related hospital activity in the relatively young and relatively old age groups. The proportion of time spent in hospital for non-cancer-related care was much lower than that for cancer-related care and increased gradually with age but was generally constant regardless of time since diagnosis. Conclusion: The person-time approach used in this study is more revealing than a simple enumeration of cancer survivors and hospital admissions. Hospital activity among cancer survivors is highest soon after diagnosis. The effect of age on the amount of hospital activity is different for each type of

  7. Structure of coronavirus hemagglutinin-esterase offers insight into corona and influenza virus evolution

    PubMed Central

    Zeng, Qinghong; Langereis, Martijn A.; van Vliet, Arno L. W.; Huizinga, Eric G.; de Groot, Raoul J.

    2008-01-01

    The hemagglutinin-esterases (HEs) are a family of viral envelope glycoproteins that mediate reversible attachment to O-acetylated sialic acids by acting both as lectins and as receptor-destroying enzymes (RDEs). Related HEs occur in influenza C, toro-, and coronaviruses, apparently as a result of relatively recent lateral gene transfer events. Here, we report the crystal structure of a coronavirus (CoV) HE in complex with its receptor. We show that CoV HE arose from an influenza C-like HE fusion protein (HEF). In the process, HE was transformed from a trimer into a dimer, whereas remnants of the fusion domain were adapted to establish novel monomer–monomer contacts. Whereas the structural design of the RDE-acetylesterase domain remained unaltered, the HE receptor-binding domain underwent remodeling to such extent that the ligand is now bound in opposite orientation. This is surprising, because the architecture of the HEF site was preserved in influenza A HA over a much larger evolutionary distance, a switch in receptor specificity and extensive antigenic variation notwithstanding. Apparently, HA and HEF are under more stringent selective constraints than HE, limiting their exploration of alternative binding-site topologies. We attribute the plasticity of the CoV HE receptor-binding site to evolutionary flexibility conferred by functional redundancy between HE and its companion spike protein S. Our findings offer unique insights into the structural and functional consequences of independent protein evolution after interviral gene exchange and open potential avenues to broad-spectrum antiviral drug design. PMID:18550812

  8. Notch Signaling in Inflammation-Induced Preterm Labor

    PubMed Central

    Jaiswal, Mukesh K.; Agrawal, Varkha; Pamarthy, Sahithi; Katara, Gajendra K.; Kulshrestha, Arpita; Gilman-Sachs, Alice; Beaman, Kenneth D.; Hirsch, Emmet

    2015-01-01

    Notch signaling plays an important role in regulation of innate immune responses and trophoblast function during pregnancy. To identify the role of Notch signaling in preterm labor, Notch receptors (Notch1-4), its ligands (DLL (Delta-like protein)-1/3/4), Jagged 1/2) and Notch-induced transcription factor Hes1 were assessed during preterm labor. Preterm labor was initiated on gestation day 14.5 by intrauterine (IU) injection of peptidoglycan (PGN) and polyinosinic:cytidylic acid (poly(I:C). Notch1, Notch2, Notch4, DLL-1 and nuclear localization of Hes1 were significantly elevated in uterus and placenta during PGN+poly(I:C)-induced preterm labor. Ex vivo, Gamma secretase inhibitor (GSI) (inhibitor of Notch receptor processing) significantly diminished the PGN+poly(I:C)-induced secretion of M1- and M2-associated cytokines in decidual macrophages, and of proinflammatory cytokines (IFN-γ, TNF-α and IL-6) and chemokines (MIP-1β) in decidual and placental cells. Conversely, angiogenesis factors including Notch ligands Jagged 1/2 and DLL-4 and VEGF were significantly reduced in uterus and placenta during PGN+poly(I:C)-induced preterm labor. In vivo GSI treatment prevents PGN+poly(I:C)-induced preterm delivery by 55.5% and increased the number of live fetuses in-utero significantly compared to respective controls 48 hrs after injections. In summary, Notch signaling is activated during PGN+poly(I:C)-induced preterm labor, resulting in upregulation of pro-inflammatory responses, and its inhibition improves in-utero survival of live fetuses. PMID:26472156

  9. X chromosome inactivation in human parthenogenetic embryonic stem cells following prolonged passaging

    PubMed Central

    QI, QUAN; DING, CHENHUI; HONG, PINGPING; YANG, GANG; XIE, YANXIN; WANG, JING; HUANG, SUNXING; HE, KE; ZHOU, CANQUAN

    2015-01-01

    The present study aimed to investigate the X chrochromosome inactivation (XCI) status in long-term cultured human parthenogenetic embryonic stem cells. One human embryonic stem (hES) cell line and 2 human parthenogenetic embryonic stem (hPES) cell lines were subjected to long-term culture in vitro (>50 passages). Karyotyping, array-based comparative genomic hybridization (aCGH), X-inactive specific transcript (XIST) RNA, immunofluorescence staining and real-time PCR were used to assess the chromosome karyotypes of these cells and the XCI status. X chromosome microdeletion was observed in the hPES-2 cells following culture for 50 passages. As early as 20 passages, XIST RNA expression was detected in the hPES-2 cells and was followed by low X-linked gene expression. The XIST RNA expression level was higher in the differentiated hPES-2 cells. The hPES-2′ cells that were subclones of hPES-2 retained the XCI status, and had low XIST and X-linked gene expression. XIST RNA expression remained at a low level in the differentiated hPES-2′ cells. The human biparental embryonic stem (hBES)-1 and hPES-1 cells did not exhibit XCI, and the differentiated hPES-1 cells had high expression levels of XIST RNA. In conclusion, the chromosome karyotypes of some hPES cell lines revealed instabilities. Similar to the hES cells, the hPES cells exhibited 3 XCI statuses. The unstable XCI status of the hPES-2 line may have been related to chromosome instability. These unstable chromosomes renedered these cells susceptible to environmental conditions and freezing processes, which may be the result of environmental adaptations. PMID:25524499

  10. Microglial proliferation in the brain of chronic alcoholics with hepatic encephalopathy

    PubMed Central

    Dennis, Claude V.; Sheahan, Pamela J.; Graeber, Manuel B.; Sheedy, Donna L.; Kril, Jillian J.; Sutherland, Greg T.

    2014-01-01

    Hepatic encephalopathy (HE) is a common complication of chronic alcoholism and patients show neurological symptoms ranging from mild cognitive dysfunction to coma and death. The HE brain is characterized by glial changes, including microglial activation, but the exact pathogenesis of HE is poorly understood. During a study investigating cell proliferation in the subventricular zone of chronic alcoholics, a single case with widespread proliferation throughout their adjacent grey and white matter was noted. This case also had concomitant HE raising the possibility that glial proliferation might be a pathological feature of the disease. In order to explore this possibility fixed postmortem human brain tissue from chronic alcoholics with cirrhosis and HE (n = 9), alcoholics without HE (n = 4) and controls (n = 4) were examined using immunohistochemistry and cytokine assays. In total, 4/9 HE cases had PCNA- and a second proliferative marker, Ki-67-positive cells throughout their brain and these cells co-stained with the microglial marker, Iba1. These cases were termed ‘proliferative HE’ (pHE). The microglia in pHEs displayed an activated morphology with hypertrophied cell bodies and short, thickened processes. In contrast, the microglia in white matter regions of the non-proliferative HE cases were less activated and appeared dystrophic. pHEs were also characterized by higher interleukin-6 levels and a slightly higher neuronal density . These findings suggest that microglial proliferation may form part of an early neuroprotective response in HE that ultimately fails to halt the course of the disease because underlying etiological factors such as high cerebral ammonia and systemic inflammation remain. PMID:24346482

  11. Deciphering the onychophoran 'segmentation gene cascade': Gene expression reveals limited involvement of pair rule gene orthologs in segmentation, but a highly conserved segment polarity gene network.

