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Sample records for kritische anlage zum htr

  1. Friction forces position the neural anlage.

    PubMed

    Smutny, Michael; Ákos, Zsuzsa; Grigolon, Silvia; Shamipour, Shayan; Ruprecht, Verena; Čapek, Daniel; Behrndt, Martin; Papusheva, Ekaterina; Tada, Masazumi; Hof, Björn; Vicsek, Tamás; Salbreux, Guillaume; Heisenberg, Carl-Philipp

    2017-04-01

    During embryonic development, mechanical forces are essential for cellular rearrangements driving tissue morphogenesis. Here, we show that in the early zebrafish embryo, friction forces are generated at the interface between anterior axial mesoderm (prechordal plate, ppl) progenitors migrating towards the animal pole and neurectoderm progenitors moving in the opposite direction towards the vegetal pole of the embryo. These friction forces lead to global rearrangement of cells within the neurectoderm and determine the position of the neural anlage. Using a combination of experiments and simulations, we show that this process depends on hydrodynamic coupling between neurectoderm and ppl as a result of E-cadherin-mediated adhesion between those tissues. Our data thus establish the emergence of friction forces at the interface between moving tissues as a critical force-generating process shaping the embryo.

  2. HTR Fuel Development in Europe

    SciTech Connect

    Languille, Alain; Conrad, R.; Haas, D.

    2002-07-01

    In the frame of the European Network HTR-TN and in the 5. EURATOM RTD Framework Programme (FP5) European programmes have been launched to consolidate advanced modular HTR technology in Europe. This paper gives an overall description and first results of this programme. The major tasks covered concern a complete recovery of the past experience on fuel irradiation behaviour in Europe, qualification of HTR fuel by irradiating of fuel elements in the HFR reactor, understanding of fuel behaviour with the development of a fuel particle code and finally a recover of the fuel fabrication capability. (authors)

  3. Changes in expression of human serine protease HtrA1, HtrA2 and HtrA3 genes in benign and malignant thyroid tumors.

    PubMed

    Zurawa-Janicka, Dorota; Kobiela, Jarosław; Galczynska, Natalia; Stefaniak, Tomasz; Lipinska, Barbara; Lachinski, Andrzej; Skorko-Glonek, Joanna; Narkiewicz, Joanna; Proczko-Markuszewska, Monika; Sledzinski, Zbigniew

    2012-11-01

    Human HtrA proteins are serine proteases involved in essential physiological processes. HtrA1 and HtrA3 function as tumor suppressors and inhibitors of the TGF-β signaling pathway. HtrA2 regulates mitochondrial homeostasis and plays a pivotal role in the induction of apoptosis. The aim of the study was to determine whether the HtrA proteins are involved in thyroid carcinogenesis. We used the immunoblotting technique to estimate protein levels of HtrA1, HtrA2, long and short variants of HtrA3 (HtrA3-L and HtrA3-S) and TGF-β1 in tissues of benign and malignant thyroid lesions, and control groups. We found that the levels of HtrA2 and HtrA3-S were higher in thyroid malignant tumors compared to normal tissues and benign tumors. The HtrA3-L level was increased in malignant tumor tissues compared to benign tumor tissues and control tissues from patients with benign lesions, and elevated in normal tissues from patients with thyroid carcinoma compared to normal tissues from patients with benign lesions. We also compared levels of HtrA proteins in follicular thyroid carcinoma (FTC) and papillary thyroid carcinoma (PTC) and found that these types of carcinoma differed in the expression of HtrA3-S and HtrA1. These results indicate the implication of HtrA proteins in thyroid carcinogenesis suggest that HtrA3 variants may play different roles in cancer development, and that the increased HtrA3-L levels in thyroid tissue could be correlated with the development of malignant lesions. The TGF-β1 levels in tumor tissues were not significantly altered compared to control tissues.

  4. 5-HTR3 and 5-HTR4 located on the mitochondrial membrane and functionally regulated mitochondrial functions

    PubMed Central

    Wang, Qingyi; Zhang, Huiyuan; Xu, Hao; Guo, Dongqing; Shi, Hui; Li, Yuan; Zhang, Weiwei; Gu, Yuchun

    2016-01-01

    5-HT has been reported to possess significant effects on cardiac activities, but activation of 5-HTR on the cell membrane failed to illustrate the controversial cardiac reaction. Because 5-HT constantly comes across the cell membrane via 5-HT transporter (5-HTT) into the cytoplasm, whether 5-HTR is functional present on the cellular organelles is unknown. Here we show 5-HTR3 and 5-HTR4 were located in cardiac mitochondria, and regulated mitochondrial activities and cellular functions. Knock down 5-HTR3 and 5-HTR4 in neonatal cardiomyocytes resulted in significant increase of cell damage in response to hypoxia, and also led to alternation in heart beating. Activation of 5-HTR4 attenuated mitochondrial Ca2+ uptake under the both normoxic and hypoxic conditions, whereas 5-HTR3 augmented Ca2+ uptake only under hypoxia. 5-HTR3 and 5-HTR4 exerted the opposite effects on the mitochondrial respiration: 5-HTR3 increased RCR (respiration control ratio), but 5-HTR4 reduced RCR. Moreover, activation of 5-HTR3 and 5-HTR4 both significantly inhibited the opening of mPTP. Our results provided the first evidence that 5-HTR as a GPCR and an ion channel, functionally expressed in mitochondria and participated in the mitochondria function and regulation to maintain homeostasis of mitochondrial [Ca2+], ROS, and ATP generation efficiency in cardiomyocytes in response to stress and O2 tension. PMID:27874067

  5. The Renewal of HTR Development in Europe

    SciTech Connect

    Hittner, Dominique

    2002-07-01

    The European HTR-Technology Network (HTR-TN), created in 2000, presently groups 20 organisations from European nuclear research and industry for developing the technologies of direct-cycle modular HTRs, which presently raise a large world-wide interest, because of their high potential for economic competitiveness, natural resource sparing, safety and minimisation of the waste impacts, in line with the goals of sustainable development of Generation IV. All aspects of HTR technologies are addressed by HTR-TN, from the reactor physics to the development of materials, fuel and components. Most of this activity is supported by the European Commission in the frame of its 5. EURATOM Framework Programme. The first results of HTR-TN programme are given: the analysis of the reactor physics international benchmark on the commissioning tests of HTTR (Japan), the long term behaviour of spent HTR fuel in geologic disposal conditions, the preparation of a very high burnup fuel irradiation and the development of fabrication processes for producing high performance coated particles, etc. (authors)

  6. HTR1B and HTR2C in autism spectrum disorders in Brazilian families.

    PubMed

    Orabona, G M; Griesi-Oliveira, K; Vadasz, E; Bulcão, V L S; Takahashi, V N V O; Moreira, E S; Furia-Silva, M; Ros-Melo, A M S; Dourado, F; Matioli, S R; Matioli, R; Otto, P; Passos-Bueno, M R

    2009-01-23

    Autism spectrum disorders (ASD) is a group of behaviorally defined neurodevelopmental disabilities characterized by multiple genetic etiologies and a complex presentation. Several studies suggest the involvement of the serotonin system in the development of ASD, but only few have investigated serotonin receptors. We have performed a case-control and a family-based study with 9 polymorphisms mapped to two serotonin receptor genes (HTR1B and HTR2C) in 252 Brazilian male ASD patients of European ancestry. These analyses showed evidence of undertransmission of the HTR1B haplotypes containing alleles -161G and -261A at HTR1B gene to ASD (P=0.003), but no involvement of HTR2C to the predisposition to this disease. Considering the relatively low level of statistical significance and the power of our sample, further studies are required to confirm the association of these serotonin-related genes and ASD.

  7. Evaluation of association of common variants in HTR1A and HTR5A with schizophrenia and executive function

    PubMed Central

    Guan, Fanglin; Lin, Huali; Chen, Gang; Li, Lu; Chen, Teng; Liu, Xinshe; Han, Jiuqiang; Li, Tao

    2016-01-01

    The 5-HT1A receptor (HTR1A) and the 5-HT5A receptor (HTR5A) are key 5-HT receptors with distinct inhibitory functions. Studies have been conducted to investigate the association of a few HTR1A polymorphisms with schizophrenia, producing conflicting results, and the relationship between HTR5A and schizophrenia has not yet been well investigated. We aimed to examine the association of HTR1A and HTR5A with schizophrenia and executive function. The study included a discovery stage with 1,115 patients and 2,289 controls and a replication stage with 2,128 patients and 3,865 controls. A total of 30 common SNPs in HTR1A and HTR5A were genotyped in the discovery stage, and significantly associated SNPs were genotyped in the replication stage. We identified that two SNPs (rs878567 in HTR1A and rs1800883 in HTR5A) were significantly associated with schizophrenia in both datasets, and similar results were observed in imputation and haplotype association analyses. Moreover, we found that SNP rs1800883 significantly interacted with executive function when processing the perseverative error of Wisconsin Card Sorting Test in patients. Our results provide further supportive evidence of the effect of HTR1A and HTR5A on the etiology of schizophrenia and suggest that the selected genetic variations in HTR5A may be involved in impaired executive function. PMID:27897266

  8. Das Smart Meter Gateway - Der kritische Erfolgsfaktor für die Digitalisierung der Energiewende

    NASA Astrophysics Data System (ADS)

    Abs, Paul-Vincent

    Der kritische Erfolgsfaktor der Digitalisierung in der Energiewirtschaft liegt im bevorstehenden hohen Investitionsvolumen und der Beherrschung der neuen Technik. Ein Weg zur Kostenminimierung liegt im Zusammenschluss der einzelnen Akteure, um gemeinsame Skalenvorteile zu heben. Auch bieten neue Geschäftsmodelle die Möglichkeit, zusätzliche Erlöse zu generieren.

  9. HTR7 mediates serotonergic acute and chronic itch

    PubMed Central

    Morita, Takeshi; McClain, Shannan P.; Batia, Lyn M.; Pellegrino, Maurizio; Wilson, Sarah R.; Kienzler, Michael A.; Lyman, Kyle; Olsen, Anne Sofie Braun; Wong, Justin F.; Stucky, Cheryl L.; Brem, Rachel B.; Bautista, Diana M.

    2015-01-01

    SUMMARY Chronic itch is a prevalent and debilitating condition for which few effective therapies are available. We harnessed the natural variation across genetically distinct mouse strains to identify transcripts co-regulated with itch behavior. This survey led to the discovery of the serotonin receptor, HTR7, as a key mediator of serotonergic itch. Activation of HTR7 promoted opening of the ion channel TRPA1, which in turn triggered itch behaviors. In addition, acute itch triggered by serotonin or a selective serotonin reuptake inhibitor required both HTR7 and TRPA1. Aberrant serotonin signaling has long been linked to a variety of human chronic itch conditions, including atopic dermatitis. In a mouse model of atopic dermatitis, mice lacking HTR7 or TRPA1 displayed reduced scratching and skin lesion severity. These data highlight a role for HTR7 in acute and chronic itch, and suggest that HTR7 antagonists may be useful for treating a variety of pathological itch conditions. PMID:26074006

  10. Associations of serotonin receptor gene HTR3A, HTR3B, and HTR3A haplotypes with bipolar disorder in Chinese patients.

    PubMed

    Jian, J; Li, C; Xu, J; Qiao, D; Mi, G; Chen, X; Tang, M

    2016-09-16

    Single nucleotide polymorphisms (SNPs) in HTR3A and HTR3B have been reported to be associated with bipolar disorder in European and Japanese populations. We explored the roles of 21 tag SNPs in HTR3A and HTR3B in susceptibility to bipolar disorder in a Chinese cohort. Twenty-one Tag SNPs were genotyped in a study consisting of 130 patients with bipolar disorder, who visited Shandong Mental Health Center between June 2013 and May 2014, and 109 healthy individuals as controls. All of the tag SNPs were genotyped using Sequenom MassArray matrix-assisted laser desorption/ionization time of flight spectrometry. Plink 1.07, Haploview 4.2, and SPSS 20.0 were used for the analysis of the genotypes and the associations of the haplotypes with bipolar disorder. Association analyses of tag SNPs detected significant associations with the A allele in HTR3A rs1176719 (P = 0.030) and the C allele in HTR3A rs1176713 (P = 0.048). Haplotype-based association analyses indicated a statistically significant (P = 0.035) five-SNP haplotype (rs1062613:C, rs11604247:C, rs1176722:G, rs2276302:A, rs1176719:G) of linkage disequilibrium in block 3. Analysis of our small Chinese sample revealed a significant association of HTR3A with bipolar disorder, but yielded no evidence of an association between HTR3B and bipolar disorder. Furthermore, evidence for an association was found for a haplotype of HTR3A. Studies with larger Chinese samples are needed to verify our findings.

  11. Graphite Oxidation Simulation in HTR Accident Conditions

    SciTech Connect

    El-Genk, Mohamed

    2012-10-19

    Massive air and water ingress, following a pipe break or leak in steam-generator tubes, is a design-basis accident for high-temperature reactors (HTRs). Analysis of these accidents in both prismatic and pebble bed HTRs requires state-of-the-art capability for predictions of: 1) oxidation kinetics, 2) air helium gas mixture stratification and diffusion into the core following the depressurization, 3) transport of multi-species gas mixture, and 4) graphite corrosion. This project will develop a multi-dimensional, comprehensive oxidation kinetics model of graphite in HTRs, with diverse capabilities for handling different flow regimes. The chemical kinetics/multi-species transport model for graphite burning and oxidation will account for temperature-related changes in the properties of graphite, oxidants (O2, H2O, CO), reaction products (CO, CO2, H2, CH4) and other gases in the mixture (He and N2). The model will treat the oxidation and corrosion of graphite in geometries representative of HTR core component at temperatures of 900°C or higher. The developed chemical reaction kinetics model will be user-friendly for coupling to full core analysis codes such as MELCOR and RELAP, as well as computational fluid dynamics (CFD) codes such as CD-adapco. The research team will solve governing equations for the multi-dimensional flow and the chemical reactions and kinetics using Simulink, an extension of the MATLAB solver, and will validate and benchmark the model's predictions using reported experimental data. Researchers will develop an interface to couple the validated model to a commercially available CFD fluid flow and thermal-hydraulic model of the reactor , and will perform a simulation of a pipe break in a prismatic core HTR, with the potential for future application to a pebble-bed type HTR.

  12. The salt-sensitive structure and zinc inhibition of Borrelia burgdorferi protease BbHtrA.

    PubMed

    Russell, Theresa M; Tang, Xiaoling; Goldstein, Jason M; Bagarozzi, Dennis; Johnson, Barbara J B

    2016-02-01

    HtrA serine proteases are highly conserved and essential ATP-independent proteases with chaperone activity. Bacteria express a variable number of HtrA homologues that contribute to the virulence and pathogenicity of bacterial pathogens. Lyme disease spirochetes possess a single HtrA protease homologue, Borrelia burgdorferi HtrA (BbHtrA). Previous studies established that, like the human orthologue HtrA1, BbHtrA is proteolytically active against numerous extracellular proteins in vitro. In this study, we utilized size exclusion chromatography and blue native polyacrylamide gel electrophoresis (BN-PAGE) to demonstrate BbHtrA oligomeric structures that were substrate independent and salt sensitive. Examination of the influence of transition metals on the activity of BbHtrA revealed that this protease is inhibited by Zn(2+) > Cu(2+) > Mn(2+). Extending this analysis to two other HtrA proteases, E. coli DegP and HtrA1, revealed that all three HtrA proteases were reversibly inhibited by ZnCl2 at all micro molar concentrations examined. Commercial inhibitors for HtrA proteases are not available and physiologic HtrA inhibitors are unknown. Our observation of conserved zinc inhibition of HtrA proteases will facilitate structural and functional studies of additional members of this important class of proteases. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

  13. Structural and functional analysis of human HtrA3 protease and its subdomains

    DOE PAGES

    Glaza, Przemyslaw; Osipiuk, Jerzy; Wenta, Tomasz; ...

    2015-06-25

    Human HtrA3 protease, which induces mitochondria-mediated apoptosis, can be a tumor suppressor and a potential therapeutic target in the treatment of cancer. However, there is little information about its structure and biochemical properties. HtrA3 is composed of an N-terminal domain not required for proteolytic activity, a central serine protease domain and a C-terminal PDZ domain. HtrA3S, its short natural isoform, lacks the PDZ domain which is substituted by a stretch of 7 C-terminal amino acid residues, unique for this isoform. This paper presents the crystal structure of the HtrA3 protease domain together with the PDZ domain (ΔN-HtrA3), showing that themore » protein forms a trimer whose protease domains are similar to those of human HtrA1 and HtrA2. The ΔN-HtrA3 PDZ domains are placed in a position intermediate between that in the flat saucer-like HtrA1 SAXS structure and the compact pyramidal HtrA2 X-ray structure. The PDZ domain interacts closely with the LB loop of the protease domain in a way not found in other human HtrAs. ΔN-HtrA3 with the PDZ removed (ΔN-HtrA3-ΔPDZ) and an N-terminally truncated HtrA3S (ΔN-HtrA3S) were fully active at a wide range of temperatures and their substrate affinity was not impaired. This indicates that the PDZ domain is dispensable for HtrA3 activity. As determined by size exclusion chromatography, ΔN-HtrA3 formed stable trimers while both ΔN-HtrA3-ΔPDZ and ΔN-HtrA3S were monomeric. This suggests that the presence of the PDZ domain, unlike in HtrA1 and HtrA2, influences HtrA3 trimer formation. The unique C-terminal sequence of ΔN-HtrA3S appeared to have little effect on activity and oligomerization. Additionally, we examined the cleavage specificity of ΔN-HtrA3. Results reported in this paper provide new insights into the structure and function of ΔN-HtrA3, which seems to have a unique combination of features among human HtrA proteases.« less

  14. Structural and functional analysis of human HtrA3 protease and its subdomains

    SciTech Connect

    Glaza, Przemyslaw; Osipiuk, Jerzy; Wenta, Tomasz; Zurawa-Janicka, Dorota; Jarzab, Miroslaw; Lesner, Adam; Banecki, Bogdan; Skorko-Glonek, Joanna; Joachimiak, Andrzej; Lipinska, Barbara; van Raaij, Mark J.

    2015-06-25

    Human HtrA3 protease, which induces mitochondria-mediated apoptosis, can be a tumor suppressor and a potential therapeutic target in the treatment of cancer. However, there is little information about its structure and biochemical properties. HtrA3 is composed of an N-terminal domain not required for proteolytic activity, a central serine protease domain and a C-terminal PDZ domain. HtrA3S, its short natural isoform, lacks the PDZ domain which is substituted by a stretch of 7 C-terminal amino acid residues, unique for this isoform. This paper presents the crystal structure of the HtrA3 protease domain together with the PDZ domain (ΔN-HtrA3), showing that the protein forms a trimer whose protease domains are similar to those of human HtrA1 and HtrA2. The ΔN-HtrA3 PDZ domains are placed in a position intermediate between that in the flat saucer-like HtrA1 SAXS structure and the compact pyramidal HtrA2 X-ray structure. The PDZ domain interacts closely with the LB loop of the protease domain in a way not found in other human HtrAs. ΔN-HtrA3 with the PDZ removed (ΔN-HtrA3-ΔPDZ) and an N-terminally truncated HtrA3S (ΔN-HtrA3S) were fully active at a wide range of temperatures and their substrate affinity was not impaired. This indicates that the PDZ domain is dispensable for HtrA3 activity. As determined by size exclusion chromatography, ΔN-HtrA3 formed stable trimers while both ΔN-HtrA3-ΔPDZ and ΔN-HtrA3S were monomeric. This suggests that the presence of the PDZ domain, unlike in HtrA1 and HtrA2, influences HtrA3 trimer formation. The unique C-terminal sequence of ΔN-HtrA3S appeared to have little effect on activity and oligomerization. Additionally, we examined the cleavage specificity of ΔN-HtrA3. Results reported in this paper provide new insights into the structure and function of ΔN-HtrA3, which seems to have a unique combination of features among human HtrA proteases.

  15. The HTR1A and HTR1B receptor genes influence stress-related information processing.

    PubMed

    Mekli, Krisztina; Payton, Antony; Miyajima, Fabio; Platt, Hazel; Thomas, Emma; Downey, Darragh; Lloyd-Williams, Kathryn; Chase, Diana; Toth, Zoltan G; Elliott, Rebecca; Ollier, William E; Anderson, Ian M; Deakin, J F William; Bagdy, Gyorgy; Juhasz, Gabriella

    2011-01-01

    The serotonergic system has been widely implicated in stress related psychiatric disorders such as depression and anxiety. We investigated the possible association between depression and anxiety scores and SNPs within the HTR1A and HTR1B genes in a population sample (n=1387). There was no direct SNP-phenotype association, but in interaction with recent stressful life events rs6295 G, rs878567 T alleles and rs6296 C alleles were associated with significantly higher symptom scores. A subset of control subjects (n=101) took part in a computerised face emotion processing task. Healthy rs6295 GG carriers did not show an affective bias to perceive more negative emotions but reacted more quickly to fearful faces. Thus we conclude that the serotonin-1A receptor conveys vulnerability to these psychiatric disorders by modulating threat-related information processing. Our results extend previous findings of an interaction between stressful life events and the serotonin transporter gene to two other genes in the serotonergic pathway and emphasise the possible role of increased threat-related information processing as an intermediate phenotype.

  16. Graphite as a structural material in HTR plants

    NASA Astrophysics Data System (ADS)

    Theymann, W.; Schmidt, A.

    1990-04-01

    Graphite has been selected as a structural material in HTR plants because of its favourable characteristics. The low ductility and the low tensile strength of this material as well as its behaviour under the impact of fast neutron irradiation require special construction directives and design criteria. It is demonstrated that by an appropriate structural design it is possible to separate the tasks and functions of the individual graphite components, which permits a classification of each component into one of three classes of qualitity assurance. Adequate stress criteria have been developed for the graphite internals of HTR based on probabilistic methods.

  17. HtrA2 suppresses autoimmune arthritis and regulates activation of STAT3

    PubMed Central

    Lee, Seung Hoon; Moon, Young-Mee; Seo, Hyeon-Beom; Kim, Se-Young; Kim, Eun-Kyung; Yi, Junyeong; Nam, Min-Kyung; Min, Jun-Ki; Park, Sung-Hwan; Rhim, Hyangshuk; Cho, Mi-La

    2016-01-01

    Rheumatoid arthritis (RA) is an autoimmune disease that is related to the induction of T helper (Th)17 cells, which secrete interleukin-17, and activation of the signal transducer and activator of transcription (STAT) 3. The expression of high-temperature requirement protein A (HtrA) 2, a serine protease involved in apoptosis, was decreased in RA patients nonresponsive to drug treatment of RA. The aim of this study was to determine whether overexpression of HtrA2 has a therapeutic effect on RA. Th17 differentiation, osteoclastogenesis, and lymphocyte activation are increased in motor neuron degeneration (mnd)2 mice, which lack HtrA2 activity because of a missense mutation (Ser276Cys) in the protease domain of HtrA2. The inhibitor of HtrA2 also increased Th17 differentiation. On the other hand, HtrA2 induced cleavage of STAT3 and overexpression of HtrA2 attenuated CIA in a mouse model. HtrA2 overexpression inhibited plaque development as well as the differentiation of Th17 in ApoE−/− mice after immunization with proteoglycans to induce a hyperlipidemia-based RA animal model. The therapeutic function of HtrA2 in inflammatory diseases is linked with Th17 development and the STAT3 pathway in splenocytes. These results suggest that HtrA2 participates in immunomodulatory activity where the upregulation of HtrA2 may shed light on therapeutic approaches to RA and hyperlipidemia. PMID:28008946

  18. Reactor Physics Characterization of the HTR Module with UCO Fuel

    SciTech Connect

    Gerhard Strydom

    2011-01-01

    ABSTRACT The HTR Module [1] is a graphite-moderated, helium cooled pebble bed High Temperature Reactor (HTR) design that has been extensively used as a reference template for the former South African and current Chinese HTR [2] programs. This design utilized spherical fuel elements packed into a dynamic pebble bed, consisting of TRISO coated uranium oxide (UO2) fuel kernels with a U-235 enrichment of 7.8% and a Heavy Metal loading of 7 grams per pebble. The main objective of this study is to compare several important reactor physics and core design parameters for the HTR Module and an identical design utilizing UCO fuel kernels. Fuel kernels of this type are currently being tested in the Idaho National Laboratory’s (INL) Advanced Test Reactor (ATR) as part of the larger Next Generation Nuclear Plant (NGNP) project. The PEBBED-THERMIX [3] code, which was developed specifically for the analysis of pebble bed HTRs, was used to compare the coupled neutronic and thermal fluid performance of the two designs.

  19. Protoporphyrins Enhance Oligomerization and Enzymatic Activity of HtrA1 Serine Protease

    PubMed Central

    Jo, Hakryul; Patterson, Victoria; Stoessel, Sean; Kuan, Chia-Yi; Hoh, Josephine

    2014-01-01

    High temperature requirement protein A1 (HtrA1), a secreted serine protease of the HtrA family, is associated with a multitude of human diseases. However, the exact functions of HtrA1 in these diseases remain poorly understood. We seek to unravel the mechanisms of HtrA1 by elucidating its interactions with chemical or biological modulators. To this end, we screened a small molecule library of 500 bioactive compounds to identify those that alter the formation of extracellular HtrA1 complexes in the cell culture medium. An initial characterization of two novel hits from this screen showed that protoporphyrin IX (PPP-IX), a precursor in the heme biosynthetic pathway, and its metalloporphyrin (MPP) derivatives fostered the oligomerization of HtrA1 by binding to the protease domain. As a result of the interaction with MPPs, the proteolytic activity of HtrA1 against Fibulin-5, a specific HtrA1 substrate in age-related macular degeneration (AMD), was increased. This physical interaction could be abolished by the missense mutations of HtrA1 found in patients with cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL). Furthermore, knockdown of HtrA1 attenuated apoptosis induced by PPP-IX. These results suggest that PPP-IX, or its derivatives, and HtrA1 may function as co-factors whereby porphyrins enhance oligomerization and the protease activity of HtrA1, while active HtrA1 elevates the pro-apoptotic actions of porphyrin derivatives. Further analysis of this interplay may shed insights into the pathogenesis of diseases such as AMD, CARASIL and protoporphyria, as well as effective therapeutic development. PMID:25506911

  20. Corrosion of metallic materials in HTR-helium environments

    NASA Astrophysics Data System (ADS)

    Graham, L. W.

    1990-04-01

    The HTR core system has been successfully developed to provide helium coolant temperatures up to 1200 K giving the potential to exploit a wide variety of commercial applications. The present paper summarises the main corrosion effects found in HTR-helium environments, especially those which could change the mechanical properties of component materials. It is shown that corrosion phenomena derive from the presence of low concentrations of the impurities H 2O, CO, CH 4 and H 2 in the helium which can produce oxidation, decarburisation and carburisation depending on the relative and absolute partial pressures of the impurities and the temperature. Major corrosion effects can be avoided by control of the environment but the large heat to heat variations in behaviour found in materials exposed to nominally protective regimes at the higher temperatures of application requires further study.

  1. EBSD investigation of SiC for HTR fuel particles

    NASA Astrophysics Data System (ADS)

    Helary, D.; Dugne, O.; Bourrat, X.; Jouneau, P. H.; Cellier, F.

    2006-05-01

    Electron back-scattering diffraction (EBSD) can be successfully performed on SiC coatings for HTR fuel particles. EBSD grain maps obtained from thick and thin unirradiated samples are presented, along with pole figures showing textures and a chart showing the distribution of grain aspect ratios. This information is of great interest, and contributes to improving the process parameters and ensuring the reproducibility of coatings.

  2. Comparison of irradiation behaviour of HTR graphite grades

    NASA Astrophysics Data System (ADS)

    Heijna, M. C. R.; de Groot, S.; Vreeling, J. A.

    2017-08-01

    The INNOGRAPH irradiations were executed in the High Flux Reactor (HFR) in Petten by NRG supported by the European Framework programs HTR-M, RAPHAEL, and ARCHER to generate data on the irradiation behaviour of graphite grades for High Temperature Reactor (HTR) application available at that time. Samples of the graphite grades NBG-10, NBG-17, NBG-18, NBG-20, NBG-25, PCEA, PPEA, PCIB, and IG-110 have been irradiated at 750 °C and 950 °C. The inherent scatter induced by the probabilistic material behaviour of graphite requires uncertainty and scatter induced by test conditions and post-irradiation examination to be minimized. The INNOGRAPH irradiations supplied an adequate number of irradiated samples to enable accurate determination of material properties and their evolution under irradiation. This allows comparison of different graphite grades and a qualitative assessment of their appropriateness for HTR applications, as a basis of selection, design and core component lifetime. The results indicate that coarse grained graphite grades exhibit more favourable behaviour for application in HTRs due to their low dimensional anisotropy and fracture propagation resilience.

  3. Laser ultrasonics evaluation and testing of coated HTR nuclear fuel

    NASA Astrophysics Data System (ADS)

    Amziane, Ahmed; Amari, Mohamed; Mounier, Denis; Breteau, Jean-Marc; Joly, Nicolas; Edely, Mathieu; Larcher, Maxime; Noiré, Paul; Banchet, Julien; Tisseur, David; Gusev, Vitalyi

    2011-05-01

    Laser ultrasonics was applied to the manufacturing control of the quality and integrity (no failure) of coated spherical particles designed for High Temperature Reactors (HTR). The coating of the nuclear fuel kernel is designed to prevent from the diffusion of fission products outside the particle during reactor operation. The quality assessment of the coating layers is of major importance. Using laser ultrasonics, we determined the vibration eigenmodes of dummy HTR particles. The vibration spectrum of a HTR particle provides a non-destructive method of evaluating some important mechanical parameters of the coating. Moreover, without damaging the particle, the laser ultrasonics technique allows to test the presence of a crack in the SiC layer, through the observation of the particle vibration spectrum, which is significantly changed, compared to that of a defect-free particle. We applied nanosecond acoustic pulses, i.e., high frequency laser-generated ultrasound, to measure the acoustic velocity of longitudinal waves the SiC layer. This technique provides an alternative method of evaluation of the Young modulus of the SiC layer. We measure the velocity of surface acoustic waves (SAW) on a pyrocarbon layer cross-section and we demonstrated that the anisotropy of the internal pyrocarbon layer can be evaluated by laser ultrasonics.

  4. [Effect of embryonic anlage allografts of the rat spinal cord on growth of regenerating fibers of the recipient nerve].

    PubMed

    Petrova, E S; Isaeva, E N

    2014-01-01

    A comparative study of the effect of tissue and suspension allografts of an embryonic spinal cord on regeneration of nerve fibers of impaired (by application of a ligature) sciatic nerve in rats was conducted. It was demonstrated that unlike tissue grafts that reach a large volume 21 and 60 days after transplantation, suspension grafts do not inhibit the growth of axons of the recipient to the periphery. It was established that introduction of a suspension of dissociated cells of the spinal cord embryonic anlages (but not fragments of these anlages) into the impaired sciatic nerve in rats results in an increase in the amount of myelinated regenerating nerve fibers of the recipient 60 days after the operation.

  5. The trimeric serine protease HtrA1 forms a cage-like inhibition complex with an anti-HtrA1 antibody.

    PubMed

    Ciferri, Claudio; Lipari, Michael T; Liang, Wei-Ching; Estevez, Alberto; Hang, Julie; Stawicki, Scott; Wu, Yan; Moran, Paul; Elliott, Mike; Eigenbrot, Charles; Katschke, Kenneth J; van Lookeren Campagne, Menno; Kirchhofer, Daniel

    2015-12-01

    High temperature requirement A1 (HtrA1) is a trypsin-fold serine protease implicated in the progression of age-related macular degeneration (AMD). Our interest in an antibody therapy to neutralize HtrA1 faces the complication that the target adopts a trimeric arrangement, with three active sites in close proximity. In the present study, we describe antibody 94, obtained from a human antibody phage display library, which forms a distinct macromolecular complex with HtrA1 and inhibits the enzymatic activity of recombinant and native HtrA1 forms. Using biochemical methods and negative-staining EM we were able to elucidate the molecular composition of the IgG94 and Fab94 complexes and the associated inhibition mechanism. The 246-kDa complex between the HtrA1 catalytic domain trimer (HtrA1_Cat) and Fab94 had a propeller-like organization with one Fab bound peripherally to each protomer. Low-resolution EM structures and epitope mapping indicated that the antibody binds to the surface-exposed loops B and C of the catalytic domain, suggesting an allosteric inhibition mechanism. The HtrA1_Cat-IgG94 complex (636 kDa) is a cage-like structure with three centrally located IgG94 molecules co-ordinating two HtrA1_Cat trimers and the six active sites pointing into the cavity of the cage. In both complexes, all antigen-recognition regions (paratopes) are found to bind one HtrA1 protomer and all protomers are bound by a paratope, consistent with the complete inhibition of enzyme activity. Therefore, in addition to its potential therapeutic usefulness, antibody 94 establishes a new paradigm of multimeric serine protease inhibition.

  6. Cardiac Specific Overexpression of Mitochondrial Omi/HtrA2 Induces Myocardial Apoptosis and Cardiac Dysfunction

    PubMed Central

    Wang, Ke; Yuan, Yuexing; Liu, Xin; Lau, Wayne Bond; Zuo, Lin; Wang, Xiaoliang; Ma, Lu; Jiao, Kun; Shang, Jianyu; Wang, Wen; Ma, Xinliang; Liu, Huirong

    2016-01-01

    Myocardial apoptosis is a significant problem underlying ischemic heart disease. We previously reported significantly elevated expression of cytoplasmic Omi/HtrA2, triggers cardiomyocytes apoptosis. However, whether increased Omi/HtrA2 within mitochondria itself influences myocardial survival in vivo is unknown. We aim to observe the effects of mitochondria-specific, not cytoplasmic, Omi/HtrA2 on myocardial apoptosis and cardiac function. Transgenic mice overexpressing cardiac-specific mitochondrial Omi/HtrA2 were generated and they had increased myocardial apoptosis, decreased systolic and diastolic function, and decreased left ventricular remodeling. Transiently or stably overexpression of mitochondria Omi/HtrA2 in H9C2 cells enhance apoptosis as evidenced by elevated caspase-3, -9 activity and TUNEL staining, which was completely blocked by Ucf-101, a specific Omi/HtrA2 inhibitor. Mechanistic studies revealed mitochondrial Omi/HtrA2 overexpression degraded the mitochondrial anti-apoptotic protein HAX-1, an effect attenuated by Ucf-101. Additionally, transfected cells overexpressing mitochondrial Omi/HtrA2 were more sensitive to hypoxia and reoxygenation (H/R) induced apoptosis. Cyclosporine A (CsA), a mitochondrial permeability transition inhibitor, blocked translocation of Omi/HtrA2 from mitochondrial to cytoplasm, and protected transfected cells incompletely against H/R-induced caspase-3 activation. We report in vitro and in vivo overexpression of mitochondrial Omi/HtrA2 induces cardiac apoptosis and dysfunction. Thus, strategies to directly inhibit Omi/HtrA2 or its cytosolic translocation from mitochondria may protect against heart injury. PMID:27924873

  7. Characterisation of worldwide Helicobacter pylori strains reveals genetic conservation and essentiality of serine protease HtrA

    PubMed Central

    Tegtmeyer, Nicole; Moodley, Yoshan; Yamaoka, Yoshio; Pernitzsch, Sandy Ramona; Schmidt, Vanessa; Traverso, Francisco Rivas; Schmidt, Thomas P.; Rad, Roland; Yeoh, Khay Guan; Bow, Ho; Torres, Javier; Gerhard, Markus; Schneider, Gisbert; Wessler, Silja

    2015-01-01

    Summary HtrA proteases and chaperones exhibit important roles in periplasmic protein quality control and stress responses. The genetic inactivation of htrA has been described for many bacterial pathogens. However, in some cases such as the gastric pathogen H elicobacter pylori, HtrA is secreted where it cleaves the tumour‐suppressor E‐cadherin interfering with gastric disease development, but the generation of htrA mutants is still lacking. Here, we show that the htrA gene locus is highly conserved in worldwide strains. HtrA presence was confirmed in 992 H . pylori isolates in gastric biopsy material from infected patients. Differential RNA‐sequencing (dRNA‐seq) indicated that htrA is encoded in an operon with two subsequent genes, HP1020 and HP1021. Genetic mutagenesis and complementation studies revealed that HP1020 and HP1021, but not htrA, can be mutated. In addition, we demonstrate that suppression of HtrA proteolytic activity with a newly developed inhibitor is sufficient to effectively kill H . pylori, but not other bacteria. We show that H elicobacter  htrA is an essential bifunctional gene with crucial intracellular and extracellular functions. Thus, we describe here the first microbe in which htrA is an indispensable gene, a situation unique in the bacterial kingdom. HtrA can therefore be considered a promising new target for anti‐bacterial therapy. PMID:26568477

  8. Characterisation of worldwide Helicobacter pylori strains reveals genetic conservation and essentiality of serine protease HtrA.

    PubMed

    Tegtmeyer, Nicole; Moodley, Yoshan; Yamaoka, Yoshio; Pernitzsch, Sandy Ramona; Schmidt, Vanessa; Traverso, Francisco Rivas; Schmidt, Thomas P; Rad, Roland; Yeoh, Khay Guan; Bow, Ho; Torres, Javier; Gerhard, Markus; Schneider, Gisbert; Wessler, Silja; Backert, Steffen

    2016-03-01

    HtrA proteases and chaperones exhibit important roles in periplasmic protein quality control and stress responses. The genetic inactivation of htrA has been described for many bacterial pathogens. However, in some cases such as the gastric pathogen Helicobacter pylori, HtrA is secreted where it cleaves the tumour-suppressor E-cadherin interfering with gastric disease development, but the generation of htrA mutants is still lacking. Here, we show that the htrA gene locus is highly conserved in worldwide strains. HtrA presence was confirmed in 992 H. pylori isolates in gastric biopsy material from infected patients. Differential RNA-sequencing (dRNA-seq) indicated that htrA is encoded in an operon with two subsequent genes, HP1020 and HP1021. Genetic mutagenesis and complementation studies revealed that HP1020 and HP1021, but not htrA, can be mutated. In addition, we demonstrate that suppression of HtrA proteolytic activity with a newly developed inhibitor is sufficient to effectively kill H. pylori, but not other bacteria. We show that Helicobacter htrA is an essential bifunctional gene with crucial intracellular and extracellular functions. Thus, we describe here the first microbe in which htrA is an indispensable gene, a situation unique in the bacterial kingdom. HtrA can therefore be considered a promising new target for anti-bacterial therapy.

  9. Evolution of mitochondrial cell death pathway: Proapoptotic role of HtrA2/Omi in Drosophila

    SciTech Connect

    Igaki, Tatsushi; Suzuki, Yasuyuki; Tokushige, Naoko; Aonuma, Hiroka; Takahashi, Ryosuke . E-mail: ryosuket@kuhp.kyoto-u.ac.jp; Miura, Masayuki . E-mail: miura@mol.f.u-tokyo.ac.jp

    2007-05-18

    Despite the essential role of mitochondria in a variety of mammalian cell death processes, the involvement of mitochondrial pathway in Drosophila cell death has remained unclear. To address this, we cloned and characterized DmHtrA2, a Drosophila homolog of a mitochondrial serine protease HtrA2/Omi. We show that DmHtrA2 normally resides in mitochondria and is up-regulated by UV-irradiation. Upon receipt of apoptotic stimuli, DmHtrA2 is translocated to extramitochondrial compartment; however, unlike its mammalian counterpart, the extramitochondrial DmHtrA2 does not diffuse throughout the cytosol but stays near the mitochondria. RNAi-mediated knock-down of DmHtrA2 in larvae or adult flies results in a resistance to stress stimuli. DmHtrA2 specifically cleaves Drosophila inhibitor-of-apoptosis protein 1 (DIAP1), a cellular caspase inhibitor, and induces cell death both in vitro and in vivo as potent as other fly cell death proteins. Our observations suggest that DmHtrA2 promotes cell death through a cleavage of DIAP1 in the vicinity of mitochondria, which may represent a prototype of mitochondrial cell death pathway in evolution.

  10. HtrA Is Important for Stress Resistance and Virulence in Haemophilus parasuis

    PubMed Central

    Zhang, Luhua; Li, Ying; Wen, Yiping; Lau, Gee W.; Huang, Xiaobo; Wu, Rui; Yan, Qigui; Huang, Yong; Zhao, Qin; Ma, Xiaoping

    2016-01-01

    Haemophilus parasuis is an opportunistic pathogen that causes Glässer's disease in swine, with polyserositis, meningitis, and arthritis. The high-temperature requirement A (HtrA)-like protease, which is involved in protein quality control, has been reported to be a virulence factor in many pathogens. In this study, we showed that HtrA of H. parasuis (HpHtrA) exhibited both chaperone and protease activities. Finally, nickel import ATP-binding protein (NikE), periplasmic dipeptide transport protein (DppA), and outer membrane protein A (OmpA) were identified as proteolytic substrates for HpHtrA. The protease activity reached its maximum at 40°C in a time-dependent manner. Disruption of the htrA gene from strain SC1401 affected tolerance to temperature stress and resistance to complement-mediated killing. Furthermore, increased autoagglutination and biofilm formation were detected in the htrA mutant. In addition, the htrA mutant was significantly attenuated in virulence in the murine model of infection. Together, these data demonstrate that HpHtrA plays an important role in the virulence of H. parasuis. PMID:27217419

  11. DNA Methylation Analysis of HTR2A Regulatory Region in Leukocytes of Autistic Subjects.

    PubMed

    Hranilovic, Dubravka; Blazevic, Sofia; Stefulj, Jasminka; Zill, Peter

    2016-02-01

    Disturbed brain and peripheral serotonin homeostasis is often found in subjects with autism spectrum disorder (ASD). The role of the serotonin receptor 2A (HTR2A) in the regulation of central and peripheral serotonin homeostasis, as well as its altered expression in autistic subjects, have implicated the HTR2A gene as a major candidate for the serotonin disturbance seen in autism. Several studies, yielding so far inconclusive results, have attempted to associate autism with a functional SNP -1438 G/A (rs6311) in the HTR2A promoter region, while possible contribution of epigenetic mechanisms, such as DNA methylation, to HTR2A dysregulation in autism has not yet been investigated. In this study, we compared the mean DNA methylation within the regulatory region of the HTR2A gene between autistic and control subjects. DNA methylation was analysed in peripheral blood leukocytes using bisulfite conversion and sequencing of the HTR2A region containing rs6311 polymorphism. Autistic subjects of rs6311 AG genotype displayed higher mean methylation levels within the analysed region than the corresponding controls (P < 0.05), while there was no statistically significant difference for AA and GG carriers. Our study provides preliminary evidence for increased HTR2A promoter methylation in leukocytes of a portion of adult autistic subjects, indicating that epigenetic mechanisms might contribute to HTR2A dysregulation observed in individuals with ASD. © 2015 International Society for Autism Research, Wiley Periodicals, Inc.

  12. Pathogenic Role of Serine Protease HtrA2/Omi in Neurodegenerative Diseases.

    PubMed

    Goo, Hui-Gwan; Rhim, Hyangshuk; Kang, Seongman

    2017-01-01

    High-temperature-requirement A2 (HtrA2)/Omi/PARK13 is a serine protease with extensive homology to the Escherichia coli HtrAs that are required for bacterial survival at high temperatures. The HtrA2 protein is a key modulator of mitochondrial molecular quality control but under stressful conditions it is released into the cytosol, where it promotes cell death by various pathways, including caspase-dependent pathway and ER stress-mediated apoptosis. Recently, the HtrA2 protein has received great attention for its potential role in neurodegeneration. Here, we review the current knowledge and pathophysiological functions of the HtrA2 protein in neurodegenerative disorders such as Parkinson's and Alzheimer's disease. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  13. Placental HTR2A methylation is associated with infant neurobehavioral outcomes

    PubMed Central

    Paquette, Alison G; Lesseur, Corina; Armstrong, David A; Koestler, Devin C; Appleton, Allison A; Lester, Barry M; Marsit, Carmen J

    2013-01-01

    The serotonin receptor, HTR2A, exhibits placental expression and function and can be controlled through DNA methylation. The relationship between methylation of HTR2A in the placenta and neurodevelopmental outcomes, evaluated using the NICU Network Neurobehavioral Scales (NNNS), was assessed in newborn infants (n = 444). HTR2A methylation was significantly higher in males and marginally higher in infants whose mothers reported tobacco use during pregnancy. Controlling for confounding variables, HTR2A methylation was negatively associated with infant quality of movement (p = 0.05) and positively associated with infant attention (p = 0.0001). These results suggest that methylation of the HTR2A gene can be biologically and environmentally modulated and is associated with key measures of neurodevelopment. PMID:23880519

  14. HTR Spherical Super Lattice Model for Equilibrium Fuel Cycle Analysis

    SciTech Connect

    Gray S. Cahng

    2005-09-01

    Advanced High Temperature gas-cooled Reactors (HTR) currently being developed (GFR, VHTR - Very High Temperature gas-cooled Reactor, PBMR, and GT-MHR) are able to achieve a simplification of safety through reliance on innovative features and passive systems. One of the innovative features in these HTRs is reliance on ceramic-coated fuel particles to retain the fission products even under extreme accident conditions. The effect of the random fuel kernel distribution in the fuel pebble / block is addressed through the use of the Dancoff correction factor in the resonance treatment. In addition, the Dancoff correction factor is a function of burnup and fuel kernel packing factor, which requires that the Dancoff correction factor be updated during Equilibrium Fuel Cycle (EqFC) analysis. Although HTR fuel is rather homogeneously dispersed in the fuel graphite matrix, the heterogeneity effects in between fuel kernels and pebbles cannot be ignored. The double-heterogeneous lattice model recently developed at the Idaho National Engineering and Environmental Laboratory (INEEL) contains tens of thousands of cubic fuel kernel cells, which makes it very difficult to deplete the fuel, kernel by kernel (KbK), for the EqFC analysis. In addition, it is not possible to preserve the cubic size and packing factor in a spherical fuel pebble. To avoid these difficulties, a newly developed and validated HTR pebble-bed Kernel-by-Kernel spherical (KbK-sph) model, has been developed and verified in this study. The objective of this research is to introduce the KbK-sph model and super whole Pebble lattice model (PLM). The verified double-heterogeneous KbK-sph and pebble homogeneous lattice model (HLM) are used for the fuel burnup chracteristics analysis and important safety parameters validation. This study summarizes and compares the KbK-sph and HLM burnup analyzed results. Finally, we discus the Monte-Carlo coupling with a fuel depletion and buildup code - Origen-2 as a fuel burnup

  15. HTR Spherical Super Lattice Model for Equilibrium Fuel Cycle Analysis

    SciTech Connect

    Gray S. Cahng

    2005-09-01

    Advanced High Temperature gas-cooled Reactors (HTR) currently being developed (GFR, VHTR - Very High Temperature gas-cooled Reactor, PBMR, and GT-MHR) are able to achieve a simplification of safety through reliance on innovative features and passive systems. One of the innovative features in these HTRs is reliance on ceramic-coated fuel particles to retain the fission products even under extreme accident conditions. The effect of the random fuel kernel distribution in the fuel pebble / block is addressed through the use of the Dancoff correction factor in the resonance treatment. In addition, the Dancoff correction factor is a function of burnup and fuel kernel packing factor, which requires that the Dancoff correction factor be updated during Equilibrium Fuel Cycle (EqFC) analysis. Although HTR fuel is rather homogeneously dispersed in the fuel graphite matrix, the heterogeneity effects in between fuel kernels and pebbles cannot be ignored. The double-heterogeneous lattice model recently developed at the Idaho National Engineering and Environmental Laboratory (INEEL) contains tens of thousands of cubic fuel kernel cells, which makes it very difficult to deplete the fuel, kernel by kernel (KbK), for the EqFC analysis. In addition, it is not possible to preserve the cubic size and packing factor in a spherical fuel pebble. To avoid these difficulties, a newly developed and validated HTR pebble-bed Kernel-by-Kernel spherical (KbK-sph) model, has been developed and verified in this study. The objective of this research is to introduce the KbK-sph model and super whole Pebble lattice model (PLM). The verified double-heterogeneous KbK-sph and pebble homogeneous lattice model (HLM) are used for the fuel burnup chracteristics analysis and important safety parameters validation. This study summarizes and compares the KbK-sph and HLM burnup analyzed results. Finally, we discus the Monte-Carlo coupling with a fuel depletion and buildup code - Origen-2 as a fuel burnup

  16. HtrA3 Is Downregulated in Cancer Cell Lines and Significantly Reduced in Primary Serous and Granulosa Cell Ovarian Tumors.

    PubMed

    Singh, Harmeet; Li, Ying; Fuller, Peter J; Harrison, Craig; Rao, Jyothsna; Stephens, Andrew N; Nie, Guiying

    2013-01-01

    Objective. The high temperature requirement factor A3 (HtrA3) is a serine protease homologous to bacterial HtrA. Four human HtrAs have been identified. HtrA1 and HtrA3 share a high degree of domain organization and are downregulated in a number of cancers, suggesting a widespread loss of these proteases in cancer. This study examined how extensively the HtrA (HtrA1-3) proteins are downregulated in commonly used cancer cell lines and primary ovarian tumors.Methods. RT-PCR was applied to various cancer cell lines (n=17) derived from the ovary, endometrium, testes, breast, prostate, and colon, and different subtypes of primary ovarian tumors [granulosa cell tumors (n=19), mucinous cystadenocarcinomas (n=6), serous cystadenocarcinomas (n=8)] and normal ovary (n = 9). HtrA3 protein was localized by immunohistochemistry.Results. HtrA3 was extensively downregulated in the cancer cell lines examined including the granulosa cell tumor-derived cell lines. In primary ovarian tumors, the HtrA3 was significantly lower in serous cystadenocarcinoma and granulosa cell tumors. In contrast, HtrA1 and HtrA2 were expressed in all samples with no significant differences between the control and tumors. In normal postmenopausal ovary, HtrA3 protein was localized to lutenizing stromal cells and corpus albicans. In serous cystadenocarcinoma, HtrA3 protein was absent in the papillae but detected in the mesenchymal cyst wall.Conclusion. HtrA3 is more extensively downregulated than HtrA1-2 in cancer cell lines. HtrA3, but not HtrA1 or HtrA2, was decreased in primary ovarian serous cystadenocarcinoma and granulosa cell tumors. This study provides evidence that HtrA3 may be the most relevant HtrA associated with ovarian malignancy.

  17. HtrA3 stromal expression is correlated with tumor budding in stage II colorectal cancer.

    PubMed

    Forse, Catherine L; Rahimi, Mahdi; Diamandis, Eleftherios P; Assarzadegan, Naziheh; Dawson, Heather; Grin, Andrea; Kennedy, Erin; O'Connor, Brenda; Messenger, David E; Riddell, Robert H; Kirsch, Richard; Karagiannis, George S

    2017-08-01

    Tumor budding is a well-established adverse prognostic factor in colorectal carcinoma (CRC). It may represent a form of epithelial-to-mesenchymal transition (EMT), although the underlying mechanisms remain unclear. High-temperature requirement A3 (HtrA3) is an inhibitor of the bone morphogenetic protein pathway, the suppression of which has been linked to EMT. Since HtrA3 is highly expressed in the desmoplastic stroma at the CRC invasive front, we sought to evaluate the relationship between tumor budding and HtrA3 expression in 172 stage II CRC resection specimens. All tumors were evaluated for tumor budding, with the highest budding slide selected for pan-keratin (CK) and HtrA3 immunohistochemistry. Representative areas of tumor core and invasive front, including budding and non-budding areas, were marked on CK stained slides, and then evaluated on HtrA3 stained slides. HtrA3 expression in tumor cells (tHtrA3) and peritumoral stroma (sHtrA3) was assessed for staining percentage and intensity (the product yielding a final score). Tumors with high-grade tumor budding (HGTB) showed increased expression of sHtrA3 in budding areas compared to non-budding areas at the invasive front (P<0.001). In addition, sHtrA3 expression at the invasive front was significantly higher in HGTB tumors compared to minimally budding tumors (P<0.05). tHtrA3 expression at the invasive front was significantly associated with high histological grade (P<0.05). Higher sHtrA3 expression in the tumor core (but not invasive front) was significantly associated with decreased 5-year overall survival on univariate analysis (P<0.05), but not multivariate analysis. HtrA3 expression in the peritumoral stroma of patients with stage II CRC is associated with HGTB and may be a novel marker of poor outcome. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Primary structure and functional analysis of the soluble transducer protein HtrXI in the archaeon Halobacterium salinarium.