    PubMed

    Janssen, Ralf; Budd, Graham E

    2013-10-01

    The hallmark of the arthropods is their segmented body, although origin of segmentation, however, is unresolved. In order to shed light on the origin of segmentation we investigated orthologs of pair rule genes (PRGs) and segment polarity genes (SPGs) in a member of the closest related sister-group to the arthropods, the onychophorans. Our gene expression data analysis suggests that most of the onychophoran PRGs do not play a role in segmentation. One possible exception is the even-skipped (eve) gene that is expressed in the posterior end of the onychophoran where new segments are likely patterned, and is also expressed in segmentation-gene typical transverse stripes in at least a number of newly formed segments. Other onychophoran PRGs such as runt (run), hairy/Hes (h/Hes) and odd-skipped (odd) do not appear to have a function in segmentation at all. Onychophoran PRGs that act low in the segmentation gene cascade in insects, however, are potentially involved in segment-patterning. Most obvious is that from the expression of the pairberry (pby) gene ortholog that is expressed in a typical SPG-pattern. Since this result suggested possible conservation of the SPG-network we further investigated SPGs (and associated factors) such as Notum in the onychophoran. We find that the expression patterns of SPGs in arthropods and the onychophoran are highly conserved, suggesting a conserved SPG-network in these two clades, and indeed also in an annelid. This may suggest that the common ancestor of lophotrochozoans and ecdysozoans was already segmented utilising the same SPG-network, or that the SPG-network was recruited independently in annelids and onychophorans/arthropods. PMID:23880430

  12. Notch Signaling in Inflammation-Induced Preterm Labor.

    PubMed

    Jaiswal, Mukesh K; Agrawal, Varkha; Pamarthy, Sahithi; Katara, Gajendra K; Kulshrestha, Arpita; Gilman-Sachs, Alice; Beaman, Kenneth D; Hirsch, Emmet

    2015-01-01

    Notch signaling plays an important role in regulation of innate immune responses and trophoblast function during pregnancy. To identify the role of Notch signaling in preterm labor, Notch receptors (Notch1-4), its ligands (DLL (Delta-like protein)-1/3/4), Jagged 1/2) and Notch-induced transcription factor Hes1 were assessed during preterm labor. Preterm labor was initiated on gestation day 14.5 by intrauterine (IU) injection of peptidoglycan (PGN) and polyinosinic:cytidylic acid (poly(I:C). Notch1, Notch2, Notch4, DLL-1 and nuclear localization of Hes1 were significantly elevated in uterus and placenta during PGN+poly(I:C)-induced preterm labor. Ex vivo, Gamma secretase inhibitor (GSI) (inhibitor of Notch receptor processing) significantly diminished the PGN+poly(I:C)-induced secretion of M1- and M2-associated cytokines in decidual macrophages, and of proinflammatory cytokines (IFN-γ, TNF-α and IL-6) and chemokines (MIP-1β) in decidual and placental cells. Conversely, angiogenesis factors including Notch ligands Jagged 1/2 and DLL-4 and VEGF were significantly reduced in uterus and placenta during PGN+poly(I:C)-induced preterm labor. In vivo GSI treatment prevents PGN+poly(I:C)-induced preterm delivery by 55.5% and increased the number of live fetuses in-utero significantly compared to respective controls 48 hrs after injections. In summary, Notch signaling is activated during PGN+poly(I:C)-induced preterm labor, resulting in upregulation of pro-inflammatory responses, and its inhibition improves in-utero survival of live fetuses. PMID:26472156

  13. Chronic Gastrointestinal Nematode Infection Mutes Immune Responses to Mycobacterial Infection Distal to the Gut.

    PubMed

    Obieglo, Katja; Feng, Xiaogang; Bollampalli, Vishnu Priya; Dellacasa-Lindberg, Isabel; Classon, Cajsa; Österblad, Markus; Helmby, Helena; Hewitson, James P; Maizels, Rick M; Gigliotti Rothfuchs, Antonio; Nylén, Susanne

    2016-03-01

    Helminth infections have been suggested to impair the development and outcome of Th1 responses to vaccines and intracellular microorganisms. However, there are limited data regarding the ability of intestinal nematodes to modulate Th1 responses at sites distal to the gut. In this study, we have investigated the effect of the intestinal nematode Heligmosomoides polygyrus bakeri on Th1 responses to Mycobacterium bovis bacillus Calmette-Guérin (BCG). We found that H. polygyrus infection localized to the gut can mute BCG-specific CD4(+) T cell priming in both the spleen and skin-draining lymph nodes. Furthermore, H. polygyrus infection reduced the magnitude of delayed-type hypersensitivity (DTH) to PPD in the skin. Consequently, H. polygyrus-infected mice challenged with BCG had a higher mycobacterial load in the liver compared with worm-free mice. The excretory-secretory product from H. polygyrus (HES) was found to dampen IFN-γ production by mycobacteria-specific CD4(+) T cells. This inhibition was dependent on the TGF-βR signaling activity of HES, suggesting that TGF-β signaling plays a role in the impaired Th1 responses observed coinfection with worms. Similar to results with mycobacteria, H. polygyrus-infected mice displayed an increase in skin parasite load upon secondary infection with Leishmania major as well as a reduction in DTH responses to Leishmania Ag. We show that a nematode confined to the gut can mute T cell responses to mycobacteria and impair control of secondary infections distal to the gut. The ability of intestinal helminths to reduce DTH responses may have clinical implications for the use of skin test-based diagnosis of microbial infections. PMID:26819205

  14. Stratification of surface waters during the last glacial millennial climatic events: a key factor in subsurface and deep water mass dynamics

    NASA Astrophysics Data System (ADS)

    Wary, Mélanie; Eynaud, Frédérique; Sabine, Marjolaine; Zaragosi, Sébastien; Rossignol, Linda; Malaize, Bruno; Palis, Edouard; Zumaque, Jena; Caulle, Clémence; Penaud, Aurélie; Michel, Elisabeth; Charlier, Karine

    2015-04-01

    The last glacial period was punctuated by abrupt climatic events with extrema known as Heinrich events (HEs) and Dansgaard-Oeschger (DO) cycles. These millennial events have been the subject of many paleoreconstructions and model experiments in the past decades, but yet the hydrological processes involved remain elusive. In the present work, high resolution analyses were conducted on the 12-42 ka BP section of core MD99-2281 retrieved Southwest off Faeroes. Our multiproxy approach, coupling micropaleontological, geochemical and sedimentological analyses, allows us to track surface, subsurface, and deep hydrological processes occurring during these rapid climatic changes. Records indicate that the coldest episodes of the studied period (Greenland stadials and HEs) were characterized by a strong stratification of surface waters. This surface stratification seems to have played a key role in the dynamics of subsurface and deep water masses. Indeed, periods of high surface stratification are marked by a coupling of subsurface and deep circulations which sharply weaken at the beginning of stadials; simultaneously, surface conditions progressively deteriorate throughout these cold episodes. At the opposite, periods of decreasing surface stratification (Greenland interstadials) are characterized by a coupling of surface and deep hydrological processes with progressively milder surface conditions and gradual intensification of the deep circulation; at the same time, the vigor of the subsurface Atlantic inflow remains constantly high. In addition to this remarkable oceanographical scheme, our results also reveal different and atypical hydrological signatures for Heinrich events 2 and 3. Our findings thus bring valuable information to better understand hydrological processes occurring in a key area during the abrupt climatic shifts of the last glacial period.

  15. Chronic Gastrointestinal Nematode Infection Mutes Immune Responses to Mycobacterial Infection Distal to the Gut

    PubMed Central

    Obieglo, Katja; Feng, Xiaogang; Bollampalli, Vishnu Priya; Dellacasa-Lindberg, Isabel; Classon, Cajsa; Österblad, Markus; Helmby, Helena; Hewitson, James P.; Maizels, Rick M.