    PubMed Central

    Brooun, A; Zhang, W; Alam, M

    1997-01-01

    Signal transduction in the archaeon Halobacterium salinarium is mediated by a family of 13 soluble and membrane-bound transducers. Here, we report the primary structure and functional analysis of one of the smallest halobacterial putative transducers, HtrXI. Hydropathy plot analysis of the primary structure predicts no membrane-spanning segments in HtrXI. The fractionation of the H. salinarium proteins confirmed that HtrXI is a soluble protein. Capillary assay with an HtrXI deletion mutant and a complemented strain revealed that this soluble transducer is involved in Asp and Glu taxis. In vivo analysis of the methylesterase activity of the htrXI-1 deletion mutant suggests that HtrXI plays an important role in the adaptation of the chemotactic responses to His, Asp, and Glu, which are attractants for halobacteria. Stimulation by Asp and Glu causes demethylation of HtrXI and of another putative transducer, HtrVII. But addition of His to halobacterial cells increases HtrXI methylation together with that of other putative transducers. In the absence of HtrXI, stimulation by either Glu or His does not decrease or increase the methylation of any putative transducers. Therefire, the HtrXI transducer appears to have a complex role in chemotaxis signal transduction. PMID:9139915

  19. Insights into the Cyanobacterial Deg/HtrA Proteases

    PubMed Central

    Cheregi, Otilia; Wagner, Raik; Funk, Christiane

    2016-01-01

    Proteins are the main machinery for all living processes in a cell; they provide structural elements, regulate biochemical reactions as enzymes, and are the interface to the outside as receptors and transporters. Like any other machinery proteins have to be assembled correctly and need maintenance after damage, e.g., caused by changes in environmental conditions, genetic mutations, and limitations in the availability of cofactors. Proteases and chaperones help in repair, assembly, and folding of damaged and misfolded protein complexes cost-effective, with low energy investment compared with neo-synthesis. Despite their importance for viability, the specific biological role of most proteases in vivo is largely unknown. Deg/HtrA proteases, a family of serine-type ATP-independent proteases, have been shown in higher plants to be involved in the degradation of the Photosystem II reaction center protein D1. The objective of this review is to highlight the structure and function of their cyanobacterial orthologs. Homology modeling was used to find specific features of the SynDeg/HtrA proteases of Synechocystis sp. PCC 6803. Based on the available data concerning their location and their physiological substrates we conclude that these Deg proteases not only have important housekeeping and chaperone functions within the cell, but also are needed for remodeling the cell exterior. PMID:27252714

  20. Improved Neutronics Treatment of Burnable Poisons for the Prismatic HTR

    SciTech Connect

    Y. Wang; A. A. Bingham; J. Ortensi; C. J. Permann

    2012-10-01

    In prismatic block High Temperature Reactors (HTR), highly absorbing material such a burnable poison (BP) cause local flux depressions and large gradients in the flux across the blocks which can be a challenge to capture accurately with traditional homogenization methods. The purpose of this paper is to quantify the error associated with spatial homogenization, spectral condensation and discretization and to highlight what is needed for improved neutronics treatments of burnable poisons for the prismatic HTR. A new triangular based mesh is designed to separate the BP regions from the fuel assembly. A set of packages including Serpent (Monte Carlo), Xuthos (1storder Sn), Pronghorn (diffusion), INSTANT (Pn) and RattleSnake (2ndorder Sn) is used for this study. The results from the deterministic calculations show that the cross sections generated directly in Serpent are not sufficient to accurately reproduce the reference Monte Carlo solution in all cases. The BP treatment produces good results, but this is mainly due to error cancellation. However, the Super Cell (SC) approach yields cross sections that are consistent with cross sections prepared on an “exact” full core calculation. In addition, very good agreement exists between the various deterministic transport and diffusion codes in both eigenvalue and power distributions. Future research will focus on improving the cross sections and quantifying the error cancellation.

  1. The Developmental Basis of Epigenetic Regulation of HTR2A and Psychiatric Outcomes

    PubMed Central

    Paquette, Alison G.; Marsit, Carmen J.

    2014-01-01

    The serotonin receptor 5-HT2A (encoded by HTR2A) is an important regulator of fetal brain development and adult cognitive function. Environmental signals that induce epigenetic changes of serotonin response genes, including HTR2A, have been implicated in adverse mental health outcomes. The objective of this perspective article is to address the medical implications of HTR2A epigenetic regulation, which has been associated with both infant neurobehavioral outcomes and adult mental health. Ongoing research has identified a region of the HTR2A promoter that has been associated with a number of medical outcomes in adults and infants, including bipolar disorder, schizophrenia, chronic fatigue syndrome, borderline personality disorder, suicidality, and neurobehavioral outcomes. Epigenetic regulation of HTR2A has been studied in several different types of tissues, including the placenta. The placenta is an important source of serotonin during fetal neurodevelopment, and placental epigenetic variation of HTR2A has been associated with infant neurobehavioral outcomes, which may represent the basis of adult mental health disorders. Further analysis is needed to identify intrinsic and extrinsic factors modulate HTR2A methylation, and the mechanism by which this epigenetic variation influences fetal growth and leads to altered brain development, manifesting in psychiatric disorders. PMID:25043477

  2. Genetic variation in HTR4 and lung function: GWAS follow-up in mouse.

    PubMed

    House, John S; Li, Huiling; DeGraff, Laura M; Flake, Gordon; Zeldin, Darryl C; London, Stephanie J

    2015-01-01

    Human genome-wide association studies (GWASs) have identified numerous associations between single nucleotide polymorphisms (SNPs) and pulmonary function. Proving that there is a causal relationship between GWAS SNPs, many of which are noncoding and without known functional impact, and these traits has been elusive. Furthermore, noncoding GWAS-identified SNPs may exert trans-regulatory effects rather than impact the proximal gene. Noncoding variants in 5-hydroxytryptamine (serotonin) receptor 4 (HTR4) are associated with pulmonary function in human GWASs. To gain insight into whether this association is causal, we tested whether Htr4-null mice have altered pulmonary function. We found that HTR4-deficient mice have 12% higher baseline lung resistance and also increased methacholine-induced airway hyperresponsiveness (AHR) as measured by lung resistance (27%), tissue resistance (48%), and tissue elastance (30%). Furthermore, Htr4-null mice were more sensitive to serotonin-induced AHR. In models of exposure to bacterial lipopolysaccharide, bleomycin, and allergic airway inflammation induced by house dust mites, pulmonary function and cytokine profiles in Htr4-null mice differed little from their wild-type controls. The findings of altered baseline lung function and increased AHR in Htr4-null mice support a causal relationship between genetic variation in HTR4 and pulmonary function identified in human GWAS.

  3. Borrelia burgdorferi HtrA: evidence for two-fold proteolysis of outer membrane protein p66

    PubMed Central

    Coleman, James L.; Toledo, Alvaro; Benach, Jorge L.

    2016-01-01

    Summary In prokaryotes, members of the High Temperature Requirement A (HtrA) family of serine proteases function in the periplasm to degrade damaged or improperly folded membrane proteins. Borrelia burgdorferi, the agent of Lyme disease, codes for a single HtrA homolog. Two-dimensional electrophoresis analysis of B. burgdorferi B31A3 and a strain that over-expresses HtrA (A3HtrAOE) identified a down-regulated protein in A3HtrAOE with a mass, pI and MALDI-TOF spectrum consistent with outer membrane protein p66. P66 and HtrA from cellular lysates partitioned into detergent-resistant membranes, which contain cholesterol-glycolipid-rich membrane regions known as lipid rafts, suggesting that HtrA and p66 may reside together in lipid rafts also. This agrees with previous work from our laboratory, which showed that HtrA and p66 are constituents of B. burgdorferi outer membrane vesicles. HtrA degraded p66 in vitro and A3HtrAOE expressed reduced levels of p66 in vivo. Fluorescence confocal microscopy revealed that HtrA and p66 co-localize in the membrane. The association of HtrA and p66 establishes that they could interact efficiently and their protease/substrate relationship provides functional relevance to this interaction. A3HtrAOE also showed reduced levels of p66 transcript in comparison to wild-type B31A3, indicating that HtrA-mediated regulation of p66 may occur at multiple levels. PMID:26370492

  4. The DBA Analysis of One Control Rod Withdrawal Out of the HTR-10GT Core

    SciTech Connect

    Mingang Lang; Yujie Dong

    2006-07-01

    The 10 MW High Temperature Gas Cooled Test Reactor (HTR-10) has been built in Institute of Nuclear and New Energy Technology (INET) and has been operating successfully since the beginning of 2003. The core outlet temperature of HTR-10 is 700 deg. C. To verify the technology of gas-turbine direct cycle, INET has planned to increase its core outlet temperature to 750 deg. C and use a helium gas turbine instead of the steam generator (then the reactor is called HTR-10GT). Though HTR-10 has good intrinsic safety, the design basic accidents and beyond design basic accidents of HTR10-GT must be analyzed according to China's nuclear regulations due to changed operation parameters. THERMIX code system is used to study the accident on one control rod withdrawal out of the core by a mistake. After a control rod in the side reflector was withdrawn out at a speed of 1 cm/s by a mistake, a positive reactivity was inserted and the reactor power increased and the temperature of the core increased. When the neutron flux of power measuring range exceeded 123% and the core outlet temperature was lager than 800 deg. C, the reactor was scrammed. During the accident sequence the maximum fuel temperature was 1200.9 deg. C. It was lower than the fuel temperature limitation of 1230 deg. C. The paper compares the analysis result of HTR10-GT to those of HTR-10. The results shows that the HTR-10GT is still safe during the accident though its operating temperature is higher than HTR-10 when the fuel safety limits are the same. (authors)

  5. The nucleus accumbens 5-HTR4-CART pathway ties anorexia to hyperactivity

    PubMed Central

    Jean, A; Laurent, L; Bockaert, J; Charnay, Y; Dusticier, N; Nieoullon, A; Barrot, M; Neve, R; Compan, V

    2012-01-01

    In mental diseases, the brain does not systematically adjust motor activity to feeding. Probably, the most outlined example is the association between hyperactivity and anorexia in Anorexia nervosa. The neural underpinnings of this ‘paradox', however, are poorly elucidated. Although anorexia and hyperactivity prevail over self-preservation, both symptoms rarely exist independently, suggesting commonalities in neural pathways, most likely in the reward system. We previously discovered an addictive molecular facet of anorexia, involving production, in the nucleus accumbens (NAc), of the same transcripts stimulated in response to cocaine and amphetamine (CART) upon stimulation of the 5-HT4 receptors (5-HTR4) or MDMA (ecstasy). Here, we tested whether this pathway predisposes not only to anorexia but also to hyperactivity. Following food restriction, mice are expected to overeat. However, selecting hyperactive and addiction-related animal models, we observed that mice lacking 5-HTR1B self-imposed food restriction after deprivation and still displayed anorexia and hyperactivity after ecstasy. Decryption of the mechanisms showed a gain-of-function of 5-HTR4 in the absence of 5-HTR1B, associated with CART surplus in the NAc and not in other brain areas. NAc-5-HTR4 overexpression upregulated NAc-CART, provoked anorexia and hyperactivity. NAc-5-HTR4 knockdown or blockade reduced ecstasy-induced hyperactivity. Finally, NAc-CART knockdown suppressed hyperactivity upon stimulation of the NAc-5-HTR4. Additionally, inactivating NAc-5-HTR4 suppressed ecstasy's preference, strengthening the rewarding facet of anorexia. In conclusion, the NAc-5-HTR4/CART pathway establishes a ‘tight-junction' between anorexia and hyperactivity, suggesting the existence of a primary functional unit susceptible to limit overeating associated with resting following homeostasis rules. PMID:23233022

  6. Evaluation of Borrelia burgdorferi BbHtrA Protease as a Vaccine Candidate for Lyme Borreliosis in Mice.

    PubMed

    Ullmann, Amy J; Russell, Theresa M; Dolan, Marc C; Williams, Martin; Hojgaard, Andrias; Weiner, Zachary P; Johnson, Barbara J B

    2015-01-01

    Borrelia burgdorferi synthesizes an HtrA protease (BbHtrA) which is a surface-exposed, conserved protein within Lyme disease spirochetes with activity toward CheX and BmpD of Borrelia spp, as well as aggrecan, fibronectin and proteoglycans found in skin, joints and neural tissues of vertebrates. An antibody response against BbHtrA is observed in Lyme disease patients and in experimentally infected laboratory mice and rabbits. Given the surface location of BbHtrA on B. burgdorferi and its ability to elicit an antibody response in infected hosts, we explored recombinant BbHtrA as a potential vaccine candidate in a mouse model of tick-transmitted Lyme disease. We immunized mice with two forms of BbHtrA: the proteolytically active native form and BbHtrA ablated of activity by a serine to alanine mutation at amino acid 226 (BbHtrA(S226A)). Although inoculation with either BbHtrA or BbHtrA(S226A) produced high-titer antibody responses in C3H/HeJ mice, neither antigen was successful in protecting mice from B. burgdorferi challenge. These results indicate that the search for novel vaccine candidates against Lyme borreliosis remains a challenge.

  7. 5-Hydroxytryptamine modulates cytokine and chemokine production in LPS-primed human monocytes via stimulation of different 5-HTR subtypes.

    PubMed

    Dürk, Thorsten; Panther, Elisabeth; Müller, Tobias; Sorichter, Stephan; Ferrari, Davide; Pizzirani, Cinzia; Di Virgilio, Francesco; Myrtek, Daniel; Norgauer, Johannes; Idzko, Marco

    2005-05-01

    The neurotransmitter 5-hydroxytryptamine (5-HT), commonly known as serotonin, is released at peripheral sites from activated enterochromaffin cells, mast cells and platelets. In this study we analyzed the biological activity and intracellular signaling of 5-HT in human monocytes. By reverse transcription (RT) and PCR, messenger RNA (mRNA) expression of 5-HT receptor 1E (5-HTR(1E)), 5-HTR(2A), 5-HTR(3), 5-HTR(4) and 5-HTR(7) could be revealed. Functional studies showed that 5-HT modulates the release of IL-1beta, IL-6, IL-8/CXCL8, IL-12p40 and tumor necrosis factor-alpha (TNF-alpha), while it has no effect on the production of IL-18 and IFN-gamma in LPS-stimulated human blood monocytes. Moreover, RT and PCR revealed that 5-HT modulated mRNA levels of IL-6 and IL-8/CXCL8, but did not influence mRNA levels of IL-1beta and TNF-alpha. Pharmacological studies with isotype-selective receptor agonists allowed us to show that 5-HTR(3) subtype up-regulates the LPS-induced production of IL-1beta, IL-6 and IL-8/CXCL8, while it was not involved in TNF-alpha and IL-12p40 secretion. Furthermore, activation of the G(s)-coupled 5-HTR(4) and 5-HTR(7) subtypes increased intracellular cyclic AMP (cAMP) and secretion of IL-1beta, IL-6, IL-12p40 and IL-8/CXCL8, while, on the contrary, it inhibited LPS-induced TNF-alpha release. Interestingly, 5-HTR(1) and 5-HTR(2) agonists did not modulate the LPS-induced cytokine production in human monocytes. Our results point to a new role for 5-HT in inflammation by modulating cytokine production in monocytes via activation of 5-HTR(3), 5-HTR(4) and 5-HTR(7) subtypes.

  8. Stress Conditions Increase Vimentin Cleavage by Omi/HtrA2 Protease in Human Primary Neurons and Differentiated Neuroblastoma Cells.

    PubMed

    Lucotte, Bérangère; Tajhizi, Mehdi; Alkhatib, Dareen; Samuelsson, Eva-Britt; Wiehager, Birgitta; Schedin-Weiss, Sophia; Sundström, Erik; Winblad, Bengt; Tjernberg, Lars O; Behbahani, Homira

    2015-12-01

    Dysfunctional Omi/HtrA2, a mitochondrial serine protease, has been implicated in various neurodegenerative disorders. Despite the wealth of evidence on the roles of Omi/HtrA2 in apoptosis, little is known about its cytosolic targets, the cleavage of which could account for the observed morphological changes such as cytoskeletal reorganizations in axons. By proteomic analysis, vimentin was identified as a substrate for Omi/HtrA2 and we have reported increased Omi/HtrA2 protease activity in Alzheimer disease (AD) brain. Here, we investigated a possible link between Omi/HtrA2 and vimentin cleavage, and consequence of this cleavage on mitochondrial distribution in neurons. In vitro protease assays showed vimentin to be cleaved by Omi/HtrA2 protease, and proximity ligation assay demonstrated an increased interaction between Omi/HtrA2 and vimentin in human primary neurons upon stress stimuli. Using differentiated neuroblastoma SH-SY5Y cells, we showed that Omi/HtrA2 under several different stress conditions induces cleavage of vimentin in wild-type as well as SH-SY5Y cells transfected with amyloid precursor protein with the Alzheimer disease-associated Swedish mutation. After stress treatment, inhibition of Omi/HtrA2 protease activity by the Omi/HtrA2 specific inhibitor, Ucf-101, reduced the cleavage of vimentin in wild-type cells. Following altered vimentin filaments integrity by stress stimuli, mitochondria was redistributed in differentiated SH-SY5Y cells and human primary neurons. In summary, the findings outlined in this paper suggest a role of Omi/HtrA2 in modulation of vimentin filamentous structure in neurons. Our results provide important findings for understanding the biological role of Omi/HtrA2 activity during stress conditions, and give knowledge of interplay between Omi/HtrA2 and vimentin which might affect mitochondrial distribution in neurons.

  9. [Pineal anlage tumor in a 8-month-old boy. The first case reported in Spanish language].

    PubMed

    Rodríguez-Velasco, Alicia; Ramírez-Reyes, Alma Griselda

    2014-01-01

    INTRODUCCIÓN: el tumor pineal anlage es una neoplasia maligna muy poco frecuente. Aunque ya ha sido informado en la literatura, aún no se encuentra listado en la última clasificación de los tumores del sistema nervioso central de la Organización Mundial de la Salud publicada en 2007. Es un tumor primario de la pineal con diferenciación neuroepitelial y ectomesenquimal. Debido a su poca frecuencia, el entendimiento de su comportamiento biológico y su tratamiento apropiado son incompletos. En una búsqueda realizada en PubMed como pineal anlage tumor solo se identificaron siete casos informados entre 1989 y 2011. CASO CLÍNICO: niño de ocho meses de edad que fue llevado a consulta por aumento progresivo del perímetro cefálico y estrabismo convergente de dos meses de evolución. Se identificó tumor pineal conformado por glia, células ganglionares, neuroepitelio pigmentado y células musculares estriadas. Se realizó derivación ventrículo-peritoneal para mitigar la hidrocefalia y resección quirúrgica completa. El paciente recibió dos cursos de quimioterapia con carboplatino, ifosfamida y mesna. Al año permanecía asintomático.

  10. HTR-100 industrial nuclear power plant for generation of heat and electricity

    SciTech Connect

    Brandes, S.; Kohl, W.

    1987-11-01

    Based on their proven high-temperature reactor (HTR) with pebble-bed core, Brown, Boveri and Cie/Hochtemperatur-Reaktorbau have developed an HTR-100 plant that combines favorable capital costs and high availability. Due to the high HTR-specific standards and passive safety features, this plant is especially well suited for siting near the end user. The safety concept permits further operation of the plant or decay heat removal via the operational heat sinks in the event of maloperation and design basis accidents having a higher probability of occurrence. In the event of hypothetical accidents, the decay heat is removed from the reactor pressure vessel by radiation, conduction, and convection to a concrete cooling system operating in natural convection. As an example of the new HTR-100 plant concept, a twin-block plant design for extraction of industrial steam is presented.

  11. Optimization of milling time before and after recalcination on HTR processed strontium hexaferrite powder

    NASA Astrophysics Data System (ADS)

    Seyyed Ebrahimi, S. A.

    2006-12-01

    HTR (Hydrogen Treatment and Recalculation) process is a novel technique for heat treatment of conventional strontium hexaferrite powder in static hydrogen and then its recalcination in air. This process has a marked effect on the microstructure and on the magnetic properties of the material. In this work, this HTR-processed powder has been milled before and after recalcination, and the effect of milling on the magnetic properties has been optimized.

  12. The novel role of HtrA1 in gingivitis, chronic and aggressive periodontitis.

    PubMed

    Lorenzi, Teresa; Niţulescu, Elena Annabel; Zizzi, Antonio; Lorenzi, Maria; Paolinelli, Francesca; Aspriello, Simone Domenico; Baniţă, Monica; Crăiţoiu, Stefania; Goteri, Gaia; Barbatelli, Giorgio; Lombardi, Tommaso; Di Felice, Roberto; Marzioni, Daniela; Rubini, Corrado; Castellucci, Mario

    2014-01-01

    Proteolytic tissue degradation is a typical phenomenon in inflammatory periodontal diseases. HtrA1 (High temperature requirement A 1) has a serine protease activity and is able to degrade fibronectin whose fragments induce the expression and secretion of several matrix metalloproteinases (MMPs). The aim of this study was to investigate for the first time if HtrA1 has a role in gingivitis and in generalized forms of chronic and aggressive periodontitis. Expression of HtrA1 was investigated in 16 clinically healthy gingiva, 16 gingivitis, 14 generalized chronic periodontitis and 10 generalized aggressive periodontitis by immunohistochemistry and real-time PCR. Statistical comparisons were performed by the Kruskall-Wallis test. Significantly higher levels of HtrA1 mRNA and protein expression were observed in pathological respect to healthy tissues. In particular, we detected an increase of plasma cell HtrA1 immunostaining from gingivitis to chronic and aggressive periodontitis, with the higher intensity in aggressive disease. In addition, we observed the presence of HtrA1 in normal and pathological epithelium, with an increased expression, particularly in its superficial layer, associated with increasingly severe forms of periodontal disease. We can affirm that HtrA1 expression in plasma cells could be correlated with the destruction of pathological periodontal tissue, probably due to its ability to trigger the overproduction of MMPs and to increase the inflammatory mediators TNF-α and IL-1β by inhibition of TGF-β. Moreover, epithelial HtrA1 immunostaining suggests a participation of the molecule in the host inflammatory immune responses necessary for the control of periodontal infection.

  13. The Novel Role of HtrA1 in Gingivitis, Chronic and Aggressive Periodontitis

    PubMed Central

    Zizzi, Antonio; Lorenzi, Maria; Paolinelli, Francesca; Aspriello, Simone Domenico; Baniţă, Monica; Crăiţoiu, Ştefania; Goteri, Gaia; Barbatelli, Giorgio; Lombardi, Tommaso; Di Felice, Roberto; Marzioni, Daniela; Rubini, Corrado; Castellucci, Mario

    2014-01-01

    Proteolytic tissue degradation is a typical phenomenon in inflammatory periodontal diseases. HtrA1 (High temperature requirement A 1) has a serine protease activity and is able to degrade fibronectin whose fragments induce the expression and secretion of several matrix metalloproteinases (MMPs). The aim of this study was to investigate for the first time if HtrA1 has a role in gingivitis and in generalized forms of chronic and aggressive periodontitis. Expression of HtrA1 was investigated in 16 clinically healthy gingiva, 16 gingivitis, 14 generalized chronic periodontitis and 10 generalized aggressive periodontitis by immunohistochemistry and real-time PCR. Statistical comparisons were performed by the Kruskall-Wallis test. Significantly higher levels of HtrA1 mRNA and protein expression were observed in pathological respect to healthy tissues. In particular, we detected an increase of plasma cell HtrA1 immunostaining from gingivitis to chronic and aggressive periodontitis, with the higher intensity in aggressive disease. In addition, we observed the presence of HtrA1 in normal and pathological epithelium, with an increased expression, particularly in its superficial layer, associated with increasingly severe forms of periodontal disease. We can affirm that HtrA1 expression in plasma cells could be correlated with the destruction of pathological periodontal tissue, probably due to its ability to trigger the overproduction of MMPs and to increase the inflammatory mediators TNF-α and IL-1β by inhibition of TGF-β. Moreover, epithelial HtrA1 immunostaining suggests a participation of the molecule in the host inflammatory immune responses necessary for the control of periodontal infection. PMID:24979214

  14. Physik gestern und heute Von der Metallstange zum Hochenergielaser

    NASA Astrophysics Data System (ADS)

    Heering, Peter

    2002-05-01

    Im Mai 1752 wurde in Marly bei Paris auf Anregung des amerikanischen Forschers und Politikers Benjamin Franklin erstmals die elektrische Natur des Blitzes nachgewiesen. Damals beschrieb Franklin auch eine technische Vorrichtung, die als Schutz von Gebäuden vor Blitzschlägen dienen sollte: den Blitzableiter. Diese aus heutiger Sicht scheinbar triviale Vorrichtung wurde aber keineswegs unmittelbar akzeptiert. Und bis heute ist die Forschung zum Schutz von Einrichtungen vor Blitzschlägen nicht abgeschlossen.

  15. HtrA3 as an Early Marker for Preeclampsia: Specific Monoclonal Antibodies and Sensitive High-Throughput Assays for Serum Screening

    PubMed Central

    Dynon, Kemperly; Heng, Sophea; Puryer, Michelle; Li, Ying; Walton, Kelly; Endo, Yaeta; Nie, Guiying

    2012-01-01

    Mammalian HtrA3 (high temperature requirement A3) is a serine protease of the HtrA family. It has two isoforms [long (HtrA3-L) and short (HtrA3-S)] and is important for placental development and cancer progression. Recently, HtrA3 was identified as a potential diagnostic marker for early detection of preeclampsia, a life-threatening pregnancy-specific disorder. Currently there are no high-throughput assays available to detect HtrA3 in human serum. In this study we generated and fully tested a panel of five HtrA3 mouse monoclonal antibodies (mAbs). Three mAbs recognised both HtrA3-L and HtrA3-S and the other two detected HtrA3-L only. All five mAbs were highly specific to HtrA3 and applicable in western blotting and immunohistochemical analysis of endogenous HtrA3 proteins in the mouse and human tissues. Amplified luminescent proximity homogeneous assays-linked immunosorbent assays (AlphaLISAs), were developed to detect HtrA3 isoforms in picomolar levels in serum. The HtrA3 AlphaLISA detected significantly higher serum levels of HtrA3 in women at 13–14 weeks of gestation who subsequently developed preeclampsia compared to gestational-age matched controls. These HtrA3 mAbs are valuable for the development of immunoassays and characterisation of HtrA3 isoform-specific biology. The newly developed HtrA3 AlphaLISA assays are suitable for large scale screening of human serum. PMID:23049902

  16. Glucose uptake and catabolite repression in dominant HTR1 mutants of Saccharomyces cerevisiae.

    PubMed Central

    Ozcan, S; Freidel, K; Leuker, A; Ciriacy, M

    1993-01-01

    Growth and carbon metabolism in triosephosphate isomerase (delta tpi1) mutants of Saccharomyces cerevisiae are severely inhibited by glucose. By using this feature, we selected for secondary site revertants on glucose. We defined five complementation groups, some of which have previously been identified as glucose repression mutants. The predominant mutant type, HTR1 (hexose transport regulation), is dominant and causes various glucose-specific metabolic and regulatory defects in TPI1 wild-type cells. HTR1 mutants are deficient in high-affinity glucose uptake and have reduced low-affinity transport. Transcription of various known glucose transporter genes (HXT1, HXT3, and HXT4) was defective in HTR1 mutants, leading us to suggest that HTR mutations affect a negative factor of HXT gene expression. By contrast, transcript levels for SNF3, which encodes a component of high-affinity glucose uptake, were unaffected. We presume that HTR1 mutations affect a negative factor of HXT gene expression. Multicopy expression of HXT genes or parts of their regulatory sequences suppresses the metabolic defects of HTR1 mutants but not their derepressed phenotype at high glucose concentrations. This suggests that the glucose repression defect is not a direct result of the metabolically relevant defect in glucose transport. Alternatively, some unidentified regulatory components of the glucose transport system may be involved in the generation or transmission of signals for glucose repression. Images PMID:8366037

  17. Relaxin has anti-apoptotic effects on human trophoblast-derived HTR-8/SV neo cells.

    PubMed

    Lodhi, Romana S Z; Nakabayashi, Koji; Suzuki, Kaho; Yamada, Ai Y; Hazama, Rhoichi; Ebina, Yasuhiko; Yamada, Hideto

    2013-12-01

    The study was conducted to evaluate the effects of human relaxin on apoptosis in the human trophoblast derived HTR-8/SV neo cell line, which is a possible model of human extravillous trophoblasts (EVTs). HTR-8/SV neo cells, cultured in phenol red free RPMI1640 medium, were treated with different doses of human recombinant (rH2) relaxin in serum-deprived conditions. RT-PCR was used for evaluating relaxin receptor: RXFP1 and RXFP2 expression in HTR-8/SV neo cells. The cell death was examined by TUNEL assay. Furthermore, we investigated caspase-3, cleaved PARP and Bcl-2 expressions by Western blot analysis to recognize the translational effects of anti-apoptotic and pro-apoptotic proteins. RXFP1 and RXFP2 mRNA expression was observed in HTR-8/SV neo cells. Compared with untreated control cultures, treatment with rH2 relaxin, decreased TUNEL-positive rate in HTR-8/SV neo cells was observed. Western blot analysis revealed that treatment with rH2 relaxin decreased the expression of caspase-3 and cleaved PARP, but in contrast increased Bcl-2 expression in those cells. These results suggest that rH2 relaxin has anti-apoptotic effects on HTR8/SV neo cells by decreasing pro-apoptotic caspase-3 and cleaved PARP expression and up-regulating anti-apoptotic Bcl-2 expression.

  18. Construction and characterization of a Lactococcus lactis strain deficient in intracellular ClpP and extracellular HtrA proteases.

    PubMed

    Cortes-Perez, N G; Poquet, I; Oliveira, M; Gratadoux, J J; Madsen, S M; Miyoshi, A; Corthier, G; Azevedo, V; Langella, P; Bermúdez-Humarán, L G

    2006-09-01

    A Lactococcus lactis strain deficient in both its major proteases, intracellular (ClpP) and extracellular (HtrA), was constructed and characterized. This strain, hereafter called clpP-htrA, could be obtained only by conjugation between a clpP donor strain and an htrA recipient strain in the NZ9000 context, allowing heterologous gene expression under the control of the NICE (nisin-controlled expression) system. The clpP-htrA double mutant showed both higher stress tolerance (e.g. high temperature and ethanol resistance) and higher viability than single clpP or htrA mutant strains. In addition, the secretion rate of two heterologous proteins (staphylococcal nuclease Nuc and Nuc-E7) was also higher in clpP-htrA than in the wild-type strain. This strain should be a useful host for high-level production and quality of stable heterologous proteins.

  19. Increased hTR expression during transition from adenoma to carcinoma is not associated with promoter methylation.

    PubMed

    Nakamura, Atsuo; Suda, Takeshi; Honma, Terasu; Takahashi, Toru; Igarashi, Masato; Waguri, Nobuo; Kawai, Hirokazu; Mita, Yusaku; Aoyagi, Yutaka

    2004-09-01

    Human telomerase RNA component (hTR) expression, which increases in the majority of cancer cells with an acquisition of telomerase activity, was concomitantly evaluated with methylation status and human telomerase reverse transcriptase (hTERT) expression in colorectal cancers and precursor lesions. hTR and hTERT expressions were detected by in situ hybridization and reverse transcription following polymerase chain reaction, respectively, in 15 colonic adenomas, 19 sporadic colonic cancers at various histological stages, and 3 normal colonic mucosa samples. The methylationstatus of hTR was evaluated by methylation-specific polymerase chain reaction following restriction endonuclease digestion and direct sequencing. hTERT expression was detected in 16 of 19 cancers. hTR expression was detected in all cancers including two cases of intramucosal carcinoma. No hTR signals were detected in the normal epithelium or in the adenomas with severe atypism. CpG dinucleotides in the 5'-untranslated region of hTR were completely unmethylated from -204 to -3 and mosaically methylated from -290 to -272, irrespective of the atypism. These results suggest that hTR expression is increased at the adenoma-to-carcinoma transition stage but is not always associated with hTERT expression. Hypomethylation of the hTR promoter region is not likely to be the main mechanism regulating hTR expression.

  20. Benchmark Evaluation of HTR-PROTEUS Pebble Bed Experimental Program

    DOE PAGES

    Bess, John D.; Montierth, Leland; Köberl, Oliver; ...

    2014-10-09

    Benchmark models were developed to evaluate 11 critical core configurations of the HTR-PROTEUS pebble bed experimental program. Various additional reactor physics measurements were performed as part of this program; currently only a total of 37 absorber rod worth measurements have been evaluated as acceptable benchmark experiments for Cores 4, 9, and 10. Dominant uncertainties in the experimental keff for all core configurations come from uncertainties in the ²³⁵U enrichment of the fuel, impurities in the moderator pebbles, and the density and impurity content of the radial reflector. Calculations of keff with MCNP5 and ENDF/B-VII.0 neutron nuclear data are greatermore » than the benchmark values but within 1% and also within the 3σ uncertainty, except for Core 4, which is the only randomly packed pebble configuration. Repeated calculations of keff with MCNP6.1 and ENDF/B-VII.1 are lower than the benchmark values and within 1% (~3σ) except for Cores 5 and 9, which calculate lower than the benchmark eigenvalues within 4σ. The primary difference between the two nuclear data libraries is the adjustment of the absorption cross section of graphite. Simulations of the absorber rod worth measurements are within 3σ of the benchmark experiment values. The complete benchmark evaluation details are available in the 2014 edition of the International Handbook of Evaluated Reactor Physics Benchmark Experiments.« less

  1. Benchmark Evaluation of HTR-PROTEUS Pebble Bed Experimental Program

    SciTech Connect

    Bess, John D.; Montierth, Leland; Köberl, Oliver; Snoj, Luka

    2014-10-09

    Benchmark models were developed to evaluate 11 critical core configurations of the HTR-PROTEUS pebble bed experimental program. Various additional reactor physics measurements were performed as part of this program; currently only a total of 37 absorber rod worth measurements have been evaluated as acceptable benchmark experiments for Cores 4, 9, and 10. Dominant uncertainties in the experimental keff for all core configurations come from uncertainties in the ²³⁵U enrichment of the fuel, impurities in the moderator pebbles, and the density and impurity content of the radial reflector. Calculations of keff with MCNP5 and ENDF/B-VII.0 neutron nuclear data are greater than the benchmark values but within 1% and also within the 3σ uncertainty, except for Core 4, which is the only randomly packed pebble configuration. Repeated calculations of keff with MCNP6.1 and ENDF/B-VII.1 are lower than the benchmark values and within 1% (~3σ) except for Cores 5 and 9, which calculate lower than the benchmark eigenvalues within 4σ. The primary difference between the two nuclear data libraries is the adjustment of the absorption cross section of graphite. Simulations of the absorber rod worth measurements are within 3σ of the benchmark experiment values. The complete benchmark evaluation details are available in the 2014 edition of the International Handbook of Evaluated Reactor Physics Benchmark Experiments.

  2. Polymorphisms of the HTR1a Allele are linked to Frontal Brain Electrical Asymmetry

    PubMed Central

    Bismark, Andrew W.; Moreno, Francisco A.; Stewart, Jennifer L.; Towers, David N.; Coan, James A.; Oas, Jennifer; Erickson, Robert P.; Allen, John J.B.

    2010-01-01

    Polymorphic variations in genes related to serotonin synthesis, transport, recognition, or degradation may convey subtle changes in serotonin system architecture that may place an individual at risk for psychopathology when faced with life stressors. The relationship between three key serotonin alleles and frontal brain electrical asymmetry, a putative endophenotype of depression, was examined. Risk alleles were hypothesized to predict relatively greater right frontal brain activity regardless of current clinical state. A sample of 313 college-age individuals, spanning a range of depressive severity from no symptomotology to clinically meaningful levels, participated. Resting encephalographic (EEG) activity was recorded from 64 scalp sites on four occasions separated by at least 24 hours (two 8-min recording sessions occurring at each occasion). Alpha power asymmetry scores between homologous sites were calculated for each session and then averaged to form a trait metric of asymmetry for each pair. PCR based genotyping was conducted for the HTR1A, HTR2A, and HTTLPR genes. Variations in the HTR1A gene were related to trait EEG asymmetry, regardless of any history of depression. Compared to subjects with at least one non-risk allele, subjects with homozygous HTR1A risk alleles had significantly greater relative right frontal activity at sites F7/F8, F5/F6, & F1/F2. In conclusion, variation in HTR1A can influence trait level brain activity, which may ultimately be indicative of risk for psychopathology. PMID:20025927

  3. hTR RNA component as a marker of cellular proliferation in oral lichen planus.

    PubMed

    Flatharta, Cathal O; Flint, Stephen; Toner, Mary; Mabruk, Mohamed

    2008-01-01

    Previously, we have shown that the telomerase RNA component hTR is highly expressed in the epithelium of non-dysplastic Oral Lichen Planus (OLP) lesions (11). We concluded that it is possible that this high expression might be related to the increased cellular proliferation seen in OLP rather than being an indicator of potential malignant transformation. In the present study, and in order to confirm our finding in the previous study that hTR might be a marker for cellular proliferation in OLP, we analysed OLP biopsies known to be positive for RNA component of Telomerase (hTR) for the expression of Ki-67 as a marker for cellular proliferation. Fourteen OLP tissue biopsies known to be positive for telomerase RNA component hTR, were investigated using an immunohistochemical approach to determine the rate of cellular proliferation in OLP, looking at the expression of Ki-67 protein as a marker for cellular proliferation. A statistically significant increase was found between Ki-67 expression in OLP in comparison to normal control buccal mucosa samples. The expression of hTR component in OLP might thus be a marker for cellular proliferation.

  4. Inhibiting Helicobacter pylori HtrA protease by addressing a computationally predicted allosteric ligand binding site

    PubMed Central

    Perna, Anna Maria; Reisen, Felix; Schmidt, Thomas P.; Geppert, Tim; Pillong, Max; Weisel, Martin; Hoy, Benjamin; Simister, Philip C.; Feller, Stephan M.; Wessler, Silja; Schneider, Gisbert

    2016-01-01

    Helicobacter pylori is associated with inflammatory diseases and can cause gastric cancer and mucosa-associated lymphoma. One of the bacterium’s key proteins is high temperature requirement A (HpHtrA) protein, an extracellular serine protease that cleaves E-cadherin of gastric epithelial cells, which leads to loss of cell-cell adhesion. Inhibition of HpHtrA may constitute an intervention strategy against H. pylori infection. Guided by the computational prediction of hypothetical ligand binding sites on the surface of HpHtrA, we performed residue mutation experiments that confirmed the functional relevance of an allosteric region. We virtually screened for potential ligands addressing this surface cleft located between the catalytic and PDZ1 domains. Our receptor-based computational method represents protein surface pockets in terms of graph frameworks and retrieves small molecules that satisfy the constraints given by the pocket framework. A new chemical entity was identified that blocked E-cadherin cleavage in vitro by direct binding to HpHtrA, and efficiently blocked pathogen transmigration across the gastric epithelial barrier. A preliminary crystal structure of HpHtrA confirms the validity of a comparative “homology” model of the enzyme, which we used for the computational study. The results of this study demonstrate that addressing orphan protein surface cavities of target macromolecules can lead to new bioactive ligands. PMID:26819700

  5. Evaluation of the HTR-10 Reactor as a Benchmark for Physics Code QA

    SciTech Connect

    William K. Terry; Soon Sam Kim; Leland M. Montierth; Joshua J. Cogliati; Abderrafi M. Ougouag

    2006-09-01

    The HTR-10 is a small (10 MWt) pebble-bed research reactor intended to develop pebble-bed reactor (PBR) technology in China. It will be used to test and develop fuel, verify PBR safety features, demonstrate combined electricity production and co-generation of heat, and provide experience in PBR design, operation, and construction. As the only currently operating PBR in the world, the HTR-10 can provide data of great interest to everyone involved in PBR technology. In particular, if it yields data of sufficient quality, it can be used as a benchmark for assessing the accuracy of computer codes proposed for use in PBR analysis. This paper summarizes the evaluation for the International Reactor Physics Experiment Evaluation Project (IRPhEP) of data obtained in measurements of the HTR-10’s initial criticality experiment for use as benchmarks for reactor physics codes.

  6. The functional -1019C/G HTR1A polymorphism and mechanisms of fear.

    PubMed

    Straube, B; Reif, A; Richter, J; Lueken, U; Weber, H; Arolt, V; Jansen, A; Zwanzger, P; Domschke, K; Pauli, P; Konrad, C; Gerlach, A L; Lang, T; Fydrich, T; Alpers, G W; Ströhle, A; Wittmann, A; Pfleiderer, B; Wittchen, H-U; Hamm, A; Deckert, J; Kircher, T

    2014-12-16

    Serotonin receptor 1A gene (HTR1A) knockout mice show pronounced defensive behaviour and increased fear conditioning to ambiguous conditioned stimuli. Such behaviour is a hallmark of pathological human anxiety, as observed in panic disorder with agoraphobia (PD/AG). Thus, variations in HTR1A might contribute to neurophysiological differences within subgroups of PD/AG patients. Here, we tested this hypothesis by combining genetic with behavioural techniques and neuroimaging. In a clinical multicentre trial, patients with PD/AG received 12 sessions of manualized cognitive-behavioural therapy (CBT) and were genotyped for HTR1A rs6295. In four subsamples of this multicentre trial, exposure behaviour (n=185), defensive reactivity measured using a behavioural avoidance test (BAT; before CBT: n=245; after CBT: n=171) and functional magnetic resonance imaging (fMRI) data during fear conditioning were acquired before and after CBT (n=39). HTR1A risk genotype (GG) carriers more often escaped during the BAT before treatment. Exploratory fMRI results suggest increased activation of the amygdala in response to threat as well as safety cues before and after treatment in GG carriers. Furthermore, GG carriers demonstrated reduced effects of CBT on differential conditioning in regions including the bilateral insulae and the anterior cingulate cortex. Finally, risk genotype carriers demonstrated reduced self-initiated exposure behaviour to aversive situations. This study demonstrates the effect of HTR1A variation on defensive behaviour, amygdala activity, CBT-induced neural plasticity and normalization of defence behaviour in PD/AG. Our results, therefore, translate evidence from animal studies to humans and suggest a central role for HTR1A in differentiating subgroups of patients with anxiety disorders.

  7. The functional −1019C/G HTR1A polymorphism and mechanisms of fear

    PubMed Central

    Straube, B; Reif, A; Richter, J; Lueken, U; Weber, H; Arolt, V; Jansen, A; Zwanzger, P; Domschke, K; Pauli, P; Konrad, C; Gerlach, A L; Lang, T; Fydrich, T; Alpers, G W; Ströhle, A; Wittmann, A; Pfleiderer, B; Wittchen, H-U; Hamm, A; Deckert, J; Kircher, T

    2014-01-01

    Serotonin receptor 1A gene (HTR1A) knockout mice show pronounced defensive behaviour and increased fear conditioning to ambiguous conditioned stimuli. Such behaviour is a hallmark of pathological human anxiety, as observed in panic disorder with agoraphobia (PD/AG). Thus, variations in HTR1A might contribute to neurophysiological differences within subgroups of PD/AG patients. Here, we tested this hypothesis by combining genetic with behavioural techniques and neuroimaging. In a clinical multicentre trial, patients with PD/AG received 12 sessions of manualized cognitive-behavioural therapy (CBT) and were genotyped for HTR1A rs6295. In four subsamples of this multicentre trial, exposure behaviour (n=185), defensive reactivity measured using a behavioural avoidance test (BAT; before CBT: n=245; after CBT: n=171) and functional magnetic resonance imaging (fMRI) data during fear conditioning were acquired before and after CBT (n=39). HTR1A risk genotype (GG) carriers more often escaped during the BAT before treatment. Exploratory fMRI results suggest increased activation of the amygdala in response to threat as well as safety cues before and after treatment in GG carriers. Furthermore, GG carriers demonstrated reduced effects of CBT on differential conditioning in regions including the bilateral insulae and the anterior cingulate cortex. Finally, risk genotype carriers demonstrated reduced self-initiated exposure behaviour to aversive situations. This study demonstrates the effect of HTR1A variation on defensive behaviour, amygdala activity, CBT-induced neural plasticity and normalization of defence behaviour in PD/AG. Our results, therefore, translate evidence from animal studies to humans and suggest a central role for HTR1A in differentiating subgroups of patients with anxiety disorders. PMID:25514753

  8. Influence of Polymorphisms in the HTR3A and HTR3B Genes on Experimental Pain and the Effect of the 5-HT3 Antagonist Granisetron

    PubMed Central

    Hedenberg-Magnusson, Britt; List, Thomas; Svensson, Peter; Schalling, Martin

    2016-01-01

    The aim of this study was to investigate experimentally if 5-HT3 single nucleotide polymorphisms (SNP) contribute to pain perception and efficacy of the 5-HT3-antagonist granisetron and sex differences. Sixty healthy participants were genotyped regarding HTR3A (rs1062613) and HTR3B (rs1176744). First, pain was induced by bilateral hypertonic saline injections (HS, 5.5%, 0.2 mL) into the masseter muscles. Thirty min later the masseter muscle on one side was pretreated with 0.5 mL granisetron (1 mg/mL) and on the other side with 0.5 mL placebo (isotonic saline) followed by another HS injection (0.2 mL). Pain intensity, pain duration, pain area and pressure pain thresholds (PPTs) were assessed after each injection. HS evoked moderate pain, with higher intensity in the women (P = 0.023), but had no effect on PPTs. None of the SNPs influenced any pain variable in general, but compared to men, the pain area was larger in women carrying the C/C (HTR3A) (P = 0.015) and pain intensity higher in women with the A/C alleles (HTR3B) (P = 0.019). Pre-treatment with granisetron reduced pain intensity, duration and area to a lesser degree in women (P < 0.05), but the SNPs did not in general influence the efficacy of granisetron. Women carrying the C/T & T/T (HTR3A) genotype had less reduction of pain intensity (P = 0.041) and area (P = 0.005), and women with the C/C genotype (HTR3B) had less reduction of pain intensity (P = 0.030), duration (P = 0.030) and area compared to men (P = 0.017). In conclusion, SNPs did not influence experimental muscle pain or the effect of granisetron on pain variables in general, but there were some sex differences in pain variables that seem to be influenced by genotypes. However, due to the small sample size further research is needed before any firm conclusions can be drawn. PMID:28002447

  9. Recombinant outer membrane vesicles carrying Chlamydia muridarum HtrA induce antibodies that neutralize chlamydial infection in vitro.

    PubMed

    Bartolini, Erika; Ianni, Elvira; Frigimelica, Elisabetta; Petracca, Roberto; Galli, Giuliano; Berlanda Scorza, Francesco; Norais, Nathalie; Laera, Donatello; Giusti, Fabiola; Pierleoni, Andrea; Donati, Manuela; Cevenini, Roberto; Finco, Oretta; Grandi, Guido; Grifantini, Renata

    2013-01-01

    Outer membrane vesicles (OMVs) are spheroid particles released by all Gram-negative bacteria as a result of the budding out of the outer membrane. Since they carry many of the bacterial surface-associated proteins and feature a potent built-in adjuvanticity, OMVs are being utilized as vaccines, some of which commercially available. Recently, methods for manipulating the protein content of OMVs have been proposed, thus making OMVs a promising platform for recombinant, multivalent vaccines development. Chlamydia muridarum DO serine protease HtrA, an antigen which stimulates strong humoral and cellular responses in mice and humans, was expressed in Escherichia coli fused to the OmpA leader sequence to deliver it to the OMV compartment. Purified OMVs carrying HtrA (CM rHtrA-OMV) were analyzed for their capacity to induce antibodies capable of neutralizing Chlamydia infection of LLC-MK2 cells in vitro. CM rHtrA-OMV immunization in mice induced antibodies that neutralize Chlamydial invasion as judged by an in vitro infectivity assay. This was remarkably different from what observed with an enzymatically functional recombinant HtrA expressed in, and purified from the E. coli cytoplasm (CM rHtrA). The difference in functionality between anti-CM rHtrA and anti-CM rHtrA-OMV antibodies was associated to a different pattern of protein epitopes recognition. The epitope recognition profile of anti-CM HtrA-OMV antibodies was similar to that induced in mice during Chlamydial infection. When expressed in OMVs HtrA appears to assume a conformation similar to the native one and this results in the elicitation of functional immune responses. These data further support the potentiality of OMVs as vaccine platform.

  10. Recombinant outer membrane vesicles carrying Chlamydia muridarum HtrA induce antibodies that neutralize chlamydial infection in vitro

    PubMed Central

    Bartolini, Erika; Ianni, Elvira; Frigimelica, Elisabetta; Petracca, Roberto; Galli, Giuliano; Berlanda Scorza, Francesco; Norais, Nathalie; Laera, Donatello; Giusti, Fabiola; Pierleoni, Andrea; Donati, Manuela; Cevenini, Roberto; Finco, Oretta; Grandi, Guido; Grifantini, Renata

    2013-01-01

    Background Outer membrane vesicles (OMVs) are spheroid particles released by all Gram-negative bacteria as a result of the budding out of the outer membrane. Since they carry many of the bacterial surface-associated proteins and feature a potent built-in adjuvanticity, OMVs are being utilized as vaccines, some of which commercially available. Recently, methods for manipulating the protein content of OMVs have been proposed, thus making OMVs a promising platform for recombinant, multivalent vaccines development. Methods Chlamydia muridarum DO serine protease HtrA, an antigen which stimulates strong humoral and cellular responses in mice and humans, was expressed in Escherichia coli fused to the OmpA leader sequence to deliver it to the OMV compartment. Purified OMVs carrying HtrA (CM rHtrA-OMV) were analyzed for their capacity to induce antibodies capable of neutralizing Chlamydia infection of LLC-MK2 cells in vitro. Results CM rHtrA-OMV immunization in mice induced antibodies that neutralize Chlamydial invasion as judged by an in vitro infectivity assay. This was remarkably different from what observed with an enzymatically functional recombinant HtrA expressed in, and purified from the E. coli cytoplasm (CM rHtrA). The difference in functionality between anti-CM rHtrA and anti-CM rHtrA-OMV antibodies was associated to a different pattern of protein epitopes recognition. The epitope recognition profile of anti-CM HtrA-OMV antibodies was similar to that induced in mice during Chlamydial infection. Conclusions When expressed in OMVs HtrA appears to assume a conformation similar to the native one and this results in the elicitation of functional immune responses. These data further support the potentiality of OMVs as vaccine platform. PMID:24009891

  11. Development of a Reliable Fuel Depletion Methodology for the HTR-10 Spent Fuel Analysis

    SciTech Connect

    Chung, Kiwhan; Beddingfield, David H.; Geist, William H.; Lee, Sang-Yoon

    2012-07-03

    A technical working group formed in 2007 between NNSA and CAEA to develop a reliable fuel depletion method for HTR-10 based on MCNPX and to analyze the isotopic inventory and radiation source terms of the HTR-10 spent fuel. Conclusions of this presentation are: (1) Established a fuel depletion methodology and demonstrated its safeguards application; (2) Proliferation resistant at high discharge burnup ({approx}80 GWD/MtHM) - Unfavorable isotopics, high number of pebbles needed, harder to reprocess pebbles; (3) SF should remain under safeguards comparable to that of LWR; and (4) Diversion scenarios not considered, but can be performed.

  12. Campylobacter jejuni serine protease HtrA plays an important role in heat tolerance, oxygen resistance, host cell adhesion, invasion, and transmigration.

    PubMed

    Boehm, Manja; Lind, Judith; Backert, Steffen; Tegtmeyer, Nicole

    2015-03-01

    Campylobacter jejuni is an important pathogen of foodborne illness. Transmigration across the intestinal epithelial barrier and invasion are considered as primary reasons for tissue damage triggered by C. jejuni. Using knockout mutants, it was shown that the serine protease HtrA may be important for stress tolerance and physiology of C. jejuni. HtrA is also secreted in the extra-cellular environment, where it can cleave junctional host cell proteins such as E-cadherin. Aim of the present study was to establish a genetic complementation system in two C. jejuni strains in order to introduce the wild-type htrA gene in trans, test known htrA phenotypes, and provide the basis to perform further mutagenesis. We confirm that reexpression of the htrA wild-type gene in ΔhtrA mutants restored the following phenotypes: 1) C. jejuni growth at high temperature (44 °C), 2) growth under high oxygen stress conditions, 3) expression of proteolytically active HtrA oligomers, 4) secretion of HtrA into the supernatant, 5) cell attachment and invasion, and 6) transmigration across polarized epithelial cells. These results establish a genetic complementation system for htrA in C. jejuni, exclude polar effects in the ΔhtrA mutants, confirm important HtrA properties, and permit the discovery and dissection of new functions.

  13. Campylobacter jejuni serine protease HtrA plays an important role in heat tolerance, oxygen resistance, host cell adhesion, invasion, and transmigration

    PubMed Central

    Lind, Judith; Backert, Steffen; Tegtmeyer, Nicole

    2015-01-01

    Campylobacter jejuni is an important pathogen of foodborne illness. Transmigration across the intestinal epithelial barrier and invasion are considered as primary reasons for tissue damage triggered by C. jejuni. Using knockout mutants, it was shown that the serine protease HtrA may be important for stress tolerance and physiology of C. jejuni. HtrA is also secreted in the extra­cellular environment, where it can cleave junctional host cell proteins such as E-cadherin. Aim of the present study was to establish a genetic complementation system in two C. jejuni strains in order to introduce the wild-type htrA gene in trans, test known htrA phenotypes, and provide the basis to perform further mutagenesis. We confirm that reexpression of the htrA wild-type gene in ΔhtrA mutants restored the following phenotypes: 1) C. jejuni growth at high temperature (44 °C), 2) growth under high oxygen stress conditions, 3) expression of proteolytically active HtrA oligomers, 4) secretion of HtrA into the supernatant, 5) cell attachment and invasion, and 6) transmigration across polarized epithelial cells. These results establish a genetic complementation system for htrA in C. jejuni, exclude polar effects in the ΔhtrA mutants, confirm important HtrA properties, and permit the discovery and dissection of new functions. PMID:25883795

  14. Release of an HtrA-Like Protease from the Cell Surface of Thermophilic Brevibacillus sp. WF146 via Substrate-Induced Autoprocessing of the N-terminal Membrane Anchor

    PubMed Central

    Zhu, Fengtao; Yang, Xing; Wu, Yan; Wang, Yasi; Tang, Xiao-Feng; Tang, Bing

    2017-01-01

    High-temperature requirement A (HtrA)-like proteases participate in protein quality control in prokaryotes and eukaryotes by degrading damaged proteins; however, little is known about HtrAs produced by thermophiles. HtrAw is an HtrA-like protease of thermophilic Brevibacillus sp. WF146. The intact form of HtrAw (iHtrAw) consisting of a transmembrane segment-containing N-terminal domain, a trypsin-like protease domain, and a C-terminal PDZ domain was produced in Escherichia coli. Purified iHtrAw itself is unable to cleave the N-terminal domain, but requires protein substrates to autoprocess the N-terminal domain intermolecularly, yielding a short form (sHtrAw). Mutation at the substrate-binding site in the PDZ domain affects the conversion of iHtrAw to sHtrAw. Deletion analysis revealed that the N-terminal domain is not necessary for enzyme folding, activity, and thermostability. Compared with other known HtrAs, HtrAw contains an additional Ca2+-binding Dx[DN]xDG motif important for enzyme stability and/or activity. When produced in an htrA/htrB double deletion mutant of Bacillus subtilis, iHtrAw localized predominantly to the cell pellet, and the amount of sHtrAw in the culture supernatant increased at elevated temperatures. Moreover, HtrAw increased the heat resistance of the B. subtilis mutant. In strain WF146, HtrAw exists in both a cell-associated intact form and a cell-free short form; an increase in growth temperature enhanced HtrAw production and the amount of cell-free short form. Release of the short form of HtrAw from the membrane may have the advantage of allowing the enzyme to freely access and degrade damaged proteins surrounding the bacterium living at high temperatures. PMID:28377763

  15. Release of an HtrA-Like Protease from the Cell Surface of Thermophilic Brevibacillus sp. WF146 via Substrate-Induced Autoprocessing of the N-terminal Membrane Anchor.