    2016-01-01

    Helminth infections have been suggested to impair the development and outcome of Th1 responses to vaccines and intracellular microorganisms. However, there are limited data regarding the ability of intestinal nematodes to modulate Th1 responses at sites distal to the gut. In this study, we have investigated the effect of the intestinal nematode Heligmosomoides polygyrus bakeri on Th1 responses to Mycobacterium bovis bacillus Calmette–Guérin (BCG). We found that H. polygyrus infection localized to the gut can mute BCG-specific CD4+ T cell priming in both the spleen and skin-draining lymph nodes. Furthermore, H. polygyrus infection reduced the magnitude of delayed-type hypersensitivity (DTH) to PPD in the skin. Consequently, H. polygyrus–infected mice challenged with BCG had a higher mycobacterial load in the liver compared with worm-free mice. The excretory–secretory product from H. polygyrus (HES) was found to dampen IFN-γ production by mycobacteria-specific CD4+ T cells. This inhibition was dependent on the TGF-βR signaling activity of HES, suggesting that TGF-β signaling plays a role in the impaired Th1 responses observed coinfection with worms. Similar to results with mycobacteria, H. polygyrus–infected mice displayed an increase in skin parasite load upon secondary infection with Leishmania major as well as a reduction in DTH responses to Leishmania Ag. We show that a nematode confined to the gut can mute T cell responses to mycobacteria and impair control of secondary infections distal to the gut. The ability of intestinal helminths to reduce DTH responses may have clinical implications for the use of skin test–based diagnosis of microbial infections. PMID:26819205

  16. Regulation of nitrogenase gene expression by transcript stability in the cyanobacterium Anabaena variabilis.

    PubMed

    Pratte, Brenda S; Thiel, Teresa

    2014-10-01

    The nitrogenase gene cluster in cyanobacteria has been thought to comprise multiple operons; however, in Anabaena variabilis, the promoter for the first gene in the cluster, nifB1, appeared to be the primary promoter for the entire nif cluster. The structural genes nifHDK1 were the most abundant transcripts; however, their abundance was not controlled by an independent nifH1 promoter, but rather, by RNA processing, which produced a very stable nifH1 transcript and a moderately stable nifD1 transcript. There was also no separate promoter for nifEN1. In addition to the nifB1 promoter, there were weak promoters inside the nifU1 gene and inside the nifE1 gene, and both promoters were heterocyst specific. In an xisA mutant, which effectively separated promoters upstream of an 11-kb excision element in nifD1 from the downstream genes, the internal nifE1 promoter was functional. Transcription of the nif1 genes downstream of the 11-kb element, including the most distant genes, hesAB1 and fdxH1, was reduced in the xisA mutant, indicating that the nifB1 promoter contributed to their expression. However, with the exception of nifK1 and nifE1, which had no expression, the downstream genes showed low to moderate levels of transcription in the xisA mutant. The hesA1 gene also had a promoter, but the fdxH gene had a processing site just upstream of the gene. The processing of transcripts at sites upstream of nifH1 and fdxH1 correlated with increased stability of these transcripts, resulting in greater amounts than transcripts that were not close to processing sites. PMID:25092030

  17. Concerted Activity of IgG1 Antibodies and IL-4/IL-25-Dependent Effector Cells Trap Helminth Larvae in the Tissues following Vaccination with Defined Secreted Antigens, Providing Sterile Immunity to Challenge Infection

    PubMed Central

    Hewitson, James P.; Filbey, Kara J.; Esser-von Bieren, Julia; Camberis, Mali; Schwartz, Christian; Murray, Janice; Reynolds, Lisa A.; Blair, Natalie; Robertson, Elaine; Harcus, Yvonne; Boon, Louis; Huang, Stanley Ching-Cheng; Yang, Lihua; Tu, Yizheng; Miller, Mark J.; Voehringer, David; Le Gros, Graham; Harris, Nicola; Maizels, Rick M.

    2015-01-01

    Over 25% of the world's population are infected with helminth parasites, the majority of which colonise the gastrointestinal tract. However, no vaccine is yet available for human use, and mechanisms of protective immunity remain unclear. In the mouse model of Heligmosomoides polygyrus infection, vaccination with excretory-secretory (HES) antigens from adult parasites elicits sterilising immunity. Notably, three purified HES antigens (VAL-1, -2 and -3) are sufficient for effective vaccination. Protection is fully dependent upon specific IgG1 antibodies, but passive transfer confers only partial immunity to infection, indicating that cellular components are also required. Moreover, immune mice show greater cellular infiltration associated with trapping of larvae in the gut wall prior to their maturation. Intra-vital imaging of infected intestinal tissue revealed a four-fold increase in extravasation by LysM+GFP+ myeloid cells in vaccinated mice, and the massing of these cells around immature larvae. Mice deficient in FcRγ chain or C3 complement component remain fully immune, suggesting that in the presence of antibodies that directly neutralise parasite molecules, the myeloid compartment may attack larvae more quickly and effectively. Immunity to challenge infection was compromised in IL-4Rα- and IL-25-deficient mice, despite levels of specific antibody comparable to immune wild-type controls, while deficiencies in basophils, eosinophils or mast cells or CCR2-dependent inflammatory monocytes did not diminish immunity. Finally, we identify a suite of previously uncharacterised heat-labile vaccine antigens with homologs in human and veterinary parasites that together promote full immunity. Taken together, these data indicate that vaccine-induced immunity to intestinal helminths involves IgG1 antibodies directed against secreted proteins acting in concert with IL-25-dependent Type 2 myeloid effector populations. PMID:25816012

  18. Hawaii demand-side management resource assessment. Final report, Reference Volume 3 -- Residential and commercial sector DSM analyses: Detailed results from the DBEDT DSM assessment model; Part 1, Technical potential

    SciTech Connect

    1995-04-01

    The Hawaii Demand-Side Management Resource Assessment was the fourth of seven projects in the Hawaii Energy Strategy (HES) program. HES was designed by the Department of Business, Economic Development, and Tourism (DBEDT) to produce an integrated energy strategy for the State of Hawaii. The purpose of Project 4 was to develop a comprehensive assessment of Hawaii`s demand-side management (DSM) resources. To meet this objective, the project was divided into two phases. The first phase included development of a DSM technology database and the identification of Hawaii commercial building characteristics through on-site audits. These Phase 1 products were then used in Phase 2 to identify expected energy impacts from DSM measures in typical residential and commercial buildings in Hawaii. The building energy simulation model DOE-2.1E was utilized to identify the DSM energy impacts. More detailed information on the typical buildings and the DOE-2.1E modeling effort is available in Reference Volume 1, ``Building Prototype Analysis``. In addition to the DOE-2.1E analysis, estimates of residential and commercial sector gas and electric DSM potential for the four counties of Honolulu, Hawaii, Maui, and Kauai through 2014 were forecasted by the new DBEDT DSM Assessment Model. Results from DBEDTs energy forecasting model, ENERGY 2020, were linked with results from DOE-2.1E building energy simulation runs and estimates of DSM measure impacts, costs, lifetime, and anticipated market penetration rates in the DBEDT DSM Model. Through its algorithms, estimates of DSM potential for each forecast year were developed. Using the load shape information from the DOE-2.1E simulation runs, estimates of electric peak demand impacts were developed. Numerous tables and figures illustrating the technical potential for demand-side management are included.