    PubMed

    Zhu, Fengtao; Yang, Xing; Wu, Yan; Wang, Yasi; Tang, Xiao-Feng; Tang, Bing

    2017-01-01

    High-temperature requirement A (HtrA)-like proteases participate in protein quality control in prokaryotes and eukaryotes by degrading damaged proteins; however, little is known about HtrAs produced by thermophiles. HtrAw is an HtrA-like protease of thermophilic Brevibacillus sp. WF146. The intact form of HtrAw (iHtrAw) consisting of a transmembrane segment-containing N-terminal domain, a trypsin-like protease domain, and a C-terminal PDZ domain was produced in Escherichia coli. Purified iHtrAw itself is unable to cleave the N-terminal domain, but requires protein substrates to autoprocess the N-terminal domain intermolecularly, yielding a short form (sHtrAw). Mutation at the substrate-binding site in the PDZ domain affects the conversion of iHtrAw to sHtrAw. Deletion analysis revealed that the N-terminal domain is not necessary for enzyme folding, activity, and thermostability. Compared with other known HtrAs, HtrAw contains an additional Ca(2+)-binding Dx[DN]xDG motif important for enzyme stability and/or activity. When produced in an htrA/htrB double deletion mutant of Bacillus subtilis, iHtrAw localized predominantly to the cell pellet, and the amount of sHtrAw in the culture supernatant increased at elevated temperatures. Moreover, HtrAw increased the heat resistance of the B. subtilis mutant. In strain WF146, HtrAw exists in both a cell-associated intact form and a cell-free short form; an increase in growth temperature enhanced HtrAw production and the amount of cell-free short form. Release of the short form of HtrAw from the membrane may have the advantage of allowing the enzyme to freely access and degrade damaged proteins surrounding the bacterium living at high temperatures.

  16. Identification of E-cadherin signature motifs functioning as cleavage sites for Helicobacter pylori HtrA

    NASA Astrophysics Data System (ADS)

    Schmidt, Thomas P.; Perna, Anna M.; Fugmann, Tim; Böhm, Manja; Jan Hiss; Haller, Sarah; Götz, Camilla; Tegtmeyer, Nicole; Hoy, Benjamin; Rau, Tilman T.; Neri, Dario; Backert, Steffen; Schneider, Gisbert; Wessler, Silja

    2016-03-01

    The cell adhesion protein and tumour suppressor E-cadherin exhibits important functions in the prevention of gastric cancer. As a class-I carcinogen, Helicobacter pylori (H. pylori) has developed a unique strategy to interfere with E-cadherin functions. In previous studies, we have demonstrated that H. pylori secretes the protease high temperature requirement A (HtrA) which cleaves off the E-cadherin ectodomain (NTF) on epithelial cells. This opens cell-to-cell junctions, allowing bacterial transmigration across the polarised epithelium. Here, we investigated the molecular mechanism of the HtrA-E-cadherin interaction and identified E-cadherin cleavage sites for HtrA. Mass-spectrometry-based proteomics and Edman degradation revealed three signature motifs containing the [VITA]-[VITA]-x-x-D-[DN] sequence pattern, which were preferentially cleaved by HtrA. Based on these sites, we developed a substrate-derived peptide inhibitor that selectively bound and inhibited HtrA, thereby blocking transmigration of H. pylori. The discovery of HtrA-targeted signature sites might further explain why we detected a stable 90 kDa NTF fragment during H. pylori infection, but also additional E-cadherin fragments ranging from 105 kDa to 48 kDa in in vitro cleavage experiments. In conclusion, HtrA targets E-cadherin signature sites that are accessible in in vitro reactions, but might be partially masked on epithelial cells through functional homophilic E-cadherin interactions.

  17. Architecture and regulation of HtrA-family proteins involved in protein quality control and stress response.

    PubMed

    Hansen, Guido; Hilgenfeld, Rolf

    2013-03-01

    Protein quality control is vital for all living cells and sophisticated molecular mechanisms have evolved to prevent the excessive accumulation of unfolded proteins. High-temperature requirement A (HtrA) proteases have been identified as important ATP-independent quality-control factors in most species. HtrA proteins harbor a serine-protease domain and at least one peptide-binding PDZ domain to ensure efficient removal of misfolded or damaged proteins. One distinctive property of HtrAs is their ability to assemble into complex oligomers. Whereas all examined HtrAs are capable of forming pyramidal 3-mers, higher-order complexes consisting of up to 24 molecules have been reported. Tight control of chaperone and protease function is of pivotal importance in preventing deleterious HtrA-protease activity. In recent years, structural biology provided detailed insights into the molecular basis of the regulatory mechanisms, which include unique intramolecular allosteric signaling cascades and the dynamic switching of oligomeric states of HtrA proteins. Based on these results, functional models for many family members have been developed. The HtrA protein family represents a remarkable example of how structural and functional diversity is attained from the assembly of simple molecular building blocks.

  18. HtrA2 regulates beta-amyloid precursor protein (APP) metabolism through endoplasmic reticulum-associated degradation.

    PubMed

    Huttunen, Henri J; Guénette, Suzanne Y; Peach, Camilla; Greco, Christopher; Xia, Weiming; Kim, Doo Yeon; Barren, Cory; Tanzi, Rudolph E; Kovacs, Dora M

    2007-09-21

    Alzheimer disease-associated beta-amyloid peptide is generated from its precursor protein APP. By using the yeast two-hybrid assay, here we identified HtrA2/Omi, a stress-responsive chaperone-protease as a protein binding to the N-terminal cysteinerich region of APP. HtrA2 coimmunoprecipitates exclusively with immature APP from cell lysates as well as mouse brain extracts and degrades APP in vitro. A subpopulation of HtrA2 localizes to the cytosolic side of the endoplasmic reticulum (ER) membrane where it contributes to ER-associated degradation of APP together with the proteasome. Inhibition of the proteasome results in accumulation of retrotranslocated forms of APP and increased association of APP with HtrA2 and Derlin-1 in microsomal membranes. In cells lacking HtrA2, APP holoprotein is stabilized and accumulates in the early secretory pathway correlating with elevated levels of APP C-terminal fragments and increased Abeta secretion. Inhibition of ER-associated degradation (either HtrA2 or proteasome) promotes binding of APP to the COPII protein Sec23 suggesting enhanced trafficking of APP out of the ER. Based on these results we suggest a novel function for HtrA2 as a regulator of APP metabolism through ER-associated degradation.

  19. The HtrA protease from Streptococcus pneumoniae digests both denatured proteins and the competence-stimulating peptide.

    PubMed

    Cassone, Marco; Gagne, Alyssa L; Spruce, Lynn A; Seeholzer, Steven H; Sebert, Michael E

    2012-11-09

    The HtrA protease of Streptococcus pneumoniae functions both in a general stress response role and as an error sensor that specifically represses genetic competence when the overall level of biosynthetic errors in cellular proteins is low. However, the mechanism through which HtrA inhibits development of competence has been unknown. We found that HtrA digested the pneumococcal competence-stimulating peptide (CSP) and constituted the primary extracytoplasmic CSP-degrading activity in cultures of S. pneumoniae. Mass spectrometry demonstrated that cleavage predominantly followed residue Phe-8 of the CSP-1 isoform of the peptide within its central hydrophobic patch, and in competition assays, both CSP-1 and CSP-2 interacted with HtrA with similar efficiencies. More generally, analysis of β-casein digestion and of digestion within HtrA itself revealed a preference for substrates with non-polar residues at the P1 site. Consistent with a specificity for exposed hydrophobic residues, competition from native BSA only weakly inhibited digestion of CSP, but denaturation converted BSA into a strong competitive inhibitor of such proteolysis. Together these findings support a model in which digestion of CSP by HtrA is reduced in the presence of other unfolded proteins that serve as alternative targets for degradation. Such competition may provide a mechanism by which HtrA functions in a quality control capacity to monitor the frequency of biosynthetic errors that result in protein misfolding.

  20. HtrA1 may regulate the osteogenic differentiation of human periodontal ligament cells by TGF-β1.

    PubMed

    Li, Ran; Zhang, Qi

    2015-04-01

    Periodontal ligament cells (PDLCs) in periodontal ligament (PDL) can differentiate into osteoblasts, while physiologically PDL remains non-mineralized space although located two hard tissues. But the exact mechanism of which is still unclear. High-temperature requirement protein A1 (HtrA1) is a key mineralization regulator and could inhibit the osteogenesis by transforming growth factor-β (TGF-β) signaling. However, the role of HtrA1 in PDLCs osteogenic differentiation has yet to be clarified. We assume HtrA1 may play an important role in maintaining the balance of PDL mineralization, and may regulate human periodontal ligament cells (hPDLCs) osteogenic differentiation by TGF-β1. Firstly we confirmed the mRNA expression of HtrA1 and TGF-β1 in hPDLCs by RT-PCR, then QDs-based immunofluorescence demonstrated the co-localization of them in the cytoplasm, and co-immunoprecipitation further confirmed the interaction between them. Lentivirus-mediated HtrA1 overexpression enhanced the osteogenic differentiation of hPDLCs, as well as up-regulation of TGF-β1. In contrast, knockdown of HtrA1 suppressed the osteogenic differentiation with down-regulation of TGF-β1. These findings suggested that HtrA1 plays a positive role in hPDLCs osteogenic differentiation and may regulate this process by TGF-β1.

  1. Effects of anti-phospholipid antibodies on a human trophoblast cell line (HTR-8/SVneo).

    PubMed

    Jovanović, Milica; Bozić, Milica; Kovacević, Tamara; Radojcić, Ljiljana; Petronijević, Milos; Vićovac, Ljiljana

    2010-01-01

    Antibodies to phospholipids (aPL) have been shown to adversely affect trophoblast invasion in vivo and in vitro. HTR-8/SVneo cells derived from first trimester of pregnancy extravillous trophoblast were studied. Matrigel invasion assay, cytochemistry and cell-based enzyme-linked immunosorbant assay (ELISA) with aPL or normal IgG was used. Our data show that aPL at 100 microg/ml decrease invasiveness of HTR-8/SVneo cells to 60% of control (p<0.01), and this was also shown for primary cytotrophoblast (to 15.5% of control, p<0.001). aPL treatment caused a significant decrease in integrin alpha(1), alpha(5), and beta(1) proteins (86%, 84%, and 87%, respectively). We conclude that HTR-8/SVneo cell culture is a suitable model to study mechanisms of action of aPL on trophoblast, which in HTR-8/SVneo cells inhibit invasion by decreasing integrins alpha(5), alpha(1), and beta(1).

  2. True but not false memories are associated with the HTR2A gene.

    PubMed

    Zhu, Bi; Chen, Chuansheng; Loftus, Elizabeth F; Moyzis, Robert K; Dong, Qi; Lin, Chongde

    2013-11-01

    Previous research reported that serotonin receptor 2A gene (HTR2A) polymorphisms were associated with memory. However, it is unknown whether these genetic variants were associated with both true and false memories. The current study of 336 Han Chinese subjects tested 30 single nucleotide polymorphisms (SNPs) within the HTR2A gene for potential associations with true and false memories. False memories were assessed using the Deese-Roediger-McDermott (DRM) paradigm, in which people falsely remember semantically related (but unpresented) words. We found that 11 SNPs within the HTR2A gene were associated with true memory (p=0.000076-0.043). The associations between true memory and seven adjacent SNPs (i.e., rs1923888, rs1745837, rs9567739, rs3742279, rs655888, rs655854, and rs2296972) were still significant after multiple testing corrections. Haplotype-based association analysis revealed that, true memory was positively associated with haplotype A-C-C-G-C-T-A for these seven adjacent SNPs (p=0.000075), which was still significant after multiple testing correction. Only one SNP rs655854 was associated with false memory (p=0.023), and it was not significant after multiple testing correction. This study replicates, in an Asian population, that genetic variation in HTR2A is associated with episodic memory, and also suggests that this association is restricted to true memory.

  3. Vom Himmelsmythos zum Weltgesetz. Eine Kulturgeschichte der Astronomie.

    NASA Astrophysics Data System (ADS)

    Bialas, V.

    Contents: I: Die Betrachtung des Himmels im Zeichen des Mythos. 1. Astronomische Spuren der Vorzeit. 2. Naturvölker und Ethnoastronomie. 3. Hochkulturen in Asien und Amerika. 4. Das alte Ägypten. 5. Mesopotamien. 6. Die mythisch-religiöse Erfahrung der Welt und die frühe Astronomie. II: Vom Mythos zum Logos. 7. Antikes Griechenland. 8. Römische Antike und frühes Christentum (ca. 200 v. Chr. - 500 n. Chr.). 9. Astronomie unter dem Zeichen des Islam. 10. Europäisches Mittelalter. 11. Die Astronomie als kulturelles Erbe der Menschheit. III: Die Selbstdifferenzierung des Logos. 12. Das Buch der Natur wird aufgeschlagen. 13. Klassische Astronomie und philosophische Aufklärung (ca. 1700 - 1850). 14. Neue Wege der Kosmosforschung in Astrophysik und Kosmologie (ca. 1850 - 1950). 15. Schlußbetrachtung: Die Frage nach dem Weltbild in verunsicherter Zeit.

  4. SHAPE analysis of the htrA RNA thermometer from Salmonella enterica.

    PubMed

    Choi, Edric K; Ulanowicz, Kelsey A; Nguyen, Yen Anh H; Frandsen, Jane K; Mitton-Fry, Rachel M

    2017-10-01

    RNA thermometers regulate expression of some genes involved in virulence of pathogenic bacteria such as Yersinia, Neisseria, and Salmonella They often function through temperature-dependent conformational changes that alter accessibility of the ribosome-binding site. The 5'-untranslated region (UTR) of the htrA mRNA from Salmonella enterica contains a very short RNA thermometer. We have systematically characterized the structure and dynamics of this thermometer at single-nucleotide resolution using SHAPE (selective 2'-hydroxyl acylation analyzed by primer extension) assays. Our results confirm that the htrA thermometer adopts the predicted hairpin conformation at low temperatures, with conformational change occurring over a physiological temperature regime. Detailed SHAPE melting curves for individual nucleotides suggest that the thermometer unfolds in a cooperative fashion, with nucleotides from both upper and lower portions of the stem gaining flexibility at a common transition temperature. Intriguingly, analysis of an extended htrA 5' UTR sequence revealed not only the presence of the RNA thermometer, but also an additional, stable upstream structure. We generated and analyzed point mutants of the htrA thermometer, revealing elements that modulate its stability, allowing the hairpin to melt under the slightly elevated temperatures experienced during the infection of a warm-blooded host. This work sheds light on structure-function relationships in htrA and related thermometers, and it also illustrates the utility of SHAPE assays for detailed study of RNA thermometer systems. © 2017 Choi et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  5. Evidence of a conserved role for Chlamydia HtrA in the replication phase of the chlamydial developmental cycle.

    PubMed

    Patel, Pooja; De Boer, Leonore; Timms, Peter; Huston, Wilhelmina May

    2014-08-01

    Identification of the HtrA inhibitor JO146 previously enabled us to demonstrate an essential function for HtrA during the mid-replicative phase of the Chlamydia trachomatis developmental cycle. Here we extend our investigations to other members of the Chlamydia genus. C. trachomatis isolates with distinct replicative phase growth kinetics showed significant loss of viable infectious progeny after HtrA was inhibited during the replicative phase. Mid-replicative phase addition of JO146 was also significantly detrimental to Chlamydia pecorum, Chlamydia suis and Chlamydia cavie. These data combined indicate that HtrA has a conserved critical role during the replicative phase of the chlamydial developmental cycle. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  6. A reform in the helium purification system of the HTR-10: gamma dose rate measurement and suggestions for decommissioning

    SciTech Connect

    Feng Xie; Hong Li; Jianzhu Cao; Suyuan Yu; Liguo Zhang; Wenqian Li; Sheng Fang

    2013-07-01

    A reform will be implemented in the helium purification system of the 10 MW High Temperature Gas-cooled Test Reactor (HTR-10) in China. The measurement of the gamma dose rates of facilities, including valves, pipes, dust filter, etc., in the purification system of the HTR-10, has been performed. The results indicated that most radiation nuclides are concentrated in the dust filter and facilities at the entrance of the helium purification system upstream of the dust filter. Other facilities have the same gamma dose rate level as the background. Based on the previous study and experiences in AVR, the measurement results can be understood that the radioactive dust carried by the helium gas was filtered by the dust filter. It provides important insights for the decontamination and decommissioning of facilities in the primary loop, especially in the helium purification system of the HTR-10 as well as the High Temperature Reactor-Pebble bed Modules (HTR-PM). (authors)

  7. Structure and variation of three canine genes involved in serotonin binding and transport: the serotonin receptor 1A gene (htr1A), serotonin receptor 2A gene (htr2A), and serotonin transporter gene (slc6A4).

    PubMed

    van den Berg, L; Kwant, L; Hestand, M S; van Oost, B A; Leegwater, P A J

    2005-01-01

    Aggressive behavior is the most frequently encountered behavioral problem in dogs. Abnormalities in brain serotonin metabolism have been described in aggressive dogs. We studied canine serotonergic genes to investigate genetic factors underlying canine aggression. Here, we describe the characterization of three genes of the canine serotonergic system: the serotonin receptor 1A and 2A gene (htr1A and htr2A) and the serotonin transporter gene (slc6A4). We isolated canine bacterial artificial chromosome clones containing these genes and designed oligonucleotides for genomic sequencing of coding regions and intron-exon boundaries. Golden retrievers were analyzed for DNA sequence variations. We found two nonsynonymous single nucleotide polymorphisms (SNPs) in the coding sequence of htr1A; one SNP close to a splice site in htr2A; and two SNPs in slc6A4, one in the coding sequence and one close to a splice site. In addition, we identified a polymorphic microsatellite marker for each gene. Htr1A is a strong candidate for involvement in the domestication of the dog. We genotyped the htr1A SNPs in 41 dogs of seven breeds with diverse behavioral characteristics. At least three SNP haplotypes were found. Our results do not support involvement of the gene in domestication.

  8. Distinct 3D Architecture and Dynamics of the Human HtrA2(Omi) Protease and Its Mutated Variants

    PubMed Central

    Gieldon, Artur; Zurawa-Janicka, Dorota; Jarzab, Miroslaw; Wenta, Tomasz; Golik, Przemyslaw; Dubin, Grzegorz; Lipinska, Barbara; Ciarkowski, Jerzy

    2016-01-01

    HtrA2(Omi) protease controls protein quality in mitochondria and plays a major role in apoptosis. Its HtrA2S306A mutant (with the catalytic serine routinely disabled for an X-ray study to avoid self-degradation) is a homotrimer whose subunits contain the serine protease domain (PD) and the regulatory PDZ domain. In the inactive state, a tight interdomain interface limits penetration of both PDZ-activating ligands and PD substrates into their respective target sites. We successfully crystalized HtrA2V226K/S306A, whose active counterpart HtrA2V226K has had higher proteolytic activity, suggesting higher propensity to opening the PD-PDZ interface than that of the wild type HtrA2. Yet, the crystal structure revealed the HtrA2V226K/S306A architecture typical of the inactive protein. To get a consistent interpretation of crystallographic data in the light of kinetic results, we employed molecular dynamics (MD). V325D inactivating mutant was used as a reference. Our simulations demonstrated that upon binding of a specific peptide ligand NH2-GWTMFWV-COOH, the PDZ domains open more dynamically in the wild type protease compared to the V226K mutant, whereas the movement is not observed in the V325D mutant. The movement relies on a PDZ vs. PD rotation which opens the PD-PDZ interface in a lid-like (budding flower-like in trimer) fashion. The noncovalent hinges A and B are provided by two clusters of interfacing residues, harboring V325D and V226K in the C- and N-terminal PD barrels, respectively. The opening of the subunit interfaces progresses in a sequential manner during the 50 ns MD simulation. In the systems without the ligand only minor PDZ shifts relative to PD are observed, but the interface does not open. Further activation-associated events, e.g. PDZ-L3 positional swap seen in any active HtrA protein (vs. HtrA2), were not observed. In summary, this study provides hints on the mechanism of activation of wtHtrA2, the dynamics of the inactive HtrA2V325D, but does not

  9. Interleukin-6 stimulates cell migration, invasion and integrin expression in HTR-8/SVneo cell line.

    PubMed

    Jovanović, M; Vićovac, L

    2009-04-01

    Interleukin-6 (IL-6) is present in human endometrium throughout menstrual cycle and in pregnancy. Trophoblast also expresses IL-6. IL-6R and its associated signal transducer gp130 were found in trophoblast as well. IL-6 is generally assumed to be relevant for trophoblast invasion. This study was undertaken to determine influence of endogenous and externally added IL-6 on invasion and migration of first trimester of pregnancy trophoblast in vitro. Integrins alpha(5)beta(1) and alpha(1)beta(1) have been shown to play an important role in trophoblast invasion and the effect of IL-6 on the expression of these integrin subunits was studied. We are showing that in both isolated first trimester of pregnancy cytotrophoblast (CTB) and HTR-8/SVneo cell line IL-6 and IL-6R are present. The effect on migration was studied using cell wounding and migration test on HTR-8/SVneo cells. Effect of IL-6 and function blocking anti-IL-6 antibody in Matrigel invasion tests was studied on both cell types. The effect of IL-6 on integrin subunit expression was determined by cell-based ELISA and Western blot on HTR-8/SVneo cells. The results obtained show that exogenous IL-6 has stimulatory effect on cell migration in HTR-8/SVneo and invasion by both cell types. Function blocking anti-IL-6 inhibited unstimulated invasion by isolated first trimester cytotrophoblast and both cell migration and invasion in unstimulated HTR-8/SVneo. Integrin alpha(5) expression was stimulated by IL-6 to 134% (p<0.05), alpha(1) to 135% (p<0.005), and beta(1) to 134% (p<0.001) of control in cell-based ELISA, but also in Western blot. The data obtained show for the first time sensitivity of extravillous trophoblast cell line HTR-8/SVneo to IL-6, in addition to isolated first trimester cytotrophoblast. We conclude that both exogenous and endogenous IL-6 stimulate trophoblast cell migration and invasion, which may be partly attributable to stimulation of expression of the studied integrin subunits.

  10. Human telomerase reverse transcriptase binds to a pre-organized hTR in vivo exposing its template

    PubMed Central

    Zemora, Georgeta; Handl, Stefan; Waldsich, Christina

    2016-01-01

    Telomerase is a specialized reverse transcriptase that is responsible for telomere length maintenance. As in other organisms, the minimal components required for an active human telomerase are the template-providing telomerase RNA (hTR) and the enzymatic entity telomerase reverse transcriptase (hTERT). Here, we explored the structure of hTR and the hTERT-induced conformational changes within hTR in living cells. By employing an in vivo DMS chemical probing technique, we showed that the pseudoknot and associated triple helical scaffold form stably in vivo independently of hTERT. In fact, the dimethyl-sulfate (DMS) modification pattern suggests that hTR alone is capable of adopting a conformation that is suited to interact with hTERT. However, in the absence of hTERT the template region of hTR is only weakly accessible to DMS-modifications. The predominant change after binding of hTERT to hTR is the exposure of the template region. PMID:26481359

  11. Human telomerase reverse transcriptase binds to a pre-organized hTR in vivo exposing its template.

    PubMed

    Zemora, Georgeta; Handl, Stefan; Waldsich, Christina

    2016-01-08

    Telomerase is a specialized reverse transcriptase that is responsible for telomere length maintenance. As in other organisms, the minimal components required for an active human telomerase are the template-providing telomerase RNA (hTR) and the enzymatic entity telomerase reverse transcriptase (hTERT). Here, we explored the structure of hTR and the hTERT-induced conformational changes within hTR in living cells. By employing an in vivo DMS chemical probing technique, we showed that the pseudoknot and associated triple helical scaffold form stably in vivo independently of hTERT. In fact, the dimethyl-sulfate (DMS) modification pattern suggests that hTR alone is capable of adopting a conformation that is suited to interact with hTERT. However, in the absence of hTERT the template region of hTR is only weakly accessible to DMS-modifications. The predominant change after binding of hTERT to hTR is the exposure of the template region.

  12. Methylphenidate to adolescent rats drives enduring changes of accumbal Htr7 expression: implications for impulsive behavior and neuronal morphology.

    PubMed

    Leo, D; Adriani, W; Cavaliere, C; Cirillo, G; Marco, E M; Romano, E; di Porzio, U; Papa, M; Perrone-Capano, C; Laviola, G

    2009-04-01

    Methylphenidate (MPH) administration to adolescent rodents produces persistent region-specific changes in brain reward circuits and alterations of reward-based behavior. We show that these modifications include a marked increment of serotonin (5-hydroxy-tryptamine) receptor type 7 (Htr7) expression and synaptic contacts, mainly in the nucleus accumbens, and a reduction of basal behavioral impulsivity. We show that neural and behavioral consequences are functionally related: administration of a selective Htr7 antagonist fully counteracts the MPH-reduced impulsive behavior and enhances impulsivity when administered alone in naive rats. Agonist-induced activation of endogenous Htr7 significantly increases neurite length in striatal neuron primary cultures, thus suggesting plastic remodeling of neuronal morphology. The mixed Htr (1a/7) agonist, 8-OH-DPAT, reduces impulsive behavior in adolescent rats and in naive adults, whose impulsivity is enhanced by the Htr7 antagonist. In summary, behavioral pharmacology experiments show that Htr7 mediates self-control behavior, and brain primary cultures experiments indicate that this receptor may be involved in the underlying neural plasticity, through changes in neuronal cytoarchitecture.

  13. HtrA2 deficiency causes mitochondrial uncoupling through the F1F0-ATP synthase and consequent ATP depletion

    PubMed Central

    Plun-Favreau, H; Burchell, V S; Holmström, K M; Yao, Z; Deas, E; Cain, K; Fedele, V; Moisoi, N; Campanella, M; Miguel Martins, L; Wood, N W; Gourine, A V; Abramov, A Y

    2012-01-01

    Loss of the mitochondrial protease HtrA2 (Omi) in mice leads to mitochondrial dysfunction, neurodegeneration and premature death, but the mechanism underlying this pathology remains unclear. Using primary cultures from wild-type and HtrA2-knockout mice, we find that HtrA2 deficiency significantly reduces mitochondrial membrane potential in a range of cell types. This depolarisation was found to result from mitochondrial uncoupling, as mitochondrial respiration was increased in HtrA2-deficient cells and respiratory control ratio was dramatically reduced. HtrA2-knockout cells exhibit increased proton translocation through the ATP synthase, in combination with decreased ATP production and truncation of the F1 α-subunit, suggesting the ATP synthase as the source of the proton leak. Uncoupling in the HtrA2-deficient mice is accompanied by altered breathing pattern and, on a cellular level, ATP depletion and vulnerability to chemical ischaemia. We propose that this vulnerability may ultimately cause the neurodegeneration observed in these mice. PMID:22739987

  14. Recombinant Deg/HtrA proteases from Synechocystis sp. PCC 6803 differ in substrate specificity, biochemical characteristics and mechanism

    PubMed Central

    Huesgen, Pitter F.; Miranda, Helder; Lam, XuanTam; Perthold, Manuela; Schuhmann, Holger; Adamska, Iwona; Funk, Christiane

    2011-01-01

    Cyanobacteria require efficient protein-quality-control mechanisms to survive under dynamic, often stressful, environmental conditions. It was reported that three serine proteases, HtrA (high temperature requirement A), HhoA (HtrA homologue A) and HhoB (HtrA homologue B), are important for survival of Synechocystis sp. PCC 6803 under high light and temperature stresses and might have redundant physiological functions. In the present paper, we show that all three proteases can degrade unfolded model substrates, but differ with respect to cleavage sites, temperature and pH optima. For recombinant HhoA, and to a lesser extent for HtrA, we observed an interesting shift in the pH optimum from slightly acidic to alkaline in the presence of Mg2+ and Ca2+ ions. All three proteases formed different homo-oligomeric complexes with and without substrate, implying mechanistic differences in comparison with each other and with the well-studied Escherichia coli orthologues DegP (degradation of periplasmic proteins P) and DegS. Deletion of the PDZ domain decreased, but did not abolish, the proteolytic activity of all three proteases, and prevented substrate-induced formation of complexes higher than trimers by HtrA and HhoA. In summary, biochemical characterization of HtrA, HhoA and HhoB lays the foundation for a better understanding of their overlapping, but not completely redundant, stress-resistance functions in Synechocystis sp. PCC 6803. PMID:21332448

  15. Omi/HtrA2 promotes cell death by binding and degrading the anti-apoptotic protein ped/pea-15.

    PubMed

    Trencia, Alessandra; Fiory, Francesca; Maitan, Maria Alessandra; Vito, Pasquale; Barbagallo, Alessia Paola Maria; Perfetti, Anna; Miele, Claudia; Ungaro, Paola; Oriente, Francesco; Cilenti, Lucia; Zervos, Antonis S; Formisano, Pietro; Beguinot, Francesco

    2004-11-05

    ped/pea-15 is a ubiquitously expressed 15-kDa protein featuring a broad anti-apoptotic function. In a yeast two-hybrid screen, the pro-apoptotic Omi/HtrA2 mitochondrial serine protease was identified as a specific interactor of the ped/pea-15 death effector domain. Omi/HtrA2 also bound recombinant ped/pea-15 in vitro and co-precipitated with ped/pea-15 in 293 and HeLa cell extracts. In these cells, the binding of Omi/HtrA2 to ped/pea-15 was induced by UVC exposure and followed the mitochondrial release of Omi/HtrA2 into the cytoplasm. Upon UVC exposure, cellular ped/pea-15 protein expression levels decreased. This effect was prevented by the ucf-101 specific inhibitor of the Omi/HtrA2 proteolytic activity, in a dose-dependent fashion. In vitro incubation of ped/pea-15 with Omi/HtrA2 resulted in ped/pea-15 degradation. In intact cells, the inhibitory action of ped/pea-15 on UVC-induced apoptosis progressively declined at increasing Omi/HtrA2 expression. This further effect of Omi/HtrA2 was also inhibited by ucf-101. In addition, ped/pea-15 expression blocked Omi/HtrA2 co-precipitation with the caspase inhibitor protein XIAP and caspase 3 activation. Thus, in part, apoptosis following Omi/HtrA2 mitochondrial release is mediated by reduction in ped/pea-15 cellular levels. The ability of Omi/HtrA2 to relieve XIAP inhibition on caspases is modulated by the relative levels of Omi/HtrA2 and ped/pea-15.

  16. Circadian variation of cell proliferation in HTR-8/SVneo cell line.

    PubMed

    Lunghi, Laura; Frigato, Elena; Ferretti, Maria Enrica; Biondi, Carla; Bertolucci, Cristiano

    2011-12-01

    Circadian clock controls several physiological processes such as cell proliferation. Extravillous trophoblast proliferation is a tightly regulated function playing a fundamental role in maternal vessel remodeling. We recently demonstrated that clock genes Per2 and Dec1 as well as the clock-controlled genes Dbp and Vegf are rhythmically expressed in human extravillous trophoblast-derived HTR-8/SVneo cells. Analyzing the time course of HTR-8/SVneo cell proliferation, a circadian variation in cell number was found. Moreover, we showed a rhythmic expression of mRNAs for Wee1 and stathmin, two genes involved in cell cycle progression. We suggest that circadian clockwork may orchestrate the functionality of the several factors involved in the control of human trophoblast functions that are fundamental for a successfully pregnancy outcome.

  17. Preliminary Reactor Physics Assessment of the HTR Module with 14% Enriched UCO Fuel

    SciTech Connect

    Gerhard Strydom; Hans D. Gougar

    2013-03-01

    The high temperature reactor (HTR) Module (Lohnert, 1990) is a graphite-moderated, helium cooled pebble bed design that has been extensively used as a reference template for the former South African (Matzner, 2004) and current Chinese (Zhang et al., 2009) HTR programs. This design utilizes spherical fuel elements packed into a dynamic pebble bed, consisting of tri-structural isotropic (TRISO) coated uranium oxide (UO2) 500 µm fuel kernels with a U-235 enrichment of 7.8% and a heavy metal loading of 7 g per pebble. This fuel type was previously qualified for use in Germany for pebble bed HTRs, as well as undergoing re-qualification in South Africa for the PBMR project. It is also the fuel type being tested for use in the high temperature reactor (HTR-PM) under construction in China. In the United States, however, a different TRISO fuel form is the subject of a qualification program. The U.S. experience with HTRs has been focused upon the batch-fueled prismatic reactor in which TRISO particles are embedded in cylindrical compacts and stacked inside the graphite blocks which comprise the core. Under this type of operating regime, a smaller TRISO with a different composition and enrichment performs better than the fuel historically used in PBRs. Fuel kernels and compacting techniques more suited to prismatic core duty are currently being developed and qualified under the U.S. Department of Energy's Advanced Gas Reactor (AGR) fuel development program and in support of the Next Generation Nuclear Plant project. Interest in the pebble bed concept remains high, however, and a study was undertaken by the authors to assess the viability of using AGR fuel in a pebble bed reactor. Using the German HTR Module as the reference plant, key neutronic and thermal-hydraulic parameters were compared between the nominal design and one fueled with the fuel that is the focus of the AGR program.

  18. A family-based association study of the HTR1B gene in eating disorders.

    PubMed

    Hernández, Sandra; Camarena, Beatriz; González, Laura; Caballero, Alejandro; Flores, Griselda; Aguilar, Alejandro

    2016-01-01

    To explore the association of three polymorphisms of the serotonin receptor 1Dβ gene (HTR1B) in the etiology of eating disorders and their relationship with clinical characteristics. We analyzed the G861C, A-161T, and A1180G polymorphisms of the HTR1B gene through a family-based association test (FBAT) in 245 nuclear families. The sample was stratified into anorexia nervosa (AN) spectrum and bulimia nervosa (BN) spectrum. In addition, we performed a quantitative FBAT analysis of anxiety severity, depression severity, and Yale-Brown-Cornell Eating Disorders Scale (YBC-EDS) in the AN and BN-spectrum groups. FBAT analysis of the A-161T polymorphism found preferential transmission of allele A-161 in the overall sample. This association was stronger when the sample was stratified by spectrums, showing transmission disequilibrium between the A-161 allele and BN spectrum (z = 2.871, p = 0.004). Quantitative trait analysis showed an association between severity of anxiety symptoms and the C861 allele in AN-spectrum participants (z = 2.871, p = 0.004). We found no associations on analysis of depression severity or preoccupation and ritual scores in AN or BN-spectrum participants. Our preliminary findings suggest a role of the HTR1B gene in susceptibility to development of BN subtypes. Furthermore, this gene might have an impact on the severity of anxiety in AN-spectrum patients.

  19. Testing of HTR UO2 TRISO fuels in AVR and in material test reactors

    NASA Astrophysics Data System (ADS)

    Kania, Michael J.; Nabielek, Heinz; Verfondern, Karl; Allelein, Hans-Josef

    2013-10-01

    The German High Temperature Reactor Fuel Development Program successfully developed, licensed and manufactured many thousands of spherical fuel elements that were used to power the experimental AVR reactor and the commercial THTR reactor. In the 1970s, this program extended the performance envelope of HTR fuels by developing and qualifying the TRISO-coated particle system. Irradiation testing in real-time AVR tests and accelerated MTR tests demonstrated the superior manufacturing process of this fuel and its irradiation performance. In the 1980s, another program direction change was made to a low enriched UO2 TRISO-coated particle system coupled with high-quality manufacturing specifications designed to meet new HTR plant design needs. These needs included requirements for inherent safety under normal operation and accident conditions. Again, the German fuel development program met and exceeded these challenges by manufacturing and qualifying the low-enriched UO2 TRISO-fuel system for HTR systems with steam generation, gas-turbine systems and very high temperature process heat applications. Fuel elements were manufactured in production scale facilities that contained near defect free UO2 TRISO coated particles, homogeneously distributed within a graphite matrix with very low levels of uranium contamination. Good irradiation performance for these elements was demonstrated under normal operating conditions to 12% FIMA and under accident conditions not exceeding 1600 °C.

  20. Htr2a expression responds rapidly to environmental stimuli in an Egr3-dependent manner

    PubMed Central

    Maple, Amanda M.; Zhao, Xiuli; Elizalde, Diana I.; McBride, Andrew K.; Gallitano, Amelia L

    2015-01-01

    Pharmacologic and genetic findings have implicated the serotonin 2A receptor (5-HT2AR) in the etiology of schizophrenia. Recent studies have shown reduced 5-HT2AR levels in schizophrenia patients, yet the cause of this difference is unknown. Environmental factors, such as stress, also influence schizophrenia risk, yet little is known about how environment may affect this receptor. To determine if acute stress alters 5-HT2AR expression, we examined the effect of sleep deprivation on cortical Htr2a mRNA in mice. We found that 6 hours of sleep deprivation induces a 2-fold increase in Htr2a mRNA, a more rapid effect than has been previously reported. This effect requires the immediate early gene early growth response 3 (Egr3), as sleep deprivation failed to induce Htr2a expression in Egr3−/− mice. These findings provide a functional link between two schizophrenia candidate genes and an explanation of how environment may influence a genetic predisposition for schizophrenia. PMID:25857407

  1. Different Roles of COMT and HTR2A Genotypes in Working Memory Subprocesses

    PubMed Central

    Kondo, Hirohito M.; Nomura, Michio; Kashino, Makio

    2015-01-01

    Working memory is linked to the functions of the frontal areas, in which neural activity is mediated by dopaminergic and serotonergic tones. However, there is no consensus regarding how the dopaminergic and serotonergic systems influence working memory subprocesses. The present study used an imaging genetics approach to examine the interaction between neurochemical functions and working memory performance. We focused on functional polymorphisms of the catechol-O-methyltransferase (COMT) Val158Met and serotonin 2A receptor (HTR2A) -1438G/A genes, and devised a delayed recognition task to isolate the encoding, retention, and retrieval processes for visual information. The COMT genotypes affected recognition accuracy, whereas the HTR2A genotypes were associated with recognition response times. Activations specifically related to working memory were found in the right frontal and parietal areas, such as the middle frontal gyrus (MFG), inferior frontal gyrus (IFG), anterior cingulate cortex (ACC), and inferior parietal lobule (IPL). MFG and ACC/IPL activations were sensitive to differences between the COMT genotypes and between the HTR2A genotypes, respectively. Structural equation modeling demonstrated that stronger connectivity in the ACC-MFG and ACC-IFG networks is related to better task performance. The behavioral and fMRI results suggest that the dopaminergic and serotonergic systems play different roles in the working memory subprocesses and modulate closer cooperation between lateral and medial frontal activations. PMID:25974269

  2. Upregulation of mitochondrial protease HtrA2/Omi contributes to manganese-induced neuronal apoptosis in rat brain striatum.

    PubMed

    Jiang, J K; Ma, X; Wu, Q Y; Qian, W B; Wang, N; Shi, S S; Han, J L; Zhao, J Y; Jiang, S Y; Wan, C H

    2014-05-30

    Manganese (Mn) is an essential trace element that is required for normal brain functioning. However, excessive intake of Mn has been known to lead to neuronal loss and clinical symptoms resembling idiopathic Parkinson's disease (IPD), whose precise molecular mechanism remains largely elusive. In the study, we established a Mn-exposed rat model and identified a mitochondrial protease, the mature form of high temperature requirement A2 (HtrA2/Omi), which was significantly upregulated in rat brain striatum after Mn exposure. Western blot and immunohistochemical analyses revealed that the expression of mature HtrA2 was remarkably increased following Mn exposure. In addition, immunofluorescence assay demonstrated that overexposure to Mn could lead to significant elevation in the number of HtrA2-positive neurons. Accordingly, the expression of X-linked inhibitor of apoptosis protein (XIAP), a well-characterized target of HtrA2-mediated proteolysis, was progressively decreased following Mn exposure, and was correlated with increased level of active caspase-3. Further, we showed that Mn exposure decreased the viability and induced apparent apoptosis of NFG-differentiated PC12 cells. Importantly, the expression of HtrA2 was progressively increased, whereas the level of cellular XIAP was reduced during Mn-induced apoptosis. In addition, blockage of HtrA2 activity with UCF-101 restored Mn-induced reduction in XIAP expression. Finally, we observed that UCF-101 treatment ameliorated Mn-induced apoptosis in PC12 cells. Collectively, these findings suggested that upregulated HtrA2 played a role in Mn-induced neuronal death in brain striatum.

  3. Proteomic approaches to identify substrates of the three Deg/HtrA proteases of the cyanobacterium Synechocystis sp. PCC 6803.

    PubMed

    Tam, Lam X; Aigner, Harald; Timmerman, Evy; Gevaert, Kris; Funk, Christiane

    2015-06-15

    The family of Deg/HtrA proteases plays an important role in quality control of cellular proteins in a wide range of organisms. In the genome of the cyanobacterium Synechocystis sp. PCC 6803, a model organism for photosynthetic research and renewable energy products, three Deg proteases are encoded, termed HhoA, HhoB and HtrA. In the present study, we compared wild-type (WT) Synechocystis cells with the single insertion mutants ΔhhoA, ΔhhoB and ΔhtrA. Protein expression of the remaining Deg/HtrA proteases was strongly affected in the single insertion mutants. Detailed proteomic studies using DIGE (difference gel electrophoresis) and N-terminal COFRADIC (N-terminal combined fractional diagonal chromatography) revealed that inactivation of a single Deg protease has similar impact on the proteomes of the three mutants; differences to WT were observed in enzymes involved in the major metabolic pathways. Changes in the amount of phosphate permease system Pst-1 were observed only in the insertion mutant ΔhhoB. N-terminal COFRADIC analyses on cell lysates of ΔhhoB confirmed changed amounts of many cell envelope proteins, including the phosphate permease systems, compared with WT. In vitro COFRADIC studies were performed to identify the specificity profiles of the recombinant proteases rHhoA, rHhoB or rHtrA added to the Synechocystis WT proteome. The combined in vivo and in vitro N-terminal COFRADIC datasets propose RbcS as a natural substrate for HhoA, PsbO for HhoB and HtrA and Pbp8 for HtrA. We therefore suggest that each Synechocystis Deg protease protects the cell through different, but connected mechanisms. © The Authors Journal compilation © 2015 Biochemical Society.

  4. Polymorphisms in HTR2A and DRD4 Predispose to Smoking and Smoking Quantity

    PubMed Central

    Pérez-Rubio, Gloria; Ramírez-Venegas, Alejandra; Noé Díaz, Valeri; García Gómez, Leonor; Elvira Fabián, Karina; García Carmona, Salvador; López-Flores, Luis A.; Ambrocio-Ortiz, Enrique; Contreras Romero, Rocío; Alcantar-Ayala, Noé; Sansores, Raúl H.

    2017-01-01

    Background Genes encoding the receptors involved in the dopaminergic and serotonergic pathways are potential candidates in the mechanisms of nicotine addiction. Aims To identify genetic variants in the promoter regions and exons of the DRD4 and HTR2A genes associated with tobacco smoking and the degree of nicotine addiction in Mexican mestizos. Methods The study included 438 non-smokers (NS) and 1,157 current smokers, ranked based on their consumption of cigarettes per day (cpd): 574 heavy smokers (HS, >20 cpd) and 583 light smokers (LS, 1–10 cpd). Genotyping was performed for 4 and 8 single nucleotide polymorphisms (SNPs) in the DRD4 and HTR2A genes, respectively. Results The C allele of rs1800955 in DRD4 was found to be associated with cigarette smoking in the HS vs. NS and LS vs. NS comparisons (p = 2.34E-03 and p = 1.13E-03, respectively); the association was maintained in the homozygous CC genotype (p = 5.00E-04 and p = 2.00E-04, respectively). The T allele of rs6313 in HTR2A was significantly associated with cigarette smoking and a greater degree of nicotine addiction (p = 4.77E-03, OR = 1.55); the association was maintained in the homozygous genotype (TT) (p = 4.90E-03, OR = 1.96). The A allele of rs6313 was associated with cigarette smoking in the HS vs. NS comparison (p = 1.53E-02, OR = 1.36); the risk was nearly doubled in the homozygous AA genotype (p = 1.30E-03, OR = 1.83) compared with the heterozygous GA genotype (OR = 1.38). Conclusions Among Mexican mestizos, the C allele of rs1800955 in the DRD4 gene and the A allele of rs6311 in the HTR2A gene are associated with cigarette smoking, whereas the T allele of rs6313 in HTR2A is associated with cigarette smoking and the degree of nicotine addiction. PMID:28103253

  5. Polymorphisms in HTR2A and DRD4 Predispose to Smoking and Smoking Quantity.

    PubMed

    Pérez-Rubio, Gloria; Ramírez-Venegas, Alejandra; Noé Díaz, Valeri; García Gómez, Leonor; Elvira Fabián, Karina; García Carmona, Salvador; López-Flores, Luis A; Ambrocio-Ortiz, Enrique; Contreras Romero, Rocío; Alcantar-Ayala, Noé; Sansores, Raúl H; Falfán-Valencia, Ramcés

    2017-01-01

    Genes encoding the receptors involved in the dopaminergic and serotonergic pathways are potential candidates in the mechanisms of nicotine addiction. To identify genetic variants in the promoter regions and exons of the DRD4 and HTR2A genes associated with tobacco smoking and the degree of nicotine addiction in Mexican mestizos. The study included 438 non-smokers (NS) and 1,157 current smokers, ranked based on their consumption of cigarettes per day (cpd): 574 heavy smokers (HS, >20 cpd) and 583 light smokers (LS, 1-10 cpd). Genotyping was performed for 4 and 8 single nucleotide polymorphisms (SNPs) in the DRD4 and HTR2A genes, respectively. The C allele of rs1800955 in DRD4 was found to be associated with cigarette smoking in the HS vs. NS and LS vs. NS comparisons (p = 2.34E-03 and p = 1.13E-03, respectively); the association was maintained in the homozygous CC genotype (p = 5.00E-04 and p = 2.00E-04, respectively). The T allele of rs6313 in HTR2A was significantly associated with cigarette smoking and a greater degree of nicotine addiction (p = 4.77E-03, OR = 1.55); the association was maintained in the homozygous genotype (TT) (p = 4.90E-03, OR = 1.96). The A allele of rs6313 was associated with cigarette smoking in the HS vs. NS comparison (p = 1.53E-02, OR = 1.36); the risk was nearly doubled in the homozygous AA genotype (p = 1.30E-03, OR = 1.83) compared with the heterozygous GA genotype (OR = 1.38). Among Mexican mestizos, the C allele of rs1800955 in the DRD4 gene and the A allele of rs6311 in the HTR2A gene are associated with cigarette smoking, whereas the T allele of rs6313 in HTR2A is associated with cigarette smoking and the degree of nicotine addiction.

  6. Potential Applications for Nuclear Energy besides Electricity Generation: AREVA Global Perspective of HTR Potential Market

    SciTech Connect

    Soutworth, Finis; Gauthier, Jean-Claude; Lecomte, Michel; Carre, Franck

    2007-07-01

    Energy supply is increasingly showing up as a major issue for electricity supply, transportation, settlement, and process heat industrial supply including hydrogen production. Nuclear power is part of the solution. For electricity supply, as exemplified in Finland and France, the EPR brings an immediate answer; HTR could bring another solution in some specific cases. For other supply, mostly heat, the HTR brings a solution inaccessible to conventional nuclear power plants for very high or even high temperature. As fossil fuels costs increase and efforts to avoid generation of Greenhouse gases are implemented, a market for nuclear generated process heat will develop. Following active developments in the 80's, HTR have been put on the back burner up to 5 years ago. Light water reactors are widely dominating the nuclear production field today. However, interest in the HTR technology was renewed in the past few years. Several commercial projects are actively promoted, most of them aiming at electricity production. ANTARES is today AREVA's response to the cogeneration market. It distinguishes itself from other concepts with its indirect cycle design powering a combined cycle power plant. Several reasons support this design choice, one of the most important of which is the design flexibility to adapt readily to combined heat and power applications. From the start, AREVA made the choice of such flexibility with the belief that the HTR market is not so much in competition with LWR in the sole electricity market but in the specific added value market of cogeneration and process heat. In view of the volatility of the costs of fossil fuels, AREVA's choice brings to the large industrial heat applications the fuel cost predictability of nuclear fuel with the efficiency of a high temperature heat source free of greenhouse gases emissions. The ANTARES module produces 600 MWth which can be split into the required process heat, the remaining power drives an adapted prorated

  7. Knockdown of Litopenaeus vannamei HtrA2, an up-regulated gene in response to WSSV infection, leading to delayed shrimp mortality.

    PubMed

    Peepim, Termsri; Phiwsaiya, Kornsunee; Charoensapsri, Walaiporn; Khunrae, Pongsak; Senapin, Saengchan; Rattanarojpong, Triwit

    2016-02-10

    HtrA2 is an apoptosis-activating gene that enhances the apoptotic process by preventing the formation of the IAP-caspase complex, thereby freeing caspase to trigger the apoptosis pathway. In this study, we presented the full-length cDNA sequence of HtrA2 from Litopenaeus vannamei (LvHtrA2). The full-length LvHtrA2 was 1335 bp, encoding 444 amino acids. This deduced amino acid sequence contained five conserved domains: a mitochondrial targeting signal (MTS), a transmembrane (TM) domain, an IAP-binding motif (IBM), a trimerization motif, a serine protease domain, and a PDZ domain normally found in the HtrA2 proteins of other organisms. A phylogenetic analysis revealed that LvHtrA2 clustered with the HtrA2 from other invertebrates and was closely related to Penaeus monodon HtrA2 (PmHtrA2). RT-PCR with RNA extracts from L. vannamei revealed that LvHtrA2 expression was found in several tissues, including the lymphoid organs, the haemocytes, the hepatopancreas, the gill, and the stomach, with different expression levels. When determining the role of LvHtrA2 in WSSV infection, it was found that LvHtrA2 transcription was early up-regulated in the WSSV-infected shrimp at 8h post-infection (p.i.) and expression still remained high at 48 h p.i.. It also demonstrated that dsRNA specific to LvHtrA2 reduced the cumulative mortality in the WSSV-infected shrimp compared with the control group. Additionally, depletion of the LvHtrA2 transcripts reduced expression levels for caspase-3 (Cap-3) gene in shrimp. This result could suggest that LvHtrA2 may involved in apoptosis mediated mortality rather than providing immune protection during WSSV infection. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. High Temperature Reactor (HTR) Deep Burn Core and Fuel Analysis: Design Selection for the Prismatic Block Reactor

    SciTech Connect

    Francesco Venneri; Chang-Keun Jo; Jae-Man Noh; Yonghee Kim; Claudio Filippone; Jonghwa Chang; Chris Hamilton; Young-Min Kim; Ji-Su Jun; Moon-Sung Cho; Hong-Sik Lim; MIchael A. Pope; Abderrafi M. Ougouag; Vincent Descotes; Brian Boer

    2010-09-01

    The Deep Burn (DB) Project is a U.S. Department of Energy sponsored feasibility study of Transuranic Management using high burnup fuel in the high temperature helium cooled reactor (HTR). The DB Project consists of seven tasks: project management, core and fuel analysis, spent fuel management, fuel cycle integration, TRU fuel modeling, TRU fuel qualification, and HTR fuel recycle. In the Phase II of the Project, we conducted nuclear analysis of TRU destruction/utilization in the HTR prismatic block design (Task 2.1), deep burn fuel/TRISO microanalysis (Task 2.3), and synergy with fast reactors (Task 4.2). The Task 2.1 covers the core physics design, thermo-hydraulic CFD analysis, and the thermofluid and safety analysis (low pressure conduction cooling, LPCC) of the HTR prismatic block design. The Task 2.3 covers the analysis of the structural behavior of TRISO fuel containing TRU at very high burnup level, i.e. exceeding 50% of FIMA. The Task 4.2 includes the self-cleaning HTR based on recycle of HTR-generated TRU in the same HTR. Chapter IV contains the design and analysis results of the 600MWth DB-HTR core physics with the cycle length, the average discharged burnup, heavy metal and plutonium consumptions, radial and axial power distributions, temperature reactivity coefficients. Also, it contains the analysis results of the 450MWth DB-HTR core physics and the analysis of the decay heat of a TRU loaded DB-HTR core. The evaluation of the hot spot fuel temperature of the fuel block in the DB-HTR (Deep-Burn High Temperature Reactor) core under full operating power conditions are described in Chapter V. The investigated designs are the 600MWth and 460MWth DB-HTRs. In Chapter VI, the thermo-fluid and safety of the 600MWth DB-HTRs has been analyzed to investigate a thermal-fluid design performance at the steady state and a passive safety performance during an LPCC event. Chapter VII describes the analysis results of the TRISO fuel microanalysis of the 600MWth and 450

  9. The HtrA2 Drosophila model of Parkinson's disease is suppressed by the pro-survival Bcl-2 Buffy.

    PubMed

    M'Angale, P Githure; Staveley, Brian E

    2017-01-01

    Mutations in High temperature requirement A2 (HtrA2), also designated PARK13, which lead to the loss of its protease activity, have been associated with Parkinson's disease (PD). HtrA2 is a mitochondrial protease that translocates to the cytosol upon the initiation of apoptosis where it participates in the abrogation of inhibitors of apoptosis (IAP) inhibition of caspases. Here, we demonstrate that the loss of the HtrA2 function in the dopaminergic neurons of Drosophila melanogaster results in PD-like phenotypes, and we attempt to restore the age-dependent loss in locomotor ability by co-expressing the sole pro-survival Bcl-2 homologue Buffy. The inhibition of HtrA2 in the dopaminergic neurons of Drosophila resulted in shortened lifespan and impaired climbing ability, and the overexpression of Buffy rescued the reduction in lifespan and the age-dependent loss of locomotor ability. In supportive experiments, the inhibition of HtrA2 in the Drosophila eye results in eye defects, marked by reduction in ommatidia number and increased disruption of the ommatidial array; phenotypes that are suppressed by the overexpression of Buffy.

  10. Hepatitis B virus X protein modifies invasion, proliferation and the inflammatory response in an HTR-8/SVneo cell model.

    PubMed

    Cui, Hong; Li, Qiu-Ling; Chen, Jing; Na, Quan; Liu, Cai-Xia

    2015-10-01

    Mother-to-infant transmission of hepatitis B virus (HBV) can occur as an intrauterine, intrapartum or postpartum infection. In the present study, we induced a multifunctional viral regulator of HBV gene products, HBx, and its different fragments to overexpress in a tropho-blast cell line, HTR-8/SVneo. We then identified the biological effects of HBx on HTR-8/SVneo cells. Our results indicated that HBx inhibited apoptosis and induced invasion as detected using Annexin V/propidium iodide (PI) double staining and Transwell assay, respectively. Furthermore, we carried out western blot analysis to analyze the possible related signaling pathway. We confirmed that HBx and its different fragments can activate the Smad signaling pathway, accompanied by downregulation of E-cadherin, and upregulation of vimentin and N-cadherin. TGF‑β1 was used as a control to activate the Smad signaling pathway in HTR-8/SVneo cells. HBx activated the Smad signaling pathway in the HTR-8/SVneo cells. After the signaling pathway was activated, reduced apoptosis, higher invasive ability and enhanced inflammatory response were observed in the HTR-8/SVneo cells.

  11. Behavior of the P1.HTR mastocytoma cell line implanted in the chorioallantoic membrane of chick embryos.

    PubMed

    Avram, S F; Cimpean, A M; Raica, M

    2013-01-01

    The P1.HTR cell line includes highly transfectable cells derived from P815 mastocytoma cells originating from mouse breast tissue. Despite its widespread use in immunogenic studies, no data are available about the behavior of P1.HTR cells in the chick embryo chorioallantoic membrane model. The objective of the present investigation was to study the effects of P1.HTR cells implanted on the chorioallantoic membrane of chick embryos. We inoculated P1.HTR cells into the previously prepared chick embryo chorioallantoic membrane and observed the early and late effects of these cells by stereomicroscopy, histochemistry and immunohistochemistry. A highly angiotropic and angiogenic effect occurred early after inoculation and a tumorigenic potential with the development of mastocytoma keeping well mast cells immunophenotype was detected later during the development. The P1.HTR mastocytoma cell line is a good tool for the development of the chick embryo chorioallantoic membrane mastocytoma model and also for other studies concerning the involvement of blood vessels. The chick embryo chorioallantoic membrane model of mastocytoma retains the mast cell immunophenotype under experimental conditions and could be used as an experimental tool for in vivo preliminary testing of antitumor and antivascular drugs.

  12. Human Ku70/80 interacts directly with hTR, the RNA component of human telomerase.

    PubMed

    Ting, Nicholas S Y; Yu, Yaping; Pohorelic, Brant; Lees-Miller, Susan P; Beattie, Tara L

    2005-01-01

    Maintenance of telomere integrity requires the dynamic interplay between telomerase, telomere-associated proteins and DNA repair proteins. These interactions are vital to suppress DNA damage responses and changes in chromosome dynamics that can result in aneuploidy or other transforming aberrations. The interaction between the DNA repair protein Ku and the RNA component of telomerase (TLC1) in Saccharomyces cerevisiae has been shown to be important for maintaining telomere length. Here, we sought to determine whether this interaction was conserved in higher eukaryotes. Although there is no sequence similarity between TLC1 and the RNA component (hTR) of human telomerase, we show that human Ku70/80 interacts with hTR both in vitro and in a cellular context. Specifically, Ku70/80 interacts with a 47 nt region of the 3' end of hTR, which resembles the stem-loop region of the yeast Ku70/80 binding domain on TLC1. Furthermore, utilizing immunoprecipitation/RT-PCR experiments, we show that Ku interacts with hTR in cell lines deficient in the human telomerase reverse transcriptase protein (hTERT), suggesting that this interaction does not require hTERT. These data suggest that Ku interacts directly with hTR, independent of hTERT, providing evidence for the conservation of the interaction between Ku and telomerase RNA among various species and provide significant insight into how Ku is involved in telomere maintenance in higher eukaryotes.

  13. Human recombinant H2 relaxin induces AKT and GSK3β phosphorylation and HTR-8/SVneo cell proliferation.

    PubMed

    Astuti, Yoni; Nakabayashi, Koji; Deguchi, Masashi; Ebina, Yasuhiko; Yamada, Hideto

    2015-03-24

    Relaxin is essential for trophoblast development during pregnancy. Evidence shows that relaxin increases trophoblast cell migration capacity. Here, we show the effect of relaxin on protein kinase B (AKT) activation and glycogen synthase kinase 3-beta (GSK3β) inactivation as well as on the proliferation of HTR-8/SVneo cells, a model of human extravillous trophoblast (EVT). HTR-8/SVneo cells were treated with different doses of human recombinant (rH2) relaxin in serum-deprived conditions and treated for increasing time with 1 ng/mL of rH2 relaxin. Western blot analysis was performed to detect pAKT, AKT, pGSK3β, GSK3β, and actin expression. Proliferation of HTR-8/SVneo cells was analyzed by MTS assay. rH2 relaxin treatment increased the ratio of pAKT/AKT, pGSK3β/GSK3β, and proliferation in HTR-8/SVneo cells. Furthermore, AKT and GSK3β activation by rH2 relaxin was inhibited by a phosphoinositide 3-kinase (PI3K) inhibitor. This study suggests that rH2 relaxin induces AKT and GSK3β phosphorylation as well as proliferation in HTR-8/SVneo cells.

  14. Telomerase redefined: integrated regulation of hTR and hTERT for telomere maintenance and telomerase activity.

    PubMed

    Cairney, C J; Keith, W N

    2008-01-01

    Telomerase activity is dependent on the expression of 2 main core component genes, hTERT, which encodes the catalytic component and hTR (also called TERC), which encodes the RNA component. The correlation between telomerase activity and carcinogenesis has made this molecule of great interest in cancer research, however in order to fully understand the regulation of telomerase the mechanisms controlling both telomerase genes need to be studied. Some of these mechanisms of regulation have begun to emerge, however many more remain to be deciphered. For many years hTERT has been regarded as the limiting component of telomerase and much of the research in this field has focussed on its regulation, however it was clear from an early stage that hTR expression was also tightly regulated in normal cells and disease. More recently evidence from biochemistry, promoter studies and mouse models has been steadily increasing for a role for hTR as a limiting and essential component for telomerase activity and telomere maintenance. Perhaps the time has come to redefine our view of telomerase regulation. Knowledge of the mechanisms controlling both telomerase genes in normal systems and cancer may aid our understanding of the role of telomerase in carcinogenesis or highlight potential areas for therapeutic intervention. Here we review the essential requirement of hTR for telomere maintenance and telomerase activity in normal tissues and disease and focus on recent advances in our understanding of hTR regulation in relation to hTERT.

  15. Human Ku70/80 interacts directly with hTR, the RNA component of human telomerase

    PubMed Central

    Ting, Nicholas S. Y.; Yu, Yaping; Pohorelic, Brant; Lees-Miller, Susan P.; Beattie, Tara L.

    2005-01-01

    Maintenance of telomere integrity requires the dynamic interplay between telomerase, telomere-associated proteins and DNA repair proteins. These interactions are vital to suppress DNA damage responses and changes in chromosome dynamics that can result in aneuploidy or other transforming aberrations. The interaction between the DNA repair protein Ku and the RNA component of telomerase (TLC1) in Saccharomyces cerevisiae has been shown to be important for maintaining telomere length. Here, we sought to determine whether this interaction was conserved in higher eukaryotes. Although there is no sequence similarity between TLC1 and the RNA component (hTR) of human telomerase, we show that human Ku70/80 interacts with hTR both in vitro and in a cellular context. Specifically, Ku70/80 interacts with a 47 nt region of the 3′ end of hTR, which resembles the stem–loop region of the yeast Ku70/80 binding domain on TLC1. Furthermore, utilizing immunoprecipitation/RT–PCR experiments, we show that Ku interacts with hTR in cell lines deficient in the human telomerase reverse transcriptase protein (hTERT), suggesting that this interaction does not require hTERT. These data suggest that Ku interacts directly with hTR, independent of hTERT, providing evidence for the conservation of the interaction between Ku and telomerase RNA among various species and provide significant insight into how Ku is involved in telomere maintenance in higher eukaryotes. PMID:15824061

  16. Cloning, expression and characterisation of an HtrA-like serine protease produced in vivo by Mycobacterium leprae.

    PubMed

    Ribeiro-Guimarães, Michelle Lopes; Marengo, Eliana Blini; Tempone, Antonio Jorge; Amaral, Julio Jablonski; Klitzke, Clécio F; Silveira, Erika K Xavier da; Portaro, Fernanda Calheta Vieira; Pessolani, Maria Cristina Vidal

    2009-12-01

    Members of the high temperature requirement A (HtrA) family of chaperone proteases have been shown to play a role in bacterial pathogenesis. In a recent report, we demonstrated that the gene ML0176, which codes for a predicted HtrA-like protease, a gene conserved in other species of mycobacteria, is transcribed by Mycobacterium leprae in human leprosy lesions. In the present study, the recombinant ML0176 protein was produced and its enzymatic properties investigated. M. lepraerecombinant ML0176 was able to hydrolyse a variety of synthetic and natural peptides. Similar to other HtrA proteins, this enzyme displayed maximum proteolytic activity at temperatures above 40 degrees C and was completely inactivated by aprotinin, a protease inhibitor with high selectivity for serine proteases. Finally, analysis of M. leprae ML0176 specificity suggested a broader cleavage preference than that of previously described HtrAs homologues. In summary, we have identified an HtrA-like protease in M. lepraethat may constitute a potential new target for the development of novel prophylactic and/or therapeutic strategies against mycobacterial infections.

  17. Analysis of the Link between the Redox State and Enzymatic Activity of the HtrA (DegP) Protein from Escherichia coli

    PubMed Central

    Koper, Tomasz; Polit, Agnieszka; Sobiecka-Szkatula, Anna; Wegrzyn, Katarzyna; Scire, Andrea; Figaj, Donata; Kadzinski, Leszek; Zarzecka, Urszula; Zurawa-Janicka, Dorota; Banecki, Bogdan; Lesner, Adam; Tanfani, Fabio; Lipinska, Barbara; Skorko-Glonek, Joanna

    2015-01-01

    Bacterial HtrAs are proteases engaged in extracytoplasmic activities during stressful conditions and pathogenesis. A model prokaryotic HtrA (HtrA/DegP from Escherichia coli) requires activation to cleave its substrates efficiently. In the inactive state of the enzyme, one of the regulatory loops, termed LA, forms inhibitory contacts in the area of the active center. Reduction of the disulfide bond located in the middle of LA stimulates HtrA activity in vivo suggesting that this S-S bond may play a regulatory role, although the mechanism of this stimulation is not known. Here, we show that HtrA lacking an S-S bridge cleaved a model peptide substrate more efficiently and exhibited a higher affinity for a protein substrate. An LA loop lacking the disulfide was more exposed to the solvent; hence, at least some of the interactions involving this loop must have been disturbed. The protein without S-S bonds demonstrated lower thermal stability and was more easily converted to a dodecameric active oligomeric form. Thus, the lack of the disulfide within LA affected the stability and the overall structure of the HtrA molecule. In this study, we have also demonstrated that in vitro human thioredoxin 1 is able to reduce HtrA; thus, reduction of HtrA can be performed enzymatically. PMID:25710793

  18. Analysis of the link between the redox state and enzymatic activity of the HtrA (DegP) protein from Escherichia coli.

    PubMed

    Koper, Tomasz; Polit, Agnieszka; Sobiecka-Szkatula, Anna; Wegrzyn, Katarzyna; Scire, Andrea; Figaj, Donata; Kadzinski, Leszek; Zarzecka, Urszula; Zurawa-Janicka, Dorota; Banecki, Bogdan; Lesner, Adam; Tanfani, Fabio; Lipinska, Barbara; Skorko-Glonek, Joanna

    2015-01-01

    Bacterial HtrAs are proteases engaged in extracytoplasmic activities during stressful conditions and pathogenesis. A model prokaryotic HtrA (HtrA/DegP from Escherichia coli) requires activation to cleave its substrates efficiently. In the inactive state of the enzyme, one of the regulatory loops, termed LA, forms inhibitory contacts in the area of the active center. Reduction of the disulfide bond located in the middle of LA stimulates HtrA activity in vivo suggesting that this S-S bond may play a regulatory role, although the mechanism of this stimulation is not known. Here, we show that HtrA lacking an S-S bridge cleaved a model peptide substrate more efficiently and exhibited a higher affinity for a protein substrate. An LA loop lacking the disulfide was more exposed to the solvent; hence, at least some of the interactions involving this loop must have been disturbed. The protein without S-S bonds demonstrated lower thermal stability and was more easily converted to a dodecameric active oligomeric form. Thus, the lack of the disulfide within LA affected the stability and the overall structure of the HtrA molecule. In this study, we have also demonstrated that in vitro human thioredoxin 1 is able to reduce HtrA; thus, reduction of HtrA can be performed enzymatically.

  19. Re-examination of the role of the Brucella melitensis HtrA stress response protease in virulence in pregnant goats.

    PubMed

    Roop, R M; Phillips, R W; Hagius, S; Walker, J V; Booth, N J; Fulton, W T; Edmonds, M D; Elzer, P H

    2001-09-03

    Based on previously reported studies describing the experimental infection of pregnant goats with B. melitensis strain RWP5, we proposed that the HtrA protease plays an important role in the virulence of B. melitensis in its natural ruminant host. Subsequent studies, however, have shown that RWP5 is actually an htrA cycL double mutant. In order to definitively evaluate the role of the B. melitensis htrA in virulence, we constructed an authentic htrA mutant and examined this strain in pregnant goats. The findings of these studies indicate that the contribution of the htrA gene product to the virulence of B. melitensis in its natural host is not as great as was previously proposed.

  20. Genome wide expression profile in human HTR-8/Svneo trophoblastic cells in response to overexpression of placental alkaline phosphatase gene.

    PubMed

    Bellazi, L; Mornet, E; Meurice, G; Pata-Merci, N; De Mazancourt, P; Dieudonné, M-N

    2011-10-01

    During pregnancy, placental growth allows the adaptation of the feto-maternal unit to fetal requirements. Placental alkaline phosphatase (PLAP) is a phosphomonoesterase produced increasingly until term by the placenta and also ectopically in some tumors. To precise the role of this enzyme in the placenta, we analyzed the genome wide expression profile of HTR-8/Svneo trophoblastic cells after overexpression of the alkaline phosphatase gene (ALPP). We showed that ALPP overexpression mainly altered expression of genes implicated in cellular growth and proliferation. These results were confirmed by the study of cellular effects in HTR-8/Svneo cells overexpressing ALPP and in HTR-8/Svneo cells in which ALPP expression was suppressed by siRNA. We showed that PLAP exerts a positive effect on DNA replication and acts as a proliferative factor in trophoblastic cells.

  1. Evidence of sexual dimorphism of HTR1B gene on major adult ADHD comorbidities.

    PubMed

    Müller, Diana; Grevet, Eugenio H; Panzenhagen, Alana C; Cupertino, Renata B; da Silva, Bruna S; Kappel, Djenifer B; Mota, Nina R; Blaya-Rocha, Paula; Teche, Stefania P; Vitola, Eduardo S; Rohde, Luis A; Contini, Verônica; Rovaris, Diego L; Schuch, Jaqueline B; Bau, Claiton H D

    2017-09-08

    Attention-deficit/hyperactivity disorder (ADHD) is a very common psychiatric disorder across the life cycle and frequently presents comorbidities. Since ADHD is highly heritable, several studies have focused in the underlying genetic factors involved in its etiology. One of the major challenges in this search is the phenotypic heterogeneity, which could be partly attributable to the sexual dimorphism frequently seen in psychiatric disorders. Taking into account the well-known sexual dimorphic effect observed in serotonergic system characteristics, we differentially tested the influence of HTR1B SNPs (rs11568817, rs130058, rs6296 and rs13212041) on ADHD susceptibility and on its major comorbidities according to sex. The sample comprised 564 adults with ADHD diagnosed according to DSM-IV criteria and 635 controls. There was no association of any HTR1B SNPs tested in relation to ADHD susceptibility. As for the comorbidities evaluated, after correction for multiple tests, significant associations were observed for both rs11568817 and rs130058 with substance use disorders (Pcorr = 0.009 and Pcorr = 0.018, respectively) and for rs11568817 with nicotine dependence (Pcorr = 0.025) in men with ADHD. In women with ADHD, the same rs11568817 was associated with generalized anxiety disorder (Pcorr = 0.031). The observed effects of rs11568817 G allele presence conferring risk to either substance use disorders or generalized anxiety disorder according to sex, suggest an overall scenario where a higher transcriptional activity of HTR1B, resulting from the presence of this allele, is related to externalizing behaviors in men and internalizing behaviors in women. These results are consistent with and expand previous evidence of sexual dimorphism of the serotoninergic system. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. The effects of progesterone on apoptosis in the human trophoblast-derived HTR-8/SV neo cells.

    PubMed

    Liu, Jin; Matsuo, Hiroya; Laoag-Fernandez, Jovelle B; Xu, Qin; Maruo, Takeshi

    2007-12-01

    Progesterone (P4) is frequently used in the treatment of threatened abortion, prevention of recurrent miscarriage and threatened preterm labor. However, little is known about the molecular mechanism of P4 in the regulation of extravillous trophoblasts' (EVTs) function. This study was designed to examine the presence of progesterone receptor (PR) in the human trophoblast-derived HTR-8/SV neo cell line, which is a possible model of EVTs, and the effects of P4 on apoptosis in those cells. The HTR-8/SV neo cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 microg/ml streptomycin. When the cell the population reached 50% confluency, the cells were stepped down to serum-free conditions in the presence or absence of graded concentrations of P4 (1, 10 and 100 ng/ml) for 48 h. The cultured cells were used for RT-PCR, terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL) assay, immunocytochemistry and western blot analyses. Immunocytochemistry and western blot analyses revealed that PR was evident in HTR-8/SV neo cells. Compared with untreated cultures, treatment with P4 (10 and 100 ng/ml) resulted in significant decreases in the TUNEL-positive rate, Fas, Fas ligand (Fas-L), caspase-8, caspase-3 and poly (ADP-ribose) polymerase (PARP) expression in HTR-8/SV neo cells, and a significant increase in Bcl-2 expression in those cells. Consistently, Fas mRNA expression in those cells was significantly inhibited by the treatment with 10 ng/ml P4 compared with untreated cultures. This study suggests that PR exists in HTR-8/SV neo cells and that P4 inhibits apoptosis by down-regulating Fas, Fas-L, caspase-8, caspase-3 and PARP expression as well as up-regulating Bcl-2 expression in HTR-8/SV neo cells.

  3. Common variants of HTR3 genes are associated with obsessive-compulsive disorder and its phenotypic expression

    PubMed Central

    Kim, Hae Won; Kang, Jee In; Lee, Sang-Hyuk; An, Suk Kyoon; Sohn, Sung Yun; Hwang, Eun Hee; Lee, Su Young; Kim, Se Joo

    2016-01-01

    Evidence from literature supports the existence of associations between serotonin-related genetic variants and obsessive-compulsive disorder (OCD), but few studies have explored the involvement of serotonin receptor type 3 genes (HTR3) in OCD. To identify whether HTR3 variability affects an individual’s susceptibility to OCD, we examined 10 HTR3 variants in 596 individuals with OCD and 599 controls. A significant difference existed in the genotypic distribution of the HTR3B variant rs1176744 between individuals with OCD and controls (odds ratio [OR] = 0.74, 95% confidence interval [CI] = 0.60–0.91, P = 0.0043). A protective haplotype in HTR3B was also associated with OCD (OR = 0.77, CI = 0.63–0.95, permutated P = 0.0179). Analyses of OCD sub-phenotypes demonstrated significant associations between rs3758987 and early onset OCD in male subjects (OR = 0.49, CI = 0.31–0.79, P = 0.0031) and among rs6766410, rs6443930, and the cleaning dimension in female subjects (OR = 0.36, CI = 0.18–0.69, P = 0.0016 and OR = 0.47, CI = 0.29–0.79, P = 0.0030, respectively). Additionally, rs6766410 was related to contamination-based disgust in OCD (P = 0.0044). These results support that common HTR3 variants are involved in OCD and some of its clinical phenotypes. PMID:27616601

  4. Structural And Functional Analysis of the Ligand Specificity of the HtrA2/OmI PDZ Domain

    SciTech Connect

    Zhang, Y.; Appleton, B.A.; Wu, P.; Wiesmann, C.; Sidhu, S.S.

    2009-06-04

    The mitochondrial serine protease HtrA2/Omi helps to maintain mitochondrial function by handling misfolded proteins in the intermembrane space. In addition, HtrA2/Omi has been implicated as a proapoptotic factor upon release into the cytoplasm during the cell death cascade. The protein contains a C-terminal PDZ domain that packs against the protease active site and inhibits proteolytic activity. Engagement of the PDZ domain by peptide ligands has been shown to activate the protease and also has been proposed to mediate substrate recognition. We report a detailed structural and functional analysis of the human HtrA2/Omi PDZ domain using peptide libraries and affinity assays to define specificity, X-ray crystallography to view molecular details of PDZ-ligand interactions, and alanine-scanning mutagenesis to probe the peptide-binding groove. We show that the HtrA2/Omi PDZ domain recognizes both C-terminal and internal stretches of extended, hydrophobic polypeptides. High-affinity ligand recognition requires contacts with up to five hydrophobic side chains by distinct sites on the PDZ domain. However, no particular residue type is absolutely required at any position, and thus, the HtrA2/Omi PDZ domain appears to be a promiscuous module adapted to recognize unstructured, hydrophobic polypeptides. This type of specificity is consistent with the biological role of HtrA2/Omi in mitochondria, which requires the recognition of diverse, exposed stretches of hydrophobic sequences in misfolded proteins. The findings are less consistent with, but do not exclude, a role for the PDZ domain in targeting the protease to specific substrates during apoptosis.

  5. Modulation of mitochondrial function and morphology by interaction of Omi/HtrA2 with the mitochondrial fusion factor OPA1

    SciTech Connect

    Kieper, Nicole; Holmstroem, Kira M.; Ciceri, Dalila; Fiesel, Fabienne C.; Wolburg, Hartwig; Ziviani, Elena; Whitworth, Alexander J.; Martins, L. Miguel; Kahle, Philipp J.; Krueger, Rejko

    2010-04-15

    Loss of Omi/HtrA2 function leads to nerve cell loss in mouse models and has been linked to neurodegeneration in Parkinson's and Huntington's disease. Omi/HtrA2 is a serine protease released as a pro-apoptotic factor from the mitochondrial intermembrane space into the cytosol. Under physiological conditions, Omi/HtrA2 is thought to be involved in protection against cellular stress, but the cytological and molecular mechanisms are not clear. Omi/HtrA2 deficiency caused an accumulation of reactive oxygen species and reduced mitochondrial membrane potential. In Omi/HtrA2 knockout mouse embryonic fibroblasts, as well as in Omi/HtrA2 silenced human HeLa cells and Drosophila S2R+ cells, we found elongated mitochondria by live cell imaging. Electron microscopy confirmed the mitochondrial morphology alterations and showed abnormal cristae structure. Examining the levels of proteins involved in mitochondrial fusion, we found a selective up-regulation of more soluble OPA1 protein. Complementation of knockout cells with wild-type Omi/HtrA2 but not with the protease mutant [S306A]Omi/HtrA2 reversed the mitochondrial elongation phenotype and OPA1 alterations. Finally, co-immunoprecipitation showed direct interaction of Omi/HtrA2 with endogenous OPA1. Thus, we show for the first time a direct effect of loss of Omi/HtrA2 on mitochondrial morphology and demonstrate a novel role of this mitochondrial serine protease in the modulation of OPA1. Our results underscore a critical role of impaired mitochondrial dynamics in neurodegenerative disorders.

  6. Application of the wheat-germ cell-free translation system to produce high temperature requirement A3 (HtrA3) proteases.

    PubMed

    Singh, Harmeet; Makino, Shin-ichi; Endo, Yaeta; Li, Ying; Stephens, Andrew N; Nie, Guiying

    2012-01-01

    Mammalian high temperature requirement A3 (HtrA3) is a serine protease of the HtrA family. It is an important factor for placental development and a tumor suppressor. The biochemical properties of HtrA3 are uncharacterized. One critical step in biochemical characterization is overexpressing and purifying the full-length recombinant protein. However, utility of cell-based expression systems is limited for a protease because of autocleavage. The wheat-germ cell-free translation system is highly efficient at producing "difficult" eukaryotic multidomain proteins and is easily modifiable for protein synthesis at different temperatures. In this study, we evaluated the potential of the wheat-germ cell-free translation system for producing human HtrA3. HtrA3 underwent autocleavage when synthesized at 17 °C. When the synthesis temperature was lowered to 4 °C, full-length HtrA3 was successfully produced and proteolytically active. Catalytic site serine substitution with alanine (S305A) stabilized HtrA3 while abolishing its protease activity. This mutant was readily synthesized and stable at 17 °C. When used with glutathione S-transferase (GST) pull-down assay, S305A HtrA3 was a valuable bait in searching for endogenous HtrA3 binding proteins. Thus, we demonstrated the unique utility of the wheat-germ cell-free translation system for producing and characterizing human HtrA3. These strategies will be likely applicable to a wide range of proteases.

  7. EVALUATION OF THE INITIAL CRITICAL CONFIGURATION OF THE HTR-10 PEBBLE-BED REACTOR

    SciTech Connect

    William K. Terry

    2005-11-01

    This report describes the evaluation of data from the initial criticality measurement of the HTR-10 pebble-bed reactor at the Institute of Nuclear Energy Technology in China to determine whether the data are of sufficient quality to use as benchmarks for reactor physics computer codes intended for pebble-bed reactor analysis. The evaluation applied the INL pebble-bed reactor physics code PEBBED to perform an uncertainty analysis on the core critical height. The overall uncertainty in k-effective was slightly over 0.5%, which is considered adequate for an experimental benchmark.

  8. Spherical fuel elements for advanced HTR manufacture and qualification by irradiation testing

    NASA Astrophysics Data System (ADS)

    Mehner, A.-W.; Heit, W.; Röllig, K.; Ragoss, H.; Müller, H.

    1990-04-01

    The reference fuel cycle for future pebble bed HTRs uses low enriched uranium fuel. The spherical fuel element for these HTRs is a 60 mm diameter sphere containing TRISO-coated particles with UO 2 kernels. Qualification of this fuel was performed by production and quality control experience, irradiation testing and accident simulation experiments. The results of the qualification programme fully support the new safety concepts of advanced HTR designs. Further work concentrates on consolidating performance data sets and on quantifying the endurance limits of reference fuel elements under normal and accident conditions.

  9. Identification of a new RNA.RNA interaction site for human telomerase RNA (hTR): structural implications for hTR accumulation and a dyskeratosis congenita point mutation.

    PubMed

    Ren, Xiaojun; Gavory, Gérald; Li, Haitao; Ying, Liming; Klenerman, David; Balasubramanian, Shankar

    2003-11-15

    The enzyme telomerase is a ribonucleoprotein that has a critical role in the maintenance of stable telomeres in organisms that possess linear chromosomes. Using a recently developed single molecule fluorescence coincidence method, we have studied the RNA component of telomerase (hTR) and directly observed multimerisation of hTR in solution. RNA mutagenesis and blocking oligonucleotides were employed to identify the single-stranded internal loop J7b/8a as an important and specific hTR.hTR interaction site. This observation was confirmed by studies on a model RNA fragment (hTR(380-444)), comprising part of the H/ACA domain, the internal loop J7b/8a and the CR7 domain, that was found to dimerise. Substitution mutagenesis within the proposed RNA.RNA interaction site of hTR(380-444) resulted in a loss of dimerisation potential and insertion of the dyskeratosis congenita mutation C408G led to a significant reduction in dimer formation. Together, these results suggest that this RNA.RNA interaction site may be functionally relevant.

  10. Chlamydia Serine Protease Inhibitor, targeting HtrA, as a New Treatment for Koala Chlamydia infection

    PubMed Central

    Lawrence, Amba; Fraser, Tamieka; Gillett, Amber; Tyndall, Joel D. A.; Timms, Peter; Polkinghorne, Adam; Huston, Wilhelmina M.

    2016-01-01

    The koala, an iconic marsupial native to Australia, is a threatened species in many parts of the country. One major factor in the decline is disease caused by infection with Chlamydia. Current therapeutic strategies to treat chlamydiosis in the koala are limited. This study examines the effectiveness of an inhibitor, JO146, which targets the HtrA serine protease for treatment of C. pecorum and C. pneumoniae in vitro and ex vivo with the aim of developing a novel therapeutic for koala Chlamydia infections. Clinical isolates from koalas were examined for their susceptibility to JO146. In vitro studies demonstrated that treatment with JO146 during the mid-replicative phase of C. pecorum or C. pneumoniae infections resulted in a significant loss of infectious progeny. Ex vivo primary koala tissue cultures were used to demonstrate the efficacy of JO146 and the non-toxic nature of this compound on peripheral blood mononuclear cells and primary cell lines established from koala tissues collected at necropsy. Our results suggest that inhibition of the serine protease HtrA could be a novel treatment strategy for chlamydiosis in koalas. PMID:27530689

  11. Benchmark Evaluation of the HTR-PROTEUS Absorber Rod Worths (Core 4)

    SciTech Connect

    John D. Bess; Leland M. Montierth

    2014-06-01

    PROTEUS was a zero-power research reactor at the Paul Scherrer Institute (PSI) in Switzerland. The critical assembly was constructed from a large graphite annulus surrounding a central cylindrical cavity. Various experimental programs were investigated in PROTEUS; during the years 1992 through 1996, it was configured as a pebble-bed reactor and designated HTR-PROTEUS. Various critical configurations were assembled with each accompanied by an assortment of reactor physics experiments including differential and integral absorber rod measurements, kinetics, reaction rate distributions, water ingress effects, and small sample reactivity effects [1]. Four benchmark reports were previously prepared and included in the March 2013 edition of the International Handbook of Evaluated Reactor Physics Benchmark Experiments (IRPhEP Handbook) [2] evaluating eleven critical configurations. A summary of that effort was previously provided [3] and an analysis of absorber rod worth measurements for Cores 9 and 10 have been performed prior to this analysis and included in PROTEUS-GCR-EXP-004 [4]. In the current benchmark effort, absorber rod worths measured for Core Configuration 4, which was the only core with a randomly-packed pebble loading, have been evaluated for inclusion as a revision to the HTR-PROTEUS benchmark report PROTEUS-GCR-EXP-002.

  12. HTR Fuel Waste Management: TRISO separation and acid-graphite intercalation compounds preparation

    NASA Astrophysics Data System (ADS)

    Guittonneau, Fabrice; Abdelouas, Abdesselam; Grambow, Bernd

    2010-12-01

    Considering the need to reduce waste production and greenhouse emissions and still keeping high energy efficiency, various 4th generation nuclear energy systems have been proposed. As far as graphite-moderated reactors are concerned (future high temperature fast or thermal reactors), one of the key issues is the large volumes of irradiated graphite encountered. With the objective to reduce volume of waste in the HTR concept, it is very important to be able to separate the fuel from low level activity graphite representing a large volume. The separated TRISO particles can then be reprocessed for waste separation or disposed off in geological repository. In addition, preparation of acid-GICs from the separated graphite may constitute a way to recycle this waste. We used HTR-type compact fuel with ZrO 2 TRISO particles to test two separation methods: low (H 2SO 4 + H 2O 2) and high (H 2SO 4 + HNO 3) temperature acid treatments. In both cases the TRISO separation was complete but some TRISO layers oxidized at high temperature. At low temperature, the desegregation of graphite grains is facilitated by intercalation of sulfuric acid between the graphene layers. The acid-GIC obtained consists of pure phases of high quality suggesting their potential industrial recycling.

  13. GABA A receptor π subunit promotes apoptosis of HTR-8/SVneo trophoblastic cells: Implications in preeclampsia.

    PubMed

    Lu, Junjie; Zhang, Qian; Tan, Dongmei; Luo, Wenping; Zhao, Hai; Ma, Jing; Liang, Hao; Tan, Yi

    2016-07-01

    Gamma-aminobutyric acid (GABA) functions primarily as an inhibitory neurotransmitter through its receptors in the mature central nervous system. The GABA type A receptor π subunit (GABRP) has been identified in the tissues of the reproductive system, particularly in the uterus. In addition, we have previously detected GABRP expression in both human and mouse placentas. To examine the role of GABRP in trophoblastic cell invasion, we constructed a pIRES2-GABRP-EGFP plasmid which was used for the transfection of a human placental cell line derived from first trimester extravillous trophoblasts (HTR-8/SVneo). The number of invaded cells was decreased by GABRP overexpression. Notably, the decrease in the invasive cell number may be due to the increased apoptosis of the HTR-8/SVneo cells following GABRP transfection, which was further confirmed by flow cytometry, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. Based on the increased apoptosis of trophoblastic cells in pregnancies complicated by preeclampsia (PE) and the fact that GABRP promotes the apoptosis of trophoblastic cells, we hypothesized that GABRP expression is increased in the placental tissues from patients with PE compared with that in the normal groups and this hypothesis was confirmed by RT-qPCR and immunohistochemical analysis. Taken together, these findings imply that GABRP plays an important role in placentation and this pathway may be a promising molecular target for the development of novel therapeutic strategies for PE.

  14. DEM simulation of particle mixing for optimizing the overcoating drum in HTR fuel fabrication

    NASA Astrophysics Data System (ADS)

    Liu, Malin; Lu, Zhengming; Liu, Bing; Shao, Youlin

    2013-06-01

    The rotating drum was used for overcoating coated fuel particles in HTR fuel fabrication process. All the coated particles should be adhered to equal amount of graphite powder, which means that the particle should be mixed quickly in both radial and axial directions. This paper investigated the particle flow dynamics and mixing behavior in different regimes using the discrete element method (DEM). By varying the rotation speed, different flow regimes such as slumping, rolling, cascading, cataracting, centrifuging were produced. The mixing entropy based on radial and axial grid was introduced to describe the radial and axial mixing behaviors. From simulation results, it was found that the radial mixing can be achieved in the cascading regime more quickly than the slumping, rolling and centrifuging regimes, but the traditional rotating drum without internal components can not achieve the requirements of axial mixing and should be improved. Three different structures of internal components are proposed and simulated. The new V-shaped deflectors were found to achieve a quick axial mixing behavior and uniform axial distribution in the rotating drum based on simulation results. At last, the superiority was validated by experimental results, and the new V-shaped deflectors were used in the industrial production of the overcoating coated fuel particles in HTR fuel fabrication process.

  15. The association between telomerase activity and expression of its RNA component (hTR) in breast cancer patients: the importance of DNase treatment.

    PubMed

    Hosseini-Asl, Saied; Modarressi, Mohammad H; Atri, Morteza; Salhab, Mohamed; Mokbel, Kefah; Mehdipour, Parvin

    2006-06-02

    Telomerase is a ribonucleoprotein enzyme that compensates for the telomere length shortening which occurs during the cell cycle. Telomerase activity has been detected in most tumours but not in somatic cells. However, hTR; the RNA component of telomerase; has been reported to be universally expressed in both cancerous and non-cancerous tissues. Tumour samples from 50 patients with primary invasive breast cancer were collected. The TRAP assay was used to detect telomerase activity. RT-PCR on cDNA and DNased cDNA samples and control groups was used to detect the expression of hTR, GAPDH and PGM1 genes. Seventy-two percent of samples showed telomerase activity. DNA contamination was detected in 36 (72%) of RNA samples. Without performing DNase treatment, 49 (98%) of all samples showed hTR expression, but with the application of this strategy, hTR expression decreased from 98% to 64%. A significant association (p < 0.001) between hTR expression and telomerase activity was observed. Among the 32 hTR positive samples, 30 had telomerase activity and among the 18 hTR negative samples, telomerase activity was observed in 6 cases. Thus the application of this strategy could provide an applicable tool to use instead of the TRAP assay thus facilitating telomerase research in cancer genetic investigations.

  16. The association between telomerase activity and expression of its RNA component (hTR) in breast cancer patients: the importance of DNase treatment

    PubMed Central

    Hosseini-Asl, Saied; Modarressi, Mohammad H; Atri, Morteza; Salhab, Mohamed; Mokbel, Kefah; Mehdipour, Parvin

    2006-01-01

    Telomerase is a ribonucleoprotein enzyme that compensates for the telomere length shortening which occurs during the cell cycle. Telomerase activity has been detected in most tumours but not in somatic cells. However, hTR; the RNA component of telomerase; has been reported to be universally expressed in both cancerous and non-cancerous tissues. Tumour samples from 50 patients with primary invasive breast cancer were collected. The TRAP assay was used to detect telomerase activity. RT-PCR on cDNA and DNased cDNA samples and control groups was used to detect the expression of hTR, GAPDH and PGM1 genes. Seventy-two percent of samples showed telomerase activity. DNA contamination was detected in 36 (72%) of RNA samples. Without performing DNase treatment, 49 (98%) of all samples showed hTR expression, but with the application of this strategy, hTR expression decreased from 98% to 64%. A significant association (p < 0.001) between hTR expression and telomerase activity was observed. Among the 32 hTR positive samples, 30 had telomerase activity and among the 18 hTR negative samples, telomerase activity was observed in 6 cases. Thus the application of this strategy could provide an applicable tool to use instead of the TRAP assay thus facilitating telomerase research in cancer genetic investigations. PMID:16749934

  17. Different dark conformations function in color-sensitive photosignaling by the sensory rhodopsin I-HtrI complex.

    PubMed

    Sasaki, Jun; Phillips, Brian J; Chen, Xinpu; Van Eps, Ned; Tsai, Ah-Lim; Hubbell, Wayne L; Spudich, John L

    2007-06-01

    The haloarchaeal phototaxis receptor sensory rhodopsin I (SRI) in complex with its transducer HtrI delivers an attractant signal from excitation with an orange photon and a repellent signal from a second near-UV photon excitation. Using a proteoliposome system with purified SRI in complex with its transducer HtrI, we identified by site-directed fluorescence labeling a site (Ser(155)) on SRI that is conformationally active in signal relay to HtrI. Using site-directed spin labeling of Ser(155)Cys with a nitroxide side chain, we detected a change in conformation following one-photon excitation such that the spin probe exhibits a splitting of the outer hyperfine extrema (2A'(zz)) significantly smaller than that of the electron paramagnetic resonance spectrum in the dark state. The dark conformations of five mutant complexes that do not discriminate between orange and near-UV excitation show shifts to lower or higher 2A'(zz) values correlated with the alterations in their motility behavior to one- and two-photon stimuli. These data are interpreted in terms of a model in which the dark complex is populated by two conformers in the wild type, one that inhibits the CheA kinase (A) and the other that activates it (R), shifted in the dark by mutations and shifted in the wild-type SRI-HtrI complex in opposite directions by one-photon and two-photon reactions.

  18. Serotonin receptor gene (HTR2A) T102C polymorphism modulates individuals' perspective taking ability and autistic-like traits.

    PubMed

    Gong, Pingyuan; Liu, Jinting; Blue, Philip R; Li, She; Zhou, Xiaolin

    2015-01-01

    Previous studies have indicated that empathic traits, such as perspective taking, are associated with the levels of serotonin in the brain and with autism spectrum conditions. Inspired by the finding that the serotonin receptor 2A gene (HTR2A) modulates the availability of serotonin, this study investigated to what extent HTR2A modulates individuals' perspective taking ability and autistic-like traits. To examine the associations of the functional HTR2A polymorphism T102C (rs6313) with individuals' perspective taking abilities and autistic-like traits, we differentiated individuals according to this polymorphism and measured empathic and autistic-like traits with Interpersonal Reactivity Index (IRI) and Autism-Spectrum Quotient (AQ) scale in 523 Chinese people. The results indicated that this polymorphism was significantly associated with the scores on Perspective Taking and Personal Distress subscales of IRI, and Communication subscale of AQ. Individuals with a greater number of the C alleles were less likely to spontaneously adopt the point of view of others, more likely to be anxious when observing the pain endured by others, and more likely to have communication problems. Moreover, the genotype effect on communication problems was mediated by individuals' perspective taking ability. These findings provide evidence that the HTR2A T102C polymorphism is a predictor of individual differences in empathic and autistic-like traits and highlight the role of the gene in the connection between perspective taking and autistic-like traits.

  19. Serotonin receptor gene (HTR2A) T102C polymorphism modulates individuals’ perspective taking ability and autistic-like traits

    PubMed Central

    Gong, Pingyuan; Liu, Jinting; Blue, Philip R.; Li, She; Zhou, Xiaolin

    2015-01-01

    Previous studies have indicated that empathic traits, such as perspective taking, are associated with the levels of serotonin in the brain and with autism spectrum conditions. Inspired by the finding that the serotonin receptor 2A gene (HTR2A) modulates the availability of serotonin, this study investigated to what extent HTR2A modulates individuals’ perspective taking ability and autistic-like traits. To examine the associations of the functional HTR2A polymorphism T102C (rs6313) with individuals’ perspective taking abilities and autistic-like traits, we differentiated individuals according to this polymorphism and measured empathic and autistic-like traits with Interpersonal Reactivity Index (IRI) and Autism-Spectrum Quotient (AQ) scale in 523 Chinese people. The results indicated that this polymorphism was significantly associated with the scores on Perspective Taking and Personal Distress subscales of IRI, and Communication subscale of AQ. Individuals with a greater number of the C alleles were less likely to spontaneously adopt the point of view of others, more likely to be anxious when observing the pain endured by others, and more likely to have communication problems. Moreover, the genotype effect on communication problems was mediated by individuals’ perspective taking ability. These findings provide evidence that the HTR2A T102C polymorphism is a predictor of individual differences in empathic and autistic-like traits and highlight the role of the gene in the connection between perspective taking and autistic-like traits. PMID:26557070

  20. A genetic variant of HTR2C may play a role in the manifestation of Tourette syndrome.

    PubMed

    Dehning, Sandra; Müller, Norbert; Matz, Judith; Bender, Andreas; Kerle, Irina; Benninghoff, Jens; Musil, Richard; Spellmann, Ilja; Bondy, Brigitta; Möller, Hans-Juergen; Riedel, Michael; Zill, Peter

    2010-02-01

    Gilles de la Tourette syndrome (GTS) (MIM 137580) is a complex neuropsychiatric disorder probably originating from a disturbed interplay of several neurotransmitter systems in the prefrontal-limbic-basal ganglia loop. Polygenetic multifactorial inheritance has been postulated; nevertheless, no confirmed susceptible genes have been identified yet. As neuroimaging studies allude to dopaminergic and serotonergic dysfunction in GTS and serotonin as an important factor for dopamine release, genotyping of common polymorphisms in the serotonergic receptor (HTR1A: C-1019G; HTR2A: T102C, His452Tyr, A-1438G; HTR2C: C-759T, G-697C) and transporter genes (SLC6A4) was carried out in 87 patients with GTS, compared with 311 matched controls. We found a nominally significant association between both polymorphisms in the HTR2C and the GTS, which was more pronounced in male patients. Analysis of the further serotonergic polymorphisms did not reveal any significant result. A modified function of these promoter polymorphisms may contribute to the complex interplay of serotonin and dopamine and then to the manifestation of GTS.

  1. Phototaxis of Halobacterium salinarium requires a signalling complex of sensory rhodopsin I and its methyl-accepting transducer HtrI.

    PubMed Central

    Krah, M; Marwan, W; Verméglio, A; Oesterhelt, D

    1994-01-01

    Sensory rhodopsin I (SRI) is a photoreceptor that mediates phototaxis in the archaeon Halobacterium salinarium. Receptor excitation is relayed to the motility system of the cell by the methyl-accepting transducer protein HtrI. In membranes prepared from cells that lack HtrI the absorbance difference maximum of SRI was shifted from 587 to 565 nm. The thermal decay of the metastable photocycle intermediate SRI373 was measured as time-dependent recovery of the absorbance at 590 nm. In the absence of HtrI the decay was slowed down by two orders of magnitude. When SRI was overproduced in cells that contained normal levels of HtrI, the decay of SRI373 was biexponential indicating two kinetically distinct species. Spectroscopic measurements on intact cells revealed the same effect of HtrI on SRI photocycling as found in isolated membranes. By transient exposure of membranes from wild-type cells to low ionic strength, the decay of SR373 was slowed to the same value found for untreated membranes in the absence of HtrI. In parallel, the absorbance difference maximum was shifted to 565 nm indicating that a physical interaction of HtrI and SRI had been irreversibly destroyed. Overproduction of SRI in the presence of wild-type amounts of HtrI did not increase the light sensitivity of the cells to orange light step down stimulation. It is concluded that SRI and HtrI form a stable complex in the cell membrane that signals to the flagellar motor and defines absorbance maximum, photocycling rate and photochemical efficiency of SRI. PMID:8187768

  2. mRNA Expression and DNA Methylation Analysis of Serotonin Receptor 2A (HTR2A) in the Human Schizophrenic Brain

    PubMed Central

    Cheah, Sern-Yih; Lawford, Bruce R.; Young, Ross McD.; Morris, Charles P.; Voisey, Joanne

    2017-01-01

    Serotonin receptor 2A (HTR2A) is an important signalling factor implicated in cognitive functions and known to be associated with schizophrenia. The biological significance of HTR2A in schizophrenia remains unclear as molecular analyses including genetic association, mRNA expression and methylation studies have reported inconsistent results. In this study, we examine HTR2A expression and methylation and the interaction with HTR2A polymorphisms to identify their biological significance in schizophrenia. Subjects included 25 schizophrenia and 25 control post-mortem brain samples. Genotype and mRNA data was generated by transcriptome sequencing. DNA methylation profiles were generated for CpG sites within promoter-exon I region. Expression, genotype and methylation data were examined for association with schizophrenia. HTR2A mRNA levels were reduced by 14% (p = 0.006) in schizophrenia compared to controls. Three CpG sites were hypermethylated in schizophrenia (cg5 p = 0.028, cg7 p = 0.021, cg10 p = 0.017) and HTR2A polymorphisms rs6314 (p = 0.008) and rs6313 (p = 0.026) showed genetic association with schizophrenia. Differential DNA methylation was associated with rs6314 and rs6313. There was a strong correlation between HTR2A DNA methylation and mRNA expression. The results were nominally significant but did not survive the rigorous Benjamini-Hochberg correction for multiple testing. Differential HTR2A expression in schizophrenia in our study may be the result of the combined effect of multiple differentially methylated CpG sites. Epigenetic HTR2A regulation may alter brain function, which contributes to the development of schizophrenia. PMID:28054990

  3. mRNA Expression and DNA Methylation Analysis of Serotonin Receptor 2A (HTR2A) in the Human Schizophrenic Brain.

    PubMed

    Cheah, Sern-Yih; Lawford, Bruce R; Young, Ross McD; Morris, Charles P; Voisey, Joanne

    2017-01-04

    Serotonin receptor 2A (HTR2A) is an important signalling factor implicated in cognitive functions and known to be associated with schizophrenia. The biological significance of HTR2A in schizophrenia remains unclear as molecular analyses including genetic association, mRNA expression and methylation studies have reported inconsistent results. In this study, we examine HTR2A expression and methylation and the interaction with HTR2A polymorphisms to identify their biological significance in schizophrenia. Subjects included 25 schizophrenia and 25 control post-mortem brain samples. Genotype and mRNA data was generated by transcriptome sequencing. DNA methylation profiles were generated for CpG sites within promoter-exon I region. Expression, genotype and methylation data were examined for association with schizophrenia. HTR2A mRNA levels were reduced by 14% (p = 0.006) in schizophrenia compared to controls. Three CpG sites were hypermethylated in schizophrenia (cg5 p = 0.028, cg7 p = 0.021, cg10 p = 0.017) and HTR2A polymorphisms rs6314 (p = 0.008) and rs6313 (p = 0.026) showed genetic association with schizophrenia. Differential DNA methylation was associated with rs6314 and rs6313. There was a strong correlation between HTR2A DNA methylation and mRNA expression. The results were nominally significant but did not survive the rigorous Benjamini-Hochberg correction for multiple testing. Differential HTR2A expression in schizophrenia in our study may be the result of the combined effect of multiple differentially methylated CpG sites. Epigenetic HTR2A regulation may alter brain function, which contributes to the development of schizophrenia.

  4. Mice Lacking the Serotonin Htr2B Receptor Gene Present an Antipsychotic-Sensitive Schizophrenic-Like Phenotype.

    PubMed

    Pitychoutis, Pothitos M; Belmer, Arnauld; Moutkine, Imane; Adrien, Joëlle; Maroteaux, Luc

    2015-11-01

    Impulsivity and hyperactivity share common ground with numerous mental disorders, including schizophrenia. Recently, a population-specific serotonin 2B (5-HT2B) receptor stop codon (ie, HTR2B Q20*) was reported to segregate with severely impulsive individuals, whereas 5-HT2B mutant (Htr2B(-/-)) mice also showed high impulsivity. Interestingly, in the same cohort, early-onset schizophrenia was more prevalent in HTR2B Q*20 carriers. However, the putative role of 5-HT2B receptor in the neurobiology of schizophrenia has never been investigated. We assessed the effects of the genetic and the pharmacological ablation of 5-HT2B receptors in mice subjected to a comprehensive series of behavioral test screenings for schizophrenic-like symptoms and investigated relevant dopaminergic and glutamatergic neurochemical alterations in the cortex and the striatum. Domains related to the positive, negative, and cognitive symptom clusters of schizophrenia were affected in Htr2B(-/-) mice, as shown by deficits in sensorimotor gating, in selective attention, in social interactions, and in learning and memory processes. In addition, Htr2B(-/-) mice presented with enhanced locomotor response to the psychostimulants dizocilpine and amphetamine, and with robust alterations in sleep architecture. Moreover, ablation of 5-HT2B receptors induced a region-selective decrease of dopamine and glutamate concentrations in the dorsal striatum. Importantly, selected schizophrenic-like phenotypes and endophenotypes were rescued by chronic haloperidol treatment. We report herein that 5-HT2B receptor deficiency confers a wide spectrum of antipsychotic-sensitive schizophrenic-like behavioral and psychopharmacological phenotypes in mice and provide first evidence for a role of 5-HT2B receptors in the neurobiology of psychotic disorders.

  5. Lack of association between serotonin-2A receptor gene (HTR2A) polymorphisms and tardive dyskinesia in schizophrenia.

    PubMed

    Basile, V S; Ozdemir, V; Masellis, M; Meltzer, H Y; Lieberman, J A; Potkin, S G; Macciardi, F M; Petronis, A; Kennedy, J L

    2001-03-01

    Tardive dyskinesia (TD) is a disabling neurological side effect associated with long-term treatment with typical antipsychotics. Family studies and animal models lend evidence for hereditary predisposition to TD. The newer atypical antipsychotics pose a minimal risk for TD which is in part attributed to their ability to block the serotonin-2A (5-HT(2A)) receptor. 5-HT(2A) receptors were also identified in the basal ganglia; a brain region that plays a critical role in antipsychotic-induced movement disorders. We tested the significance of variation in the 5-HT(2A) receptor gene (HTR2A) in relation to the TD phenotype. Three polymorphisms in HTR2A, one silent (C102T), one that alters the amino acid sequence (his452tyr) and one in the promoter region (A-1437G) were investigated in 136 patients refractory or intolerant to treatment with typical antipsychotics and with a DSM-IIIR diagnosis of schizophrenia. We did not find any significant difference in allele, genotype or haplotype frequencies of polymorphisms in HTR2A among patients with or without TD (P > 0.05). Further analysis using the ANCOVA statistic with a continuous measure of the TD phenotype (Abnormal Involuntary Movement Scale (AIMS) score) found that the AIMS scores were not significantly influenced by HTR2A polymorphisms, despite controlling for potential confounders such as age, gender and ethnicity (P > 0.05). Theoretically, central serotonergic function can be subject to genetic control at various other mechanistic levels including the rate of serotonin synthesis (tryptophane hydroxylase gene), release, reuptake (serotonin transporter gene) and degradation (monoamine oxidase gene). Analyses of these other serotonergic genes are indicated. In summary, polymorphisms in HTR2A do not appear to influence the risk for TD. Further studies evaluating in tandem multiple candidate genes relevant for the serotonergic system are warranted to dissect the genetic basis of the complex TD phenotype.

  6. The structures of Arabidopsis Deg5 and Deg8 reveal new insights into HtrA proteases

    SciTech Connect

    Sun, Wei; Gao, Feng; Fan, Haitian; Shan, Xiaoyue; Sun, Renhua; Liu, Lin; Gong, Weimin

    2013-05-01

    The crystal structures of Arabidopsis Deg5 and Deg8 have been determined to resolutions of 2.6 and 2.0 Å, respectively, revealing novel structural features of HtrA proteases. Plant Deg5 and Deg8 are two members of the HtrA proteases, a family of oligomeric serine endopeptidases that are involved in a variety of protein quality-control processes. These two HtrA proteases are located in the thylakoid lumen and participate in high-light stress responses by collaborating with other chloroplast proteins. Deg5 and Deg8 degrade photodamaged D1 protein of the photosystem II reaction centre, allowing its in situ replacement. Here, the crystal structures of Arabidopsis thaliana Deg5 (S266A) and Deg8 (S292A) are reported at 2.6 and 2.0 Å resolution, respectively. The Deg5 trimer contains two calcium ions in a central channel, suggesting a link between photodamage control and calcium ions in chloroplasts. Previous structures of HtrA proteases have indicated that their regulation usually requires C-terminal PDZ domain(s). Deg5 is unique in that it contains no PDZ domain and the trimeric structure of Deg5 (S266A) reveals a novel catalytic triad conformation. A similar triad conformation is observed in the hexameric structure of the single PDZ-domain-containing Deg8 (S292A). These findings suggest a novel activation mechanism for plant HtrA proteases and provide structural clues to their function in light-stress response.