  19. United Kingdom National Ophthalmology Database Study: Diabetic Retinopathy; Report 1: prevalence of centre-involving diabetic macular oedema and other grades of maculopathy and retinopathy in hospital eye services

    PubMed Central

    Keenan, T D L; Johnston, R L; Donachie, P H J; Sparrow, J M; Stratton, I M; Scanlon, P

    2013-01-01

    Aims To report estimates of the prevalence of diabetic retinopathy (DR) and maculopathy grades for a large cohort of patients managed by the UK hospital eye service (HES). Methods Anonymised data were extracted from 30 UK NHS hospital trusts using a single ophthalmic electronic medical record (EMR) for the period from April 2000 to November 2010 to create the National Ophthalmology Database (NOD). From 2007, the EMR facilitated capture of a nationally agreed-upon standardised data set (DR Structured Assessment) relating to the presence or absence of clinical signs of DR and maculopathy. An algorithm in the software automatically calculated the Early Treatment of Diabetic Retinopathy Study grades of retinopathy and maculopathy. Results Between 2007 and 2010, 307 538 patients had data on the NOD, with 76 127 (24.8%) patients having been recorded as having diabetes. The proportion of patients with diabetes who had a structured assessment increased from 50.7% (2007) to 86.8% (2010). In each NHS year, 12.6–20.6% of eyes with structured assessments had no DR; 59.6–67.3% had non-proliferative DR; and 18.3–20.9% had active or regressed proliferative DR. Clinically significant macular oedema was present in 15.8–18.1% of eyes, and in 8.7–10.0% of eyes, this involved the central macula. Conclusion This study provides contemporary estimates of the prevalence of retinopathy and maculopathy grades in a large cohort of patients with diabetes managed by the UK HES. Centre-involving diabetic macular oedema, potentially amenable to anti-VEGF therapy, is present in the eyes of almost 10% of these patients. This information is useful for clinicians, health-care economists, and commissioners involved in planning and delivering diabetic eye services. PMID:24051410

  20. Active form Notch4 promotes the proliferation and differentiation of 3T3-L1 preadipocytes

    SciTech Connect

    Lai, Peng-Yeh; Tsai, Chong-Bin; Department of Ophthalmology, Chiayi Christian Hospital, Chiayi 600, Taiwan, ROC ; Tseng, Min-Jen

    2013-01-18

    Highlights: ► Notch4IC modulates the ERK pathway and cell cycle to promote 3T3-L1 proliferation. ► Notch4IC facilitates 3T3-L1 differentiation by up-regulating proadipogenic genes. ► Notch4IC promotes proliferation during the early stage of 3T3-L1 adipogenesis. ► Notch4IC enhances differentiation during subsequent stages of 3T3-L1 adipogenesis. -- Abstract: Adipose tissue is composed of adipocytes, which differentiate from precursor cells in a process called adipogenesis. Many signal molecules are involved in the transcriptional control of adipogenesis, including the Notch pathway. Previous adipogenic studies of Notch have focused on Notch1 and HES1; however, the role of other Notch receptors in adipogenesis remains unclear. Q-RT-PCR analyses showed that the augmentation of Notch4 expression during the differentiation of 3T3-L1 preadipocytes was comparable to that of Notch1. To elucidate the role of Notch4 in adipogenesis, the human active form Notch4 (N4IC) was transiently transfected into 3T3-L1 cells. The expression of HES1, Hey1, C/EBPδ and PPARγ was up-regulated, and the expression of Pref-1, an adipogenic inhibitor, was down-regulated. To further characterize the effect of N4IC in adipogenesis, stable cells expressing human N4IC were established. The expression of N4IC promoted proliferation and enhanced differentiation of 3T3-L1 cells compared with those of control cells. These data suggest that N4IC promoted proliferation through modulating the ERK pathway and the cell cycle during the early stage of 3T3-L1 adipogenesis and facilitated differentiation through up-regulating adipogenic genes such as C/EBPα, PPARγ, aP2, LPL and HSL during the middle and late stages of 3T3-L1 adipogenesis.

  1. Helium effects on microstructural change in RAFM steel under irradiation: Reaction rate theory modeling

    NASA Astrophysics Data System (ADS)

    Watanabe, Y.; Morishita, K.; Nakasuji, T.; Ando, M.; Tanigawa, H.

    2015-06-01

    Reaction rate theory analysis has been conducted to investigate helium effects on the formation kinetics of interstitial type dislocation loops (I-loops) and helium bubbles in reduced-activation-ferritic/martensitic steel during irradiation, by focusing on the nucleation and growth processes of the defect clusters. The rate theory model employs the size and chemical composition dependence of thermal dissociation of point defects from defect clusters. In the calculations, the temperature and the production rate of Frenkel pairs are fixed to be T = 723 K and PV = 10-6 dpa/s, respectively. And then, only the production rate of helium atoms was changed into the following three cases: PHe = 0, 10-7 and 10-5 appm He/s. The calculation results show that helium effect on I-loop formation quite differs from that on bubble formation. As to I-loops, the loop formation hardly depends on the existence of helium, where the number density of I-loops is almost the same for the three cases of PHe. This is because helium atoms trapped in vacancies are easily emitted into the matrix due to the recombination between the vacancies and SIAs, which induces no pronounced increase or decrease of vacancies and SIAs in the matrix, leading to no remarkable impact on the I-loop nucleation. On the other hand, the bubble formation depends much on the existence of helium, in which the number density of bubbles for PHe = 10-7 and 10-5 appm He/s is much higher than that for PHe = 0. This is because helium atoms trapped in a bubble increase the vacancy binding energy, and suppress the vacancy dissociation from the bubble, resulting in a promotion of the bubble nucleation. And then, the helium effect on the promotion of bubble nucleation is very strong, even the number of helium atoms in a bubble is not so large.

  2. Estrogen Receptor β-Selective Agonists Stimulate Calcium Oscillations in Human and Mouse Embryonic Stem Cell-Derived Neurons

    PubMed Central

    Zhang, Lili; Blackman, Brigitte E.; Schonemann, Marcus D.; Zogovic-Kapsalis, Tatjana; Pan, Xiaoyu; Tagliaferri, Mary; Harris, Heather A.; Cohen, Isaac; Reijo Pera, Renee A.; Mellon, Synthia H.; Weiner, Richard I.; Leitman, Dale C.

    2010-01-01

    Estrogens are used extensively to treat hot flashes in menopausal women. Some of the beneficial effects of estrogens in hormone therapy on the brain might be due to nongenomic effects in neurons such as the rapid stimulation of calcium oscillations. Most studies have examined the nongenomic effects of estrogen receptors (ER) in primary neurons or brain slices from the rodent brain. However, these cells can not be maintained continuously in culture because neurons are post-mitotic. Neurons derived from embryonic stem cells could be a potential continuous, cell-based model to study nongenomic actions of estrogens in neurons if they are responsive to estrogens after differentiation. In this study ER-subtype specific estrogens were used to examine the role of ERα and ERβ on calcium oscillations in neurons derived from human (hES) and mouse embryonic stem cells. Unlike the undifferentiated hES cells the differentiated cells expressed neuronal markers, ERβ, but not ERα. The non-selective ER agonist 17β-estradiol (E2) rapidly increased [Ca2+]i oscillations and synchronizations within a few minutes. No change in calcium oscillations was observed with the selective ERα agonist 4,4′,4″-(4-Propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT). In contrast, the selective ERβ agonists, 2,3-bis(4-Hydroxyphenyl)-propionitrile (DPN), MF101, and 2-(3-fluoro-4-hydroxyphenyl)-7-vinyl-1,3 benzoxazol-5-ol (ERB-041; WAY-202041) stimulated calcium oscillations similar to E2. The ERβ agonists also increased calcium oscillations and phosphorylated PKC, AKT and ERK1/2 in neurons derived from mouse ES cells, which was inhibited by nifedipine demonstrating that ERβ activates L-type voltage gated calcium channels to regulate neuronal activity. Our results demonstrate that ERβ signaling regulates nongenomic pathways in neurons derived from ES cells, and suggest that these cells might be useful to study the nongenomic mechanisms of estrogenic compounds. PMID:20668547

  3. High-dose methylprednisolone treatment of laser-induced retinal injury exacerbates acute inflammation and long-term scarring