  7. Combustion of HTR fuel: Fission product release and dissolution of plutonium contained in the ash

    NASA Astrophysics Data System (ADS)

    Zimmer, E.; Brodda, B. G.; Kirchner, H.; Kroth, K.

    1985-04-01

    Single spent spherical HTR fuel elements were subjected to combustion between 800 and 1100°C. The elements were loaded with either UO 2 or (Th, U)O 2 pyrocarbon coated fuel kernels. The UO 2 loaded elements released a large fraction of their cesium and ruthenium inventory during combustion, whereas the release of the (Th, U)O 2 loaded element was significantly lower. The ash of the UO 2 elements was dissolved in nitric acid to simulate Purex reprocessing. About 5% of the bred plutonium was indissoluble and remained in the residue. The spent (Th, U)O 2 fuel was broken into pieces after combustion and dissolved five times faster than fresh fuel in fluoride containing nitric acid.

  8. The function of the DegP (HtrA) protein: Protease versus chaperone.

    PubMed

    Chang, Zengyi

    2016-11-01

    The DegP (or HtrA) is a highly conserved family of proteins functioning in all living organisms. It was initially identified as a protease functioning in the periplasmic space of the Gram-negative bacterial cells. It was later reported to also exhibit chaperone activity and thus has been designated as a bifunctional protein. However, recent studies demonstrated that in living cells it more likely functions only as a protease with hardly detectable chaperone activities. In this review, I will summarize the evidences clarifying that DegP more likely only functions as a protease rather than as a chaperone in cells. © 2016 IUBMB Life, 68(11):904-907, 2016. © 2016 International Union of Biochemistry and Molecular Biology.

  9. Preliminary analysis of graphite dust releasing behavior in accident for HTR

    SciTech Connect

    Peng, W.; Yang, X. Y.; Yu, S. Y.; Wang, J.

    2012-07-01

    The behavior of the graphite dust is important to the safety of High Temperature Gas-cooled Reactors. This study investigated the flow of graphite dust in helium mainstream. The analysis of the stresses acting on the graphite dust indicated that gas drag played the absolute leading role. Based on the understanding of the importance of gas drag, an experimental system is set up for the research of dust releasing behavior in accident. Air driven by centrifugal fan is used as the working fluid instead of helium because helium is expensive, easy to leak which make it difficult to seal. The graphite particles, with the size distribution same as in HTR, are added to the experiment loop. The graphite dust releasing behavior at the loss-of-coolant accident will be investigated by a sonic nozzle. (authors)

  10. Regulation of HtrA2 on WT1 gene expression under imatinib stimulation and its effects on the cell biology of K562 cells.

    PubMed

    Zhang, Lixia; Li, Yan; Li, Xiaoyan; Zhang, Qing; Qiu, Shaowei; Zhang, Qi; Wang, Min; Xing, Haiyan; Rao, Qing; Tian, Zheng; Tang, Kejing; Wang, Jianxiang; Mi, Yingchang

    2017-09-01

    The aim of the present study was to investigate the regulation of Wilms Tumor 1 (WT1) by serine protease high-temperature requirement protein A2 (HtrA2), a member of the Htr family, in K562 cells. In addition, the study aimed to observe the effect of this regulation on cell biological functions and its associated mechanisms. Expression of WT1 and HtrA2 mRNA, and proteins following imatinib and the HtrA2 inhibitor 5-[5-(2-nitrophenyl) furfuryl iodine]-1, 3-diphenyl-2-thiobarbituric acid (UCF-101) treatment was detected with reverse transcription-quantitative polymerase chain reaction and western blot analysis. Subsequent to treatment with drugs and UCF-101, the proliferative function of K562 cells was detected using MTT assays, and the rate of apoptosis was detected using Annexin V with propidium iodide flow cytometry in K562 cells. The protein levels in the signaling pathway were analyzed using western blotting following treatment with imatinib and UCF-101. In K562 cells, imatinib treatment activated HtrA2 gene at a transcription level, while the WT1 gene was simultaneously downregulated. Following HtrA2 inhibitor (UCF-101) treatment, the downregulation of WT1 increased gradually. At the protein level, imatinib induced the increase in HtrA2 protein level and concomitantly downregulated WT1 protein level. Subsequent to HtrA2 inhibition by UCF-101, the WT1 protein level decreased temporarily, but eventually increased. Imatinib induced apoptosis in K562 cells, but this effect was attenuated by the HtrA2 inhibitor UCF-101, resulting in the upregulation of the WT1 protein level. However; UCF-101 did not markedly change the proliferation inhibition caused by imatinib. Imatinib activated the p38 mitogen activated protein kinase (p38 MAPK) signaling pathway in K562 cells, and UCF-101 affected the activation of imatinib in the p38 MAPK signaling pathway. Imatinib inhibited the extracellular signal-related kinase (ERK1/2) pathway markedly and persistently, but UCF-101

  11. Effects of transforming growth factor-β1 on the proliferation and invasion of the HTR-8/SVneo cell line.

    PubMed

    Zuo, Yanzhen; Fu, Zhihua; Hu, Yatao; Li, Yuhong; Xu, Qian; Sun, Dayong; Tan, Yusi

    2014-11-01

    Transforming growth factor-β1 (TGF-β1) is involved in the regulation of trophoblast cell proliferation and invasion. However, the mechanism underlying this process remains unknown, which is predominantly due to the difficulty in obtaining and maintaining primary trophoblast cells in culture over a long period of time. The HTR-8/SVneo cell line is an immortalized trophoblast cell line, which has been reported to exhibit a number of similar characteristics to those of parental trophoblast cells. Therefore, the cell line has been a useful tool for the investigation of placental function and tumor progression. In the present study, the HTR-8/SVneo cell line was used as a model to investigate the TGF-β1/SMAD signaling pathway in the proliferation and invasion of trophoblast cells. The proliferation and invasion ability of HTR-8/SVneo cells was determined using the MTT and Transwell assays, respectively. In addition, reverse transcription polymerase chain reactions were performed to detect the mRNA expression of a panel of known downstream mediators of TGF-β1, including TGF-β receptor I (TβRI), SMAD4, SMAD3, SMAD7 and tissue inhibitor of metalloproteinases-1 (TIMP-1). The results indicated that TGF-β1 promotes the proliferation and invasion of the HTR-8/SVneo cell line at passage 90. Furthermore, the expression of TβRI, SMAD3 and SMAD4 were reduced following treatment with TGF-β1, while the expression of SMAD7 was increased and the expression of TIMP-1 remained unchanged following TGF-β1 treatment. These observations indicated that the effects of TGF-β1 on the proliferation and invasion of the HTR-8/SVneo cell line at passage 90 were different from those of parental trophoblasts, which is in contrast to the results of previous studies. It was concluded that the HTR-8/SVneo cell lines, which have been grown for over 90 passages, do not accurately represent parental trophoblast cells in studies of the TGF-β/SMAD signaling pathway.

  12. Next-Generation Bacillus anthracis Live Attenuated Spore Vaccine Based on the htrA(-) (High Temperature Requirement A) Sterne Strain.

    PubMed

    Chitlaru, Theodor; Israeli, Ma'ayan; Bar-Haim, Erez; Elia, Uri; Rotem, Shahar; Ehrlich, Sharon; Cohen, Ofer; Shafferman, Avigdor

    2016-01-06

    Anthrax is a lethal disease caused by the gram-positive spore-producing bacterium Bacillus anthracis. Live attenuated vaccines, such as the nonencapsulated Sterne strain, do not meet the safety standards mandated for human use in the Western world and are approved for veterinary purposes only. Here we demonstrate that disrupting the htrA gene, encoding the chaperone/protease HtrA (High Temperature Requirement A), in the virulent Bacillus anthracis Vollum strain results in significant virulence attenuation in guinea pigs, rabbits and mice, underlying the universality of the attenuated phenotype associated with htrA knockout. Accordingly, htrA disruption was implemented for the development of a Sterne-derived safe live vaccine compatible with human use. The novel B. anthracis SterneΔhtrA strain secretes functional anthrax toxins but is 10-10(4)-fold less virulent than the Sterne vaccine strain depending on animal model (mice, guinea pigs, or rabbits). In spite of this attenuation, double or even single immunization with SterneΔhtrA spores elicits immune responses which target toxaemia and bacteremia resulting in protection from subcutaneous or respiratory lethal challenge with a virulent strain in guinea pigs and rabbits. The efficacy of the immune-protective response in guinea pigs was maintained for at least 50 weeks after a single immunization.

  13. HtrA3 is regulated by 15-deoxy-{Delta}12,14-prostaglandin J2 independently of PPAR{gamma} in clear cell renal cell carcinomas

    SciTech Connect

    Theoleyre, Sandrine; Mottier, Stephanie; Masson, Damien; Denis, Marc G.

    2010-04-09

    Peroxisome proliferator-activated receptor gamma (PPAR{gamma}) ligands have been shown to possess anti-proliferative effects in many types of cancer. In clear cell renal cell carcinoma (CCRCC), the targets involved in these effects are not known. In this study, we demonstrated that, in CCRCC cell lines, the endogenous PPAR{gamma} ligand 15-deoxy-{Delta}12,14-prostaglandin J2 (15dPGJ2) induces the expression, both at the mRNA and the protein levels, of the HtrA3 gene. This gene belongs to the High-Temperature Requirement Factor A family of serine proteases that repress signaling by TGF-{beta} family members and inhibit cell migration. Rosiglitazone or ciglitazone, synthetic PPAR{gamma} agonists, did not induce HtrA3 expression, and the PPAR{gamma} antagonist GW9662 did not prevent 15dPGJ2 induction, suggesting that the up-regulation of HtrA3 by 15dPGJ2 is independent of PPAR{gamma}. The MEK/ERK inhibitor PD98059 dramatically repressed HtrA3 induction. Altogether, these data indicate that 15dPGJ2 is able to stimulate the expression of HtrA3 through an indirect mechanism involving the MEK/ERK pathway but independent of PPAR{gamma}. Our results provide a better understanding of the mechanisms involved in the regulation of HtrA3, a potential tumor suppressor gene.

  14. Prediction of extracellular proteases of the human pathogen Helicobacter pylori reveals proteolytic activity of the Hp1018/19 protein HtrA.

    PubMed

    Löwer, Martin; Weydig, Christiane; Metzler, Dirk; Reuter, Andreas; Starzinski-Powitz, Anna; Wessler, Silja; Schneider, Gisbert

    2008-01-01

    Exported proteases of Helicobacter pylori (H. pylori) are potentially involved in pathogen-associated disorders leading to gastric inflammation and neoplasia. By comprehensive sequence screening of the H. pylori proteome for predicted secreted proteases, we retrieved several candidate genes. We detected caseinolytic activities of several such proteases, which are released independently from the H. pylori type IV secretion system encoded by the cag pathogenicity island (cagPAI). Among these, we found the predicted serine protease HtrA (Hp1019), which was previously identified in the bacterial secretome of H. pylori. Importantly, we further found that the H. pylori genes hp1018 and hp1019 represent a single gene likely coding for an exported protein. Here, we directly verified proteolytic activity of HtrA in vitro and identified the HtrA protease in zymograms by mass spectrometry. Overexpressed and purified HtrA exhibited pronounced proteolytic activity, which is inactivated after mutation of Ser205 to alanine in the predicted active center of HtrA. These data demonstrate that H. pylori secretes HtrA as an active protease, which might represent a novel candidate target for therapeutic intervention strategies.

  15. Duplication of centromeric histone H3 (HTR12) gene in Arabidopsis halleri and A. lyrata, plant species with multiple centromeric satellite sequences.

    PubMed

    Kawabe, Akira; Nasuda, Shuhei; Charlesworth, Deborah

    2006-12-01

    Arabidopsis halleri and lyrata have three different major centromeric satellite sequences, a unique finding for a diploid Arabidopsis species. Since centromeric histones coevolve with centromeric satellites, these proteins would be predicted to show signs of selection when new centromere satellites have recently arisen. We isolated centromeric protein genes from A. halleri and lyrata and found that one of them, HTR12 (CENP-A), is duplicated, while CENP-C is not. Phylogenetic analysis indicates that the HTR12 duplication occurred after these species diverged from A. thaliana. Genetic mapping shows that HTR12 copy B has the same genomic location as the A. thaliana gene; the other copy (A, at the other end of the same chromosome) is probably the new copy. To test for selection since the duplication, we surveyed diversity at both HTR12 loci within A. lyrata. Overall, there is no strong evidence for an "evolutionary arms race" causing multiple replacement substitutions. The A. lyrata HTR12B sequences fall into three classes of haplotypes, apparently maintained for a long time, but they all encode the same amino acid sequence. In contrast, HTR12A has low diversity, but many variants are amino acid replacements, possibly due to independent selective sweeps within populations of the species.

  16. Prediction of Extracellular Proteases of the Human Pathogen Helicobacter pylori Reveals Proteolytic Activity of the Hp1018/19 Protein HtrA

    PubMed Central

    Löwer, Martin; Weydig, Christiane; Metzler, Dirk; Reuter, Andreas; Starzinski-Powitz, Anna

    2008-01-01

    Exported proteases of Helicobacter pylori (H. pylori) are potentially involved in pathogen-associated disorders leading to gastric inflammation and neoplasia. By comprehensive sequence screening of the H. pylori proteome for predicted secreted proteases, we retrieved several candidate genes. We detected caseinolytic activities of several such proteases, which are released independently from the H. pylori type IV secretion system encoded by the cag pathogenicity island (cagPAI). Among these, we found the predicted serine protease HtrA (Hp1019), which was previously identified in the bacterial secretome of H. pylori. Importantly, we further found that the H. pylori genes hp1018 and hp1019 represent a single gene likely coding for an exported protein. Here, we directly verified proteolytic activity of HtrA in vitro and identified the HtrA protease in zymograms by mass spectrometry. Overexpressed and purified HtrA exhibited pronounced proteolytic activity, which is inactivated after mutation of Ser205 to alanine in the predicted active center of HtrA. These data demonstrate that H. pylori secretes HtrA as an active protease, which might represent a novel candidate target for therapeutic intervention strategies. PMID:18946507

  17. Benzo(a)pyrene inhibits migration and invasion of extravillous trophoblast HTR-8/SVneo cells via activation of the ERK and JNK pathway.

    PubMed

    Liu, Liyuan; Wang, Yingxiong; Shen, Cha; He, Junlin; Liu, Xueqing; Ding, Yubin; Gao, Rufei; Chen, Xuemei

    2016-07-01

    Benzo(a)pyrene (BaP) is a persistent organic pollutant (POP) that is a serious threat to human health. Numerous studies have shown that BaP causes adverse effects in pregnancy, but the mechanism remains unclear. The moderate invasion of trophoblast cells into the endometrium is an important factor during successful embryo implantation. The aim of this study was to investigate the effect and mechanism of BaP on the invasion and migration of trophoblast cells. HTR-8/SVneo cells were treated with different concentrations (1, 5, 10, 25, 50 and 100 μM) of BaP. The invasion and migration of HTR-8/SVneo cells were observed after BaP treatment. The protein levels related to migration and invasion was detected by Western blot. The results confirmed that BaP inhibits the migration and invasion of extravillous trophoblast HTR-8/SVneo cells. Further investigations indicated that the protein levels of MMP-2, MMP-9 and E-cadherin in HTR-8/SVneo cells were changed by BaP treatment. Moreover, the data demonstrated that BaP activated the MAPK signaling pathway. Pretreatment with specific inhibitors of MAPK rescued BaP-induced change in the migration and invasion of HTR-8/SVneo cells. Taken together, our results indicated that BaP inhibits invasion and the migration of HTR-8/SVneo cells, which might cause a failure in early pregnancy. Copyright © 2015 John Wiley & Sons, Ltd.

  18. The protease inhibitor JO146 demonstrates a critical role for CtHtrA for Chlamydia trachomatis reversion from penicillin persistence

    PubMed Central

    Ong, Vanissa A.; Marsh, James W.; Lawrence, Amba; Allan, John A.; Timms, Peter; Huston, Wilhelmina M.

    2013-01-01

    The Chlamydia trachomatis serine protease HtrA (CtHtrA) has recently been demonstrated to be essential during the replicative phase of the chlamydial developmental cycle. A chemical inhibition strategy (serine protease inhibitor JO146) was used to demonstrate this essential role and it was found that the chlamydial inclusions diminish in size and are lost from the cell after CtHtrA inhibition without formation of viable elementary bodies. The inhibitor (JO146) was used in this study to investigate the role of CtHtrA for penicillin persistence and heat stress conditions for Chlamydia trachomatis. JO146 addition during penicillin persistence resulted in only minor reductions (~1 log) in the final viable infectious yield after persistent Chlamydia were reverted from persistence. However, JO146 treatment during the reversion and recovery from penicillin persistence was completely lethal for Chlamydia trachomatis. JO146 was completely lethal when added either during heat stress conditions, or during the recovery from heat stress conditions. These data together indicate that CtHtrA has essential roles during some stress environments (heat shock), recovery from stress environments (heat shock and penicillin persistence), as well as the previously characterized essential role during the replicative phase of the chlamydial developmental cycle. Thus, CtHtrA is an essential protease with both replicative phase and stress condition functions for Chlamydia trachomatis. PMID:24392355

  19. Induced rates of mitotic crossing over and possible mitotic gene conversion per wing anlage cell in Drosophila melanogaster by X rays and fission neutrons

    SciTech Connect

    Ayaki, T.; Fujikawa, K.; Ryo, H.; Itoh, T.; Kondo, S. )

    1990-09-01

    As a model for chromosome aberrations, radiation-induced mitotic recombination of mwh and flr genes in Drosophila melanogaster strain (mwh +/+ flr) was quantitatively studied. Fission neutrons were five to six times more effective than X rays per unit dose in producing either crossover-mwh/flr twins and mwh singles-or flr singles, indicating that common processes are involved in the production of crossover and flr singles. The X-ray-induced rate/wing anlage cell/Gy for flr singles was 1 X 10(-5), whereas that of crossover was 2 x 10(-4); the former and the latter rate are of the same order of magnitude as those of gene conversion and crossover in yeast, respectively. Thus, we conclude that proximal-marker flr singles induced in the transheterozygote are gene convertants. Using the model based on yeast that recombination events result from repair of double-strand breaks or gaps, we propose that mitotic recombination in the fly is a secondary result of recombinational DNA repair. Evidence for recombinational misrepair in the fly is given. The relative ratio of radiation-induced mitotic crossover to spontaneous meiotic crossover is one order of magnitude higher in the fly than in yeast and humans.

  20. Association of serotonin transporter (SLC6A4) & receptor (5HTR1A, 5HTR2A) polymorphisms with response to treatment with escitalopram in patients with major depressive disorder: A preliminary study

    PubMed Central

    Basu, Aniruddha; Chadda, R.K.; Sood, Mamta; Kaur, Harpreet; Kukreti, Ritushree

    2015-01-01

    Background & objectives: Genetic factors have potential of predicting response to antidepressants in patients with major depressive disorder (MDD). In this study, an attempt was made to find an association between response to escitalopram in patients with MDD, and serotonin transporter (SLC6A4) and receptor (5HTR1A, 5HTR2A) polymorphisms. Methods: Fifty five patients diagnosed as suffering from MDD, were selected for the study. The patients were treated with escitalopram over a period of 6-8 wk. Severity of depression, response to treatment and side effects were assessed using standardised instruments. Genetic variations from HTR1A (rs6295), HTR2A (rs6311 and rs6313) and SLC6A4 (44 base-pair insertion/deletion at 5-HTTLPR) were genotyped. The genetic data of the responders and non-responders were compared to assess the role of genetic variants in therapeutic outcome. Results: Thirty six (65.5%) patients responded to treatment, and 19 (34.5%) had complete remission. No association was observed for genotype and allelic frequencies of single nucleotide polymorphisms (SNPs) among remitter/non-remitter and responder/non-responder groups, and six most common side-effects, except memory loss which was significantly associated with rs6311 (P =0.03). Interpretation & conclusions: No significant association was found between the SNPs analysed and response to escitalopram in patients with MDD though a significant association was seen between the side effect of memory loss and rs6311. Studies with larger sample are required to find out genetic basis of antidepressant response in Indian patients. PMID:26261165

  1. Loss of HtrA2/Omi activity in non-neuronal tissues of adult mice causes premature aging.

    PubMed

    Kang, S; Louboutin, J-P; Datta, P; Landel, C P; Martinez, D; Zervos, A S; Strayer, D S; Fernandes-Alnemri, T; Alnemri, E S

    2013-02-01

    mnd2 mice die prematurely as a result of neurodegeneration 30-40 days after birth due to loss of the enzymatic activity of the mitochondrial quality control protease HtrA2/Omi. Here, we show that transgenic expression of human HtrA2/Omi in the central nervous system of mnd2 mice rescues them from neurodegeneration and prevents their premature death. Interestingly, adult transgenic mnd2 mice develop accelerated aging phenotypes, such as premature weight loss, hair loss, reduced fertility, curvature of the spine, heart enlargement, increased autophagy, and death by 12-17 months of age. These mice also have elevated levels of clonally expanded mitochondrial DNA (mtDNA) deletions in their tissues. Our results provide direct genetic evidence linking mitochondrial protein quality control to mtDNA deletions and aging in mammals.

  2. Decommissioning for the containment vessel of HTR-PM: Monte Carlo simulation and long-time predictions

    SciTech Connect

    Wenqian Li; Hong Li; Jianzhu Cao; Feng Xie; Sheng Fang; Ming Li

    2013-07-01

    A high temperature gas cooled reactor-pebble bed module (HTR-PM) is currently under design and construction in China. Calculations of induced radioactivity in the concrete containment vessel of HTR-PM were performed in order to predict the decommissioning problems. The main activation products and the ambient dose equivalent rate corresponding to 40 years of operations followed by 30 days/1 year/10 years of decay, were calculated by using the Monte Carlo particle transport code FLUKA. Finally, according to the Chinese National Standard and on the basis of the knowledge gained from this study, the classifications of the radioactive solid wastes were discussed, and some suggestions for the working duration of the employees were given. (authors)

  3. HtrC is involved in proteolysis of YpeB during germination of Bacillus anthracis and Bacillus subtilis spores.

    PubMed

    Bernhards, Casey B; Chen, Yan; Toutkoushian, Hannah; Popham, David L

    2015-01-01

    Bacterial endospores can remain dormant for decades yet can respond to nutrients, germinate, and resume growth within minutes. An essential step in the germination process is degradation of the spore cortex peptidoglycan wall, and the SleB protein in Bacillus species plays a key role in this process. Stable incorporation of SleB into the spore requires the YpeB protein, and some evidence suggests that the two proteins interact within the dormant spore. Early during germination, YpeB is proteolytically processed to a stable fragment. In this work, the primary sites of YpeB cleavage were identified in Bacillus anthracis, and it was shown that the stable products are comprised of the C-terminal domain of YpeB. Modification of the predominant YpeB cleavage sites reduced proteolysis, but cleavage at other sites still resulted in loss of full-length YpeB. A B. anthracis strain lacking the HtrC protease did not generate the same stable YpeB products. In B. anthracis and Bacillus subtilis htrC mutants, YpeB was partially stabilized during germination but was still degraded at a reduced rate by other, unidentified proteases. Purified HtrC cleaved YpeB to a fragment similar to that observed in vivo, and this cleavage was stimulated by Mn(2+) or Ca(2+) ions. A lack of HtrC did not stabilize YpeB or SleB during spore formation in the absence of the partner protein, indicating other proteases are involved in their degradation during sporulation.

  4. Bacteriocin Activity of Streptococcus pneumoniae Is Controlled by the Serine Protease HtrA via Posttranscriptional Regulation▿

    PubMed Central

    Dawid, Suzanne; Sebert, Michael E.; Weiser, Jeffrey N.

    2009-01-01

    The blp locus of a type 6A strain of Streptococcus pneumoniae encodes a two-peptide bacteriocin, pneumocin MN, which mediates intraspecies competition during mouse nasopharyngeal colonization. This locus is regulated by a quorum-sensing mechanism consisting of a dedicated two-component regulatory system and a peptide pheromone. Like most clinical isolates, this type 6A strain can be separated into opaque and transparent colony variants, each playing a different role during pneumococcal infection. In this study, we show that the blp locus is differentially regulated at the posttranscriptional level in pneumococcal opacity variants. Transparent and opaque variants produce equivalent amounts of blpMNPO transcript when stimulated with a synthetic pheromone, but transparent variants have no pneumocin MN-mediated inhibitory activity while opaque variants produce large zones of inhibitory activity. The differential regulation in opacity variants is driven by the two-component regulatory system CiaRH via its regulation of the serine protease HtrA. Transparent mutants deficient in CiaH or HtrA show increased pneumocin MN-mediated inhibition. In addition, these mutants demonstrate alterations in their dose response to a synthetic peptide pheromone, suggesting that HtrA activity impacts pneumocin MN production at the level of signaling. This, in addition to its known effects on competence, suggests that HtrA is a pleiotropic regulator whose protease activity affects several important bacterial pathways. The complex regulation of pneumocins may allow the pneumococcus to reserve the secretion of active peptides for situations where the benefit of their inhibitory activity outweighs the cost of their production. PMID:19103930

  5. Epigenetic and genetic variants in the HTR1B gene and clinical improvement in children and adolescents treated with fluoxetine.

    PubMed

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

    2017-04-03

    The serotonin 1B receptor (5-HT1B) is important to both the pathogenesis of major depressive disorder and the antidepressant effects of selective serotonin reuptake inhibitors. Although fluoxetine has been shown to be effective and safe in children and adolescents, not all patients experience a proper clinical response, which has led to further study into the main factors involved in this inter-individual variability. Our aim was to study the effect of epigenetic and genetic factors that could affect 5-hydroxytryptamine receptor 1B (HTR1B) gene expression, and thereby response to fluoxetine. A total of 83 children and adolescents were clinically assessed 12weeks after of initiating an antidepressant treatment with fluoxetine for the first time. We evaluated the influence of single nucleotide polymorphisms (SNPs) specifically located in transcription factor binding sites (TFBSs) on their clinical improvement. A combined genetic analysis considering the significant SNPs together with the functional variant rs130058 previously associated in our population was also performed. Moreover, we assessed, for the first time in the literature, whether methylation levels of the HTR1B promoter region could be associated with the pharmacological response. Two, rs9361233 and rs9361235, were significantly associated with clinical improvement after treatment with fluoxetine. The heterozygous genotype combination analysis showed a negative correlation with clinical improvement. The lowest improvement was experienced by patients who were heterozygous for all three SNPs. Moreover, a negative correlation was found between clinical improvement and the average methylation level of the HTR1B promoter. These results give new evidence for the role of epigenetic and genetic factors which could modulate HTR1B expression in the pharmacological response to antidepressants.

  6. HtrC Is Involved in Proteolysis of YpeB during Germination of Bacillus anthracis and Bacillus subtilis Spores

    PubMed Central

    Bernhards, Casey B.; Chen, Yan; Toutkoushian, Hannah

    2014-01-01

    Bacterial endospores can remain dormant for decades yet can respond to nutrients, germinate, and resume growth within minutes. An essential step in the germination process is degradation of the spore cortex peptidoglycan wall, and the SleB protein in Bacillus species plays a key role in this process. Stable incorporation of SleB into the spore requires the YpeB protein, and some evidence suggests that the two proteins interact within the dormant spore. Early during germination, YpeB is proteolytically processed to a stable fragment. In this work, the primary sites of YpeB cleavage were identified in Bacillus anthracis, and it was shown that the stable products are comprised of the C-terminal domain of YpeB. Modification of the predominant YpeB cleavage sites reduced proteolysis, but cleavage at other sites still resulted in loss of full-length YpeB. A B. anthracis strain lacking the HtrC protease did not generate the same stable YpeB products. In B. anthracis and Bacillus subtilis htrC mutants, YpeB was partially stabilized during germination but was still degraded at a reduced rate by other, unidentified proteases. Purified HtrC cleaved YpeB to a fragment similar to that observed in vivo, and this cleavage was stimulated by Mn2+ or Ca2+ ions. A lack of HtrC did not stabilize YpeB or SleB during spore formation in the absence of the partner protein, indicating other proteases are involved in their degradation during sporulation. PMID:25384476

  7. AP-1 transcription factors, mucin-type molecules and MMPs regulate the IL-11 mediated invasiveness of JEG-3 and HTR-8/SVneo trophoblastic cells.

    PubMed

    Suman, Pankaj; Godbole, Geeta; Thakur, Ravi; Morales-Prieto, Diana M; Modi, Deepak N; Markert, Udo R; Gupta, Satish K

    2012-01-01

    This study examines the IL-11 mediated activation of downstream signaling and expression of effector molecules to resolve the controversies associated with the IL-11 mediated regulation of the invasiveness of two commonly used trophoblastic cell models viz. JEG-3 and HTR-8/SVneo cells. It has been reported that IL-11 increases the invasiveness of JEG-3 cells while, reduces the invasiveness of HTR-8/SVneo cells. Invasion assay performed simultaneously for both the cell lines confirmed the above findings. In addition, HTR-8/SVneo cells showed a higher basal invasiveness than JEG-3 cells. Western blot showed the IL-11 mediated activation of STAT3(tyr705) and STAT1(tyr701) in both the cell lines. However, IL-11 activated the ERK1/2 phosphorylation in JEG-3 cells but, inhibited it in HTR-8/SVneo cells. Within 10 min of IL-11 treatment, p-STAT3(tyr705) was localized inside the nucleus of both the cell lines but, there was enhanced co-localization of protein inhibitor of activated STAT1/3 (PIAS1/3) and p-STAT3(tyr705) in HTR-8/SVneo cells and not in JEG-3 cells. This could be reason for the poor responsiveness of STAT3 responsive genes like mucin 1 (MUC1) in HTR-8/SVneo cells and not in JEG-3 cells. Further, microarray analysis of the IL-11 treated cells revealed differential responsiveness of JEG-3 as compared to HTR-8/SVneo cells. Several family of genes like activator protein-1 (AP-1) transcription factors (Jun and Fos), mucin-type molecules, MMP23B etc showed enhanced expression in IL-11 treated JEG-3 cells while, there was no response or decrease in their expression in IL-11 treated HTR-8/SVneo cells. Expression of these molecules was confirmed by quantitative RT-PCR. In addition, HTR-8/SVneo cells also showed a significant decrease in the expression of MMP2, MMP3 and MMP9 upon IL-11 treatment. Hence, IL-11 mediated differential activation of signaling and expression of effector molecules is responsible for the differential invasive response of JEG-3 and HTR-8/SVneo

  8. AP-1 Transcription Factors, Mucin-Type Molecules and MMPs Regulate the IL-11 Mediated Invasiveness of JEG-3 and HTR-8/SVneo Trophoblastic Cells

    PubMed Central

    Suman, Pankaj; Godbole, Geeta; Thakur, Ravi; Morales-Prieto, Diana M.; Modi, Deepak N.; Markert, Udo R.; Gupta, Satish K.

    2012-01-01

    This study examines the IL-11 mediated activation of downstream signaling and expression of effector molecules to resolve the controversies associated with the IL-11 mediated regulation of the invasiveness of two commonly used trophoblastic cell models viz. JEG-3 and HTR-8/SVneo cells. It has been reported that IL-11 increases the invasiveness of JEG-3 cells while, reduces the invasiveness of HTR-8/SVneo cells. Invasion assay performed simultaneously for both the cell lines confirmed the above findings. In addition, HTR-8/SVneo cells showed a higher basal invasiveness than JEG-3 cells. Western blot showed the IL-11 mediated activation of STAT3(tyr705) and STAT1(tyr701) in both the cell lines. However, IL-11 activated the ERK1/2 phosphorylation in JEG-3 cells but, inhibited it in HTR-8/SVneo cells. Within 10 min of IL-11 treatment, p-STAT3(tyr705) was localized inside the nucleus of both the cell lines but, there was enhanced co-localization of protein inhibitor of activated STAT1/3 (PIAS1/3) and p-STAT3(tyr705) in HTR-8/SVneo cells and not in JEG-3 cells. This could be reason for the poor responsiveness of STAT3 responsive genes like mucin 1 (MUC1) in HTR-8/SVneo cells and not in JEG-3 cells. Further, microarray analysis of the IL-11 treated cells revealed differential responsiveness of JEG-3 as compared to HTR-8/SVneo cells. Several family of genes like activator protein-1 (AP-1) transcription factors (Jun and Fos), mucin-type molecules, MMP23B etc showed enhanced expression in IL-11 treated JEG-3 cells while, there was no response or decrease in their expression in IL-11 treated HTR-8/SVneo cells. Expression of these molecules was confirmed by quantitative RT-PCR. In addition, HTR-8/SVneo cells also showed a significant decrease in the expression of MMP2, MMP3 and MMP9 upon IL-11 treatment. Hence, IL-11 mediated differential activation of signaling and expression of effector molecules is responsible for the differential invasive response of JEG-3 and HTR-8/SVneo

  9. Inventory of telomerase components in human cells reveals multiple subpopulations of hTR and hTERT

    PubMed Central

    Xi, Linghe; Cech, Thomas R.

    2014-01-01

    Telomerase is the ribonucleoprotein (RNP) enzyme that elongates telomeric DNA to compensate for the attrition occurring during each cycle of DNA replication. Knowing the levels of telomerase in continuously dividing cells is important for understanding how much telomerase is required for cell immortality. In this study, we measured the endogenous levels of the human telomerase RNP and its two key components, human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT). We estimate ∼240 telomerase monomers per cell for HEK 293T and HeLa, a number similar to that of telomeres in late S phase. The subunits were in excess of RNPs (e.g. ∼1150 hTR and ∼500 hTERT molecules per HeLa cell), suggesting the existence of unassembled components. This hypothesis was tested by overexpressing individual subunits, which increased total telomerase activity as measured by the direct enzyme assay. Thus, there are subpopulations of both hTR and hTERT not assembled into telomerase but capable of being recruited. We also determined the specific activity of endogenous telomerase and of overexpressed super-telomerase both to be ∼60 nt incorporated per telomerase per minute, with Km(dGTP) ∼17 μM, indicating super-telomerase is as catalytically active as endogenous telomerase and is thus a good model for biochemical studies. PMID:24990373

  10. Rapid blockade of telomerase activity and tumor cell growth by the DPL lipofection of ribbon antisense to hTR.

    PubMed

    Bajpai, Arun K; Park, Jeong-Hoh; Moon, Ik-Jae; Kang, Hyungu; Lee, Yun-Han; Doh, Kyung-Oh; Suh, Seong-Il; Chang, Byeong-Churl; Park, Jong-Gu

    2005-09-29

    Ribbon antisense (RiAS) to the hTR RNA, a component of the telomerase complex, was employed to inhibit telomerase activity and cancer cell growth. The antisense molecule, hTR-RiAS, combined with enhanced cellular uptake was shown to effectively inhibit telomerase activity and cause rapid cell death in various cancer cell lines. When cancer cells were treated with hTR-RiAS, the level of hTR RNA was reduced by more than 90% accompanied with reduction in telomerase activity. When checked for cancer cell viability, cancer cell lines treated with hTR-RiAS using DNA+Peptide+Lipid complex showed 70-80% growth inhibition in 3 days. The reduced cell viability was due to apoptosis as the percentage of cells exhibiting the sub-G0 arrest and DNA fragmentation increased after antisense treatment. Further, when subcutaneous tumors of a colon cancer cell line (SW480) were treated intratumorally with hTR-RiAS, tumor growth was markedly suppressed with almost total ablation of hTR RNA in the tumor tissue. Cells in the tumor tissue were also found to undergo apoptosis after hTR-RiAS treatment. These results suggest that hTR-RiAS is an effective anticancer reagent, with a potential for broad efficacy to diverse malignant tumors.

  11. Characterization of interactions between PinX1 and human telomerase subunits hTERT and hTR.

    PubMed

    Banik, Soma S R; Counter, Christopher M

    2004-12-10

    The addition of telomeric repeats to chromosome ends by the enzyme telomerase is a highly orchestrated process. Although much is known regarding telomerase catalytic activity in vitro, less is known about how this activity is regulated in vivo to ensure proper telomere elongation. One protein that appears to be involved in negatively regulating telomerase function in vivo is PinX1 because overexpression of PinX1 inhibits telomerase activity and causes telomere shortening. To understand the nature of this repression, we characterized the interactions among PinX1 and the core components of telomerase, the human telomerase reverse transcriptase (hTERT) and associated human telomerase RNA (hTR). We now show that in vitro PinX1 binds directly to the hTERT protein subunit, primarily to the hTR-binding domain, as well as to the hTR subunit. However, in a cellular context, the association of PinX1 with hTR is dependent on the presence of hTERT. Taken together, we suggest that PinX1 represses telomerase activity in vivo by binding to the assembled hTERT.hTR complex.

  12. Inventory of telomerase components in human cells reveals multiple subpopulations of hTR and hTERT.

    PubMed

    Xi, Linghe; Cech, Thomas R

    2014-07-01

    Telomerase is the ribonucleoprotein (RNP) enzyme that elongates telomeric DNA to compensate for the attrition occurring during each cycle of DNA replication. Knowing the levels of telomerase in continuously dividing cells is important for understanding how much telomerase is required for cell immortality. In this study, we measured the endogenous levels of the human telomerase RNP and its two key components, human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT). We estimate ∼ 240 telomerase monomers per cell for HEK 293T and HeLa, a number similar to that of telomeres in late S phase. The subunits were in excess of RNPs (e.g. ∼ 1150 hTR and ∼ 500 hTERT molecules per HeLa cell), suggesting the existence of unassembled components. This hypothesis was tested by overexpressing individual subunits, which increased total telomerase activity as measured by the direct enzyme assay. Thus, there are subpopulations of both hTR and hTERT not assembled into telomerase but capable of being recruited. We also determined the specific activity of endogenous telomerase and of overexpressed super-telomerase both to be ∼ 60 nt incorporated per telomerase per minute, with Km(dGTP) ∼ 17 μM, indicating super-telomerase is as catalytically active as endogenous telomerase and is thus a good model for biochemical studies.

  13. Impact of the HTR3A gene with early life trauma on emotional brain networks and depressed mood.

    PubMed

    Gatt, Justine M; Williams, Leanne M; Schofield, Peter R; Dobson-Stone, Carol; Paul, Robert H; Grieve, Stuart M; Clark, C Richard; Gordon, Evian; Nemeroff, Charles B

    2010-08-01

    The risk for mental illnesses such as depression is increasingly conceptualized as the product of gene-environment interactions and their impact on brain structure and function. The role of serotonin 3A receptor gene (HTR3A -42C>T polymorphism) and its interaction with early life stress (ELS) was investigated in view of the receptor's localization to brain regions central to emotion processing. Fronto-limbic grey matter (GM) loss was measured using magnetic resonance imaging and assessed using voxel-based morphometry analysis in 397 nonclinical individuals from the Brain Resource International Database. Negative mood symptoms were also assessed. The HTR3A CC genotype group, compared to the T carriers, demonstrated comparative loss to GM in hippocampal structures, which extended to the frontal cortices for those CC genotype individuals also exposed to ELS. Elevations in depressed mood were also evident. These findings suggest that the HTR3A CC genotype may be associated with alterations in brain structures central to emotion processing, particularly when exposed to stress, and further highlight the potential role of the serotonin system in the pathophysiology of affective disorders. In contrast, those individuals with the T allele, in particular the TT genotype, may be more protected from such alterations combined with minimal exposure to ELS events. 2010 Wiley-Liss, Inc.

  14. Up-regulation of lymphocyte antigen 6 complex expression in side-population cells derived from a human trophoblast cell line HTR-8/SVneo.

    PubMed

    Inagaki, Tetsunori; Kusunoki, Soshi; Tabu, Kouichi; Okabe, Hitomi; Yamada, Izumi; Taga, Tetsuya; Matsumoto, Akemi; Makino, Shintaro; Takeda, Satoru; Kato, Kiyoko

    2016-01-01

    The continual proliferation and differentiation of trophoblasts are critical for the maintenance of pregnancy. It is well known that the tissue stem cells are associated with the development of tissues and pathologies. It has been demonstrated that side-population (SP) cells identified by fluorescence-activated cell sorting (FACS) are enriched with stem cells. The SP cells in HTR-8/SVneo cells derived from human primary trophoblast cells were isolated by FACS. HTR-8/SVneo-SP cell cultures generated both SP and non-SP (NSP) subpopulations. In contrast, NSP cell cultures produced NSP cells and failed to produce SP cells. These SP cells showed self-renewal capability by serial colony-forming assay. Microarray expression analysis using a set of HTR-8/SVneo-SP and -NSP cells revealed that SP cells overexpressed several stemness genes including caudal type homeobox2 (CDX2) and bone morphogenic proteins (BMPs), and lymphocyte antigen 6 complex locus D (LY6D) gene was the most highly up-regulated in HTR-8/SVneo-SP cells. LY6D gene reduced its expression in the course of a 7-day cultivation in differentiation medium. SP cells tended to reduce its fraction by treatment of LY6D siRNA indicating that LY6D had potential to maintain cell proliferation of HTR-8/SVneo-SP cells. On ontology analysis, epithelial-mesenchymal transition (EMT) pathway was involved in the up-regulated genes on microarray analysis. HTR-SVneo-SP cells showed enhanced migration. This is the first report that LY6D was important for the maintenance of HTR-8/SVneo-SP cells. EMT was associated with the phenotype of these SP cells.

  15. The LA loop as an important regulatory element of the HtrA (DegP) protease from Escherichia coli: structural and functional studies.

    PubMed

    Figaj, Donata; Gieldon, Artur; Polit, Agnieszka; Sobiecka-Szkatula, Anna; Koper, Tomasz; Denkiewicz, Milena; Banecki, Bogdan; Lesner, Adam; Ciarkowski, Jerzy; Lipinska, Barbara; Skorko-Glonek, Joanna

    2014-05-30

    Bacterial HtrAs are serine proteases engaged in extracytoplasmic protein quality control and are required for the virulence of several pathogenic species. The proteolytic activity of HtrA (DegP) from Escherichia coli, a model prokaryotic HtrA, is stimulated by stressful conditions; the regulation of this process is mediated by the LA, LD, L1, L2, and L3 loops. The precise mechanism of action of the LA loop is not known due to a lack of data concerning its three-dimensional structure as well as its mode of interaction with other regulatory elements. To address these issues we generated a theoretical model of the three-dimensional structure of the LA loop as per the resting state of HtrA and subsequently verified its correctness experimentally. We identified intra- and intersubunit contacts that formed with the LA loops; these played an important role in maintaining HtrA in its inactive conformation. The most significant proved to be the hydrophobic interactions connecting the LA loops of the hexamer and polar contacts between the LA' (the LA loop on an opposite subunit) and L1 loops on opposite subunits. Disturbance of these interactions caused the stimulation of HtrA proteolytic activity. We also demonstrated that LA loops contribute to the preservation of the integrity of the HtrA oligomer and to the stability of the monomer. The model presented in this work explains the regulatory role of the LA loop well; it should also be applicable to numerous Enterobacteriaceae pathogenic species as the amino acid sequences of the members of this bacterial family are highly conserved.

  16. Secretion Chaperones PrsA2 and HtrA Are Required for Listeria monocytogenes Replication following Intracellular Induction of Virulence Factor Secretion

    PubMed Central

    Ahmed, Jana K.

    2016-01-01

    The Gram-positive bacterium Listeria monocytogenes transitions from an environmental organism to an intracellular pathogen following its ingestion by susceptible mammalian hosts. Bacterial replication within the cytosol of infected cells requires activation of the central virulence regulator PrfA followed by a PrfA-dependent induction of secreted virulence factors. The PrfA-induced secreted chaperone PrsA2 and the chaperone/protease HtrA contribute to the folding and stability of select proteins translocated across the bacterial membrane. L. monocytogenes strains that lack both prsA2 and htrA exhibit near-normal patterns of growth in broth culture but are severely attenuated in vivo. We hypothesized that, in the absence of PrsA2 and HtrA, the increase in PrfA-dependent protein secretion that occurs following bacterial entry into the cytosol results in misfolded proteins accumulating at the bacterial membrane with a subsequent reduction in intracellular bacterial viability. Consistent with this hypothesis, the introduction of a constitutively activated allele of prfA (prfA*) into ΔprsA2 ΔhtrA strains was found to essentially inhibit bacterial growth at 37°C in broth culture. ΔprsA2 ΔhtrA strains were additionally found to be defective for cell invasion and vacuole escape in selected cell types, steps that precede full PrfA activation. These data establish the essential requirement for PrsA2 and HtrA in maintaining bacterial growth under conditions of PrfA activation. In addition, chaperone function is required for efficient bacterial invasion and rapid vacuole lysis within select host cell types, indicating roles for PrsA2/HtrA prior to cytosolic PrfA activation and the subsequent induction of virulence factor secretion. PMID:27481256

  17. The effects of a HTR2B stop codon and testosterone on energy metabolism and beta cell function among antisocial Finnish males.

    PubMed

    Tikkanen, Roope; Saukkonen, Tero; Fex, Malin; Bennet, Hedvig; Rautiainen, Marja-Riitta; Paunio, Tiina; Koskinen, Mika; Panarsky, Rony; Bevilacqua, Laura; Sjöberg, Rickard L; Tiihonen, Jari; Virkkunen, Matti

    2016-10-01

    Herein, we examined insulin resistance (IR), insulin sensitivity (IS), beta cell activity, and glucose metabolism in subjects with antisocial personality disorder (ASPD), and whether the serotonin 2B (5-HT2B) receptor and testosterone have a role in energy metabolism. A cohort of subjects belonging to a founder population that included 98 ASPD males, aged 25-30, was divided into groups based on the presence of a heterozygous 5-HT2B receptor loss-of-function gene mutation (HTR2B Q20*; n = 9) or not (n = 89). Serum glucose and insulin levels were measured in a 5 h oral glucose tolerance test (75 g) and indices describing IR, IS, and beta cell activity were calculated. Body mass index (BMI) was also determined. Concentrations of the serotonin metabolite 5-hydroxyindoleacetic acid were measured in cerebrospinal fluid, and testosterone levels from serum. An IR-like state comprising high IR, low IS, and high beta cell activity indices was observed among ASPD subjects without the HTR2B Q20* allele. By contrast, being an ASPD HTR2B Q20* carrier appeared to be preventive of these pathophysiologies. The HTR2B Q20* allele and testosterone predicted lower BMI independently, but an interaction between HTR2B Q20* and testosterone lead to increased insulin sensitivity among HTR2B Q20* carriers with low testosterone levels. The HTR2B Q20* allele also predicted reduced beta cell activity and enhanced glucose metabolism. Reduced 5-HT2B receptor function at low or normal testosterone levels may be protective of obesity. Results were observed among Finnish males having an antisocial personality disorder, which limits the generality. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. The proteases HtrA2/Omi and UCH-L1 regulate TNF-induced necroptosis

    PubMed Central

    2013-01-01

    Background In apoptosis, proteolysis by caspases is the primary mechanism for both initiation and execution of programmed cell death (PCD). In contrast, the impact of proteolysis on the regulation and execution of caspase-independent forms of PCD (programmed necrosis, necroptosis) is only marginally understood. Likewise, the identity of the involved proteases has remained largely obscure. Here, we have investigated the impact of proteases in TNF-induced necroptosis. Results The serine protease inhibitor TPKC protected from TNF-induced necroptosis in multiple murine and human cells systems whereas inhibitors of metalloproteinases or calpain/cysteine and cathepsin proteases had no effect. A screen for proteins labeled by a fluorescent TPCK derivative in necroptotic cells identified HtrA2/Omi (a serine protease previously implicated in PCD) as a promising candidate. Demonstrating its functional impact, pharmacological inhibition or genetic deletion of HtrA2/Omi protected from TNF-induced necroptosis. Unlike in apoptosis, HtrA2/Omi did not cleave another protease, ubiquitin C-terminal hydrolase (UCH-L1) during TNF-induced necroptosis, but rather induced monoubiquitination indicative for UCH-L1 activation. Correspondingly, pharmacologic or RNA interference-mediated inhibition of UCH-L1 protected from TNF-induced necroptosis. We found that UCH-L1 is a mediator of caspase-independent, non-apoptotic cell death also in diseased kidney podocytes by measuring cleavage of the protein PARP-1, caspase activity, cell death and cell morphology. Indicating a role of TNF in this process, podocytes with stably downregulated UCH-L1 proved resistant to TNF-induced necroptosis. Conclusions The proteases HtrA2/Omi and UCH-L1 represent two key components of TNF-induced necroptosis, validating the relevance of proteolysis not only for apoptosis, but also for caspase-independent PCD. Since UCH-L1 clearly contributes to the non-apoptotic death of podocytes, interference with the necroptotic

  19. Temperature modeling for analysis and design of the sintering furnance in HTR fuel type of ball

    SciTech Connect

    Saragi, Elfrida; Setiadji, Moch

    2013-09-09

    One of the factors that determine the safety of the operation of the sintering furnace fuel HTR ball is the temperature distribution in the ceramic tube furnace. The temperature distribution must be determined at design stage. The tube has a temperature of 1600 °C at one end and about 40 °C at the other end. The outside of the tube was cooled by air through natural convection. The tube is a furnace ceramic tube which its geometry are 0.08, 0.09 and 0.5 m correspondingly for the inner tube diameter, outer tube diameter and tube length. The temperature distribution of the tube is determined by the natural convection coefficient (NCF), which is difficult to be calculated manually. The determination of NCF includes the Grasshoff, Prandtl, and Nusselt numbers which is a function of the temperature difference between the surrounding air with the ceramic tube. If the temperature vary along the tube, the complexity of the calculations increases. Thus the proposed modeling was performed to determine the temperature distribution along the tube and heat transfer coefficient using a self-developed software which permit the design process easier.

  20. Carbon monoxide formation in UO 2 kerneled HTR fuel particles containing oxygen getters

    NASA Astrophysics Data System (ADS)

    Proksch, E.; Strigl, A.; Nabielek, H.

    1986-06-01

    Mass spectrometric measurements of CO in irradiated UO 2 kerneled HTR fuel particles containing various oxygen getters are summarized and evaluated. Uranium carbide addition in the 3 to 15% range reduces the CO release by factors between 25 and 80, up to burn-up levels as high as 70% FIMA. Unintentional gettering by SiC in TRISO coated particles with failed inner pyrocarbon layers results in CO reduction factors between 15 and 110. For ZrC, only somewhat ambiguous results have been obtained; most likely, ZrC results in CO reduction by a factor of about 40. Ce 2O 3 and La 2O 3 seem to be somewhat less effective than the three carbides; for Ce 2O 3, reduction factors between 3 and 15 have been found. However, these results are possibly incorrect due to premature oxidation of the getter already during fabrication. Addition of SiO 2 + Al 2O 3 has no influence on CO release at all.

  1. Temperature modeling for analysis and design of the sintering furnance in HTR fuel type of ball

    NASA Astrophysics Data System (ADS)

    Saragi, Elfrida; Setiadji, Moch

    2013-09-01

    One of the factors that determine the safety of the operation of the sintering furnace fuel HTR ball is the temperature distribution in the ceramic tube furnace. The temperature distribution must be determined at design stage. The tube has a temperature of 1600 °C at one end and about 40 °C at the other end. The outside of the tube was cooled by air through natural convection. The tube is a furnace ceramic tube which its geometry are 0.08, 0.09 and 0.5 m correspondingly for the inner tube diameter, outer tube diameter and tube length. The temperature distribution of the tube is determined by the natural convection coefficient (NCF), which is difficult to be calculated manually. The determination of NCF includes the Grasshoff, Prandtl, and Nusselt numbers which is a function of the temperature difference between the surrounding air with the ceramic tube. If the temperature vary along the tube, the complexity of the calculations increases. Thus the proposed modeling was performed to determine the temperature distribution along the tube and heat transfer coefficient using a self-developed software which permit the design process easier.

  2. Production of CO during burnup of (Th, U)O 2 kerneled HTR fuel particles

    NASA Astrophysics Data System (ADS)

    Proksch, Emil; Strigl, Anton; Nabielek, Heinz

    1985-10-01

    The CO content of irradiated (Th, U)O 2 kerneled HTR fuel particles has been measured by mass spectrometry. An evaluation of all the data thus obtained showed that the oxygen release, O/f (atoms per fission), during irradiation is governed by thermodynamic equilibrium; O/f is a function of the irradiation temperature T(K), the initial Th/U-235 ratio N, and the burnup F (fissions per initial heavy metal atom). Within the limits of 1073 < T < 2273, 4 < N <50, and 0.04 < F < 0.17, the oxygen release can be represented by the expression log 10O/f = 0.96 - 4420/T + 0.4 log 10N + 0.3 log 10F . The attainment of equilibrium proceeds rather slowly; at 1473 K it takes about 130 h to reach 99% of the equilibrium value. Coated particles which had undergone large fission-product losses showed significantly increased oxygen release values.

  3. Effects of Homogeneous Geometry Models in Simulating the Fuel Balls in HTR-10

    NASA Astrophysics Data System (ADS)

    Wang, Meng-Jen; Peir, Jinn-Jer; Chao, Der-Sheng; Liang, Jenq-Horng

    In this study, the core geometry of HTR-10 was simulated using four different models including: (1) model 1 - an explicit double heterogeneous geometry, (2) model 2 - a mixing of UO2 kernel and four layers in each TRISO particle into one, (3) model 3 - a mixing of 8,335 TRISO particles and the inner graphite matrix in each fuel ball into one, and (4) model 4 - a mixing of the outer graphite shell, 8,335 TRISO particles, and the inner graphite matrix in each fuel ball into one. The associated initial core computations were performed using the MCNP version 1.51 computer code. The experimental fuel loading height of 123 cm was employed for each model. The results revealed that the multiplication factors ranged from largest to smallest with model 1, model 2, model 3, and model 4. The neutron spectrum in the fuel region of each models varied from the hardest to the softest are model 1, model 2, model 3, and model 4 while the averaged neutron spectrum in fuel ball from hardest to softest are model 4, model 3, model 2, and model 1. In addition, the CPU execution times extended from longest to shortest with model 1, model 2, model 3, and model 4.

  4. Omi/HtrA2 protease mediates cisplatin-induced cell death in renal cells.

    PubMed

    Cilenti, Lucia; Kyriazis, George A; Soundarapandian, Mangala M; Stratico, Valerie; Yerkes, Adam; Park, Kwon Moo; Sheridan, Alice M; Alnemri, Emad S; Bonventre, Joseph V; Zervos, Antonis S

    2005-02-01

    Omi/HtrA2 is a mitochondrial proapoptotic serine protease that is able to induce both caspase-dependent and caspase-independent cell death. After apoptotic stimuli, Omi is released to the cytoplasm where it binds and cleaves inhibitor of apoptosis proteins. In this report, we investigated the role of Omi in renal cell death following cisplatin treatment. Using primary mouse proximal tubule cells, as well as established renal cell lines, we show that the level of Omi protein is upregulated after treatment with cisplatin. This upregulation is followed by the release of Omi from mitochondria to the cytoplasm and degradation of XIAP. Reducing the endogenous level of Omi protein using RNA interference renders renal cells resistant to cisplatin-induced cell death. Furthermore, we show that the proteolytic activity of Omi is necessary and essential for cisplatin-induced cell death in this system. When renal cells are treated with Omi's specific inhibitor, ucf-101, they become significantly resistant to cisplatin-induced cell death. Ucf-101 was also able to minimize cisplatin-induced nephrotoxic injury in animals. Our results demonstrate that Omi is a major mediator of cisplatin-induced cell death in renal cells and suggest a way to limit renal injury by specifically inhibiting its proteolytic activity.

  5. High Temperature Reactor (HTR) Deep Burn Core and Fuel Analysis: Design Selection for the Prismatic Block Reactor With Results from FY-2011 Activities

    SciTech Connect

    Michael A. Pope

    2011-10-01

    The Deep Burn (DB) Project is a U.S. Department of Energy sponsored feasibility study of Transuranic Management using high burnup fuel in the high temperature helium cooled reactor (HTR). The DB Project consists of seven tasks: project management, core and fuel analysis, spent fuel management, fuel cycle integration, TRU fuel modeling, TRU fuel qualification, and HTR fuel recycle. In the Phase II of the Project, we conducted nuclear analysis of TRU destruction/utilization in the HTR prismatic block design (Task 2.1), deep burn fuel/TRISO microanalysis (Task 2.3), and synergy with fast reactors (Task 4.2). The Task 2.1 covers the core physics design, thermo-hydraulic CFD analysis, and the thermofluid and safety analysis (low pressure conduction cooling, LPCC) of the HTR prismatic block design. The Task 2.3 covers the analysis of the structural behavior of TRISO fuel containing TRU at very high burnup level, i.e. exceeding 50% of FIMA. The Task 4.2 includes the self-cleaning HTR based on recycle of HTR-generated TRU in the same HTR. Chapter IV contains the design and analysis results of the 600MWth DB-HTR core physics with the cycle length, the average discharged burnup, heavy metal and plutonium consumptions, radial and axial power distributions, temperature reactivity coefficients. Also, it contains the analysis results of the 450MWth DB-HTR core physics and the analysis of the decay heat of a TRU loaded DB-HTR core. The evaluation of the hot spot fuel temperature of the fuel block in the DB-HTR (Deep-Burn High Temperature Reactor) core under full operating power conditions are described in Chapter V. The investigated designs are the 600MWth and 460MWth DB-HTRs. In Chapter VI, the thermo-fluid and safety of the 600MWth DB-HTRs has been analyzed to investigate a thermal-fluid design performance at the steady state and a passive safety performance during an LPCC event. Chapter VII describes the analysis results of the TRISO fuel microanalysis of the 600MWth and 450

  6. Assignment of the 5HT7 receptor gene (HTR7) to chromosome 10q and exclusion of genetic linkage with Tourette syndrome

    SciTech Connect

    Gelernter, J.; Rao, P.A.; Pauls, D.L.

    1995-03-20

    A novel serotonin receptor designated 5HT7 (genetic locus HTR7) was cloned in 1993. This receptor has interesting properties related to ligand affinity and CNS distribution that render HTR7 a very interesting candidate gene for neuropsychiatric disorders. We mapped this gene, first by physical methods and then by genetic linkage. First, we made a tentative assignment to chromosome 10, based on hybridization of an HTR7 probe to a Southern blot of DNA from somatic cell hybrids. We then identified a genetic polymorphism at the HTR7 locus. We identified one extended pedigree where the polymorphism segregated. Using the LEPED computer program for pairwise linkage analysis, we confirmed the assignment of the gene to chromosome 10, specifically 10q21-q24, based on a lod score of 5.37 at 0% recombination between HTR7 and D10S20 (a chromosome 10 reference marker). Finally, we excluded genetic linkage between this locus and Tourette syndrome under a reasonable set of assumptions. 15 refs., 1 fig., 1 tab.

  7. HTR8/SVneo Cells Display Trophoblast Progenitor Cell-Like Characteristics Indicative of Self-Renewal, Repopulation Activity, and Expression of “Stemness-” Associated Transcription Factors

    PubMed Central

    Weber, Maja; Knoefler, Ilka; Schleussner, Ekkehard; Markert, Udo R.; Fitzgerald, Justine S.

    2013-01-01

    Introduction. JEG3 is a choriocarcinoma—and HTR8/SVneo a transformed extravillous trophoblast—cell line often used to model the physiologically invasive extravillous trophoblast. Past studies suggest that these cell lines possess some stem or progenitor cell characteristics. Aim was to study whether these cells fulfill minimum criteria used to identify stem-like (progenitor) cells. In summary, we found that the expression profile of HTR8/SVneo (CDX2+, NOTCH1+, SOX2+, NANOG+, and OCT-) is distinct from JEG3 (CDX2+ and NOTCH1+) as seen only in human-serum blocked immunocytochemistry. This correlates with HTR8/SVneo's self-renewal capacities, as made visible via spheroid formation and multi-passagability in hanging drops protocols paralleling those used to maintain embryoid bodies. JEG3 displayed only low propensity to form and reform spheroids. HTR8/SVneo spheroids migrated to cover and seemingly repopulate human chorionic villi during confrontation cultures with placental explants in hanging drops. We conclude that HTR8/SVneo spheroid cells possess progenitor cell traits that are probably attained through corruption of “stemness-” associated transcription factor networks. Furthermore, trophoblastic cells are highly prone to unspecific binding, which is resistant to conventional blocking methods, but which can be alleviated through blockage with human serum. PMID:23586024

  8. A Brucella melitensis high-temperature-requirement A (htrA) deletion mutant is attenuated in goats and protects against abortion.

    PubMed

    Phillips, R W; Elzer, P H; Robertson, G T; Hagius, S D; Walker, J V; Fatemi, M B; Enright, F M; Roop, R M

    1997-01-01

    It has been previously demonstrated that a Brucella melitensis high-temperature-requirement A (htrA) deletion mutant is more susceptible to oxidative killing in vitro than the parental strain and is attenuated in mice. To evaluate the contribution of the B melitensis HtrA protease to virulence in ruminants, the capacity of the B melitensis htrA mutant RWP5 to produce abortion in goats was compared to that of the virulent parental strain 16M. Experimental infection with strain 16M caused abortion in eight of 12 pregnant nannies, while none of the 12 nannies inoculated with RWP5 aborted. Furthermore, intramuscular injection of fetuses in utero with RWP5 led to colonisation of the fetus with subsequent colonisation of the nanny, but no abortion was observed. Nannies vaccinated with RWP5 showed complete protection against abortion when challenged with 16M during the third trimester of pregnancy. However, these animals were not protected from colonisation by 16M. The results presented here clearly indicate that the B melitensis htrA gene product contributes to pathogenesis in goats, but the utility of B melitensis htrA mutants as vaccines in this host appears to be limited.

  9. Quantitative relationship between functionally active telomerase and major telomerase components (hTERT and hTR) in acute leukaemia cells.

    PubMed

    Ohyashiki, J H; Hisatomi, H; Nagao, K; Honda, S; Takaku, T; Zhang, Y; Sashida, G; Ohyashiki, K

    2005-05-23

    Functionally active telomerase is affected at various steps including transcriptional and post-transcriptional levels of major telomerase components (hTR and human telomerase reverse transcriptase (hTERT)). We therefore developed a rapid and sensitive method to quantify hTERT and its splicing variants as well as the hTR by a Taqman real-time reverse transcriptase-polymerase chain reaction to determine whether their altered expression may contribute to telomere attrition in vivo or not. Fresh leukaemia cells obtained from 38 consecutive patients were used in this study. The enzymatic level of telomerase activity measured by TRAP assay was generally associated with the copy numbers of full-length hTERT+alpha+beta mRNA (P=0.0024), but did not correlate with hTR expression (P=0.6753). In spite of high copy numbers of full-length hTERT mRNA, telomerase activity was low in some cases correlating with low copy numbers of hTR, raising the possibility that alteration of the hTR : hTERT ratio may affect functionally active telomerase activity in vivo. The spliced nonactive hTERT mRNA tends to be lower in patients with high telomerase activity, suggesting that this epiphenomenon may play some role in telomerase regulation. An understanding of the complexities of telomerase gene regulation in biologically heterogeneous leukaemia cells may offer new therapeutic approaches to the treatment of acute leukaemia.

  10. Telomerase inhibition strategies by siRNAs against either hTR or hTERT in oral squamous cell carcinoma.

    PubMed

    Li, Y; Li, M; Yao, G; Geng, N; Xie, Y; Feng, Y; Zhang, P; Kong, X; Xue, J; Cheng, S; Zhou, J; Xiao, L

    2011-05-01

    Human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) are considered effective molecular targets for current anticancer therapy. In this study, we investigated the therapeutic effects of targeting hTR and hTERT individually or in combination by recombinant adenovirus-delivered small interfering RNA (siRNA) in oral squamous cell carcinoma (OSCC) Tca8113. Further, we screened the optimal strategy for RNA interference. Our results show that these different recombinant adenoviruses specifically reduced the levels of hTR mRNA, hTERT mRNA, hTERT protein and telomerase activity in Tca8113 cells. Moreover, they successfully inhibited xenograft tumor growth in nude mice. The potency of their antitumor activities was ranked as follows: anti-hTR >anti-hTR+anti-hTERT >anti-hTERT. Therefore, we demonstrated that the siRNA-expressing recombinant adenoviruses were an effective anticancer tool for treatment of OSCC. Furthermore, the anticancer effect of solely targeting hTR was more direct and efficient, compared with the effect of targeting hTR and hTERT in combination, or hTERT exclusively. The mechanism of this anticancer effect in OSCC was not only related to the inhibition of cell proliferation and the induction of cell apoptosis, but might also involve the inhibition of tumor angiogenesis.

  11. The LD loop as an important structural element required for transmission of the allosteric signal in the HtrA (DegP) protease from Escherichia coli.

    PubMed

    Figaj, Donata; Gieldon, Artur; Bartczak, Marlena; Koper, Tomasz; Zarzecka, Urszula; Lesner, Adam; Lipinska, Barbara; Skorko-Glonek, Joanna

    2016-09-01

    High-temperature requirement A (HtrA; DegP) from Escherichia coli, an important element of the extracytoplasmic protein quality-control system, is a member of the evolutionarily conserved family of serine proteases. The characteristic feature of this protein is its allosteric mode of activation. The regulatory loops, L3, L2, L1 and LD, play a crucial role in the transmission of the allosteric signal. Yet, the role of LD has not been fully elucidated. Therefore, we undertook a study to explain the role of the individual LD residues in inducing and maintaining the proteolytic activity of HtrA. We investigated the influence of amino acid substitutions located within the LD loop on the kinetics of a model substrate cleavage as well as on the dynamics of the oligomeric structure of HtrA. We found that the mutations that were expected to disturb the loop's structure and/or interactions with the remaining regulatory loops severely diminished the proteolytic activity of HtrA. The opposite effect, that is, increased activity, was observed for G174S substitution, which was predicted to strengthen the interactions mediated by LD. HtrAG174S protein had an equilibrium shifted toward the active enzyme and formed preferentially high-order oligomeric forms.

  12. Prediction of psychosis onset in Alzheimer disease: the role of depression symptom severity and the HTR2A T102C polymorphism.

    PubMed

    Wilkosz, Patricia A; Kodavali, Chowdari; Weamer, Elise A; Miyahara, Sachiko; Lopez, Oscar L; Nimgaonkar, Vishwajit L; DeKosky, Steven T; Sweet, Robert A

    2007-12-05

    Psychotic symptoms in Alzheimer disease (AD + P) identify a heritable phenotype associated with a more severe course. We recently found an association of AD + P with depression symptom severity. Reports have shown an association of a serotonin-2A receptor (HTR2A) gene T102C polymorphism with AD + P and with depression during AD. We examined the interaction of this common genetic polymorphism with depression and increased psychosis risk. Subjects with possible or probable AD or mild cognitive impairment (MCI) without psychosis at study entry were genotyped for the HTR2A T102C polymorphism and reassessed every 6 months until psychosis onset. Psychotic and depressive symptoms were rated using the CERAD behavioral rating scale (CBRS). Cox proportional hazard models with time-dependent covariates were used to examine associations with psychosis onset. A total of 324 Caucasian subjects completed at least one follow-up exam. Depressive symptom severity was a strong predictor of psychosis onset. Neither psychosis onset nor depression severity was associated with the HTR2A genotype. Genotype interacted with depression severity to moderate the risk of AD + P onset. This did not result from an interaction of HTR2A genotype with antidepressant use. Psychosis onset in AD is strongly associated with severity of depressive symptoms, an association that may be modified by HTR2A genotype.

  13. Prediction of Psychosis Onset in Alzheimer Disease: The Role of Depression Symptom Severity and the HTR2A T102C Polymorphism

    PubMed Central

    Wilkosz, Patricia A.; Kodavali, Chowdari; Weamer, Elise A.; Miyahara, Sachiko; Lopez, Oscar L.; Nimgaonkar, Vishwajit L.; DeKosky, Steven T.; Sweet, Robert A.

    2009-01-01

    Background Psychotic symptoms in Alzheimer Disease (AD+P) identify a heritable phenotype associated with a more severe course. We recently found an association of AD+P with depression symptom severity. Reports have shown an association of a serotonin-2A receptor (HTR2A) gene T102C polymorphism with AD+P and with depression during AD. We examined the interaction of this common genetic polymorphism with depression and increased psychosis risk. Methods Subjects with possible or probable AD or Mild Cognitive Impairment (MCI) without psychosis at study entry were genotyped for the HTR2A T102C polymorphism and reassessed every 6 months until psychosis onset. Psychotic and depressive symptoms were rated using the CERAD Behavioral Rating Scale (CBRS). Cox proportional hazard models with time dependent covariates were used to examine associations with psychosis onset. Results A total of 324 Caucasian subjects completed at least one follow-up exam. Depressive symptom severity was a strong predictor of psychosis onset. Neither psychosis onset nor depression severity was associated with the HTR2A genotype. Genotype interacted with depression severity to moderate the risk of AD+P onset. This did not result from an interaction of HTR2A genotype with antidepressant use. Conclusion Psychosis onset in AD is strongly associated with severity of depressive symptoms, an association that may be modified by HTR2A genotype. PMID:17525976

  14. Comparative analysis of the invasion-associated genes expression pattern in first trimester trophoblastic (HTR-8/SVneo) and JEG-3 choriocarcinoma cells.

    PubMed

    Suman, P; Gupta, S K

    2012-10-01

    Several cellular models of trophoblast have been proposed to understand their invasion. We had reported that JEG-3 and HTR-8/SVneo cells show differential invasive behavior in response to IL-11 treatment. So, the present study aims to compare the expression of invasion-associated molecules in these two cell lines by performing cDNA microarray followed by quantitative RT-PCR. We have observed that HTR-8/SVneo cells have significantly higher invasiveness than JEG-3 cells, which might be due to higher expression of proteases and signaling intermediates of JAK/STAT and MAPK signaling pathways. Like extravillous trophoblasts (EVTs), a higher expression of functionally significant proteases like MMP1, MMP2, MMP9, PLAU etc in HTR-8/SVneo cells, project them as a close mimic of EVTs under in vitro conditions.

  15. Influence of HTR core inlet and outlet temperatures on hydrogen generation efficiency using the sulfur-iodine water-splitting cycle

    SciTech Connect

    Buckingham, Robert; Brown, Lloyd; Russ, Ben; Lovera, Patrick; Carles, Philippe; Borgard, Jean-Marc; Yvon, Pascal

    2012-04-15

    The performance of hydrogen production via thermochemical cycles is typically evaluated using thermal efficiency. In this study, the sulfur-iodine cycle with heat supplied by a high-temperature reactor (HTR) is analyzed. Two cases are examined: one flow sheet designed by General Atomics in the United States, the other by Commissariat a l'energie atomique et aux energies alternatives in France. In each case, HTR helium inlet and outlet temperatures are specified. Differences in these temperature specifications lead to process variations between the limy sheets and in how the hydrogen processes interface with the nuclear heat source. Two principal conclusions result from the analysis. First, the thermal efficiency tends to plateau above a certain outlet helium temperature. This is a characteristic effect of the method of Ozturk et al. for sulfuric acid decomposition. Second, it is clear that it is impractical to discuss efficiencies for the hydrogen process that are independent of defined operating parameters of the HTR. (authors)

  16. No mutations in the serotonin related TPH1 and HTR1B genes in patients with monogenic sclerosing bone disorders.

    PubMed

    Boudin, Eveline; Jennes, Karen; de Freitas, Fenna; Tegay, David; Mortier, Geert; Van Hul, Wim

    2013-07-01

    Since the identification of LRP5 as the causative gene for the osteoporosis pseudoglioma syndrome (OPPG) as well as the high bone mass (HBM) phenotype, LRP5 and the Wnt/β-catenin signaling have been extensively studied for their role in the differentiation and proliferation of osteoblasts, in the apoptosis of osteoblasts and osteocytes and in the response of bone to mechanical loading. However, more recently the direct effect of LRP5 on osteoblasts and bone formation has been questioned. Gene expression studies showed that mice lacking lrp5 have increased expression of tph1, the rate limiting enzyme for the production of serotonin in the gut. Furthermore mice lacking either tph1 or htr1B, the receptor for serotonin on the osteoblasts, were reported to have an increased bone mass due to increased bone formation. This led to the still controversial hypothesis that LRP5 influences bone formation indirectly by regulating the expression of thp1 and as a consequence influencing the production of serotonin in the gut. Based on these data we decided to evaluate the role of TPH1 and HTR1B in the development of craniotubular hyperostoses, a group of monogenic sclerosing bone dysplasias. We screened the coding regions of both genes in 53 patients lacking a mutation in the known causative genes LRP5, LRP4 and SOST. We could not find disease-causing coding variants in neither of the tested genes and therefore, we cannot provide support for an important function of TPH1 and HTR1B in the pathogenesis of sclerosing bone dysplasias in our tested patient cohort.

  17. HTR1B gene variants associate with the susceptibility of Raynauds' phenomenon in workers exposed hand-arm vibration.

    PubMed

    Chen, Qingsong; Lang, Li; Xiao, Bin; Lin, Hansheng; Yang, Aichu; Li, Hongling; Tang, Shichuan; Huang, Hanlin

    2016-10-05

    To explore whether polymorphic variants of the HTR1B gene are associated with the susceptibility of Raynauds' Phenomenon (RP) coursed by vibration. 148 subjects exposed to vibration for more than 2 years were classified into either induced white finger (VWF) group (n = 72), or non-VWF group (n = 76). Vibration exposure levels were measured and assessed following ISO 5349-1:2001 protocol. All workers were genotyped by sequencing for the single nucleotide polymorphisms (SNPs) in the 5'-flanking and coding region of HTR1B. Genetic characteristics and linkage disequilibrium (LD) were analyzed with Haploview. Serum serotonin levels of each subject were detected using ELISA. The association between the susceptibility of vascular damage and genotype was analyzed via logistic regression. 7 known SNPs were obtained and their allele frequencies were inserted into the Hardy-Weinberg equilibrium. rs6297 variant genotype had an increased risk of VWF compared with wild genotype (OR = 2.14, 95% CI = 1.04- 4.58, P < 0.05). rs6298 mutant type (AG+GG) was found to have a significant interaction on vibration exposure LN(CEI), accounting for VWF occurrence. LN(5-HT) level is significantly different between the VWF group (x¯±s= 1.99±1.09 ng/mL) and the non-VWF group (x¯±s= 2.72±1.47 ng/mL). Serotonin levels may affect the progression of secondary RP. Polymorphic variants of the HTR1B gene are associated with the susceptibility of secondary RP in vibration-exposed occupational populations of Chinese Han people.

  18. Inhibition of telomerase RNA (hTR) in cervical cancer by adenovirus-delivered siRNA.

    PubMed

    Li, Y; Li, H; Yao, G; Li, W; Wang, F; Jiang, Z; Li, M

    2007-08-01

    Small interfering RNA (siRNA) has become a powerful tool for selectively silencing gene expression in cultured mammalian cells. In this study, a 67-bp oligonucleotide encoding human telomerase RNA (hTR) was introduced into pSIREN, a shuttle vector for construction of recombinant adenovirus. Then the U6-RNA promoter and siRNA-encoding insert were cut out from the pSIREN and subcloned into pAdeno-X to construct the plasmid pAd-hTR. After the pAd-hTR was transfected into a mammalian cell line HEK-293, adenovirus carrying the hTR-targeting siRNA (Ad-hTR-siRNA) was obtained. We performed a series of experiments to demonstrate silencing of the telomerase mediated by Ad-hTR-siRNA in HeLa cells. Compared with control virus (Ad-NT-siRNA), Ad-hTR-siRNA significantly reduced both hTR mRNA level (by 70.21%) and telomerase activity (by 58.87%) in HeLa cells. Moreover, it induced apoptosis in HeLa cells. Treatment of subcutaneous tumor xenografted with Ad-hTR-siRNA could slow down tumor growth, at least partially due to the induction of apoptosis (P<0.05) in vivo. Taken together, our results demonstrated efficient and specific knockdown of telomerase in HeLa cell line by the hTR siRNA, and indicated the prospect of applying this siRNA expressing recombinant adenovirus system in cancer gene therapy.

  19. Lack of telomerase gene expression in alternative lengthening of telomere cells is associated with chromatin remodeling of the hTR and hTERT gene promoters.

    PubMed

    Atkinson, Stuart P; Hoare, Stacey F; Glasspool, Rosalind M; Keith, W Nicol

    2005-09-01

    The presence of active telomere maintenance mechanisms in immortal cells allows the bypass of senescence by maintaining telomere length. In most immortal cell lines and tumors, telomere maintenance is attributable to telomerase reactivation. However, a number of immortal cell lines and tumors can achieve telomere maintenance in the absence of detectable telomerase activity by the alternative lengthening of telomere (ALT) mechanism. Epigenetic mechanisms have been implicated in the regulation of telomerase expression. We show that specific modifications within the chromatin environment of the hTR and hTERT promoters correlate with expression of hTR and hTERT in ALT, normal and telomerase-positive tumor cell lines. Lack of expression of hTR and hTERT in ALT cell lines is associated with histone H3 and H4 hypoacetylation and methylation of Lys9 histone H3. Conversely, hTR and hTERT expression in telomerase-positive cell lines is associated with hyperacetylation of H3 and H4 and methylation of Lys4 H3. Methylation of Lys20 H4 was not linked to gene expression but instead was specific to the hTR and hTERT promoters of ALT cells. This may provide an insight into the differences between ALT and telomerase-positive cells as well as a novel marker for the ALT phenotype. Treatment of normal and ALT cells with 5-azadeoxycytidine in combination with Trichostatin A caused chromatin remodeling of both promoters and reactivation of hTR and hTERT expression in ALT and normal cell lines. This data establishes a definite link between the chromatin environment of the telomerase gene promoters and transcriptional activity.

  20. Mild hyperoxia limits hTR levels, telomerase activity, and telomere length maintenance in hTERT-transduced bone marrow endothelial cells.

    PubMed

    Napier, Christine E; Veas, Laura A; Kan, Chin-Yi; Taylor, Lisa M; Yuan, Jun; Wen, Victoria W; James, Alexander; O'Brien, Tracey A; Lock, Richard B; MacKenzie, Karen L

    2010-10-01

    Reactivation of telomerase in endothelial cells (ECs) may be an effective approach to the treatment of vascular disorders associated with telomere attrition and EC senescence. However, overexpression of human telomerase reverse transcriptase (hTERT) does not prevent net telomere loss in ECs grown in standard culture medium with exposure to atmospheric oxygen (21% O(2)). Since these culture conditions are hyperoxic relative to normal tissue in vivo, where oxygen tension is estimated to be 1%-6%, we examined the effects of reduced exposure to oxidative stress (OS) on telomere length maintenance in hTERT-transduced bone marrow endothelial (BMhTERT) cells. Propagation of BMhTERT cells in the free radical scavenger, tert-butylhydroxylamine (tBN), and/or in 5% O(2) increased telomerase enzyme activity and facilitated telomere length maintenance. The enhancement of telomerase activity correlated with higher levels of the telomerase RNA component (hTR). We also investigated the role of the telomere binding protein, TRF1, in telomere length regulation under alternate OS conditions. Inhibition of TRF1 function had no effect on telomere length in BMhTERT cells grown under standard culture conditions. However, alleviation of OS by growth in tBN plus 5% O(2), elevated hTR levels, enhanced telomerase enzyme activity, and enabled progressive telomere lengthening. The direct impact of hTR levels on telomerase-mediated telomere lengthening was demonstrated by overexpression of hTR. BMhTERT cells transduced with hTR exhibited very high telomerase enzyme activity and underwent dramatic telomere lengthening under standard culture conditions. Overall, these results demonstrate that hTR levels are reduced by mild hyperoxia and limit telomerase-mediated telomere lengthening in hTERT-transduced ECs.

  1. A large-scale genetic association study to evaluate the contribution of Omi/HtrA2 (PARK13) to Parkinson's disease.

    PubMed

    Krüger, Rejko; Sharma, Manu; Riess, Olaf; Gasser, Thomas; Van Broeckhoven, Christine; Theuns, Jessie; Aasly, Jan; Annesi, Grazia; Bentivoglio, Anna Rita; Brice, Alexis; Djarmati, Ana; Elbaz, Alexis; Farrer, Matthew; Ferrarese, Carlo; Gibson, J Mark; Hadjigeorgiou, Georgios M; Hattori, Nobutaka; Ioannidis, John P A; Jasinska-Myga, Barbara; Klein, Christine; Lambert, Jean-Charles; Lesage, Suzanne; Lin, Juei-Jueng; Lynch, Timothy; Mellick, George D; de Nigris, Francesa; Opala, Grzegorz; Prigione, Alessandro; Quattrone, Aldo; Ross, Owen A; Satake, Wataru; Silburn, Peter A; Tan, Eng King; Toda, Tatsushi; Tomiyama, Hiroyuki; Wirdefeldt, Karin; Wszolek, Zbigniew; Xiromerisiou, Georgia; Maraganore, Demetrius M

    2011-03-01

    High-profile studies have provided conflicting results regarding the involvement of the Omi/HtrA2 gene in Parkinson's disease (PD) susceptibility. Therefore, we performed a large-scale analysis of the association of common Omi/HtrA2 variants in the Genetic Epidemiology of Parkinson's disease (GEO-PD) consortium. GEO-PD sites provided clinical and genetic data including affection status, gender, ethnicity, age at study, age at examination (all subjects); age at onset and family history of PD (patients). Genotyping was performed for the five most informative SNPs spanning the Omi/HtrA2 gene in approximately 2-3 kb intervals (rs10779958, rs2231250, rs72470544, rs1183739, rs2241028). Fixed as well as random effect models were used to provide summary risk estimates of Omi/HtrA2 variants. The 20 GEO-PD sites provided data for 6378 cases and 8880 controls. No overall significant associations for the five Omi/HtrA2 SNPs and PD were observed using either fixed effect or random effect models. The summary odds ratios ranged between 0.98 and 1.08 and the estimates of between-study heterogeneity were not large (non-significant Q statistics for all 5 SNPs; I(2) estimates 0-28%). Trends for association were seen for participants of Scandinavian descent for rs2241028 (OR 1.41, p=0.04) and for rs1183739 for age at examination (cut-off 65 years; OR 1.17, p=0.02), but these would not be significant after adjusting for multiple comparisons and their Bayes factors were only modest. This largest association study performed to define the role of any gene in the pathogenesis of Parkinson's disease revealed no overall strong association of Omi/HtrA2 variants with PD in populations worldwide.

  2. Lack of association between the T-->C 267 serotonin 5-HT6 receptor gene (HTR6) polymorphism and prediction of response to clozapine in schizophrenia.

    PubMed

    Masellis, M; Basile, V S; Meltzer, H Y; Lieberman, J A; Sevy, S; Goldman, D A; Hamblin, M W; Macciardi, F M; Kennedy, J L

    2001-01-15

    The affinity of clozapine for 5-HT2A, 5-HT2C, 5-HT6, 5-HT7, and 5-HT1A receptors has been suggested to contribute to various aspects of its complex clinical actions. This study examined the hypothesis that genetic variation in 5-HT1A, 5-HT6, and 5-HT7 receptor genes is involved in the variability observed in response to clozapine. We employed a pharmacogenetic approach in a group (n=185) of schizophrenia patients that have been clinically well characterized for clozapine response. Polymorphisms in the 5-HT6 (HTR6), 5-HT1A (HTR1A) and 5-HT7 (HTR7) receptor genes were genotyped. No evidence for either an allelic or genotypic association of the T-->C 267 HTR6 polymorphism with response to clozapine was found in our sample (allele: chi(2)=0.06, 1 df, P=0.80; genotype: chi(2)=1.21, 2 df, P=0.55). The pro16leu HTR1A polymorphism was not observed in our sample; all individuals genotyped were pro/pro 16 homozygotes. With respect to the pro279leu HTR7 polymorphism, one Caucasian male responder to clozapine was observed to be heterozygous (pro/leu 279 genotype). This individual was clinically similar to the other clozapine responders. Overall, our findings do not support a role for the T-->C 267 polymorphism of the 5-HT6 receptor gene in response to clozapine, although replication is required to confirm this finding.

  3. Serine protease Omi/HtrA2 targets WARTS kinase to control cell proliferation.

    PubMed

    Kuninaka, S; Iida, S-I; Hara, T; Nomura, M; Naoe, H; Morisaki, T; Nitta, M; Arima, Y; Mimori, T; Yonehara, S; Saya, H

    2007-04-12

    The serine protease Omi/HtrA2 was initially regarded as a proapoptotic molecule that proteolyses several proteins to induce cell death. Recent studies, however, indicate that loss of Omi protease activity increases susceptibility to stress-induced cell death. These complicated findings suggest that the protease activity of Omi is involved not only in apoptosis but also in cellular homeostasis. However, the targets which Omi uses to mediate this novel process are unknown. Previously, we showed that WARTS (WTS)/large tumor-suppressor 1 mitotic kinase interacts with the protein/discs-large protein/zonula (PDZ) domain of Omi and promotes its protease activity. We now report that WTS is a substrate for Omi protease activity, thus it is not only a regulator but also a downstream target of this protease. Interaction with Omi PDZ domain is required for WTS to be proteolysed. When caspase-9-deficient mouse embryonic fibroblasts (MEFs) were treated with staurosporine, WTS was proteolysed by activated endogenous Omi without induction of cell death. Therefore, protease activity of Omi and proteolysis of WTS are not necessarily required for cell death. We found that depletion of Omi from HeLa cells results in accelerated cell proliferation despite no significant change in the duration of mitosis. The depletion of WTS showed the same effect on S phase progression. Therefore, WTS proteolytic fragment(s) generated by Omi may act as an inhibitor of G1/S progression. Our data reveal a role for Omi-mediated processing of WTS in negative regulation of cell cycle progression at interphase, suggesting a novel function of Omi other than apoptosis.

  4. Gamma dose rate estimation and operation management suggestions for decommissioning the reactor pressure vessel of HTR-PM

    SciTech Connect

    Sheng Fang; Hong Li; Jianzhu Cao; Wenqian Li; Feng Xie; Jiejuan Tong

    2013-07-01

    China is now designing and constructing a high temperature gas cooled reactor-pebble bed module (HTR-PM). In order to investigate the future decommissioning approach and evaluate possible radiation dose, gamma dose rate near the reactor pressure vessel was calculated for different cooling durations using QAD-CGA program. The source term of this calculation was provided by KORIGEN program. Based on the calculated results, the spatial distribution and temporal changes of gamma dose rate near reactor pressure vessel was systematically analyzed. A suggestion on planning decommissioning operation of reactor pressure vessel of HTRPM was given based on calculated dose rate and the Chinese Standard GB18871-2002. (authors)

  5. Innovative non-destructive evaluation methods on HTR fuel at AREVA NP: towards a 100% non invasive control strategy

    SciTech Connect

    Banchet, J.; Tisseur, D.; Hermosilla Lara, S.; Piriou, M.; Bargain, R.; Guillermier, P.

    2007-07-01

    High Temperature Reactor (HTR) fuel consists in millimetric multilayered particles called TRISO, embedded, depending on the reactor design, in a pebble or cylinder-shaped graphite matrix called compact. Particles are typically composed of a 500 {mu}m fissile material kernel, a 95 {mu}m porous carbon layer called buffer, a 40 {mu}m dense pyrolytic carbon layer, a 35 {mu}m silicon carbide layer and another 40 {mu}m dense pyrolytic carbon layer. In order to ensure fuel qualification, as well as reactor safety, particles and compacts need to satisfy specifications concerning their physical characteristics and their integrity. In particular, geometrical parameters such as particle diameter and sphericity as well as layers thickness, but also layers density and the absence of structural defects such as cracks or de-cohesions need to be detected and characterized. In the past, a huge R and D work was carried out to build a TRISO particle characterization quality control plan, mainly based on particle sampling as well as destructive characterization methods. However, since then, development of industrial non-destructive evaluation techniques and devices contributed to envisage not only a non invasive control of HTR fuel, but also a 100% production control strategy. Since 2004, AREVA NP is engaged in a R and D program aiming at the development of innovative industrial nondestructive evaluation methods for HTR fuel. After investigating a number of potential techniques, some of them were selected based on their performances and/or their industrial potential. In particular, development has been carried out on high resolution X-Ray imaging allowing accurate layer thickness, layer density and structural defects characterization, X-Ray tomography offering the possibility to characterize fuel element homogeneity and determine the number of in-contact particles contained in a fuel element, infrared thermal imaging (ITI) allowing cracks detection, eddy currents (EC) enabling

  6. Quantitative Homogeneity and In-Contact Particles of High Temperature Reactors (htr) Compacts Determination via X-Ray Tomography

    NASA Astrophysics Data System (ADS)

    Lecomte, G.; Tisseur, D.; Létang, J. M.; Banchet, J.; Vitali, M. P.

    2008-02-01

    In AREVA Nuclear Power's High Temperature Reactor (HTR) design called ANTARES, fuel consists of compacts composed of few thousands millimetric quasi-spherical particles dispersed in a graphite matrix. Compact homogeneity, defined as the homogeneous particles spatial distribution in the matrix, as well as the possibility of obtaining particles in contact, need to be assessed since they condition the thermo-mechanical behavior of the nuclear fuel under irradiation. In this paper, image and data processing algorithms are developed to do so, based on X-Ray tomographic images.

  7. Assessment of safety and availability of dissimilar weldments in the water-steam circuit of HTR plants

    NASA Astrophysics Data System (ADS)

    Schneider, Klaus; Gnirss, Günther; Grünling, Hermann

    1990-04-01

    A concept for the assessment of dissimilar weldments in HTR steam piping is described. A combination of experiments and calculation shows that dissimilar metal welds between the ferritic steel X 20 CrMoV 21 1 and the austenitic alloy X 10 NiCrAlTi 32 20 can be operated for the design life of the component. The combination of reliable manufacturing and nondestructive testing assures that defects in weld joints will show only negligible crack growth. The leak before break criterion is fulfilled. The concept of assessment of dissimilar weldments is verified by the component test MINERVA.

  8. Infrared Flash Thermography Applied to the External Defect Detection on Htr Fuel Particles during the Manufacturing Process

    NASA Astrophysics Data System (ADS)

    Hermosilla-Lara, S.; Piriou, M.; Levesque, P.

    2008-02-01

    The authors propose the use of an adapted flash thermography method to perform the online detection of open cracks on high temperature reactor (HTR) fuel. The method consists, via an infrared camera, in the measurement of the particle temperature increase produced by a heating pulse on the particle external surface. Study is performed on laser notches and real cracks located in the different particle layers. It includes the influence of the defect relative position with respect to the heating source, as well as the influence of operating conditions. It is shown that this technique is able to yield good discrimination between sound and flawed particles at a rate that meets production constraints.

  9. Serine protease HtrA1 accumulates in corneal transforming growth factor beta induced protein (TGFBIp) amyloid deposits

    PubMed Central

    Karring, Henrik; Poulsen, Ebbe Toftgaard; Runager, Kasper; Thøgersen, Ida B.; Klintworth, Gordon K.; Højrup, Peter

    2013-01-01

    Purpose Specific mutations in the transforming growth factor beta induced (TGFBI) gene are associated with lattice corneal dystrophy (LCD) type 1 and its variants. In this study, we performed an in-depth proteomic analysis of human corneal amyloid deposits associated with the heterozygous A546D mutation in TGFBI. Methods Corneal amyloid deposits and the surrounding corneal stroma were procured by laser capture microdissection from a patient with an A546D mutation in TGFBI. Proteins in the captured corneal samples and healthy corneal stroma were identified with liquid chromatography-tandem mass spectrometry and quantified by calculating exponentially modified Protein Abundance Index values. Mass spectrometry data were further compared for identifying enriched regions of transforming growth factor beta induced protein (TGFBIp/keratoepithelin/βig-h3) and detecting proteolytic cleavage sites in TGFBIp. Results A C-terminal fragment of TGFBIp containing residues Y571-R588 derived from the fourth fasciclin 1 domain (FAS1–4), serum amyloid P-component, apolipoprotein A-IV, clusterin, and serine protease HtrA1 were significantly enriched in the amyloid deposits compared to the healthy cornea. The proteolytic cleavage sites in TGFBIp from the diseased cornea are in accordance with the activity of serine protease HtrA1. We also identified small amounts of the serine protease kallikrein-14 in the amyloid deposits. Conclusions Corneal amyloid caused by the A546D mutation in TGFBI involves several proteins associated with other varieties of amyloidosis. The proteomic data suggest that the sequence 571-YHIGDEILVSGGIGALVR-588 contains the amyloid core of the FAS1–4 domain of TGFBIp and point at serine protease HtrA1 as the most likely candidate responsible for the proteolytic processing of amyloidogenic and aggregated TGFBIp, which explains the accumulation of HtrA1 in the amyloid deposits. With relevance to identifying serine proteases, we also found glia-derived nexin

  10. [Development of noninvasive bladder cancer diagnosis on basis of telomerase and it's subunits hTERT and hTR detection].

    PubMed

    Glukhov, A I; Grigorieva, Y E; Gordeev, S A; Vinarov, A Z; Potoldykova, N V

    2015-01-01

    Telomerase activity (TA) and expression of genes coding it's subunits (hTERT and hTR) have been examined in tumor tissue and urine sediment samples taken from patients with bladder cancer (BC) using the modified TRAP assay (in the case of telomerase detection) and RT-PCR (in the case of hTERT and hTR expression). Results obtained in this study demonstrate possibility of noninvasive diagnosis of BC with sensitivity of 96% and specificity of 100% in the case of telomerase detection and with sensitivity of 80% and specificity of 100% in the case of hTERT detection in urine sediment samples.

  11. Detektion von fahrspuren und kreuzungen auf nichtmarkierten straen zum autonomen führen von fahrzeugen

    NASA Astrophysics Data System (ADS)

    Vacek, Stefan; Bürkle, Cornelius; Schröder, Joachim; Dillmann, Rüdiger

    Das Wissen über Position und Verlauf der Straße ist eine der wichtigsten Informationen, die zum Führen autonomer Straßenfahrzeuge benötigt wird. Die meisten Arbeiten gehen davon aus, dass Markierungen auf der Straße vorhanden sind, die die Erkennung enorm erleichtern. Üblicherweise werden die Fahrbahnränder detektiert und die Fahrspur mit Hilfe eines Kaiman-Filters geschätzt [1]. Andere Arbeiten verwenden zusätzlich die Straßenfarbe und kombinieren die verschiedenen Hinweise in einem Partikel-Filter [2]. Ein allgemeiner Überblick über Verfahren zur Fahrspurdetektion findet sich in [3].

  12. Differential effect of DDT, DDE, and DDD on COX-2 expression in the human trophoblast derived HTR-8/SVneo cells.

    PubMed

    Dominguez-Lopez, Pablo; Diaz-Cueto, Laura; Olivares, Aleida; Ulloa-Aguirre, Alfredo; Arechavaleta-Velasco, Fabian

    2012-11-01

    The purpose of this study was to investigate the effect of 1,1,1-trichloro-2,2-bis-(chlorophenyl)ethane (DDT), 1,1-bis-(chlorophenyl)-2,2-dichloroethene (DDE), and 1,1-dichloro-2,2-bis(chlorophenyl)ethane (DDD) isomers on COX-2 expression in a human trophoblast-derived cell line. Cultured HTR-8/SVneo trophoblast cells were exposed to DDT isomers and its metabolites for 24 h, and COX-2 mRNA and protein expression were assessed by RT-PCR, Western blotting, and ELISA. Prostaglandin E₂ production was also measured by ELISA. Both COX-2 mRNA and protein were detected under control (unexposed) conditions in the HTR-8/SVneo cell line. COX-2 protein expression and prostaglandin E₂ production but not COX-2 mRNA levels increased only after DDE and DDD isomers exposure. It is concluded that DDE and DDD exposure induce the expression of COX-2 protein, leading to increased prostaglandin E2 production. Interestingly, the regulation of COX-2 by these organochlorines pesticides appears to be at the translational level. © 2012 Wiley Periodicals, Inc.

  13. Optimization of Plasmid Maintenance in the Attenuated Live Vector Vaccine Strain Salmonella typhi CVD 908-htrA†

    PubMed Central

    Galen, James E.; Nair, Jay; Wang, Jin Yuang; Wasserman, Steven S.; Tanner, Michael K.; Sztein, Marcelo B.; Levine, Myron M.

    1999-01-01

    The broad objective of the research presented here is to develop a noncatalytic plasmid maintenance system for the stabilization of multicopy expression plasmids encoding foreign antigens in a Salmonella typhi live-vector vaccine strain such as CVD 908-htrA. We have enhanced the maintenance of expression plasmids at two independent levels. First, we removed dependence upon balanced-lethal maintenance systems that involve catalytic enzymes expressed from multicopy plasmids; we accomplished this through incorporation into expression plasmids of a postsegregational killing system based on the noncatalytic hok-sok plasmid addiction system from the antibiotic resistance factor pR1. We also included at least one naturally occurring plasmid partition function in our expression plasmids, which eliminates random segregation of these plasmids, thereby enhancing their inheritance and stability; to accomplish this, we incorporated either the par locus from pSC101, the parA locus from pR1, or both. We monitored the stability of optimized expression plasmids within CVD 908-htrA by quantitating expression of a variant of green fluorescent protein (GFPuv) by using flow cytometry. In this report, we demonstrate the utility of this novel plasmid maintenance system in enhancing the stability of our expression plasmids and go on to show that as the copy number of stabilized plasmids increases, the toxicity of GFPuv synthesis also increases. The implications of these observations for the rational design of immunogenic and protective bacterial live vector vaccines are discussed. PMID:10569759

  14. The Three Streptomyces lividans HtrA-Like Proteases Involved in the Secretion Stress Response Act in a Cooperative Manner

    PubMed Central

    Vicente, Rebeca L.; Gullón, Sonia; Marín, Silvia; Mellado, Rafael P.

    2016-01-01

    Overproduction of Sec-proteins in S. lividans accumulates misfolded proteins outside of the cytoplasmic membrane where the accumulated proteins interfere with the correct functioning of the secretion machinery and with the correct cell functionality, triggering the expression in S. lividans of a CssRS two-component system which regulates the degradation of the accumulated protein, the so-called secretion stress response. Optimization of secretory protein production via the Sec route requires the identification and characterisation of quality factors involved in this process. The phosphorylated regulator (CssR) interacts with the regulatory regions of three genes encoding three different HtrA-like proteases. Individual mutations in each of these genes render degradation of the misfolded protein inoperative, and propagation in high copy number of any of the three proteases encoding genes results on indiscriminate alpha-amylase degradation. None of the proteases could complement the other two deficiencies and only propagation of each single copy protease gene can restore its own deficiency. The obtained results strongly suggest that the synthesis of the three HtrA-like proteases needs to be properly balanced to ensure the effective degradation of misfolded overproduced secretory proteins and, at the same time, avoid negative effects in the secreted proteins and the secretion machinery. This is particularly relevant when considering the optimisation of Streptomyces strains for the overproduction of homologous or heterologous secretory proteins of industrial application. PMID:27977736

  15. Serotonergic genes (5-HTT and HTR1A) and separation life events: gene-by-environment interaction for panic disorder.

    PubMed

    Choe, Ah Young; Kim, Borah; Lee, Kang Soo; Lee, Ji Eun; Lee, Jun-Yeob; Choi, Tai Kiu; Lee, Sang-Hyuk

    2013-01-01

    The polymorphisms of serotonergic genes (5-HTTLPR and HTR1A rs6295) and separation life events have been studied to find an association with panic disorder, respectively. However, there are no studies that have yet evaluated the interaction effect between serotonergic genes and separation life events for panic disorder. For this study, 194 panic disorder patients and 172 healthy controls were included for genotyping and environmental factor analysis. Separation life events were assessed using the Stressful Life Events Scale and clinical interviews. To evaluate the potential endophenotypes of panic disorder, the Anxiety Sensitivity Index-revised (ASI-R), harm avoidance in the Temperament and Character Inventory (HA), and neuroticism in the Eysenck Personality Questionnaire (neuroticism) scales were administered. For 5-HTTLPR and HTR1A rs6295, there was no significant main effect of each genotype on panic disorder alone. However, the number of separation life events and their interaction with 5-HTTLPR showed a statistically significant effect on panic disorder. In addition, the interaction between 5-HTTLPR and the number of separation life events significantly affected the HA for potential endophenotypes. This study could suggest the effect of the interaction between 5-HTTLPR and separation life events on panic disorder and its potential endophenotype. Copyright © 2013 S. Karger AG, Basel.

  16. EPHB4 Regulates Human Trophoblast Cell Line HTR-8/SVneo Function: Implications for the Role of EPHB4 in Preeclampsia.

    PubMed

    Liu, Xiaoping; Hu, Ying; Zheng, Yanfang; Liu, Xiaoxia; Luo, Minglian; Liu, Weifang; Zhao, Yin; Zou, Li

    2016-09-01

    Successful pregnancy depends on well-regulated extravillous trophoblast (EVT) invasion into the uterine decidua and moderate uterine spiral artery remodeling. Ephrin receptor B4 (EPHB4) is a membrane-anchored receptor tyrosine kinase that plays an important role in various cellular functions in human normal tissue and tumors. Reportedly, EPHB4 plays important roles during placentation. Still, there is no investigation of EPHB4 modulating trophoblast function. In our study, term placentas of preeclamptic pregnancies showed a significantly increased EPHB4 expression compared to those of uncomplicated pregnancies (n = 15). Exogenous up-regulation of EPHB4 in HTR-8/SVneo cells was performed to investigate the effects of EPHB4 on cell biological behavior. The results showed that EPHB4 enhancement reduced cell proliferation and promoted trophoblast apoptosis; and inhibited cell migration, invasion, and endothelial replacement. Associated factors, such as matrix metalloproteinases, vascular endothelial growth factor, placental growth factor, and soluble Fms-like tyrosine kinase 1 were examined at transcriptional level. Furthermore, cell functional results were confirmed in a placenta-decidua coculture system, showing poor vascular remodeling. Additionally, we detected possible down-stream PI3K-Akt signal pathway involved in EPHB4-mediated function of HTR-8/SVneo cells. Our study demonstrates that EPHB4 overexpression may contribute to trophoblasts dysfunction and impair maternal artery remodeling, as is associated with the pathogenesis of preeclampsia.

  17. The Three Streptomyces lividans HtrA-Like Proteases Involved in the Secretion Stress Response Act in a Cooperative Manner.

    PubMed

    Vicente, Rebeca L; Gullón, Sonia; Marín, Silvia; Mellado, Rafael P

    2016-01-01

    Overproduction of Sec-proteins in S. lividans accumulates misfolded proteins outside of the cytoplasmic membrane where the accumulated proteins interfere with the correct functioning of the secretion machinery and with the correct cell functionality, triggering the expression in S. lividans of a CssRS two-component system which regulates the degradation of the accumulated protein, the so-called secretion stress response. Optimization of secretory protein production via the Sec route requires the identification and characterisation of quality factors involved in this process. The phosphorylated regulator (CssR) interacts with the regulatory regions of three genes encoding three different HtrA-like proteases. Individual mutations in each of these genes render degradation of the misfolded protein inoperative, and propagation in high copy number of any of the three proteases encoding genes results on indiscriminate alpha-amylase degradation. None of the proteases could complement the other two deficiencies and only propagation of each single copy protease gene can restore its own deficiency. The obtained results strongly suggest that the synthesis of the three HtrA-like proteases needs to be properly balanced to ensure the effective degradation of misfolded overproduced secretory proteins and, at the same time, avoid negative effects in the secreted proteins and the secretion machinery. This is particularly relevant when considering the optimisation of Streptomyces strains for the overproduction of homologous or heterologous secretory proteins of industrial application.

  18. HTR2 Receptors in a Songbird Premotor Cortical-Like Area Modulate Spectral Characteristics of Zebra Finch Song

    PubMed Central

    Wood, William E.; Roseberry, Thomas K.; Perkel, David J.

    2013-01-01

    Serotonin [5-hydroxytryptamine (5-HT)] is involved in modulating an array of complex behaviors including learning, depression, and circadian rhythms. Additionally, HTR2 receptors on layer V pyramidal neurons are thought to mediate the actions of psychedelic drugs; the native function of these receptors at this site, however, remains unknown. Previously, we found that activation of HTR2 receptors in the zebra finch forebrain song premotor structure the robust nucleus of the arcopallium (RA) led to increased excitation, and that endogenous 5-HT could roughly double spontaneous firing rate. Here, using in vivo single-unit recordings, we found that direct application of 5-HT to these same RA projection neurons, which are analogous to layer V cortical pyramidal neurons, caused a significant increase in the number of action potentials per song-related burst, and a dramatic decrease in signal-to-noise ratio. Injection of the serotonergic neurotoxin 5,7-dihydroxytryptamine into the third ventricle greatly reduced telencephalic 5-HT and resulted in decreased fundamental frequency of harmonic syllables as well as increased goodness of pitch. Both of these results can be explained by the observed actions of 5-HT on RA projection neurons, and both effects recovered to baseline within 2 weeks following the toxin injection. These results show that 5-HT is involved in modulating spectral properties of song, likely via effects on RA projection neurons, but that adult zebra finches can partially compensate for this deficit within 7 d. PMID:23407949

  19. Inhibition of HtrA2/Omi ameliorates heart dysfunction following ischemia/reperfusion injury in rat heart in vivo.

    PubMed

    Bhuiyan, Md Shenuarin; Fukunaga, Kohji

    2007-02-28

    High temperature requirement A2 (HtrA2)/Omi is a mitochondrial serine protease that is released into the cytosol from mitochondria and in turn promotes caspase activation by proteolyzing inhibitor of apoptosis proteins. Here we asked whether treatment with an HtrA2/Omi inhibitor, 5-[5-(2-nitrophenyl)furfuryliodine]-1,3-diphenyl-2-thiobarbituric acid (UCF-101), restores heart dysfunction following ischemia/reperfusion injury in vivo. Rats underwent a 30-min ischemia by occluding the left anterior descending artery, followed by 24 h reperfusion. UCF-101 (0.75 or 1.5 micromol/kg, i.p.) was administered 10 min before reperfusion. UCF-101 treatment significantly recovered the mean arterial blood pressure and ameliorated contractile dysfunction of the left ventricle 72 h after reperfusion with concomitant reduction of infarct size. Cardio-protection mediated by UCF-101 was correlated with reduced X-linked inhibitor of apoptosis protein (XIAP) degradation and inhibition of Caspase-9, Caspase-3, and Caspase-7 processing. Furthermore, UCF-101 prevented loss of membrane integrity by inhibiting fodrin breakdown in cardiomyocytes. UCF-101-induced cytoprotection was also correlated with reduced Fas ligand expression and inhibition of FLIP degradation following ischemia/reperfusion. These results suggest that UCF-101 rescues cardiomyocytes from ischemia/reperfusion injury by inhibiting XIAP degradation and Fas/Fas-ligand-induced apoptosis, thereby ameliorating ischemia/reperfusion-induced myocardial dysfunction.

  20. Hyperactivity in Childhood as a Predictor of School Performance in Elementary School: Modifying Effect of a Serotonin Receptor Gene (5-HTR2A)

    ERIC Educational Resources Information Center

    Pulkki-Raback, Laura; Pullmann, Helle; Hintsanen, Mirka; Alatupa, Saija; Ravaja, Niklas; Lehtimaki, Terho; Keltikangas-jarvinen, Liisa

    2010-01-01

    Introduction: Genes have been suggested to interact with predictors of school performance, but evidence is scarce. The purpose was to examine whether a hyperactive temperament leads to different school performance, depending on variability in a serotonin receptor gene (5-HTR2A). Method: The participants were a population-based sample of 909 girls…

  1. Hyperactivity in Childhood as a Predictor of School Performance in Elementary School: Modifying Effect of a Serotonin Receptor Gene (5-HTR2A)

    ERIC Educational Resources Information Center

    Pulkki-Raback, Laura; Pullmann, Helle; Hintsanen, Mirka; Alatupa, Saija; Ravaja, Niklas; Lehtimaki, Terho; Keltikangas-jarvinen, Liisa

    2010-01-01

    Introduction: Genes have been suggested to interact with predictors of school performance, but evidence is scarce. The purpose was to examine whether a hyperactive temperament leads to different school performance, depending on variability in a serotonin receptor gene (5-HTR2A). Method: The participants were a population-based sample of 909 girls…

  2. Electroporation as a strategy to promote HtrA1 gene uptake and chemotherapy efficacy in a mouse model of mesothelioma.