    NASA Astrophysics Data System (ADS)

    Schuschereba, Steven T.; Cross, Michael E.; Scales, David K.; Pizarro, Jose M.; Edsall, Peter R.; Stuck, Bruce E.; Marshall, John

    1999-06-01

    Purpose. To evaluate therapeutics for attenuating retinal laser injury. Methods. New Zealand Red rabbits (n=76) were pretreated (IV) with either a single dose of hydroxyethyl starch conjugated deferoxamine (HES-DFO, n=29) (6.1 ml/kg, 16.4 mg/ml) or methylprednisolone sodium succinate (MP, n=22) (30 mg/kg, followed by taper of 30, 20, 20, and 10 mg/kg/day for a total of 5d). Controls were untreated (n=25). Fifteen min later, animals were irradiated with a multiline cw argon laser (285 mW, 10 msec pulse durations, 16 lesions/eye). Funduscopy, fluorescein angiography, histology, and morphometry were performed at 10 min, 1h, 3h, 24h, 1 mo, and 6 mo after irradiation. Leukocytes were counted at lesion centers for retinal and choroidal compartments at 1, 3, and 24h. Results. At 3h, percent area incrase for the lesions was highest for MP (44%) and lowest for HES-DFO (16%)(p<0.05). In hemorrhagic lesions, MP treatment resulted in the highest increase of retinal neotrophils by 24h (p<0.05), and by 1 and 6 mo extensive chorio-retinal scarring occurred in nonhemorrhagic and hemorrhagic lesions. Also, no benefit was demonstrated on sparing of photoreceptors with MP treatment. Conclusions. Treatment of laser-induced retinal injury with methylprednisolone (MP) exacerbates acute inflammation and long-term chorio-retinal scarring; however, hydroxyethyl starch conjugated deferoxamine therapy ameliorates these aspects of injury. Data suggest caution in the use of MP therapy for laser injuries.

  4. Transcriptomic Changes Triggered by Hypoxia: Evidence for HIF-1α -Independent, [Na+]i/[K+]i-Mediated, Excitation-Transcription Coupling

    PubMed Central

    Koltsova, Svetlana V.; Shilov, Boris; Birulina, Julia G.; Akimova, Olga A.; Haloui, Mounsif; Kapilevich, Leonid V.; Gusakova, Svetlana V.; Tremblay, Johanne; Hamet, Pavel; Orlov, Sergei N.

    2014-01-01

    This study examines the relative impact of canonical hypoxia-inducible factor-1alpha- (HIF-1α and Na+i/K+i-mediated signaling on transcriptomic changes evoked by hypoxia and glucose deprivation. Incubation of RASMC in ischemic conditions resulted in ∼3-fold elevation of [Na+]i and 2-fold reduction of [K+]i. Using global gene expression profiling we found that Na+,K+-ATPase inhibition by ouabain or K+-free medium in rat aortic vascular smooth muscle cells (RASMC) led to the differential expression of dozens of genes whose altered expression was previously detected in cells subjected to hypoxia and ischemia/reperfusion. For further investigations, we selected Cyp1a1, Fos, Atf3, Klf10, Ptgs2, Nr4a1, Per2 and Hes1, i.e. genes possessing the highest increments of expression under sustained Na+,K+-ATPase inhibition and whose implication in the pathogenesis of hypoxia was proved in previous studies. In ouabain-treated RASMC, low-Na+, high-K+ medium abolished amplification of the [Na+]i/[K+]i ratio as well as the increased expression of all tested genes. In cells subjected to hypoxia and glucose deprivation, dissipation of the transmembrane gradient of Na+ and K+ completely eliminated increment of Fos, Atf3, Ptgs2 and Per2 mRNAs and sharply diminished augmentation expression of Klf10, Edn1, Nr4a1 and Hes1. In contrast to low-Na+, high-K+ medium, RASMC transfection with Hif-1a siRNA attenuated increments of Vegfa, Edn1, Klf10 and Nr4a1 mRNAs triggered by hypoxia but did not impact Fos, Atf3, Ptgs2 and Per2 expression. Thus, our investigation demonstrates, for the first time, that Na+i/K+i-mediated, Hif-1α- -independent excitation-transcription coupling contributes to transcriptomic changes evoked in RASMC by hypoxia and glucose deprivation. PMID:25375852

  5. Mutant Kras Promotes Hyperplasia and Alters Differentiation in the Colon Epithelium But Does Not Expand the Presumptive Stem Cell Pool

    PubMed Central

    Feng, Ying; Bommer, Guido T.; Zhao, Jenny; Green, Maranne; Sands, Evan; Zhai, Yali; Brown, Kelly; Burberry, Aaron; Cho, Kathleen R.; Fearon, Eric R.

    2011-01-01

    BACKGROUND & AIMS Adenomatous polyps are precursors to colorectal cancer (CRC), whereas hyperplastic polyps (HPPs) have a small risk of progression to CRC. Mutations in KRAS are found in ~40% of CRCs and large adenomas and a subset of HPPs. We investigated the reasons that HPPs with KRAS mutations lack malignant potential; we compared the effects of Kras/KRAS activation to those of Adenomatous polyposis coli (Apc)/APC inactivation, which promotes adenoma formation. METHODS We activated a KrasG12D mutant allele or inactivated Apc alleles in mouse colon epithelium and analyzed phenotypes and expression of selected genes and proteins. The mouse data were validated using samples of human HPPs and adenomas. Signaling pathways and factors that contribute to Kras/KRAS-induced phenotypes were studied in intestinal epithelial cells. RESULTS Activation of Kras led to hyperplasia and serrated crypt architecture akin to that observed in human HPPs. We also observed loss of Paneth cells and increases in goblet cell numbers. Abnormalities in Kras-mediated differentiation and proliferation required mitogen-activated protein kinase (MAPK) signaling and were linked to activation of the Hes1 transcription factor. Human HPPs also had activation of HES1. In contrast to Apc/APC inactivation, Kras/KRAS activation did not increase expression of crypt stem cell markers in colon epithelium or colony formation in vitro. Kras/KRAS activation was not associated with substantial induction of p16INK4a protein expression in mouse colon epithelium or human HPPs. CONCLUSIONS Although Kras/KRAS mutation promotes serrated and hyperplastic morphological features in colon epithelium, it is not able to initiate adenoma development, perhaps in part because activated Kras/KRAS signaling does not increase the number of presumptive stem cells in affected crypts. PMID:21699772

  6. Pharmacokinetics and pharmacodynamics of mepolizumab, an anti-interleukin-5 monoclonal antibody.

    PubMed

    Smith, Deborah A; Minthorn, Elisabeth A; Beerahee, Misba

    2011-04-01

    Mepolizumab is a fully humanized monoclonal antibody (IgG1/κ) targeting human interleukin-5 (IL-5), a key haematopoietin needed for eosinophil development and function. Mepolizumab blocks human IL-5 from binding to the α-chain of the IL-5 receptor complex on the eosinophil cell surface, thereby inhibiting IL-5 signalling. The pharmacokinetics of mepolizumab have been evaluated in clinical studies at doses of 0.05-10 mg/kg and at 250 mg, 750 mg and 1500 mg. Mepolizumab was eliminated slowly, with mean initial and terminal phase half-life values of approximately 2 and 20 days, respectively. Plasma clearance ranged from 0.064 to 0.163 mL/h/kg and steady-state volume of distribution ranged from 49 to 93 mL/kg. Pharmacokinetics were dose proportional and time independent. Estimates based on a two-compartment intravenous infusion model from patients with asthma or healthy subjects following single doses predicted mepolizumab plasma concentrations in multiple-dose studies involving patients with hypereosinophilic syndrome (HES), asthma or eosinophilic oesophagitis. The absolute bioavailability of mepolizumab was 64-75% following subcutaneous injection and 81% following intramuscular injection. Peripheral blood eosinophil levels decreased in healthy subjects and patients with HES, asthma, eosinophilic oesophagitis or atopic dermatitis after intravenous mepolizumab infusion and subcutaneous injection. Reductions in eosinophil counts in oesophagus, sputum, skin, bone marrow, nasal lavage fluid and/or bronchial mucosa after treatment with mepolizumab were observed in placebo-controlled studies in various indications. The relationship between percentage change from baseline in blood eosinophils and mepolizumab plasma concentrations was described by an indirect pharmacological response model. The estimated maximal decrease in eosinophil count was approximately 85% from baseline and the half-maximal inhibitory concentration (IC50) was approximately 0.45

  7. The need for and provision of intrathecal baclofen therapy for the management of spasticity in England: an assessment of the Hospital Episode Statistics database

    PubMed Central

    Narendran, Rajesh C; Duarte, Rui V; Valyi, Andrea; Eldabe, Sam

    2015-01-01

    Objectives The aim of this study was to evaluate changes in the uptake of intrathecal baclofen (ITB) following commissioning of this therapy by the National Health Service (NHS) England in April 2013. The specific objectives of this study were: (i) to explore the gap between the need for and the actual provision of ITB services; and (ii) to compare England figures with other European countries with comparable data available. Setting Data for ITB -related procedures were obtained from the Hospital Episode Statistics (HES) database from 2009/2010 to 2013/2014. Participants Patients receiving ITB for the management of spasticity. Results The available data for implantation of ITB from 2009/2010 to 2013/2014 for the treatment of spasticity due to varied aetiologies show that there has not been an increase in uptake of this therapy. The estimated need for this treatment based on the incidence and prevalence of conditions susceptible to ITB therapy is between 4.6 and 5.7 per million population. Our analysis of the data available from the HES database showed that the actual number of implants is around 3.0 per million population. The same period 2009–2014 has seen an increase in the delivery of other neuromodulation techniques including spinal cord stimulation, deep brain stimulation and sacral nerve stimulation. Conclusions There is a considerable gap between the need for and provision of ITB figures nationally. Additionally, within the same area, we have observed important differences in the ITB service delivery between the various trusts. The reasons for this can be multifactorial, including individual experience and opinions, organisational structures, resource and financial limitations. Further research analysing the efficacy and cost-effectiveness of this treatment in the UK might inform the development of Technology Appraisal Guidance for ITB, potentially leading to an improvement in service provision. PMID:26129634

  8. Progressive effects of N-myc deficiency on proliferation, neurogenesis, and morphogenesis in the olfactory epithelium.

    PubMed

    Wittmann, Walter; Schimmang, Thomas; Gunhaga, Lena

    2014-06-01

    N-myc belongs to the myc proto-oncogene family, which is involved in numerous cellular processes such as proliferation, growth, apoptosis, and differentiation. Conditional deletion of N-myc in the mouse nervous system disrupted brain development, indicating that N-myc plays an essential role during neural development. How the development of the olfactory epithelium and neurogenesis within are affected by the loss of N-myc has, however, not been determined. To address these issues, we examined an N-myc(Foxg1Cre) conditional mouse line, in which N-myc is depleted in the olfactory epithelium. First changes in N-myc mutants were detected at E11.5, with reduced proliferation and neurogenesis in a slightly smaller olfactory epithelium. The phenotype was more pronounced at E13.5, with a complete lack of Hes5-positive progenitor cells, decreased proliferation, and neurogenesis. In addition, stereological analyses revealed reduced cell size of post-mitotic neurons in the olfactory epithelium, which contributed to a smaller olfactory pit. Furthermore, we observed diminished proliferation and neurogenesis also in the vomeronasal organ, which likewise was reduced in size. In addition, the generation of gonadotropin-releasing hormone neurons was severely reduced in N-myc mutants. Thus, diminished neurogenesis and proliferation in combination with smaller neurons might explain the morphological defects in the N-myc depleted olfactory structures. Moreover, our results suggest an important role for N-myc in regulating ongoing neurogenesis, in part by maintaining the Hes5-positive progenitor pool. In summary, our results provide evidence that N-myc deficiency in the olfactory epithelium progressively diminishes proliferation and neurogenesis with negative consequences at structural and cellular levels. PMID:24376126

  9. Hesperidin inhibits collagen-induced arthritis possibly through suppression of free radical load and reduction in neutrophil activation and infiltration.

    PubMed

    Umar, Sadiq; Kumar, Anubhav; Sajad, Mir; Zargan, Jamil; Ansari, Meraj; Ahmad, Sayeed; Katiyar, Chandra Kant; Khan, Haider A

    2013-03-01

    Rheumatoid arthritis is a chronic inflammatory disease characterized by the destruction of articular cartilage and bone in a chronic phase. Pathology of rheumatoid arthritis suggests autoimmunity linked to inflammation. In our study, rheumatoid arthritis was induced in Wistar rats by intradermal injections of 100 μl of emulsion containing bovine type II collagen in complete Freund's adjuvant at the base of the tail. Disease developed about 13 ± 1 days after immunization and treatment with hesperidin (HES) at a dose of 160 mg kg(-1) body weight was given after onset of disease daily until 20th day. The effect of treatment in the rats was monitored by clinical scoring, biochemical parameters and histological evaluations in joints. A steady increase in the articular elastase, nitric oxide and lipid peroxidation was observed in joints of arthritic rats as compared to control, whereas a significant decrease in reduced glutathione, superoxide dismutase activity and catalase was observed in collagen-induced arthritis rats as compared to control group. The results from the present work indicate that the treatment with hesperidin was effective in bringing about significant changes on all the parameters studied in collagen-induced arthritis rats. These data confirm that erosive destruction of the joint cartilage in collagen-induced arthritis is due free radicals released by activated neutrophils and produced by other biochemical pathways. In the present study, an attempt has been made to amelioration of the disease process by a natural product. These results suggest that oral administration of HES could be effective for treating human RA patients. PMID:22527139

  10. Inhibition of Notch rescues the angiogenic potential impaired by cardiovascular risk factors in epicardial adipose stem cells.

    PubMed

    Bejar, Maria Teresa; Ferrer-Lorente, Raquel; Peña, Esther; Badimon, Lina

    2016-08-01

    The epicardial adipose tissue (EAT) is a reservoir of adipose-derived stem cells (ASCs), with as yet unknown effects on myocardial and coronary arteries homeostasis. The purpose of this study was to investigate the angiogenic function of epicardial ASCs and their regulation by the common cardiovascular risk factors (CVRFs) affecting heart disease. Epicardial fat was obtained from a rodent model with clustering of CVRFs [Zucker diabetic fatty (ZDF)-Lepr(fa)] rats and from their lean control (ZDF-Crl) littermates without CVRFs, ASCs were isolated, and their function was assessed by proliferation and differentiation assays, flow cytometry, gene expression, and in vivo Matrigel angiogenesis analysis. Epicardial ASCs from both groups showed adipogenic and osteogenic differentiation capacity; however, epicardial ASCs from CVRF animals had a lesser ability to form tubular structures in vitro after endothelial differentiation, as well as a reduced angiogenic potential in vivo compared to control animals. Epicardial ASCs from CVRF rats showed up-regulation of the downstream Notch signaling genes Hes7, Hey1, and Heyl compared with control animals. The inhibition of Notch signaling by conditioning epicardial ASCs from CVRF animals with a γ-secretase inhibitor induced a reduction in Hes/Hey gene expression and rescued their angiogenic function in vivo We report for the first time the impact of CVRF burden on the ASCs of EAT and that the defective function is in part caused by increased Notch signaling. Conditioning ASCs by blocking Notch signaling rescues their angiogenic potential.-Bejar, M. T., Ferrer-Lorente, R., Peña, E., Badimon, L. Inhibition of Notch rescues the angiogenic potential impaired by cardiovascular risk factors in epicardial adipose stem cells. PMID:27150622