    PubMed

    Spugnini, Enrico P; Cardillo, Irene; Fanciulli, Maurizio; Crispi, Stefania; Vincenzi, Bruno; Boccellino, Mariarosaria; Quagliuolo, Lucio; Baldi, Alfonso

    2013-06-01

    There is not a consensus on the best therapeutic approach to mesothelioma and the prognosis is still dismal. We have recently demonstrated that HtrA1 is a potential therapeutic target in mesothelioma cells. In this manuscript we describe that electroporation in a mouse mesothelioma xenograft was able to facilitate the expression of exogenous HtrA1 injected intra-lesionally in the tumors and to increase the penetration in the neoplastic cells of cisplatin given intra-peritoneally. Indeed, HtrA1 over-expression caused a significant slowing down of tumor growth; moreover, cisplatin efficacy in reducing tumor mass was amplified by electroporation and this phenomenon was even more significant when combining the electroporation of cisplatin and HtrA1. Considering that a substantial number of mesothelioma patients develop early local recurrence, even with radical resection combined with aggressive chemo- and radiotherapy, this multi-modality approach could be very effective in improving local tumor control after surgery. The identification of effective combination coupled with the development of novel equipments and electrodes will be instrumental in planning the translation of these results to humans as per correct laboratory-clinical interface.

  3. The Nrf2-ARE pathway is associated with Schisandrin b attenuating benzo(a)pyrene-Induced HTR cells damages in vitro.

    PubMed

    Dong, Qulong; Hou, Haiyan; Wu, Jun; Chen, Yaqiong

    2016-11-01

    As is ubiquitous in the environmental sources, benzo(a)pyrene (BaP) has been reported to induce reprotoxicity in previous studies. Toxicity to trophoblast cells may be one key factor, but evidences were absent. We speculated that BaP can induce cytotoxicity in human trophoblast HTR-8/SVneo (HTR) cells, and Schisandrin B (Sch B) as a potential protector can inhibit the cytotoxicity. MTS assay identified that BaP induced HTR cells death while Sch B played a cytoprotective role. And after Nrf2 interference, the ability of Sch B-induced cytoprotection was declined. Furthermore, PCR, western blot, ELISA, and SOD assays were found that Sch B significantly increased the mRNA and protein expression of Nrf2, HO1, NQO1, and SOD in the Nrf2-ARE pathway, and the extents of increase were declined after Nrf2 interference. These results demonstrated that the Nrf2-ARE pathway plays an important role in Sch B attenuating BaP-induced HTR cells damages in vitro. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1439-1449, 2016.

  4. Insulin-like growth factor binding protein 7 modulates estrogen-induced trophoblast proliferation and invasion in HTR-8 and JEG-3 cells.

    PubMed

    Liu, Zhen-Kun; Liu, Hai-Yan; Fang, Wen-Ning; Yang, Ying; Wang, Hong-Mei; Peng, Jing-Pian

    2012-05-01

    Previous research has reported that IGFBP7 functions as a tumor suppressor gene in different tumors, but its role in the trophoblast has not been elucidated. In this research, we studied the regulation mechanism of IGFBP7 in trophoblast proliferation and invasion in HTR-8 and JEG-3 cell lines. We found that IGFBP7 was abundantly expressed in normal human syncytiotrophoblast tissue samples but that this was lacking in hydatidiform moles. The proliferation and invasion capacities of HTR-8 and JEG-3 cells were significantly inhibited by recombinant IGFBP7. Estrogen (E2) stimulated the expression of IGFBP7 at a concentration of 5-10 ng/mL. This stimulation was inhibited by the estrogen receptor antagonist Fulvestrant (ICI182.780) and a TGFβ-neutralizing antibody. In conclusion, our data reveals that estrogen stimulates the expression of IGFBP7 through estrogen receptors and TGFβ. The expression of IGFBP7 could be stimulated by TGFβ in a dose-dependent manner and inhibited by IFNγ in HTR-8 and JEG-3 cells. IGFBP7 could also inhibit the phosphorylation of ERK and the expression of PCNA, MMP2 and MMP9 in HTR-8 and JEG-3 cells. These findings suggest that IGFBP7 is a key regulator of E2-induced trophoblast proliferation and invasion.

  5. No association between lithium full responders and the DRD1, DRD2, DRD3, DAT1, 5-HTTLPR and HTR2A genes in a Sardinian sample.

    PubMed

    Manchia, Mirko; Congiu, Donatella; Squassina, Alessio; Lampus, Simona; Ardau, Raffaella; Chillotti, Caterina; Severino, Giovanni; Del Zompo, Maria

    2009-09-30

    Polymorphisms within the DRD1, DRD2, DRD3, DAT1, 5-HTTLPR and HTR2A genes are being studied for association with lithium prophylaxis in a sample of 155 Sardinian unrelated probands affected by bipolar disorder (BP). No significant association was shown between the polymorphisms of the genes studied and response to lithium treatment.

  6. Gender differences in the association between HTR2C gene variants and suicidal behavior in a Mexican population: a case–control study

    PubMed Central

    Molina-Guzman, Gabriel; González-Castro, Thelma Beatriz; Hernández Díaz, Yazmín; Tovilla-Zárate, Carlos Alfonso; Juárez-Rojop, Isela E; Guzmán-Priego, Crystell Guadalupe; Genis, Alma; Pool García, Sherezada; López-Narvaez, María Lilia; Rodriguez-Perez, José Manuel

    2017-01-01

    Background The aim of this case–control study was to explore the association by gender between the HTR2C gene variants and suicidal behavior in a Mexican population. Subjects and methods A total of 183 suicide attempters and 208 healthy volunteers were included in this study. We genotyped five polymorphisms of HTR2C (rs547536, rs2192372, rs4272555, rs6318, and rs2428707), then measured the association by genotype, allele, and haplotype. Results In the female group, we found an association between two polymorphisms of the HTR2C (rs4272555 and rs2428707) and suicide attempts. The C allele of the single-nucleotide polymorphism (SNP) rs4272555 was associated with a decreased risk of suicide attempt (P=0.01, odds ratio =0.26, 95% confidence interval: 0.09–0.79), whereas the G allele of the SNP rs2428707 was associated with an increased risk of suicide attempt (P=0.01, odds ratio =3.68, 95% confidence interval: 1.24–10.90). No significant association was observed between the other polymorphisms studied (rs547536, rs2192372, rs6318) or haplotypes with suicide attempts. Conclusion These findings suggest a possible risk factor of the HTR2C gene in the pathology of suicidal behavior in Mexican population. More studies are necessary to confirm this association. PMID:28260903

  7. The Role of 5-HTR6 in Mossy Fiber Sprouting: Activating Fyn and p-ERK1/2 in Pilocarpine-Induced Chronic Epileptic Rats.

    PubMed

    Lin, Wanhui; Huang, Wenli; Chen, Shenggen; Lin, Mingxing; Huang, Qingyu; Huang, Huapin

    2017-01-01

    Our primary objective is to verify whether 5-HTR6 is involved in the development of mossy fiber sprouting (MFS), and to determine how the progression of MFS is affected by 5-HTR6. A total of 90 male adult Sprague-Dawley rats were allocated into either the control group (n=36) or the epileptic group (n=54). Status epilepticus (SE) of rats was induced by the intraperitoneal (i.p.) injection of LiCl-pilocarpine. We conducted our experiments in two stages. The first stage involves equally dividing 36 epileptic rats into three groups with treatments of none, 5-HTR6 antagonist SB-27104 (SB) and vehicle DMSO. Then behavior and electroencephalogram (EEG) of rats were monitored by video-EEG. The second stage involves dividing 126 epileptic rats into seven groups with treatments of none, 10% DMSO, SB (100 µg/kg), Fyn antagonist PP2 (50 µg/kg), p-ERK1/2 antagonist PD-98059 (30 µg/kg), SB (100 µg/ kg) + PP2 (50 µg/kg); SB (100 µg/kg) + PD-98059 (30 µg/kg). We also treated 18 rats in the control group of the first stage with 100 µg/kg 5-HTR6 agonist WAY-181187 (WAY). MFS of rats was detected through the approach of Timm's staining. Finally, expressions of 5-HTR6, Fyn, p-ERK1/2 and GAP-3 were qualified and semi-quantified via western blotting or RT-PCR. Induction of SE could stimulate formation of MFS and increased GAP-43 expressions. Expressions of 5-HTR6, Fyn and p-ERK1/2 were also up-regulated with increasing time after establishment of SE models. The development of MFS was remarkably inhibited by SB, PP2 and PD. Compared to the single antagonist, such an inhibitory effect was enhanced by SB+PD or SB+PP. Moreover, treatment of healthy rats with WAY would contribute to up-regulated Fyn and p-ERK1/2 expressions, as well as development of MFS (P < 0.05). Suppression of Fyn triggered a down-regulating trend of p-ERK1/2 (P < 0.05), however, suppressed p-ERK1/2 did not have such a significant effect on Fyn expression. HTR6 may affect the progression of MFS by activating

  8. Correlation Between Expression of High Temperature Requirement Serine Protease A1 (HtrA1) in Nucleus Pulposus and T2 Value of Magnetic Resonance Imaging.

    PubMed

    Li, Dapeng; Yue, Jiawei; Jiang, Lu; Huang, Yonghui; Sun, Jifu; Wu, Yan

    2017-04-22

    BACKGROUND Degrading enzymes play an important role in the process of disc degeneration. The objective of this study was to investigate the correlation between the expression of high temperature requirement serine protease A1 (HtrA1) in the nucleus pulposus and the T2 value of the nucleus pulposus region in magnetic resonance imaging (MRI). MATERIAL AND METHODS Thirty-six patients who had undergone surgical excision of the nucleus pulposus were examined by MRI before surgery. Pfirrmann grading of the target intervertebral disc was performed according to the sagittal T2-weighted imaging, and the T2 value of the target nucleus pulposus was measured according to the median sagittal T2 mapping. The correlation between the Pfirrmann grade and the T2 value was analyzed. The expression of HtrA1 in the nucleus pulposus was analyzed by RT-PCR and Western blot. The correlation between the expression of HtrA1 and the T2 value was analyzed. RESULTS The T2 value of the nucleus pulposus region was 33.11-167.91 ms, with an average of 86.64±38.73 ms. According to Spearman correlation analysis, there was a rank correlation between T2 value and Pfirrmann grade (P<0.0001), and the correlation coefficient (rs)=-0.93617. There was a linear correlation between the mRNA level of HtrA1 and T2 value in nucleus pulposus tissues (a=3.88, b=-0.019, F=112.63, P<0.0001), normalized regression coefficient=-0.88. There was a linear correlation between the expression level of HtrA1 protein and the T2 value in the nucleus pulposus tissues (a=3.30, b=-0.016, F=93.15, P<0.0001) and normalized regression coefficient=-0.86. CONCLUSIONS The expression of HtrA1 was strongly related to the T2 value, suggesting that HtrA1 plays an important role in the pathological process of intervertebral disc degeneration.

  9. Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells.

    PubMed

    Cataldo, Luis Rodrigo; Mizgier, María L; Bravo Sagua, Roberto; Jaña, Fabián; Cárdenas, César; Llanos, Paola; Busso, Dolores; Olmos, Pablo; Galgani, José E; Santos, José L; Cortés, Víctor A

    2017-01-01

    Pancreatic β-cells synthesize and release serotonin (5 hydroxytryptamine, 5HT); however, the role of 5HT receptors on glucose stimulated insulin secretion (GSIS) and the mechanisms mediating this function is not fully understood. The aims of this study were to determine the expression profile of 5HT receptors in murine MIN6 β-cells and to examine the effects of pharmacological activation of 5HT receptor Htr2b on GSIS and mitochondrial function. mRNA levels of 5HT receptors in MIN6 cells were quantified by RT qPCR. GSIS was assessed in MIN6 cells in response to global serotonergic activation with 5HT and pharmacological Htr2b activation or inhibition with BW723C86 or SB204741, respectively. In response to Htr2b activation also was evaluated the mRNA and protein levels of PGC1α and PPARy by RT-qPCR and western blotting and mitochondrial function by oxygen consumption rate (OCR) and ATP cellular content. We found that mRNA levels of most 5HT receptors were either very low or undetectable in MIN6 cells. By contrast, Htr2b mRNA was present at moderate levels in these cells. Preincubation (6 h) of MIN6 cells with 5HT or BW723C86 reduced GSIS and the effect of 5HT was prevented by SB204741. Preincubation with BW723C86 increased PGC1α and PPARy mRNA and protein levels and decreased mitochondrial respiration and ATP content in MIN6 cells. Our results indicate that prolonged Htr2b activation in murine β-cells decreases glucose-stimulated insulin secretion and mitochondrial activity by mechanisms likely dependent on enhanced PGC1α/PPARy expression.

  10. Streptococcus pneumoniae serine protease HtrA, but not SFP or PrtA, is a major virulence factor in pneumonia.

    PubMed

    de Stoppelaar, Sacha F; Bootsma, Hester J; Zomer, Aldert; Roelofs, Joris J T H; Hermans, Peter W M; van 't Veer, Cornelis; van der Poll, Tom

    2013-01-01

    Streptococcus (S.) pneumoniae is a common causative pathogen in pneumonia. Serine protease orthologs expressed by a variety of bacteria have been found of importance for virulence. Previous studies have identified two serine proteases in S. pneumoniae, HtrA (high-temperature requirement A) and PrtA (cell wall-associated serine protease A), that contributed to virulence in models of pneumonia and intraperitoneal infection respectively. We here sought to identify additional S. pneumoniae serine proteases and determine their role in virulence. The S. pneumoniae D39 genome contains five putative serine proteases, of which HtrA, Subtilase Family Protein (SFP) and PrtA were selected for insertional mutagenesis because they are predicted to be secreted and surface exposed. Mutant D39 strains lacking serine proteases were constructed by in-frame insertion deletion mutagenesis. Pneumonia was induced by intranasal infection of mice with wild-type or mutant D39. After high dose infection, only D39ΔhtrA showed reduced virulence, as reflected by strongly reduced bacterial loads, diminished dissemination and decreased lung inflammation. D39ΔprtA induced significantly less lung inflammation together with smaller infiltrated lung surface, but without influencing bacterial loads. After low dose infection, D39ΔhtrA again showed strongly reduced bacterial loads; notably, pneumococcal burdens were also modestly lower in lungs after infection with D39Δsfp. These data confirm the important role for HtrA in S. pneumoniae virulence. PrtA contributes to lung damage in high dose pneumonia; it does not however contribute to bacterial outgrowth in pneumococcal pneumonia. SFP may facilitate S. pneumoniae growth after low dose infection.

  11. HTR-PROTEUS PEBBLE BED EXPERIMENTAL PROGRAM CORES 9 & 10: COLUMNAR HEXAGONAL POINT-ON-POINT PACKING WITH A 1:1 MODERATOR-TO-FUEL PEBBLE RATIO

    SciTech Connect

    John D. Bess

    2014-03-01

    PROTEUS is a zero-power research reactor based on a cylindrical graphite annulus with a central cylindrical cavity. The graphite annulus remains basically the same for all experimental programs, but the contents of the central cavity are changed according to the type of reactor being investigated. Through most of its service history, PROTEUS has represented light-water reactors, but from 1992 to 1996 PROTEUS was configured as a pebble-bed reactor (PBR) critical facility and designated as HTR-PROTEUS. The nomenclature was used to indicate that this series consisted of High Temperature Reactor experiments performed in the PROTEUS assembly. During this period, seventeen critical configurations were assembled and various reactor physics experiments were conducted. These experiments included measurements of criticality, differential and integral control rod and safety rod worths, kinetics, reaction rates, water ingress effects, and small sample reactivity effects (Ref. 3). HTR-PROTEUS was constructed, and the experimental program was conducted, for the purpose of providing experimental benchmark data for assessment of reactor physics computer codes. Considerable effort was devoted to benchmark calculations as a part of the HTR-PROTEUS program. References 1 and 2 provide detailed data for use in constructing models for codes to be assessed. Reference 3 is a comprehensive summary of the HTR-PROTEUS experiments and the associated benchmark program. This document draws freely from these references. Only Cores 9 and 10 are evaluated in this benchmark report due to similarities in their construction. The other core configurations of the HTR-PROTEUS program are evaluated in their respective reports as outlined in Section 1.0. Cores 9 and 10 were evaluated and determined to be acceptable benchmark experiments.

  12. Prolonged Activation of the Htr2b Serotonin Receptor Impairs Glucose Stimulated Insulin Secretion and Mitochondrial Function in MIN6 Cells

    PubMed Central

    Cataldo, Luis Rodrigo; Mizgier, María L.; Bravo Sagua, Roberto; Jaña, Fabián; Cárdenas, César; Llanos, Paola; Busso, Dolores; Olmos, Pablo; Galgani, José E.; Santos, José L.; Cortés, Víctor A.

    2017-01-01

    Aims Pancreatic β-cells synthesize and release serotonin (5 hydroxytryptamine, 5HT); however, the role of 5HT receptors on glucose stimulated insulin secretion (GSIS) and the mechanisms mediating this function is not fully understood. The aims of this study were to determine the expression profile of 5HT receptors in murine MIN6 β-cells and to examine the effects of pharmacological activation of 5HT receptor Htr2b on GSIS and mitochondrial function. Materials and Methods mRNA levels of 5HT receptors in MIN6 cells were quantified by RT qPCR. GSIS was assessed in MIN6 cells in response to global serotonergic activation with 5HT and pharmacological Htr2b activation or inhibition with BW723C86 or SB204741, respectively. In response to Htr2b activation also was evaluated the mRNA and protein levels of PGC1α and PPARy by RT-qPCR and western blotting and mitochondrial function by oxygen consumption rate (OCR) and ATP cellular content. Results We found that mRNA levels of most 5HT receptors were either very low or undetectable in MIN6 cells. By contrast, Htr2b mRNA was present at moderate levels in these cells. Preincubation (6 h) of MIN6 cells with 5HT or BW723C86 reduced GSIS and the effect of 5HT was prevented by SB204741. Preincubation with BW723C86 increased PGC1α and PPARy mRNA and protein levels and decreased mitochondrial respiration and ATP content in MIN6 cells. Conclusions Our results indicate that prolonged Htr2b activation in murine β-cells decreases glucose-stimulated insulin secretion and mitochondrial activity by mechanisms likely dependent on enhanced PGC1α/PPARy expression. PMID:28129327

  13. Reactive Oxygen Stimulation of Interleukin-6 Release in the Human Trophoblast Cell Line HTR-8/SVneo by the Trichlorethylene Metabolite S-(1,2-Dichloro)-l-Cysteine.

    PubMed

    Hassan, Iman; Kumar, Anjana M; Park, Hae-Ryung; Lash, Lawrence H; Loch-Caruso, Rita

    2016-09-01

    Trichloroethylene (TCE) is a common environmental pollutant associated with adverse reproductive outcomes in humans. TCE intoxication occurs primarily through its biotransformation to bioactive metabolites, including S-(1,2-dichlorovinyl)-l-cysteine (DCVC). TCE induces oxidative stress and inflammation in the liver and kidney. Although the placenta is capable of xenobiotic metabolism and oxidative stress and inflammation in placenta have been associated with adverse pregnancy outcomes, TCE toxicity in the placenta remains poorly understood. We determined the effects of DCVC by using the human extravillous trophoblast cell line HTR-8/SVneo. Exposure to 10 and 20 μM DCVC for 10 h increased reactive oxygen species (ROS) as measured by carboxydichlorofluorescein fluorescence. Moreover, 10 and 20 μM DCVC increased mRNA expression and release of interleukin-6 (IL-6) after 24-h exposure, and these responses were inhibited by the cysteine conjugate beta-lyase inhibitor aminooxyacetic acid and by treatments with antioxidants (alpha-tocopherol and deferoxamine), suggesting that DCVC-stimulated IL-6 release in HTR-8/SVneo cells is dependent on beta-lyase metabolic activation and increased generation of ROS. HTR-8/SVneo cells exhibited decreased mitochondrial membrane potential at 5, 10, and 20 μM DCVC at 5, 10, and 24 h, showing that DCVC induces mitochondrial dysfunction in HTR-8/Svneo cells. The present study demonstrates that DCVC stimulated ROS generation in the human placental cell line HTR-8/SVneo and provides new evidence of mechanistic linkage between DCVC-stimulated ROS and increase in proinflammatory cytokine IL-6. Because abnormal activation of cytokines can disrupt trophoblast functions necessary for placental development and successful pregnancy, follow-up investigations relating these findings to physiologic outcomes are warranted.

  14. HTR-PROTEUS PEBBLE BED EXPERIMENTAL PROGRAM CORES 9 & 10: COLUMNAR HEXAGONAL POINT-ON-POINT PACKING WITH A 1:1 MODERATOR-TO-FUEL PEBBLE RATIO

    SciTech Connect

    John D. Bess

    2013-03-01

    PROTEUS is a zero-power research reactor based on a cylindrical graphite annulus with a central cylindrical cavity. The graphite annulus remains basically the same for all experimental programs, but the contents of the central cavity are changed according to the type of reactor being investigated. Through most of its service history, PROTEUS has represented light-water reactors, but from 1992 to 1996 PROTEUS was configured as a pebble-bed reactor (PBR) critical facility and designated as HTR-PROTEUS. The nomenclature was used to indicate that this series consisted of High Temperature Reactor experiments performed in the PROTEUS assembly. During this period, seventeen critical configurations were assembled and various reactor physics experiments were conducted. These experiments included measurements of criticality, differential and integral control rod and safety rod worths, kinetics, reaction rates, water ingress effects, and small sample reactivity effects (Ref. 3). HTR-PROTEUS was constructed, and the experimental program was conducted, for the purpose of providing experimental benchmark data for assessment of reactor physics computer codes. Considerable effort was devoted to benchmark calculations as a part of the HTR-PROTEUS program. References 1 and 2 provide detailed data for use in constructing models for codes to be assessed. Reference 3 is a comprehensive summary of the HTR-PROTEUS experiments and the associated benchmark program. This document draws freely from these references. Only Cores 9 and 10 are evaluated in this benchmark report due to similarities in their construction. The other core configurations of the HTR-PROTEUS program are evaluated in their respective reports as outlined in Section 1.0. Cores 9 and 10 were evaluated and determined to be acceptable benchmark experiments.

  15. Streptococcus pneumoniae Serine Protease HtrA, but Not SFP or PrtA, Is a Major Virulence Factor in Pneumonia

    PubMed Central

    de Stoppelaar, Sacha F.; Bootsma, Hester J.; Zomer, Aldert; Roelofs, Joris J. T. H.; Hermans, Peter W. M.; van ’t Veer, Cornelis; van der Poll, Tom

    2013-01-01

    Streptococcus (S.) pneumoniae is a common causative pathogen in pneumonia. Serine protease orthologs expressed by a variety of bacteria have been found of importance for virulence. Previous studies have identified two serine proteases in S. pneumoniae, HtrA (high-temperature requirement A) and PrtA (cell wall-associated serine protease A), that contributed to virulence in models of pneumonia and intraperitoneal infection respectively. We here sought to identify additional S. pneumoniae serine proteases and determine their role in virulence. The S. pneumoniae D39 genome contains five putative serine proteases, of which HtrA, Subtilase Family Protein (SFP) and PrtA were selected for insertional mutagenesis because they are predicted to be secreted and surface exposed. Mutant D39 strains lacking serine proteases were constructed by in-frame insertion deletion mutagenesis. Pneumonia was induced by intranasal infection of mice with wild-type or mutant D39. After high dose infection, only D39ΔhtrA showed reduced virulence, as reflected by strongly reduced bacterial loads, diminished dissemination and decreased lung inflammation. D39ΔprtA induced significantly less lung inflammation together with smaller infiltrated lung surface, but without influencing bacterial loads. After low dose infection, D39ΔhtrA again showed strongly reduced bacterial loads; notably, pneumococcal burdens were also modestly lower in lungs after infection with D39Δsfp. These data confirm the important role for HtrA in S. pneumoniae virulence. PrtA contributes to lung damage in high dose pneumonia; it does not however contribute to bacterial outgrowth in pneumococcal pneumonia. SFP may facilitate S. pneumoniae growth after low dose infection. PMID:24244609

  16. Interaction of 5-HTT and HTR1A gene polymorphisms in treatment responses to mirtazapine in patients with major depressive disorder.

    PubMed

    Chang, Hun Soo; Lee, Hwa-Young; Cha, Ji-Hyun; Won, Eun Soo; Ham, Byung-Joo; Kim, Bohye; Lee, Min-Soo

    2014-08-01

    We tested for the association of HTR1A and 5-HTT genetic polymorphisms with treatment response to mirtazapine and evaluated the interactive effect between the polymorphisms in 283 patients with major depressive disorder. Korean subjects with diagnosis of major depressive disorder using the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition Axis I disorders were recruited. Clinical symptoms were evaluated using the 17-item Hamilton Depression Rating (HAMD-17) Scale at baseline and after 1, 2, 4, 8, and 12 weeks of treatment with mirtazapine. The genetic association of 5-HTTLPR and HTR1A+272G>A with treatment response was analyzed. We found a significant association of the 12.12-repeat genotype of 5-HTT various number tandem repeat (VNTR) with a large percentage decline in HAMD-17 Scale score after 4, 8, and 12 weeks of treatment with mirtazapine. We also found that the frequency of the 12.12-repeat genotype was higher in responders than in nonresponders at week 8. The HTR1A+272GG genotype was significantly associated with a large percentage decline in HAMD-17 Scale score at 4, 8, and 12 weeks, although the genotypic frequencies were comparable between responders and nonresponders during the study period. Patients with the 12.12-repeat 5-HTT VNTR and GG of HTR1A+272G>A showed the highest HAMD-17 Scale percentage reduction during the study period and a better treatment response status after 4 weeks. These results suggest that the interaction between HTR1A+272G>A and 5-HTT VNTR is involved in the response to mirtazapine treatment and that a combination of these may be a useful marker for predicting treatment response to mirtazapine.

  17. Leaching behaviour of unirradiated high temperature reactor (HTR) UO 2-ThO 2 mixed oxides fuel particles

    NASA Astrophysics Data System (ADS)

    Alliot, Cyrille; Grambow, Bernd; Landesman, Catherine

    2005-11-01

    The dissolution of different mixed oxide (U, Th)O 2 particles under reducing conditions has been studied using a continuous flow-through reactor. The U/Th ratio seems to have no or little influence on the normalised leaching rate of thorium or uranium, The release rate of uranium from the outer surface of a Th rich matrix seems to follow the behaviour of pure UO 2 even though U is a minor component in these phases and the dissolution rate of Th is much lower. After long time U concentrations will become depleted at the solids surface and it will be expected that U release rates will become controlled by the release rates of thorium (rates at neutral pH < 10 -6 g m -2 d -1). Under reducing conditions, the matrix of HTR fuel particles presents significant intrinsic radionuclide confinement properties.

  18. Final Report on Utilization of TRU TRISO Fuel as Applied to HTR Systems Part I: Pebble Bed Reactors

    SciTech Connect

    Brian Boer; Abderrafi M. Ougouag

    2011-03-01

    significant failure is to be expected for the reference fuel particle during normal operation. It was found, however, that the sensitivity of the coating stress to the CO production in the kernel was large. The CO production is expected to be higher in DB fuel than in UO2 fuel, but its exact level has a high uncertainty. Furthermore, in the fuel performance analysis transient conditions were not yet taken into account. The effort of this task in FY 2010 has focused on the optimization of the core to maximize the pebble discharge burnup level, while retaining its inherent safety characteristics. Using generic pebble bed reactor cores, this task will perform physics calculations to evaluate the capabilities of the pebble bed reactor to perform utilization and destruction of LWR used-fuel transuranics. The task will use established benchmarked models, and will introduce modeling advancements appropriate to the nature of the fuel considered (high transuranic [TRU] content and high burn-up). Accomplishments of this work include: •Core analysis of a HTR-MODULE design loaded with Deep-Burn fuel. •Core analysis of a HTR-MODULE design loaded with Deep-Burn fuel and Uranium. •Core analysis of a HTR-MODULE design loaded with Deep-Burn fuel and Modified Open Cycle Components. •Core analysis of a HTR-MODULE design loaded with Deep-Burn fuel and Americium targets.

  19. Comparison of Homogeneous and Heterogeneous CFD Fuel Models for Phase I of the IAEA CRP on HTR Uncertainties Benchmark

    SciTech Connect

    Gerhard Strydom; Su-Jong Yoon

    2014-04-01

    Computational Fluid Dynamics (CFD) evaluation of homogeneous and heterogeneous fuel models was performed as part of the Phase I calculations of the International Atomic Energy Agency (IAEA) Coordinate Research Program (CRP) on High Temperature Reactor (HTR) Uncertainties in Modeling (UAM). This study was focused on the nominal localized stand-alone fuel thermal response, as defined in Ex. I-3 and I-4 of the HTR UAM. The aim of the stand-alone thermal unit-cell simulation is to isolate the effect of material and boundary input uncertainties on a very simplified problem, before propagation of these uncertainties are performed in subsequent coupled neutronics/thermal fluids phases on the benchmark. In many of the previous studies for high temperature gas cooled reactors, the volume-averaged homogeneous mixture model of a single fuel compact has been applied. In the homogeneous model, the Tristructural Isotropic (TRISO) fuel particles in the fuel compact were not modeled directly and an effective thermal conductivity was employed for the thermo-physical properties of the fuel compact. On the contrary, in the heterogeneous model, the uranium carbide (UCO), inner and outer pyrolytic carbon (IPyC/OPyC) and silicon carbide (SiC) layers of the TRISO fuel particles are explicitly modeled. The fuel compact is modeled as a heterogeneous mixture of TRISO fuel kernels embedded in H-451 matrix graphite. In this study, a steady-state and transient CFD simulations were performed with both homogeneous and heterogeneous models to compare the thermal characteristics. The nominal values of the input parameters are used for this CFD analysis. In a future study, the effects of input uncertainties in the material properties and boundary parameters will be investigated and reported.

  20. SLC6A14 and 5-HTR2C polymorphisms are associated with food intake and nutritional status in children.

    PubMed

    Miranda, Raquel C K; Vetter, Silvia B; Genro, Júlia P; Campagnolo, Paula D B; Mattevi, Vanessa S; Vitolo, Márcia R; Almeida, Silvana

    2015-12-01

    Serotonin plays a critical role in the regulation of food intake. The solute carrier family 6 member 14 (SLC6A14) and serotonin receptor 2C (5-HTR2C) genes are involved in the bioavailability and action of this neurotransmitter. Evaluation of the association of six polymorphisms in these genes with food intake and nutritional status in children followed to 7-8years of age. Blood samples and the biodemographic data of 344 children were collected at three development stages, in a cross-sectional study undertaken with the cohort from a randomized trial. Polymorphisms were analyzed using polymerase chain reaction-based techniques. At 7 to 8years of age, carriers of the A alleles for both the SLC6A14 rs2312054 and SLC6A14 rs12391221 polymorphisms showed higher food intake, except for the sugar-dense food (SDF) intake parameter, than T/T and C/C homozygotes, respectively. Boy carriers of the C allele of rs2071877 had a higher sum of triceps and subscapular folds than T allele carriers at 7 to 8years old. For 5-HTR2C gene variants, A allele carriers (rs3813928) and T allele carriers (rs3813929) had higher food intake at 3 to 4years old than G/G and C/C homozygotes, respectively, except for SDF. At this age, the intake of energy-dense foods was higher in carriers of the T allele (rs3813929) than in C/C homozygotes. This study provides evidence that genetic variants of these proteins might be involved in the determination of food intake and nutritional status in children. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  1. Telomerase as a prognostic marker in breast cancer: high-throughput tissue microarray analysis of hTERT and hTR.

    PubMed

    Poremba, Christopher; Heine, Bernhard; Diallo, Raihanatou; Heinecke, Achim; Wai, Daniel; Schaefer, Karl-Ludwig; Braun, Yvonne; Schuck, Andreas; Lanvers, Claudia; Bànkfalvi, Agnes; Kneif, Sören; Torhorst, Joachim; Zuber, Markus; Köchli, Ossi R; Mross, Frank; Dieterich, Holger; Sauter, Guido; Stein, Harald; Fogt, Franz; Boecker, Werner

    2002-10-01

    Telomerase activity (TA) has been shown to correlate with poor clinical outcome in various tumour entities, indicating that tumours expressing this enzyme may be more aggressive and that TA may be a useful prognostic marker. For breast cancer, however, TA is a controversial prognostic marker; whereas some studies suggest an association between TA and disease outcome, others do not find this association. This study used tissue microarrays (breast carcinoma prognosis arrays) containing 611 samples (each 0.6 mm in diameter) from the tumour centre of paraffin-embedded breast carcinomas to analyse the catalytic subunit of telomerase, human telomerase reverse-transcriptase (hTERT), and the internal RNA component (hTR), which are the core components of the telomerase holoenzyme complex. hTERT protein expression was obtained by immunohistochemistry (human anti-telomerase antibody Ab-2, Calbiochem), and hTR RNA was measured by radioactive in situ hybridization. hTERT and hTR expression were determined semi-quantitatively and graded (scores 1-4). Clinical data, such as histological subtype, pT stage, tumour diameter, pN stage, BRE grade, tumour-specific survival (in months), patient's age and others, were available for statistical analysis. A statistically significant correlation was found between tumour-specific survival (overall survival) and hTERT expression (p < 0.0001) or hTR expression (p = 0.00110). Tumours with higher scores (scores 3, 4) for hTR and/or hTERT were associated with a worse prognosis. In multivariate analysis, hTERT expression was an independent prognostic factor. Previous studies, focusing on analysis of TA in smaller numbers of fresh-frozen breast carcinomas by the TRAP assay, gave controversial results with respect to TA as a prognostic marker. Using tissue microarrays from 611 breast carcinomas, this study has demonstrated that increased expression levels of the telomerase core components, hTERT and hTR, are associated with lower overall survival

  2. Expression and Immunogenicity of a Mutant Diphtheria Toxin Molecule, CRM197, and Its Fragments in Salmonella typhi Vaccine Strain CVD 908-htrA

    PubMed Central

    Orr, Nadav; Galen, James E.; Levine, Myron M.

    1999-01-01

    Mutant diphtheria toxin molecule CRM197 and fragments thereof were expressed in attenuated Salmonella typhi CVD 908-htrA, and the constructs were tested for their ability to induce serum antitoxin. Initially, expressed proteins were insoluble, and the constructs failed to induce neutralizing antitoxin. Soluble CRM197 was expressed at low levels by utilizing the hemolysin A secretion system from Escherichia coli. PMID:10417208

  3. -759C÷T polymorphism of the HTR2C gene is not correlated with atypical antipsychotics-induced weight gain, among Romanian psychotic patients.

    PubMed

    Grădinaru, Raluca Claudia; Andreescu, Nicoleta Ioana; Nussbaum, Laura Alexandra; Farcaş, Simona Sorina; Dumitraşcu, Victor; Suciu, Liana; Puiu, Maria

    2016-01-01

    We aim to investigate whether the -759C÷T polymorphism in 5-HTR2C gene was associated with weight change and hyperinsulinemia in Romanian pediatric patients with schizophrenia and bipolar disorders. The patients under investigation were enrolled between 2009 and 2014. A total of 81 schizophrenic and bipolar-disorder patients, aged between nine to 20 years (median age 15.74±4 years), who were following an atypical antipsychotic treatment (Risperidone, Aripiprazole, Olanzapine), were enrolled from University Hospital for Child and Adolescent Psychiatry and Neurology from Timisoara, Romania. The outcomes that we measured were the changes in Body Mass Index (BMI) from baseline to different time points: three months, six months, 12 months and 18 months, and the change in insulinemia over time, after atypical antipsychotic treatment. After carrying out the 5-HTR2C 759C÷T polymorphism identification, we found that 22 patients presented the -759C÷T polymorphism in 5-HTR2C gene. Between the patients exhibiting the 5-HTR2C -759C÷T polymorphism and the patients having the wild type alleles, there was no significant statistical difference in changes of BMI from baseline to endpoints that indicates the lack of the protective effect of the T allele against atypical antipsychotics-induced weight gain. Interestingly, we found a statistically significant association between insulinemia and T alleles' carriers, after 18 months of treatment with the above-mentioned antipsychotics. Taking into consideration that atypical antipsychotics have been associated with elevated insulin levels and insulin resistance, maybe in the future the -759C÷T polymorphism would find a role in the development of a more complex algorithm for prediction of diabetes mellitus risk, in patients taking atypical antipsychotics.

  4. Leukaemia inhibitory factor mediated proliferation of HTR-8/SVneo trophoblast cells is dependent on activation of extracellular signal-regulated kinase 1/2.

    PubMed

    Prakash, Golla Jaya; Suman, Pankaj; Morales Prieto, Diana M; Markert, Udo R; Gupta, Satish K

    2011-01-01

    Leukaemia inhibitory factor (LIF) is one of the cytokines that is indispensable for embryo implantation. The aim of the present study was to investigate the role of activation of extracellular signal-regulated kinase (ERK) 1/2 in LIF-mediated proliferation of HTR-8/SVneo cells. Stimulation of HTR-8/SVneo cells with LIF (50 ng mL(-1)) resulted in an increase in cell proliferation (P < 0.05) via increased transition of cells to the G(2)/M phase of cell cycle. Stimulation with LIF resulted in the activation of both signal transducer and activator of transcription (STAT) 3 Tyr(705) and ERK1/2, but inhibition of ERK1/2 signalling by pretreatment of cells with U0126 (10 µM) for 2h resulted in abrogation of LIF-mediated increases in G(2)/M transition, with a significant decrease (P < 0.05) in absolute cell numbers compared with control. Although STAT3 silencing had no effect on LIF-dependent proliferation of HTR-8/SVneo cells, it did result in an increase in cell apoptosis, which increased further upon inhibition of ERK1/2 activation irrespective of LIF stimulation. Stimulation of cells with LIF increased the Bcl-2/Bax ratio, whereas ERK1/2 inhibition decreased the Bcl-2/Bax ratio, even after LIF stimulation. Hence, it can be inferred that ERK1/2 activation is essential for LIF-mediated increases in proliferation and that both STAT3 and ERK1/2 activation are important for the survival of HTR-8/SVneo cells.

  5. The induction of reactive oxygen species and loss of mitochondrial Omi/HtrA2 is associated with S-nitrosoglutathione-induced apoptosis in human endothelial cells

    SciTech Connect

    Liu Qibing; Liu Lulu; Lu Yingmei; Tao Rongrong; Huang Jiyun; Shioda, Norifumi; Moriguchi, Shigeki; Fukunaga, Kohji; Han Feng; Lou Yijia

    2010-05-01

    The pathophysiological relevance of S-nitrosoglutathione (GSNO)-induced endothelial cell injury remains unclear. The main objective of this study was to elucidate the molecular mechanisms of GSNO-induced oxidative stress in endothelial cells. Morphological evaluation through DAPI staining and propidium iodide (PI) flow cytometry was used to detect apoptosis. In cultured EA.hy926 endothelial cells, exposure to GSNO led to a time- and dose-dependent apoptotic cascade. When intracellular reactive oxygen species (ROS) production was measured in GSNO-treated cells with the fluorescent probes 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, we observed elevated ROS levels and a concomitant loss in mitochondrial membrane potential, indicating that GSNO-induced death signaling is mediated through a ROS-mitochondrial pathway. Importantly, we found that peroxynitrite formation and Omi/HtrA2 release from mitochondria were involved in this phenomenon, whereas changes of death-receptor dependent signaling were not detected in the same context. The inhibition of NADPH oxidase activation and Omi/HtrA2 by a pharmacological approach provided significant protection against caspase-3 activation and GSNO-induced cell death, confirming that GSNO triggers the death cascade in endothelial cells in a mitochondria-dependent manner. Taken together, our results indicate that ROS overproduction and loss of mitochondrial Omi/HtrA2 play a pivotal role in reactive nitrogen species-induced cell death, and the modulation of these pathways can be of significant therapeutic benefit.

  6. Structure-Based Design of a Br Halogen Bond at the Complex Interface of the Human Placental HtrA1 PDZ Domain with Its Heptapeptide Ligand.

    PubMed

    Dou, Shuo-Fen; Liu, Hong; Cao, Tong-Mei; Wen, Qing-Li; Li, Jie; Shao, Qing-Chun

    2016-04-01

    The shock-induced serine protease HtrA1 is a potential regulator of human placenta development during pregnancy. The protein contains a functional PDZ domain that has been solved in complex with a phage display-derived heptapeptide: Asp-6 Ser-5 Arg-4 Ile-3 Trp-2 Trp-1 Val0 . In this study, a rationally designed halogen bond was introduced to the domain-peptide complex based on its NMR structure in solution. We computationally compared the stabilization energies and hindrance effects due to the presence of different halogens X (X = F, Cl, Br, or I), using a hybrid quantum mechanics/molecular mechanics (QM/MM) approach, and found that the Br atom could considerably promote the peptide binding free energy (ΔΔG = -5.2 kcal/mol). Fluorescence assays confirmed that the peptide affinity to the HtrA1 PDZ domain was improved by approximately sevenfold upon bromination. Structural analysis identified a geometrically perfect halogen bond between the Br atom of the peptide Trp-1 residue and the carbonyl O atom of the HtrA1 Ile385 residue, with a bond length and an interaction energy of d = 3.20 Å and ΔE = -3.7 kcal/mol, respectively.

  7. Genetic variation in the serotonin transporter and HTR1B receptor predicts reduced bone formation during serotonin reuptake inhibitor treatment in older adults

    PubMed Central

    GARFIELD, LAUREN D.; MÜLLER, DANIEL J.; KENNEDY, JAMES L.; MULSANT, BENOIT H.; REYNOLDS, CHARLES F.; TEITELBAUM, STEVEN L.; CIVITELLI, ROBERTO; DIXON, DAVID; TODOROV, ALEXANDRE A.; LENZE, ERIC J.

    2014-01-01

    Objectives Studies have reported an association between serotonin reuptake inhibitors (SRIs) and accelerated bone loss. Genetic variation in the serotonin system might modulate bone metabolism changes during SRI treatment. In a clinical trial we examined functional genetic polymorphisms of serotonin transporter and receptors involved in bone metabolism to determine whether they predict changes in bone metabolism during SRI treatment. Methods In 69 adults (age ≥ 60) participating in a 12-week, open-label trial of the SRI venlafaxine for major depression, serum markers of bone formation (P1NP) and resorption (β-CTX) were assayed before and after treatment. Participants were genotyped for putative high- versus low-expressing polymorphisms in the serotonin transporter (5HTTLPR) and 1B receptor (HTR1B) genes. Results Bone formation was significantly reduced with administration of venlafaxine in participants with the high-expressing 5HTTLPR genotype and those with the low-expressing HTR1B genotype. This primarily occurred in individuals with the combination of the high-expressing 5HTTLPR genotype and the low-expressing HTR1B genotype. Conclusions These preliminary findings indicate that genetic variation in the serotonin receptors predicts changes in bone metabolism during SRI use. If these results are replicated and clinically confirmed, we will have identified a genetic subgroup at high risk for deleterious bone outcomes with the use of SRIs. PMID:24074042

  8. The human telomerase RNA component, hTR, activates the DNA-dependent protein kinase to phosphorylate heterogeneous nuclear ribonucleoprotein A1.

    PubMed

    Ting, Nicholas S Y; Pohorelic, Brant; Yu, Yaping; Lees-Miller, Susan P; Beattie, Tara L

    2009-10-01

    Telomere integrity in human cells is maintained by the dynamic interplay between telomerase, telomere associated proteins, and DNA repair proteins. These interactions are vital to suppress DNA damage responses and unfavorable changes in chromosome dynamics. The DNA-dependent protein kinase (DNA-PK) is critical for this process. Cells deficient for functional DNA-PKcs show increased rates of telomere loss, accompanied by chromosomal fusions and translocations. Treatment of cells with specific DNA-PK kinase inhibitors leads to similar phenotypes. These observations indicate that the kinase activity of DNA-PK is required for its function at telomeres possibly through phosphorylation of essential proteins needed for telomere length maintenance. Here we show that the heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is a direct substrate for DNA-PK in vitro. Phosphorylation of hnRNP A1 is stimulated not only by the presence of DNA but also by the telomerase RNA component, hTR. Furthermore, we show that hnRNP A1 is phosphorylated in vivo in a DNA-PK-dependent manner and that this phosphorylation is greatly reduced in cell lines which lack hTR. These data are the first to report that hTR stimulates the kinase activity of DNA-PK toward a known telomere-associated protein, and may provide further insights into the function of DNA-PK at telomeres.

  9. MiR-101 regulates apoptosis of trophoblast HTR-8/SVneo cells by targeting endoplasmic reticulum (ER) protein 44 during preeclampsia.

    PubMed

    Zou, Y; Jiang, Z; Yu, X; Zhang, Y; Sun, M; Wang, W; Ge, Z; De, W; Sun, L

    2014-10-01

    To investigate a possible association between miR-101 and apoptosis of human trophoblast cells mediated by endoplasmic reticulum protein 44 (ERp44) in preeclampsia (PE), we explored the expression of miR-101 in PE placentas (n=30) compared with normotensive pregnant placentas (n=30) and the correlation between miR-101 and ERp44 was also analyzed. Furthermore, both the apoptotic rate of trophoblast cells and the ER stress-induced apoptotic proteins were assayed when the HTR-8/SVneo cells were treated with miR-101 mimics or inhibitors in vitro. We found a lower expression of miR-101 and an inverse correlation between miR-101 and ERp44 protein in PE placentas. Upregulation of miR-101 expression could inhibit trophoblast HTR-8/SVneo cell apoptosis and repress ER stress-induced apoptotic proteins by targeting ERp44 in vitro, whereas inhibition of miR-101 could induce HTR-8/SVneo cell apoptosis. Our findings indicated that overexpression of miR-101 could downregulate ERp44 and suppress apoptosis in trophoblast cells during PE. Therefore, loss of miR-101 expression could contribute to ER stress-induced trophoblast cell apoptosis by targeting ERp44.

  10. MicroRNA-155 inhibits proliferation and migration of human extravillous trophoblast derived HTR-8/SVneo cells via down-regulating cyclin D1.

    PubMed

    Dai, Y; Qiu, Z; Diao, Z; Shen, L; Xue, P; Sun, H; Hu, Y

    2012-10-01

    MiR-155 is known to participate in various cellular processes by targeting gene expression. We previously revealed a link between miR-155 and perturbation of trophoblast invasion and differentiation. This study aimed to investigate the target molecule(s) of miR-155 on the influence on the proliferation and migration of trophoblast cells. Bioinformatics analysis showed that, at the 3' untranslated region (UTR) of cyclin D1, six bases are complementary to the seed region of miR-155. Luciferase assays and cyclin D1 3'UTR transfection assays validated that cyclin D1 3'UTR was the target of miR-155 in HTR-8/SVneo cells. Overexpression of miR-155 in HTR-8/SVneo cells reduced the level of cyclin D1 protein, decreased cell proliferation and invasion, and increased cell number at the G1 stage. Furthermore, the increased expression of miR-155 also regulated the protein levels of kinase inhibitory protein p27 and phosphorylated cytoskeletal protein filamin A. In conclusion, we found that cyclin D1 may be a target of miR-155 in HTR-8/SVneo cells, and demonstrated a negative regulatory role of miR-155 involved in cyclin D1/p27 pathway in proliferation and migration of the cells.

  11. MicroRNA-135b suppresses extravillous trophoblast-derived HTR-8/SVneo cell invasion by directly down regulating CXCL12 under low oxygen conditions.

    PubMed

    Tamaru, Shunsuke; Mizuno, Yosuke; Tochigi, Hideno; Kajihara, Takeshi; Okazaki, Yasushi; Okagaki, Ryugo; Kamei, Yoshimasa; Ishihara, Osamu; Itakura, Atsuo

    2015-05-29

    The expression of numerous microRNAs (miRNAs) in the trophoblasts changes under low oxygen conditions. However, little is known regarding the regulation of the trophoblast invasion by miRNAs under low oxygen conditions. The aim of this study was to identify those miRNAs and their target genes associated with the trophoblast invasion under low oxygen conditions. Culturing the extravillous trophoblast (EVT) cell line, HTR-8/SVneo, at 2% oxygen as compared to 20% oxygen suppressed trophoblast invasion that correlated with increased expression of microRNA-135b (miR-135b) and decreased expression of the its predicted target gene CXCL12. Overexpression of miR-135b suppressed CXCL12 mRNA expression and invasion of HTR-8/SVneo cells. Adding a neutralizing antibody against CXCL12 to the culture medium suppressed HTR-8/SVneo cell invasion. Reporter assays showed that the 3'UTR sequence of CXCL12 was directly targeted by miR-135b. Our results suggest that miR-135b and CXCL12 play important roles in modulating the EVT invasion under low oxygen conditions.

  12. Relationship between the rs1414334 C/G polymorphism in the HTR2C gene and smoking in patients treated with atypical antipsychotics.

    PubMed

    Rico-Gomis, José María; Palazón-Bru, Antonio; Triano-García, Irene; Mahecha-García, Luis Fabián; García-Monsalve, Ana; Navarro-Ruiz, Andrés; Villagordo-Peñalver, Berta; Martínez-Hortelano, Alicia; Gil-Guillén, Vicente Francisco

    2017-07-14

    An association has been found between the C allele of the rs1414334 polymorphism in the HTR2C gene and the metabolic syndrome in psychiatric patients. However, no study has yet evaluated whether this allele is associated with smoking. To assess this issue, therefore, we performed a cross-sectional study with a sample of 166 adult patients treated with atypical antipsychotics in 2012-2013 in a region of Spain. The primary variable was the presence of the C allele of the rs1414334 polymorphism in the HTR2C gene. Secondary variables were the number of pack-years (number of cigarettes per day x number of smoking years ÷ 20), age, gender, schizophrenia, years since diagnosis, metabolic syndrome criteria and SCORE. A stepwise binary logistic regression model was constructed to determine associations between primary and secondary variables and their area under the ROC curve (AUC) was calculated. Of the total sample, 33 patients (19.9%) had the C allele of the polymorphism analyzed. Mean cigarette consumption was 11.6 pack-years. The multivariate analysis showed the following factors as associated with the polymorphism: higher cigarette consumption, being a woman, and not having abdominal obesity. The AUC was 0.706. An association was found between increased cigarette consumption over the years and the presence of the C allele of the rs1414334 polymorphism in the HTR2C gene.

  13. The REV project -- Experiments, techniques and theoretical considerations with a view to an ILW and spent HTR fuel emplacement test at the Asse salt mine

    SciTech Connect

    Niephaus, D.

    1993-12-31

    In the Federal Republic of Germany, radioactive waste forms of pronounced decay heat generation shall be disposed of in deep vertical boreholes in the planned underground repository at Gorleben site. The disposal technique for heat generating intermediate-level waste and for spent HTR fuel is under development in the R and D project, entitled ``Intermediate-Level Waste and Spent HTR Fuel element Test Disposal in Boreholes`` (MHV Project). The project work is divided in two subprojects and has been going on since 1983. In the subproject ``Retrievable Emplacement Test`` (REV project) an emplacement test with already existing waste packages, i.e. steel drums with cladding hulls, fuel hardware and dissolver sludges from LWR-FE reprocessing and steel canisters with spent HTR pebble bed fuel will be conducted in the Asse salt mine. This paper deals with the results obtained from a long term precursory test program, the description of the installation work that has been done with a view to measure and analyze gases released into the atmospheres of the unlined emplacement boreholes, and a brief description of engineered equipment needed for handling, shipping and emplacing the waste packages.

  14. Kisspeptin-10 inhibits cell migration in vitro via a receptor-GSK3 beta-FAK feedback loop in HTR8SVneo cells.

    PubMed

    Roseweir, A K; Katz, A A; Millar, R P

    2012-05-01

    Kisspeptin inhibits cancer cell metastasis and placental trophoblast cell migration. Kisspeptin gene expression in the placenta and circulating kisspeptin levels change during normal pregnancy and they are altered in preeclampsia. We therefore assessed the effect of kisspeptin-10 on the in vitro migration of a human placental cell line derived from first trimester extravillious trophoblasts (HTR8SVneo). HTR8SVneo cells specifically bound 125I-Kisspeptin-10 but kisspeptin-10 did not induce inositol phosphate production. Cell migration was inhibited by kisspeptin-10 with a maximal inhibition at 100nM. The signaling pathways involved in inhibition of cell migration were examined. Treatment with kisspeptin-10 elicited phosphorylation of GSK3 beta at Ser9 (which inhibits activity), with a 3-fold increase at 5 min. Transient phosphorylation of ERK1/2 and p38MAPK peaked at 10min. Phosphorylation of focal adhesion kinase (FAK) at Tyr925 increased 3-fold at 10 min. Inhibition of GSK3 beta correlated with release of beta-catenin into the cytoplasm. These signaling events were differentially blocked by inhibitors of G(q/11), Src, EGFR, PI(3)K, PKC and MEK. The data suggest that kisspeptin/GPR54 EGF-receptor transactivation leads to phosphorylation of ERK1/2, causing activation of p90rsk which in turn inhibits GSK3 beta via Ser9 phosphorylation. Inactivation of GSK3 beta results in release of beta-catenin into the cytoplasm, affecting cell-cell adhesion and Tyr925 phosphorylation of FAK, which increases phosphorylation of ERK1/2 via RAS/Raf-1 creating a feedback loop to enhance the effects on migration. These findings indicate that kisspeptin-10 inhibits the migration of human placental trophoblast-derived HTR8SVneo cells by stimulating complex ERK1/2-p90rsk-GSK3 beta-FAK feedback interactions.

  15. Long Non-Coding RNA Uc.187 Is Upregulated in Preeclampsia and Modulates Proliferation, Apoptosis, and Invasion of HTR-8/SVneo Trophoblast Cells.