  11. Xiaotan Sanjie decoction attenuates tumor angiogenesis by manipulating Notch-1-regulated proliferation of gastric cancer stem-like cells

    PubMed Central

    Yan, Bing; Liu, Long; Zhao, Ying; Xiu, Li-Juan; Sun, Da-Zhi; Liu, Xuan; Lu, Ye; Shi, Jun; Zhang, Yin-Cheng; Li, Yong-Jin; Wang, Xiao-Wei; Zhou, Yu-Qi; Feng, Shou-Han; Lv, Can; Wei, Pin-Kang; Qin, Zhi-Feng

    2014-01-01

    AIM: To determine the underlying mechanisms of action and influence of Xiaotan Sanjie (XTSJ) decoction on gastric cancer stem-like cells (GCSCs). METHODS: The gastric cancer cell line MKN-45 line was selected and sorted by FACS using the cancer stem cell marker CD44; the stemness of these cells was checked in our previous study. In an in vitro study, the expression of Notch-1, Hes1, Vascular endothelial growth factor (VEGF), and Ki-67 in both CD44-positive gastric cancer stem-like cells (GCSCs) and CD44-negative cells was measured by Western blot. The effect of XTSJ serum on cell viability and on the above markers was measured by MTT assay and Western blot, respectively. In an in vivo study, the ability to induce angiogenesis and maintenance of GCSCs in CD44-positive-MKN-45- and CD44-negative-engrafted mice were detected by immunohistochemical staining using markers for CD34 and CD44, respectively. The role of XTSJ decoction in regulating the expression of Notch-1, Hes1, VEGF and Ki-67 was measured by Western blot and real-time polymerase chain reaction. RESULTS: CD44+ GCSCs showed more cell proliferation and VEGF secretion than CD44-negative cells in vitro, which were accompanied by the high expression of Notch-1 and Hes1 and positively associated with tumor growth (GCSCs vs CD44-negative cells, 2.72 ± 0.25 vs 1.46 ± 0.16, P < 0.05) and microvessel density (MVD) (GCSCs vs CD44-negative cells, 8.15 ± 0.42 vs 3.83 ± 0.49, P < 0.001) in vivo. XTSJ decoction inhibited the viability of both cell types in a dose-dependent manner in vitro. Specifically, a significant difference in the medium- (82.87% ± 6.53%) and high-dose XTSJ groups (77.43% ± 7.34%) was detected at 24 h in the CD44+ GCSCs group compared with the saline group (95.42% ± 5.76%) and the low-dose XTSJ group (90.74% ± 6.57%) (P < 0.05). However, the efficacy of XTSJ decoction was reduced in the CD44- groups; significant differences were only detected in the high-dose XTSJ group at 48 h (78.57% ± 6

  12. Investigation of high-precision Λ hypernuclear spectroscopy via the (e,e'K+) reaction

    SciTech Connect

    Kawama, Daisuke

    2011-01-01

    The study of Λ hypernuclear structure is very interesting in point of the understanding of the interaction between Λ and nucleon (Λ-N interaction) and its strange structure itself due to the containment of a Λ hyperon which has a strangeness as a new degree of freedom. In the several way to study the Lamda hypernuclei, the (e,e'K+) reaction spectroscopy is a powerful tool for the precise investigation of Λ hypernuclear structure. The purpose of the preset thesis is the establishment of the experimental design with the efficient data analysis method for the (e,e'K+) hypernuclear spectroscopic experiment in the wide mass region (from A=7 to A=52). It is very challenging to perform the (e,e'K+) spectroscopic experiment with such a heavy target, because of the huge electron background due to the bremsstrahlung process. In the experiment, it is required to obtain the necessary hypernuclear yield, suppressing the background event ratio. We achieved these requirements by newly constructing the high resolution electron spectrometer (HES) and splitter magnet (SPL) dedicated to the (e,e'K+) spectroscopic experiment. The HES consists of two quadrupole magnets and a dipole magnets (Q-Q-D) with a momentum resolution of dp/p = 3x10-4 at p = 0.84 GeV/c. It was used being vertically tilted by 6.5 degree so as to optimize signal to noise ratio and hypernuclear yield. The SPL is a dipole magnet. The experimental target was placed at the entrance of this magnet. The role of the SPL is to separate four kind of particles; scattered kaons, photons created by the bremsstrahlung, the post beam and scattered electrons. In addition, since the SPL is a part of the kaon and electron spectrometers. We designed the magnet shape carefully considering these points. The experiment was performed with 2.344 GeV/c electron beam from CEBAF at Jefferson Lab. The experimental setup consists of the HES, SPL and HKS (high momentum resolution kaon

  13. Modelling the hydrological behaviour of a coffee agroforestry basin in Costa Rica

    NASA Astrophysics Data System (ADS)

    Gómez-Delgado, F.; Roupsard, O.; Le Maire, G.; Taugourdeau, S.; Pérez, A.; van Oijen, M.; Vaast, P.; Rapidel, B.; Harmand, J. M.; Voltz, M.; Bonnefond, J. M.; Imbach, P.; Moussa, R.

    2011-01-01

    The profitability of hydropower in Costa Rica is affected by soil erosion and sedimentation in dam reservoirs, which are in turn influenced by land use, infiltration and aquifer interactions with surface water. In order to foster the provision and payment for Hydrological Environmental Services (HES), a quantitative assessment of the impact of specific land uses on the functioning of drainage-basins is required. The present paper aims to study the water balance partitioning in a volcanic coffee agroforestry micro-basin (1 km2, steep slopes) in Costa Rica, as a first step towards evaluating sediment or contaminant loads. The main hydrological processes were monitored during one year, using flume, eddy-covariance flux tower, soil water profiles and piezometers. A new Hydro-SVAT lumped model is proposed, that balances SVAT (Soil Vegetation Atmosphere Transfer) and basin-reservoir routines. The purpose of such a coupling was to achieve a trade-off between the expected performance of ecophysiological and hydrological models, which are often employed separately and at different spatial scales, either the plot or the basin. The calibration of the model to perform streamflow yielded a Nash-Sutcliffe (NS) coefficient equal to 0.89 for the year 2009, while the validation of the water balance partitioning was consistent with the independent measurements of actual evapotranspiration (R2 = 0.79, energy balance closed independently), soil water content (R2 = 0.35) and water table level (R2 = 0.84). Eight months of data from 2010 were used to validate modelled streamflow, resulting in a NS = 0.75. An uncertainty analysis showed that the streamflow modelling was precise for nearly every time step, while a sensitivity analysis revealed which parameters mostly affected model precision, depending on the season. It was observed that 64% of the incident rainfall R flowed out of the basin as streamflow and 25% as evapotranspiration, while the remaining 11% is probably explained by deep

  14. The impact of crystalloidal and colloidal infusion preparations on coronary vascular integrity, interstitial oedema and cardiac performance in isolated hearts

    PubMed Central

    2013-01-01

    Introduction Recent data suggested an interaction between plasma constituents and the endothelial glycocalyx to be relevant for vascular barrier function. This might be negatively influenced by infusion solutions, depending on ionic composition, pH and binding properties. The present study evaluated such an influence of current artificial preparations. Methods Isolated guinea pig hearts were prepared in a modified Langendorff mode and perfused with Krebs-Henseleit buffer augmented with 1g% human albumin. After equilibration the perfusion was switched to replacement of one half buffer by either isotonic saline (NaCl), ringer's acetate (Ri-Ac), 6% and 10% hydroxyethyl starch (6% and 10% HES, resp.), or 4% gelatine (Gel), the artificial colloids having been prepared in balanced solution. We analysed glycocalyx shedding, functional integrity of the vascular barrier and heart performance. Results While glycocalyx shedding was not observed, diluting albumin concentration towards 0.5g% by artificial solutions was associated with a marked functional breakdown of vascular barrier competence. This effect was biggest with isotonic saline and significantly attenuated with artificial colloids, the difference in the pressure dependent transvascular fluid filtration (basal vs. during infusion in groups NaCl, Ri-Ac, 6% HES, 10% HES and Gel, n = 6 each) being 0.31 ± 0.03 vs. 1.00 ± 0.04; 0.27 ± 0.03 vs. 0.81 ± 0.03; 0.29 ± 0.03 vs. 0.68 ± 0.02; 0.32 ± 0.03 vs. 0.59 ± 0.08 and 0.31 ± 0.04 vs. 0.61 ± 0.03 g/5min, respectively. Heart performance was directly related to pH value (7.38 ± 0.06, 7.33 ± 0.03, 7.14 ± 0.04, 7.08 ± 0.04, 7.25 ± 0.03), the change in the rate pressure product being 21,702 ± 1969 vs. 21,291 ± 2,552; 22,098 ± 2,115 vs. 14,114 ± 3,386; 20,897 ± 2,083 vs. 10,671 ± 1,948; 21,822 ± 2,470 vs. 10,047 ± 2,320 and 20,955 ± 2,296 vs. 15,951 ± 2,755 mmHg × bpm, respectively. Conclusions It appears important to maintain the pH value within a

  15. Complementary analysis of the hard and soft protein corona: sample preparation critically effects corona composition

    NASA Astrophysics Data System (ADS)

    Winzen, S.; Schoettler, S.; Baier, G.; Rosenauer, C.; Mailaender, V.; Landfester, K.; Mohr, K.

    2015-02-01

    Here we demonstrate how a complementary analysis of nanocapsule-protein interactions with and without application media allows gaining insights into the so called hard and soft protein corona. We have investigated how both human plasma and individual proteins (human serum albumin (HSA), apolipoprotein A-I (ApoA-I)) adsorb and interact with hydroxyethyl starch (HES) nanocapsules possessing different functionalities. To analyse the hard protein corona we used sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and a protein quantitation assay. No significant differences were observed with regards to the hard protein corona. For analysis of the soft protein corona we characterized the nanocapsule-protein interaction with isothermal titration calorimetry (ITC) and dynamic light scattering (DLS). DLS and ITC measurements revealed that a high amount of plasma proteins were adsorbed onto the capsules' surface. Although HSA was not detected in the hard protein corona, ITC measurements indicated the adsorption of an HSA amount similar to plasma with a low binding affinity and reaction heat. In contrast, only small amounts of ApoA-I protein adsorb to the capsules with high binding affinities. Through a comparison of these methods we have identified ApoA-I to be a component of the hard protein corona and HSA as a component of the soft corona. We demonstrate a pronounced difference in the protein corona observed depending on the type of characterization technique applied. As the biological identity of a particle is given by the protein corona it is crucial to use complementary characterization techniques to analyse different aspects of the protein corona.Here we demonstrate how a complementary analysis of nanocapsule-protein interactions with and without application media allows gaining insights into the so called hard and soft protein corona. We have investigated how both human plasma and individual proteins (human serum albumin (HSA), apolipoprotein A

  16. The Conserved MAPK Site in E(spl)-M8, an Effector of Drosophila Notch Signaling, Controls Repressor Activity during Eye Development

    PubMed Central

    Bandyopadhyay, Mohna; Bishop, Clifton P.

    2016-01-01

    The specification of patterned R8 photoreceptors at the onset of eye development depends on timely inhibition of Atonal (Ato) by the Enhancer of split (E(spl) repressors. Repression of Ato by E(spl)-M8 requires the kinase CK2 and is inhibited by the phosphatase PP2A. The region targeted by CK2 harbors additional conserved Ser residues, raising the prospect of regulation via multi-site phosphorylation. Here we investigate one such motif that meets the consensus for modification by MAPK, a well-known effector of Epidermal Growth Factor Receptor (EGFR) signaling. Our studies reveal an important role for the predicted MAPK site of M8 during R8 birth. Ala/Asp mutations reveal that the CK2 and MAPK sites ensure that M8 repression of Ato and the R8 fate occurs in a timely manner and at a specific stage (stage-2/3) of the morphogenetic furrow (MF). M8 repression of Ato is mitigated by halved EGFR dosage, and this effect requires an intact MAPK site. Accordingly, variants with a phosphomimetic Asp at the MAPK site exhibit earlier (inappropriate) activity against Ato even at stage-1 of the MF, where a positive feedback-loop is necessary to raise Ato levels to a threshold sufficient for the R8 fate. Analysis of deletion variants reveals that both kinase sites (CK2 and MAPK) contribute to ‘cis’-inhibition of M8. This key regulation by CK2 and MAPK is bypassed by the E(spl)D mutation encoding the truncated protein M8*, which potently inhibits Ato at stage-1 of R8 birth. We also provide evidence that PP2A likely targets the MAPK site. Thus multi-site phosphorylation controls timely onset of M8 repressor activity in the eye, a regulation that appears to be dispensable in the bristle. The high conservation of the CK2 and MAPK sites in the insect E(spl) proteins M7, M5 and Mγ, and their mammalian homologue HES6, suggest that this mode of regulation may enable E(spl)/HES proteins to orchestrate repression by distinct tissue-specific mechanisms, and is likely to have broader

  17. School-level economic disadvantage and obesity in middle school children in central Texas, USA: a cross-sectional study

    PubMed Central

    2015-01-01

    no obesity differences across school ED categories were found for Hispanic Spanish-speaking students, Hispanic English-speaking students (HES) attending high ED schools were 2.4 times more likely to be obese as HES students at low ED schools (p=.003). Conclusion Findings support the need to prioritize economically disadvantaged schools for obesity prevention efforts and support further exploration of school SES context in shaping children’s physical activity and dietary behaviors. PMID:26222099

  18. MicroRNA Expression Profiling of Oligodendrocyte Differentiation from Human Embryonic Stem Cells

    PubMed Central

    Letzen, Brian S.; Liu, Cyndi; Thakor, Nitish V.; Gearhart, John D.; All, Angelo H.; Kerr, Candace L.

    2010-01-01

    Background Cells of the oligodendrocyte (OL) lineage play a vital role in the production and maintenance of myelin, a multilamellar membrane which allows for saltatory conduction along axons. These cells may provide immense therapeutic potential for lost sensory and motor function in demyelinating conditions, such as spinal cord injury, multiple sclerosis, and transverse myelitis. However, the molecular mechanisms controlling OL differentiation are largely unknown. MicroRNAs (miRNAs) are considered the “micromanagers” of gene expression with suggestive roles in cellular differentiation and maintenance. Although unique patterns of miRNA expression in various cell lineages have been characterized, this is the first report documenting their expression during oligodendrocyte maturation from human embryonic stem (hES) cells. Here, we performed a global miRNA analysis to reveal and identify characteristic patterns in the multiple stages leading to OL maturation from hES cells including those targeting factors involved in myelin production. Methodology/Principal Findings We isolated cells from 8 stages of OL differentiation. Total RNA was subjected to miRNA profiling and validations preformed using real-time qRT-PCR. A comparison of miRNAs from our cultured OLs and OL progenitors showed significant similarities with published results from equivalent cells found in the rat and mouse central nervous system. Principal component analysis revealed four main clusters of miRNA expression corresponding to early, mid, and late progenitors, and mature OLs. These results were supported by correlation analyses between adjacent stages. Interestingly, the highest differentially-expressed miRNAs demonstrated a similar pattern of expression throughout all stages of differentiation, suggesting that they potentially regulate a common target or set of targets in this process. The predicted targets of these miRNAs include those with known or suspected roles in oligodendrocyte development