    PubMed

    Cao, Chunyu; Li, Jingyun; Li, Jun; Liu, Lan; Cheng, Xiaoyan; Jia, Ruizhe

    2017-06-01

    Among the preeclampsia-related long non-cording RNAs (lncRNAs) screened with a gene chip in our preliminary study, uc.187 attracted our attention because of its high conservation across different species and significant positive correlation with preeclampsia (PE). The literature and bioinformatics analysis suggested that lncRNA uc.187 might be associated with cell growth, invasion, and apoptosis. The expression of uc.187 in severe preeclamptic placentas (n = 31) and normal placentas (n = 18) was evaluated by real-time reverse transcription polymerase chain reaction (qRT-PCR). We constructed a silencing lentivirus vector (uc.187 siRNA) to explore the biological function of uc.187 in the development and progression of HTR-8/SVneo trophoblast cells in vitro. Furthermore, we utilized CCK8 analysis, a transwell invasion assay, and flow cytometry to determine the role of uc.187 in the proliferation, invasion, and apoptosis of HTR-8/SVneo trophoblast cells. The proteins related to proliferation (PCNA, Ki67), invasion (MMP-2/-9 and TIMP-1), and apoptosis (caspase-3, Bcl-2) were evaluated with a Western blot assay. The results showed that there was an obvious upregulation of uc.187 expression in preeclamptic placental tissues. In addition, uc.187 silencing enhanced cell proliferation and invasion and reduced the cellular apoptotic response. Taken together, our findings suggest for the first time that abnormal expression of lncRNA uc.187 may lead to the aberrant biological behavior of HTR-8/SVneo cells. Therefore, we propose uc.187 as a novel lncRNA molecule that might contribute to the development of PE and might represent a potential diagnostic and therapeutic target for this disease. J. Cell. Biochem. 118: 1462-1470, 2017. © 2016 Wiley Periodicals, Inc.

  16. Upregulation of long noncoding RNA SPRY4-IT1 modulates proliferation, migration, apoptosis, and network formation in trophoblast cells HTR-8SV/neo.

    PubMed

    Zou, Yanfen; Jiang, Ziyan; Yu, Xiang; Sun, Ming; Zhang, Yuanyuan; Zuo, Qing; Zhou, Jing; Yang, Nana; Han, Ping; Ge, Zhiping; De, Wei; Sun, Lizhou

    2013-01-01

    SPRY4-IT1 has been reported to have extremely high expression in normal placenta tissues. It is a Long noncoding RNA (lncRNA), which is associated with cell growth, migration, invasion, and apoptosis in melanoma. A 2.8-fold increase of SPRY4-IT1 expression was validated by Real-time reverse transcription-polymerase chain reaction (qRT-PCR) in severe preeclamptic placenta as compared with that of the normal ones (n=25) in this study. Furthermore, the role of SPRY4-IT1 in proliferation, migration, apoptosis, and network formation ability of trophoblast cells HTR-8/SVneo was assessed. Suppression of SPRY4-IT1 using siRNA treatment and its overexpression using plasmid targeting SPRY4-IT1 were performed in order to explore the biological function of SPRY4-IT1 in the development and progression of trophoblast cells HTR-8/SVneo, in vitro. The results showed that SPRY4-IT1 knockdown enhanced the cell migration and proliferation, and reduced the response of cells to apoptosis. However, exogenous SPRY4-IT1 overexpression significantly decreased the cell migration and proliferation, while increased cell apoptosis. Our study showed for the first time that aberrant expression of lncRNA SPRY4-IT1 might contribute to the abnormal condition of trophoblast cells HTR-8/SVneo. Therefore, we proposed SPRY4-IT1 as a novel lncRNA molecule, which might be associated with the pathogenesis of preeclampsia and might provide a new target for its early diagnosis and treatment.

  17. Role of GAD2 and HTR1B genes in early-onset obsessive-compulsive disorder: results from transmission disequilibrium study.

    PubMed

    Mas, S; Pagerols, M; Gassó, P; Ortiz, A; Rodriguez, N; Morer, A; Plana, M T; Lafuente, A; Lazaro, L

    2014-04-01

    One of the leading biological models of obsessive-compulsive disorder (OCD) is the frontal-striatal-thalamic model. This study undertakes an extensive exploration of the variability in genes related to the regulation of the frontal-striatal-thalamic system in a sample of early-onset OCD trios. To this end, we genotyped 266 single nucleotide polymorphisms (SNPs) in 35 genes in 84 OCD probands and their parents. Finally, 75 complete trios were included in the analysis. Twenty SNPs were overtransmitted from parents to early-onset OCD probands and presented nominal pointwise P < 0.05 values. Three of these polymorphisms achieved P < 2 × 10(-4), the significant P-value after Bonferroni corrections: rs8190748 and rs992990 localized in GAD2 and rs2000292 in HTR1B. When we stratified our sample according to gender, different trends were observed between males and females. In males, SNP rs2000292 (HTR1B) showed the lowest P-value (P = 0.0006), whereas the SNPs in GAD2 were only marginally significant (P = 0.01). In contrast, in females HTR1B polymorphisms were not significant, whereas rs8190748 (GAD2) showed the lowest P-value (P = 0.0006). These results are in agreement with several lines of evidence that indicate a role for the serotonin and γ-Aminobutyric acid (GABA) pathways in the risk of early-onset OCD and with the gender differences in OCD pathophysiology reported elsewhere. However, our results need to be replicated in studies with larger cohorts in order to confirm these associations.

  18. Association between HTR2A T102C polymorphism and major depressive disorder: a meta-analysis in the Chinese population.

    PubMed

    Lin, Chun-Xia; Hu, Zhe; Yan, Ze-Ming; Li, Wen; Chen, Yu-Sen; Zhao, Jiang-Hao; Zhang, Liang-Qing; Zhao, Bin; Zhong, Wang-Tao; Feng, Du

    2015-01-01

    Although a number of studies have been conducted on the association between HTR2A T102C polymorphism and major depressive disorder (MDD) in Chinese, this association remains elusive and controversial. To clarify the effects of HTR2A T102C polymorphism on the risk of MDD, a meta-analysis was performed in the Chinese population. Related studies were identified from PubMed, Springer Link, Ovid, Chinese Wanfang Data Knowledge Service Platform, Chinese National Knowledge Infrastructure (CNKI), and Chinese Biology Medicine (CBM) till 5 May 2015. The odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to estimate the strength of the associations. Statistical analyses were conducted with Version 10.0 STATA statistical software. A total of 12 case-control studies including 1444 MDD cases and 1445 controls were involved in this meta-analysis. Overall, no significant association with MDD risk was provided in the Chinese population (C vs. T: OR=0.97, 95% CI: 0.81-1.17, 95%; CC vs. TT: OR=0.95, 95% CI: 0.65-1.37; CC+TC vs. TT: OR=0.96, 95% CI: 0.75-1.12; CC vs. TT+TC: OR=0.94, 95% CI: 0.78-1.12). In subgroup analyses stratified by geographic area and source of controls, no significant association was found in any of the subgroups. In conclusion, this meta-analysis indicate that the HTR2A T102C polymorphism is not associated with susceptibility to MDD in Chinese population.

  19. Association between HTR2A T102C polymorphism and major depressive disorder: a meta-analysis in the Chinese population

    PubMed Central

    Lin, Chun-Xia; Hu, Zhe; Yan, Ze-Ming; Li, Wen; Chen, Yu-Sen; Zhao, Jiang-Hao; Zhang, Liang-Qing; Zhao, Bin; Zhong, Wang-Tao; Feng, Du

    2015-01-01

    Although a number of studies have been conducted on the association between HTR2A T102C polymorphism and major depressive disorder (MDD) in Chinese, this association remains elusive and controversial. To clarify the effects of HTR2A T102C polymorphism on the risk of MDD, a meta-analysis was performed in the Chinese population. Related studies were identified from PubMed, Springer Link, Ovid, Chinese Wanfang Data Knowledge Service Platform, Chinese National Knowledge Infrastructure (CNKI), and Chinese Biology Medicine (CBM) till 5 May 2015. The odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to estimate the strength of the associations. Statistical analyses were conducted with Version 10.0 STATA statistical software. A total of 12 case-control studies including 1444 MDD cases and 1445 controls were involved in this meta-analysis. Overall, no significant association with MDD risk was provided in the Chinese population (C vs. T: OR=0.97, 95% CI: 0.81-1.17, 95%; CC vs. TT: OR=0.95, 95% CI: 0.65-1.37; CC+TC vs. TT: OR=0.96, 95% CI: 0.75-1.12; CC vs. TT+TC: OR=0.94, 95% CI: 0.78-1.12). In subgroup analyses stratified by geographic area and source of controls, no significant association was found in any of the subgroups. In conclusion, this meta-analysis indicate that the HTR2A T102C polymorphism is not associated with susceptibility to MDD in Chinese population. PMID:26885016

  20. The presence of both serotonin 1A receptor (HTR1A) and dopamine transporter (DAT1) gene variants increase the risk of borderline personality disorder

    PubMed Central

    Joyce, Peter R.; Stephenson, John; Kennedy, Martin; Mulder, Roger T.; McHugh, Patrick C.

    2014-01-01

    Dysfunction in the dopaminergic and serotonergic neurotransmitter systems has been demonstrated to be important in the etiology of borderline personality disorder (BPD). We investigated the relationship of two BPD risk factors, the HTR1A promoter polymorphism -1019C > G (rs6295) and the dopamine transporter (DAT1) repeat allele, with BPD in a major depressive disorder cohort of 367 patients. Out-patients with major depressive disorder were recruited for two treatment trials and assessed for personality disorders, including BPD. DNA samples were collected and the rs6295 polymorphism was detected with a TaqMan® assay. The DAT1 repeat allele was genotyped using a modified PCR method. The impact of polymorphisms on BPD was statistically analyzed using uncontrolled logistic and multiple logistic regression models. BPD patients had higher frequencies of the DAT1 9,9 (OR = 2.67) and 9,10 (OR = 3.67) genotypes and also those homozygous HTR1A G allele (OR = 2.03). No significant interactions between HTR1A and DAT1 genotypes, were observed; however, an increased risk of BPD was observed for those patients who were either 9,10; G,G (OR = 6.64) and 9,9; C,G (OR = 5.42). Furthermore, the odds of BPD in patients exhibiting high-risk variants of these two genes differed from those of patients in low-risk groups by up to a factor of 9. Our study provides evidence implicating the importance of the serotonergic and dopaminergic systems in BPD and that the interaction between genes from different neurotransmitters may play a role in the susceptibility to BPD. PMID:24432029

  1. Genome-wide association study of theta band event-related oscillations identifies serotonin receptor gene HTR7 influencing risk of alcohol dependence.

    PubMed

    Zlojutro, Mark; Manz, Niklas; Rangaswamy, Madhavi; Xuei, Xiaoling; Flury-Wetherill, Leah; Koller, Daniel; Bierut, Laura J; Goate, Alison; Hesselbrock, Victor; Kuperman, Samuel; Nurnberger, John; Rice, John P; Schuckit, Marc A; Foroud, Tatiana; Edenberg, Howard J; Porjesz, Bernice; Almasy, Laura

    2011-01-01

    Event-related brain oscillations (EROs) represent highly heritable neuroelectrical correlates of human perception and cognitive performance that exhibit marked deficits in patients with various psychiatric disorders. We report the results of the first genome-wide association study (GWAS) of an ERO endophenotype-frontal theta ERO evoked by visual oddball targets during P300 response in 1,064 unrelated individuals drawn from a study of alcohol dependence. Forty-two SNPs of the Illumina HumanHap 1 M microarray were selected from the theta ERO GWAS for replication in family-based samples (N = 1,095), with four markers revealing nominally significant association. The most significant marker from the two-stage study is rs4907240 located within ARID protein 5A gene (ARID5A) on chromosome 2q11 (unadjusted, Fisher's combined P = 3.68 × 10⁻⁶). However, the most intriguing association to emerge is with rs7916403 in serotonin receptor gene HTR7 on chromosome 10q23 (combined P = 1.53 × 10⁻⁴), implicating the serotonergic system in the neurophysiological underpinnings of theta EROs. Moreover, promising SNPs were tested for association with diagnoses of alcohol dependence (DSM-IV), revealing a significant relationship with the HTR7 polymorphism among GWAS case-controls (P = 0.008). Significant recessive genetic effects were also detected for alcohol dependence in both case-control and family-based samples (P = 0.031 and 0.042, respectively), with the HTR7 risk allele corresponding to theta ERO reductions among homozygotes. These results suggest a role of the serotonergic system in the biological basis of alcohol dependence and underscore the utility of analyzing brain oscillations as a powerful approach to understanding complex genetic psychiatric disorders. Copyright © 2010 Wiley-Liss, Inc.

  2. The presence of both serotonin 1A receptor (HTR1A) and dopamine transporter (DAT1) gene variants increase the risk of borderline personality disorder.

    PubMed

    Joyce, Peter R; Stephenson, John; Kennedy, Martin; Mulder, Roger T; McHugh, Patrick C

    2014-01-07

    Dysfunction in the dopaminergic and serotonergic neurotransmitter systems has been demonstrated to be important in the etiology of borderline personality disorder (BPD). We investigated the relationship of two BPD risk factors, the HTR1A promoter polymorphism -1019C > G (rs6295) and the dopamine transporter (DAT1) repeat allele, with BPD in a major depressive disorder cohort of 367 patients. Out-patients with major depressive disorder were recruited for two treatment trials and assessed for personality disorders, including BPD. DNA samples were collected and the rs6295 polymorphism was detected with a TaqMan(®) assay. The DAT1 repeat allele was genotyped using a modified PCR method. The impact of polymorphisms on BPD was statistically analyzed using uncontrolled logistic and multiple logistic regression models. BPD patients had higher frequencies of the DAT1 9,9 (OR = 2.67) and 9,10 (OR = 3.67) genotypes and also those homozygous HTR1A G allele (OR = 2.03). No significant interactions between HTR1A and DAT1 genotypes, were observed; however, an increased risk of BPD was observed for those patients who were either 9,10; G,G (OR = 6.64) and 9,9; C,G (OR = 5.42). Furthermore, the odds of BPD in patients exhibiting high-risk variants of these two genes differed from those of patients in low-risk groups by up to a factor of 9. Our study provides evidence implicating the importance of the serotonergic and dopaminergic systems in BPD and that the interaction between genes from different neurotransmitters may play a role in the susceptibility to BPD.

  3. Polymorphism in the Serotonin Receptor 2a (HTR2A) Gene as Possible Predisposal Factor for Aggressive Traits

    PubMed Central

    Banlaki, Zsofia; Elek, Zsuzsanna; Nanasi, Tibor; Szekely, Anna; Nemoda, Zsofia; Sasvari-Szekely, Maria; Ronai, Zsolt

    2015-01-01

    Aggressive manifestations and their consequences are a major issue of mankind, highlighting the need for understanding the contributory factors. Still, aggression-related genetic analyses have so far mainly been conducted on small population subsets such as individuals suffering from a certain psychiatric disorder or a narrow-range age cohort, but no data on the general population is yet available. In the present study, our aim was to identify polymorphisms in genes affecting neurobiological processes that might explain some of the inter-individual variation between aggression levels in the non-clinical Caucasian adult population. 55 single nucleotide polymorphisms (SNP) were simultaneously determined in 887 subjects who also filled out the self-report Buss-Perry Aggression Questionnaire (BPAQ). Single marker association analyses between genotypes and aggression scores indicated a significant role of rs7322347 located in the HTR2A gene encoding serotonin receptor 2a following Bonferroni correction for multiple testing (p = 0.0007) both for males and females. Taking the four BPAQ subscales individually, scores for Hostility, Anger and Physical Aggression showed significant association with rs7322347 T allele in themselves, while no association was found with Verbal Aggression. Of the subscales, relationship with rs7322347 was strongest in the case of Hostility, where statistical significance virtually equaled that observed with the whole BPAQ. In conclusion, this is the first study to our knowledge analyzing SNPs in a wide variety of genes in terms of aggression in a large sample-size non-clinical adult population, also describing a novel candidate polymorphism as predisposal to aggressive traits. PMID:25658328

  4. Betamethasone, progesterone and RU-486 (mifepristone) exert similar effects on connexin expression in trophoblast-derived HTR-8/SVneo cells.

    PubMed

    Cervellati, F; Pavan, B; Lunghi, L; Manni, E; Fabbri, E; Mascoli, C; Biondi, C; Patella, A; Vesce, F

    2011-01-01

    Connexins (Cx) are membrane proteins able to influence cell trophoblast responses, such as proliferation, differentiation, migration and invasiveness. Likewise, glucocorticoids are also known to modulate many factors involved in implantation, including trophoblast gap-junction intercellular communication, although their influence on pregnancy is controversial. In order to investigate the effects of betamethasone, a synthetic glucocorticoid, on Cx and glucocorticoid receptor (GR) expression and localisation, as well as on cell proliferation, the extravillous trophoblast-derived HTR-8/SVneo cell line was used as a model. The results, confirmed by means of immunofluorescence, demonstrate that betamethasone selectively modifies GR and Cx expression, enhancing the GRα isoform without affecting GRβ, and inhibiting Cx40 expression whilst increasing that of Cx43 and Cx45. Furthermore, betamethasone was shown to exert an inhibitory action on cell proliferation. In this model the abortion drug RU-486 (mifepristone), reported to be a GR antagonist, did not counteract this effect of betamethasone. On the contrary, it induced responses similar to those of the hormone. Knowing that RU-486 is also a potent progesterone-receptor antagonist, the effect of progesterone alone and in combination with the drug on Cx expression and cell proliferation was then tested. Progesterone showed the same effect as betamethasone on Cx expression, but it did not affect proliferation. Based on these results, neither the abortion effects of RU-486 nor the protective action of betamethasone and progesterone are exerted by modulation of Cx. RU-486 did not antagonise the progesterone effect, suggesting that its abortive action does not involve alteration of trophoblast Cx expression.

  5. 5-Hydroxytryptamine-2A receptor gene (HTR 2 A) candidate polymorphism (T 102 C): Role for human platelet function under pharmacological challenge ex vivo.

    PubMed

    Ozdener, F; Gülbas, Z; Erol, K; Ozdemir, V

    2005-01-01

    Although the environmental and life-style factors influencing individual predisposition to acute myocardial infarction (AMI) have been well documented, little is known on the identity of genetic loci that may contribute to risk for AMI. Recently, genetic studies in patients with nonfatal AMI have suggested an association with the T 102 C polymorphism in the serotonin 5-HT(2A) receptor gene (HTR 2 A). Considering the significant role of the 5-HT(2A) receptor in serotonin-induced platelet responses and the contribution of platelet (patho)physiology to thromboembolic events, we postulated that the increased susceptibility to AMI in patients with the T 102 homozygosity may be attributable, in part, to altered serotonin-mediated platelet function. In a group of healthy volunteers recruited from the Eskisehir region in central Turkey (N=37), we investigated the functional consequences of HTR 2 A T 102 C polymorphism in relation to platelet pharmacodynamics ex vivo. The platelet shape change and aggregation response to serotonin were measured with use of the platelet aggregometry and expressed as aggregometer output (mm). Because the circulating catecholamine hormone epinephrine can augment platelet aggregation, pharmacodynamic response (aggregation and its inhibition by 5-HT(2A) receptor antagonist cyproheptadine) was measured in the presence of both serotonin and epinephrine, to mimic the clinical situation in patients. The mean platelet aggregation was higher by 38% in subjects with T 10 2 homozygosity (T/T genotype, N=13) when compared with the carriers of the 102 C-allele (T/C and the C/C genotypic groups, N=24) (39.5 mm+/-12.3 vs. 28.7 mm+/-16.8, respectively) (mean+/-SD) (p<0.05). On the other hand, neither the serotonin-induced platelet shape change nor the cyproheptadine inhibition of platelet aggregation was influenced by the HTR 2 A T 102 C genetic variation (p>0.05). These findings in healthy subjects may provide a mechanistic explanation for the previously

  6. Association of HTR2A T102C and A-1438G polymorphisms with susceptibility to major depressive disorder: a meta-analysis.

    PubMed

    Zhao, Xue; Sun, Liang; Sun, Ye-Huan; Ren, Cizao; Chen, Jian; Wu, Zhen-Qiang; Jiang, Yu-Hong; Lv, Xiao-Ling

    2014-12-01

    Serotonin 2A receptor (HTR2A) gene was implicated to be associated with major depressive disorder (MDD) susceptibility due to its role of key neurotransmitter in many physiologic processes. A great number of related studies reported in different populations have emerged. The results of these studies, however, have been inconsistent and thereby definite conclusions are difficult to establish. With the cumulative data in recent years, it was necessary to carry out a comprehensive analysis of previous findings. Electronic databases were systematically searched for studies published before May 2013. Pooled odds ratios (OR) and 95 % confidence interval (CI) were estimated under three different genetic models. Subgroup and sensitivity analyses were also performed. A total of 21 studies, 3,299 patients and 4,092 controls, met the selection criteria. 15 studies included HTR2A T102C polymorphism (with a total of 2,409 patients and 3,130 controls), and 9 studies included HTR2A A-1438G polymorphism (with a total of 1,510 patients and 2,281 controls). Our results showed that no significant association of MDD susceptibility with T102C polymorphism was found in allelic analysis and genotypic analysis (For T vs. C: OR = 1.06, 95 % CI = 0.95-1.18, P = 0.307; For TT + TC vs. CC: OR = 1.07, 95 % CI = 0.90-1.28, P = 0.451; For TT vs. TC + CC: OR = 1.08, 95 % CI = 0.95-1.22, P = 0.235). With respect to A-1438G polymorphism, however, carriers with A allele tend to suffer from MDD (AA + AG vs. GG: OR = 1.20, 95 % CI = 1.02-1.43, P = 0.030). When stratified by race for T102C polymorphism and A-1438G polymorphism of the HTR2A, we found no significant association. In conclusions, our study suggests that the A allele of A-1438G polymorphism might play a role in susceptibility to MDD. On the contrary, T102C polymorphism does not seem to be capable of modifying MDD risk.

  7. Effect of polysaccharides extract of rhizoma atractylodis macrocephalae on thymus, spleen and cardiac indexes, caspase-3 activity ratio, Smac/DIABLO and HtrA2/Omi protein and mRNA expression levels in aged rats.

    PubMed

    Guo, Ling; Sun, Yong Le; Wang, Ai Hong; Xu, Chong En; Zhang, Meng Yuan

    2012-10-01

    This study was designed to determine the possible protective effect of polysaccharides extract of rhizoma atractylodis macrocephalae on heart function in aged rats. Polysaccharides extract of rhizoma atractylodis macrocephalae was administered to aged rats. Results showed that thymus, spleen and cardiac indexs were significantly increased, whereas caspase-3 activity ratio, Smac/DIABLO and HtrA2/Omi protein expression, Smac/DIABLO and HtrA2/Omi mRNA expression levels were markedly reduced. It can be concluded that polysaccharides extract of rhizoma atractylodis macrocephalae may enhance immunity and improve heart function in aged rats.

  8. Blocking Epidermal Growth Factor Receptor Signaling in HTR-8/SVneo First Trimester Trophoblast Cells Results in Dephosphorylation of PKBα/AKT and Induces Apoptosis

    PubMed Central

    Bolnick, J.; Albitar, L.; Laidler, L. L.; Abdullah, R.; Leslie, K. K.

    2011-01-01

    We identified a major peptide signaling target of EGF/EGFR pathway and explored the consequences of blocking or activating this pathway in the first trimester extravillous trophoblast cells, HTR-8/SVneo. A global analysis of protein phosphorylation was undertaken using novel technology (Kinexus Kinetworks) that utilizes SDS-polyacrylamide minigel electrophoresis and multi-lane immunoblotting to permit specific and semiquantitative detection of multiple phosphoproteins. Forty-seven protein phosphorylation sites were queried, and the results reported based on relative phosphorylation at each site. EGF- and Iressa-(gefitinib, ZD1839, an inhibitor of EGFR) treated HTR-8/SVneo cells were subjected to immunoblotting and flow cytometry to confirm the phosphoprotein screen and to assess the effects of EGF versus Iressa on cell cycle and apoptosis. EGFR mediates the phosphorylation of important signaling proteins, including PKBα/AKT. This pathway is likely to be central to EGFR-mediated trophoblast survival. Furthermore, EGF treatment induces proliferation and inhibits apoptosis, while Iressa induces apoptosis. PMID:21876698

  9. BMP-2, Hypoxia, and COL1A1/HtrA1 siRNAs Favor Neo-Cartilage Hyaline Matrix Formation in Chondrocytes

    PubMed Central

    Ollitrault, David; Legendre, Florence; Drougard, Carole; Briand, Mélanie; Benateau, Hervé; Goux, Didier; Chajra, Hanane; Poulain, Laurent; Hartmann, Daniel; Vivien, Denis; Shridhar, Vijayalakshmi; Baldi, Alfonso; Mallein-Gerin, Frédéric; Boumediene, Karim; Demoor, Magali

    2015-01-01

    Osteoarthritis (OA) is an irreversible pathology that causes a decrease in articular cartilage thickness, leading finally to the complete degradation of the affected joint. The low spontaneous repair capacity of cartilage prevents any restoration of the joint surface, making OA a major public health issue. Here, we developed an innovative combination of treatment conditions to improve the human chondrocyte phenotype before autologous chondrocyte implantation. First, we seeded human dedifferentiated chondrocytes into a collagen sponge as a scaffold, cultured them in hypoxia in the presence of a bone morphogenetic protein (BMP), BMP-2, and transfected them with small interfering RNAs targeting two markers overexpressed in OA dedifferentiated chondrocytes, that is, type I collagen and/or HtrA1 serine protease. This strategy significantly decreased mRNA and protein expression of type I collagen and HtrA1, and led to an improvement in the chondrocyte phenotype index of differentiation. The effectiveness of our in vitro culture process was also demonstrated in the nude mouse model in vivo after subcutaneous implantation. We, thus, provide here a new protocol able to favor human hyaline chondrocyte phenotype in primarily dedifferentiated cells, both in vitro and in vivo. Our study also offers an innovative strategy for chondrocyte redifferentiation and opens new opportunities for developing therapeutic targets. PMID:24957638

  10. Blocking Epidermal Growth Factor Receptor Signaling in HTR-8/SVneo First Trimester Trophoblast Cells Results in Dephosphorylation of PKBα/AKT and Induces Apoptosis.

    PubMed

    Bolnick, J; Albitar, L; Laidler, L L; Abdullah, R; Leslie, K K

    2011-01-01

    We identified a major peptide signaling target of EGF/EGFR pathway and explored the consequences of blocking or activating this pathway in the first trimester extravillous trophoblast cells, HTR-8/SVneo. A global analysis of protein phosphorylation was undertaken using novel technology (Kinexus Kinetworks) that utilizes SDS-polyacrylamide minigel electrophoresis and multi-lane immunoblotting to permit specific and semiquantitative detection of multiple phosphoproteins. Forty-seven protein phosphorylation sites were queried, and the results reported based on relative phosphorylation at each site. EGF- and Iressa-(gefitinib, ZD1839, an inhibitor of EGFR) treated HTR-8/SVneo cells were subjected to immunoblotting and flow cytometry to confirm the phosphoprotein screen and to assess the effects of EGF versus Iressa on cell cycle and apoptosis. EGFR mediates the phosphorylation of important signaling proteins, including PKBα/AKT. This pathway is likely to be central to EGFR-mediated trophoblast survival. Furthermore, EGF treatment induces proliferation and inhibits apoptosis, while Iressa induces apoptosis.

  11. MiR-30e and miR-181d control radial glia cell proliferation via HtrA1 modulation.

    PubMed

    Nigro, A; Menon, R; Bergamaschi, A; Clovis, Y M; Baldi, A; Ehrmann, M; Comi, G; De Pietri Tonelli, D; Farina, C; Martino, G; Muzio, L

    2012-08-02

    The precise mechanisms by which microRNAs (miRNAs) contribute to the dynamic regulation of gene expression during the forebrain development are still partly elusive. Here we show that the depletion of miRNAs in the cerebral cortex and hippocampus, via genetic inactivation of Dicer after the onset of forebrain neurogenesis, profoundly impairs the morphological and proliferative characteristics of neural stem and progenitor cells. The cytoarchitecture and self-renewal potential of radial glial (RG) cells located within the cerebral cortex and the hippocampus were profoundly altered, thus causing a significant derangement of both the post natal dorsal sub-ventricular zone and the dentate gyrus. This effect was attributed to the High-temperature requirement A serine peptidase 1 (HtrA1) gene product whose overexpression in the developing forebrain recapitulated some of the aspects of the Dicer(-/-) phenotype. MiR-30e and miR-181d were identified as posttranscriptional negative regulators of HtrA1 by binding to its 3' untranslated region. In vivo overexpression of miR-30e and miR-181d in Dicer(-/-) forebrain rescued RG proliferation defects.

  12. BMP-2, hypoxia, and COL1A1/HtrA1 siRNAs favor neo-cartilage hyaline matrix formation in chondrocytes.

    PubMed

    Ollitrault, David; Legendre, Florence; Drougard, Carole; Briand, Mélanie; Benateau, Hervé; Goux, Didier; Chajra, Hanane; Poulain, Laurent; Hartmann, Daniel; Vivien, Denis; Shridhar, Vijayalakshmi; Baldi, Alfonso; Mallein-Gerin, Frédéric; Boumediene, Karim; Demoor, Magali; Galera, Philippe

    2015-02-01

    Osteoarthritis (OA) is an irreversible pathology that causes a decrease in articular cartilage thickness, leading finally to the complete degradation of the affected joint. The low spontaneous repair capacity of cartilage prevents any restoration of the joint surface, making OA a major public health issue. Here, we developed an innovative combination of treatment conditions to improve the human chondrocyte phenotype before autologous chondrocyte implantation. First, we seeded human dedifferentiated chondrocytes into a collagen sponge as a scaffold, cultured them in hypoxia in the presence of a bone morphogenetic protein (BMP), BMP-2, and transfected them with small interfering RNAs targeting two markers overexpressed in OA dedifferentiated chondrocytes, that is, type I collagen and/or HtrA1 serine protease. This strategy significantly decreased mRNA and protein expression of type I collagen and HtrA1, and led to an improvement in the chondrocyte phenotype index of differentiation. The effectiveness of our in vitro culture process was also demonstrated in the nude mouse model in vivo after subcutaneous implantation. We, thus, provide here a new protocol able to favor human hyaline chondrocyte phenotype in primarily dedifferentiated cells, both in vitro and in vivo. Our study also offers an innovative strategy for chondrocyte redifferentiation and opens new opportunities for developing therapeutic targets.

  13. Real-time PCR detection of Staphylococcus aureus in milk and meat using new primers designed from the heat shock protein gene htrA sequence.

    PubMed

    Chiang, Yu-Cheng; Fan, Chih-Ming; Liao, Wan-Wen; Lin, Chien-Ku; Tsen, Hau-Yang

    2007-12-01

    Staphylococcus aureus may cause foodborne disease outbreaks and staphylococcal infections and is one of the major causes of mastitis. Rapid and reliable methods for detection of this microorganism in milk and other foods are needed. In this study, we designed a primer set from the sequence of the heat shock protein gene htrA, a gene coding for high-temperature-requirement A (HtrA) protein, and used it for real-time PCR detection of S. aureus isolates: 16 reference strains and 40 strains isolated from food-poisoning cases. All strains tested generated positive results. Bacterial strains other than S. aureus, including strains of other Staphylococcus species, did not produce positive results. When this primer set was used for the real-time PCR detection of S. aureus in milk and meat samples without the preenrichment step, samples with target cell numbers greater than 10(3) CFU/ml or CFU/g could be detected, indicating the potential quantitative ability of this real-time PCR assay. With a 10-h preenrichment step, however, a detection limit of 1 CFU/ml or CFU/g could be obtained.

  14. Predicting cortisol stress responses in older individuals: influence of serotonin receptor 1A gene (HTR1A) and stressful life events.

    PubMed

    Armbruster, Diana; Mueller, Anett; Strobel, Alexander; Lesch, Klaus-Peter; Brocke, Burkhard; Kirschbaum, Clemens

    2011-06-01

    Considerable variability in the activity of the hypothalamus-pituitary-adrenal (HPA) axis in response to stress has been found in quantitative genetic studies investigating healthy individuals suggesting that at least part of this variance is due to genetic factors. Since the HPA axis is regulated by a neuronal network including amygdala, hippocampus, prefrontal cortex as well as brainstem circuits, the investigation of candidate genes that impact neurotransmitter systems related to these brain regions might further elucidate the genetic underpinnings of the stress response. However, aside from genetic risk factors, past stressful life events might also result in long-term adjustments of HPA axis reactivity. Here, we investigated the effects of the -1019 G/C polymorphism in the HTR1A gene encoding the serotonin (5-HT) receptor 1A (5-HT(1A)) and stressful life events experienced during childhood and adolescence on changes in cortisol levels in response to the Trier Social Stress Test (TSST) in a sample of healthy older adults (N=97). Regression analyses revealed a significant effect of HTR1A genotype with the G allele being associated with a less pronounced stress response. In addition, an inverse relationship between past stressful life events and cortisol release but no gene × environment interaction was detected. The results further underscore the crucial role of functional serotonergic genetic variation as well as stressful events during critical stages of development on the acute stress response later in life. Copyright © 2011 Elsevier Inc. All rights reserved.

  15. Verify Super Double-Heterogeneous Spherical Lattice Model for Equilibrium Fuel Cycle Analysis AND HTR Spherical Super Lattice Model for Equilibrium Fuel Cycle Analysis

    SciTech Connect

    Gray S. Chang

    2005-11-01

    The currently being developed advanced High Temperature gas-cooled Reactors (HTR) is able to achieve a simplification of safety through reliance on innovative features and passive systems. One of the innovative features in these HTRs is reliance on ceramic-coated fuel particles to retain the fission products even under extreme accident conditions. Traditionally, the effect of the random fuel kernel distribution in the fuel pebble / block is addressed through the use of the Dancoff correction factor in the resonance treatment. However, the Dancoff correction factor is a function of burnup and fuel kernel packing factor, which requires that the Dancoff correction factor be updated during Equilibrium Fuel Cycle (EqFC) analysis. An advanced KbK-sph model and whole pebble super lattice model (PSLM), which can address and update the burnup dependent Dancoff effect during the EqFC analysis. The pebble homogeneous lattice model (HLM) is verified by the burnup characteristics with the double-heterogeneous KbK-sph lattice model results. This study summarizes and compares the KbK-sph lattice model and HLM burnup analyzed results. Finally, we discuss the Monte-Carlo coupling with a fuel depletion and buildup code - ORIGEN-2 as a fuel burnup analysis tool and its PSLM calculated results for the HTR EqFC burnup analysis.

  16. Results of the Simulation of the HTR-Proteus Core 4.2 Using PEBBED-COMBINE: FY10 Report

    SciTech Connect

    Hans Gougar

    2010-07-01

    ABSTRACT The Idaho National Laboratory’s deterministic neutronics analysis codes and methods were applied to the computation of the core multiplication factor of the HTR-Proteus pebble bed reactor critical facility. This report is a follow-on to INL/EXT-09-16620 in which the same calculation was performed but using earlier versions of the codes and less developed methods. In that report, results indicated that the cross sections generated using COMBINE-7.0 did not yield satisfactory estimates of keff. It was concluded in the report that the modeling of control rods was not satisfactory. In the past year, improvements to the homogenization capability in COMBINE have enabled the explicit modeling of TRIS particles, pebbles, and heterogeneous core zones including control rod regions using a new multi-scale version of COMBINE in which the 1-dimensional discrete ordinate transport code ANISN has been integrated. The new COMBINE is shown to yield benchmark quality results for pebble unit cell models, the first step in preparing few-group diffusion parameters for core simulations. In this report, the full critical core is modeled once again but with cross sections generated using the capabilities and physics of the improved COMBINE code. The new PEBBED-COMBINE model enables the exact modeling of the pebbles and control rod region along with better approximation to structures in the reflector. Initial results for the core multiplication factor indicate significant improvement in the INL’s tools for modeling the neutronic properties of a pebble bed reactor. Errors on the order of 1.6-2.5% in keff are obtained; a significant improvement over the 5-6% error observed in the earlier This is acceptable for a code system and model in the early stages of development but still too high for a production code. Analysis of a simpler core model indicates an over-prediction of the flux in the low end of the thermal spectrum. Causes of this discrepancy are under investigation. New

  17. Effect of hepatitis B virus infection on trophoblast cell line (HTR-8/SVneo) and choriocarcinoma cell line (JEG3) is linked to CD133-2 (AC141) expression.

    PubMed

    Cui, Hong; Chen, Jing; Na, Quan

    2016-01-01

    Mother-to-infant transmission of hepatitis B virus (HBV) plays an important role in the chronic carrier state in China. In our studies, the response of trophoblast cell and choriocarcinoma cell to HBV infection regarding the expression of CD133-2 (AC141) was evaluated. Western blot and RT-PCR showed that a high level of CD133-2 protein and mRNA in HTR-8/SVneo cells, but a low level in JEG-3 cells. Lower proliferation and mobility, and higher apoptosis were observed in HTR-8/SVneo cells and JEG-3-CD133-2(+) cells after HBV infection than those in HTR-8-CD133-2(-) cells and JEG-3 cells. Our main finding is that CD133-negative cells (HTR-8-CD133-2(-) and JEG-3) are prone to HBV infection. In the last, our data indicated that the activation of Smad signaling pathway and the induction of epithelial-mesenchymal transition (EMT) in CD133-negative cells after HBV infection. In summary, our study demonstrated that CD133 is a key factor that mediated HBV infection to trophoblast cell and choriocarcinoma cell.

  18. HTR-Proteus Pebble Bed Experimental Program Cores 5,6,7,&8: Columnar Hexagonal Point-on-Point Packing with a 1:2 Moderator-to-Fuel Pebble Ratio

    SciTech Connect

    Bess, John D.; Sterbentz, James W.; Snoj, Luka; Lengar, Igor; Koberl, Oliver

    2015-03-01

    PROTEUS is a zero-power research reactor based on a cylindrical graphite annulus with a central cylindrical cavity. The graphite annulus remains basically the same for all experimental programs, but the contents of the central cavity are changed according to the type of reactor being investigated. Through most of its service history, PROTEUS has represented light-water reactors, but from 1992 to 1996 PROTEUS was configured as a pebble-bed reactor (PBR) critical facility and designated as HTR-PROTEUS. The nomenclature was used to indicate that this series consisted of High Temperature Reactor experiments performed in the PROTEUS assembly. During this period, seventeen critical configurations were assembled and various reactor physics experiments were conducted. These experiments included measurements of criticality, differential and integral control rod and safety rod worths, kinetics, reaction rates, water ingress effects, and small sample reactivity effects (Ref. 3). HTR-PROTEUS was constructed, and the experimental program was conducted, for the purpose of providing experimental benchmark data for assessment of reactor physics computer codes. Considerable effort was devoted to benchmark calculations as a part of the HTR-PROTEUS program. References 1 and 2 provide detailed data for use in constructing models for codes to be assessed. Reference 3 is a comprehensive summary of the HTR-PROTEUS experiments and the associated benchmark program. This document draws freely from these references. Only Cores 9 and 10 are evaluated in this benchmark report due to similarities in their construction. The other core configurations of the HTR-PROTEUS program are evaluated in their respective reports as outlined in Section 1.0. Cores 9 and 10 were evaluated and determined to be acceptable benchmark experiments.

  19. HTR-PROTEUS PEBBLE BED EXPERIMENTAL PROGRAM CORES 5, 6, 7, & 8: COLUMNAR HEXAGONAL POINT-ON-POINT PACKING WITH A 1:2 MODERATOR-TO-FUEL PEBBLE RATIO

    SciTech Connect

    John D. Bess

    2013-03-01

    PROTEUS is a zero-power research reactor based on a cylindrical graphite annulus with a central cylindrical cavity. The graphite annulus remains basically the same for all experimental programs, but the contents of the central cavity are changed according to the type of reactor being investigated. Through most of its service history, PROTEUS has represented light-water reactors, but from 1992 to 1996 PROTEUS was configured as a pebble-bed reactor (PBR) critical facility and designated as HTR-PROTEUS. The nomenclature was used to indicate that this series consisted of High Temperature Reactor experiments performed in the PROTEUS assembly. During this period, seventeen critical configurations were assembled and various reactor physics experiments were conducted. These experiments included measurements of criticality, differential and integral control rod and safety rod worths, kinetics, reaction rates, water ingress effects, and small sample reactivity effects (Ref. 3). HTR-PROTEUS was constructed, and the experimental program was conducted, for the purpose of providing experimental benchmark data for assessment of reactor physics computer codes. Considerable effort was devoted to benchmark calculations as a part of the HTR-PROTEUS program. References 1 and 2 provide detailed data for use in constructing models for codes to be assessed. Reference 3 is a comprehensive summary of the HTR-PROTEUS experiments and the associated benchmark program. This document draws freely from these references. Only Cores 9 and 10 are evaluated in this benchmark report due to similarities in their construction. The other core configurations of the HTR-PROTEUS program are evaluated in their respective reports as outlined in Section 1.0. Cores 9 and 10 were evaluated and determined to be acceptable benchmark experiments.

  20. Recruitment by the Repressor Freud-1 of Histone Deacetylase-Brg1 Chromatin Remodeling Complexes to Strengthen HTR1A Gene Repression.

    PubMed

    Souslova, Tatiana; Mirédin, Kim; Millar, Anne M; Albert, Paul R

    2016-12-02

    Five-prime repressor element under dual repression binding protein-1 (Freud-1)/CC2D1A is genetically linked to intellectual disability and implicated in neuronal development. Freud-1 represses the serotonin-1A (5-HT1A) receptor gene HTR1A by histone deacetylase (HDAC)-dependent or HDAC-independent mechanisms in 5-HT1A-negative (e.g., HEK-293) or 5-HT1A-expressing cells (SK-N-SH), respectively. To identify the underlying mechanisms, Freud-1-associated proteins were affinity-purified from HEK-293 nuclear extracts and members of the Brg1/SMARCCA chromatin remodeling and Sin3A-HDAC corepressor complexes were identified. Pull-down assays using recombinant proteins showed that Freud-1 interacts directly with the Brg1 carboxyl-terminal domain; interaction with Brg1 required the carboxyl-terminal of Freud-1. Freud-1 complexes in HEK-293 and SK-N-SH cells differed, with low levels of BAF170/SMARCC2 and BAF57/SMARCE1 in HEK-293 cells and low-undetectable BAF155/SMARCC1, Sin3A, and HDAC1/2 in SK-N-SH cells. Similarly, by quantitative chromatin immunoprecipitation, Brg1-BAF170/57 and Sin3A-HDAC complexes were observed at the HTR1A promoter in HEK-293 cells, whereas in SK-N-SH cells, Sin3A-HDAC proteins were not detected. Quantifying 5-HT1A receptor mRNA levels in cells treated with siRNA to Freud-1, Brg1, or both RNAs addressed the functional role of the Freud-1-Brg1 complex. In HEK-293 cells, 5-HT1A receptor mRNA levels were increased only when both Freud-1 and Brg1 were depleted, but in SK-N-SH cells, depletion of either protein upregulated 5-HT1A receptor RNA. Thus, recruitment by Freud-1 of Brg1, BAF155, and Sin3A-HDAC complexes appears to strengthen repression of the HTR1A gene to prevent its expression inappropriate cell types, while recruitment of the Brg1-BAF170/57 complex is permissive to 5-HT1A receptor expression. Alterations in Freud-1-Brg1 interactions in mutants associated with intellectual disability could impair gene repression leading to altered neuronal

  1. HTR-PROTEUS PEBBLE BED EXPERIMENTAL PROGRAM CORE 4: RANDOM PACKING WITH A 1:1 MODERATOR-TO-FUEL PEBBLE RATIO

    SciTech Connect

    John D. Bess; Leland M. Montierth

    2013-03-01

    In its deployment as a pebble bed reactor (PBR) critical facility from 1992 to 1996, the PROTEUS facility was designated as HTR-PROTEUS. This experimental program was performed as part of an International Atomic Energy Agency (IAEA) Coordinated Research Project (CRP) on the Validation of Safety Related Physics Calculations for Low Enriched HTGRs. Within this project, critical experiments were conducted for graphite moderated LEU systems to determine core reactivity, flux and power profiles, reaction-rate ratios, the worth of control rods, both in-core and reflector based, the worth of burnable poisons, kinetic parameters, and the effects of moisture ingress on these parameters. One benchmark experiment was evaluated in this report: Core 4. Core 4 represents the only configuration with random pebble packing in the HTR-PROTEUS series of experiments, and has a moderator-to-fuel pebble ratio of 1:1. Three random configurations were performed. The initial configuration, Core 4.1, was rejected because the method for pebble loading, separate delivery tubes for the moderator and fuel pebbles, may not have been completely random; this core loading was rejected by the experimenters. Cores 4.2 and 4.3 were loaded using a single delivery tube, eliminating the possibility for systematic ordering effects. The second and third cores differed slightly in the quantity of pebbles loaded (40 each of moderator and fuel pebbles), stacked height of the pebbles in the core cavity (0.02 m), withdrawn distance of the stainless steel control rods (20 mm), and withdrawn distance of the autorod (30 mm). The 34 coolant channels in the upper axial reflector and the 33 coolant channels in the lower axial reflector were open. Additionally, the axial graphite fillers used in all other HTR-PROTEUS configurations to create a 12-sided core cavity were not used in the randomly packed cores. Instead, graphite fillers were placed on the cavity floor, creating a funnel-like base, to discourage ordering

  2. HTR-PROTEUS PEBBLE BED EXPERIMENTAL PROGRAM CORE 4: RANDOM PACKING WITH A 1:1 MODERATOR-TO-FUEL PEBBLE RATIO

    SciTech Connect

    John D. Bess; Leland M. Montierth

    2014-03-01

    In its deployment as a pebble bed reactor (PBR) critical facility from 1992 to 1996, the PROTEUS facility was designated as HTR-PROTEUS. This experimental program was performed as part of an International Atomic Energy Agency (IAEA) Coordinated Research Project (CRP) on the Validation of Safety Related Physics Calculations for Low Enriched HTGRs. Within this project, critical experiments were conducted for graphite moderated LEU systems to determine core reactivity, flux and power profiles, reaction-rate ratios, the worth of control rods, both in-core and reflector based, the worth of burnable poisons, kinetic parameters, and the effects of moisture ingress on these parameters. One benchmark experiment was evaluated in this report: Core 4. Core 4 represents the only configuration with random pebble packing in the HTR-PROTEUS series of experiments, and has a moderator-to-fuel pebble ratio of 1:1. Three random configurations were performed. The initial configuration, Core 4.1, was rejected because the method for pebble loading, separate delivery tubes for the moderator and fuel pebbles, may not have been completely random; this core loading was rejected by the experimenters. Cores 4.2 and 4.3 were loaded using a single delivery tube, eliminating the possibility for systematic ordering effects. The second and third cores differed slightly in the quantity of pebbles loaded (40 each of moderator and fuel pebbles), stacked height of the pebbles in the core cavity (0.02 m), withdrawn distance of the stainless steel control rods (20 mm), and withdrawn distance of the autorod (30 mm). The 34 coolant channels in the upper axial reflector and the 33 coolant channels in the lower axial reflector were open. Additionally, the axial graphite fillers used in all other HTR-PROTEUS configurations to create a 12-sided core cavity were not used in the randomly packed cores. Instead, graphite fillers were placed on the cavity floor, creating a funnel-like base, to discourage ordering

  3. Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8/SVneo cell line.

    PubMed

    Takao, Tomoka; Asanoma, Kazuo; Kato, Kiyoko; Fukushima, Kotaro; Tsunematsu, Ryosuke; Hirakawa, Toshio; Matsumura, Sueo; Seki, Hiroyuki; Takeda, Satoru; Wake, Norio

    2011-01-01

    Recently, numerous studies have identified that immature cell populations including stem cells and progenitor cells can be found among "side-population" (SP) cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. These features shown by the SP cells suggested that IL7R and IL1R2 are available as markers to detect the SP cells and that vCTB progenitor cells and trophoblast stem cells were involved in the SP cell population.

  4. Association between the transcriptional levels of Htr-1a and tryptophan hydroxylase-1 in the hippocampus and the antifatigue effects of leucine on rats with postoperative fatigue.

    PubMed

    Wu, Tiantian; Chen, Jing; Zhu, Jiang; Yu, Zhen

    2014-11-01

    Leucine (Leu), a branched-chain amino acid (BCAA), is widely used in clinical practice following severe burns, gastrointestinal surgery, trauma and sepsis. In the present study, the antifatigue effects of BCAAs on a postoperative fatigue (POF) rat model, induced by 70% intestinal resection, were investigated. Leu (16.5 g/l) was administered intraperitoneally at a dose of 18 ml/kg/day. The fatigue level and antifatigue effects of Leu were evaluated by open-field testing on day 1, 3, 5 and 7 after surgery. In addition, mRNA specimens were extracted and measured using a quantitative polymerase chain reaction method. The open-field test results indicated that Leu exhibited a significant antifatigue effect. The total distance travelled and the number of times the rats passed from the outermost grids of an open-top case were greatly improved in the Leu treatment group when compared with the POF model group. With the exception of the normal group, the mRNA expression levels of Htr-1a exhibited a similar trend in all other groups, reaching a climax on day 3 and 5, while being restored to a normal level on day 7. With regard to the Leu intervention group, the mRNA expression level of Htr-1a decreased significantly on day 3 and 5 following surgery. The mRNA expression levels of tryptophan hydroxylase-1 were unchanged in this short time period; however, the levels were increased gradually in the Leu treatment group. Therefore, Leu exhibited an apparent antifatigue effect on various 5-hydroxytryptamine-associated genes.

  5. HTR-PROTEUS Pebble Bed Experimental Program Cores 1, 1A, 2, and 3: Hexagonal Close Packing with a 1:2 Moderator-to-Fuel Pebble Ratio

    SciTech Connect

    John D. Bess; Barbara H. Dolphin; James W. Sterbentz; Luka Snoj; Igor Lengar; Oliver Köberl

    2013-03-01

    In its deployment as a pebble bed reactor (PBR) critical facility from 1992 to 1996, the PROTEUS facility was designated as HTR-PROTEUS. This experimental program was performed as part of an International Atomic Energy Agency (IAEA) Coordinated Research Project (CRP) on the Validation of Safety Related Physics Calculations for Low Enriched HTGRs. Within this project, critical experiments were conducted for graphite moderated LEU systems to determine core reactivity, flux and power profiles, reaction-rate ratios, the worth of control rods, both in-core and reflector based, the worth of burnable poisons, kinetic parameters, and the effects of moisture ingress on these parameters. Four benchmark experiments were evaluated in this report: Cores 1, 1A, 2, and 3. These core configurations represent the hexagonal close packing (HCP) configurations of the HTR-PROTEUS experiment with a moderator-to-fuel pebble ratio of 1:2. Core 1 represents the only configuration utilizing ZEBRA control rods. Cores 1A, 2, and 3 use withdrawable, hollow, stainless steel control rods. Cores 1 and 1A are similar except for the use of different control rods; Core 1A also has one less layer of pebbles (21 layers instead of 22). Core 2 retains the first 16 layers of pebbles from Cores 1 and 1A and has 16 layers of moderator pebbles stacked above the fueled layers. Core 3 retains the first 17 layers of pebbles but has polyethylene rods inserted between pebbles to simulate water ingress. The additional partial pebble layer (layer 18) for Core 3 was not included as it was used for core operations and not the reported critical configuration. Cores 1, 1A, 2, and 3 were determined to be acceptable benchmark experiments.

  6. HTR-PROTEUS Pebble Bed Experimental Program Cores 1, 1A, 2, and 3: Hexagonal Close Packing with a 1:2 Moderator-to-Fuel Pebble Ratio

    SciTech Connect

    John D. Bess; Barbara H. Dolphin; James W. Sterbentz; Luka Snoj; Igor Lengar; Oliver Köberl

    2012-03-01

    In its deployment as a pebble bed reactor (PBR) critical facility from 1992 to 1996, the PROTEUS facility was designated as HTR-PROTEUS. This experimental program was performed as part of an International Atomic Energy Agency (IAEA) Coordinated Research Project (CRP) on the Validation of Safety Related Physics Calculations for Low Enriched HTGRs. Within this project, critical experiments were conducted for graphite moderated LEU systems to determine core reactivity, flux and power profiles, reaction-rate ratios, the worth of control rods, both in-core and reflector based, the worth of burnable poisons, kinetic parameters, and the effects of moisture ingress on these parameters. Four benchmark experiments were evaluated in this report: Cores 1, 1A, 2, and 3. These core configurations represent the hexagonal close packing (HCP) configurations of the HTR-PROTEUS experiment with a moderator-to-fuel pebble ratio of 1:2. Core 1 represents the only configuration utilizing ZEBRA control rods. Cores 1A, 2, and 3 use withdrawable, hollow, stainless steel control rods. Cores 1 and 1A are similar except for the use of different control rods; Core 1A also has one less layer of pebbles (21 layers instead of 22). Core 2 retains the first 16 layers of pebbles from Cores 1 and 1A and has 16 layers of moderator pebbles stacked above the fueled layers. Core 3 retains the first 17 layers of pebbles but has polyethylene rods inserted between pebbles to simulate water ingress. The additional partial pebble layer (layer 18) for Core 3 was not included as it was used for core operations and not the reported critical configuration. Cores 1, 1A, 2, and 3 were determined to be acceptable benchmark experiments.

  7. Melker Meilensteine auf dem Weg in ein naturwissenschaftliches Zeitalter - Glanzlichter der Ausstellung zum Internationalen Astronomiejahr 2009 in der Melker Stiftsbibliothek.

    NASA Astrophysics Data System (ADS)

    Beck, Paul G.; Zotti, Georg

    2009-06-01

    Das Mittelalter wird weithin als die dunkle Epoche in der Geschichte der Europäischen Wissenschaften betrachtet, und insbesondere das Leben in den Klöstern galt lange Zeit als frei von jeglichem Interesse für Naturwissenschaften abseits der Medizin. Im Mittelalter galt die Astronomie bloß als Mittel zum Zweck, um religiöse und zivile Kalender erstellen zu können. Durch den Bestand der Handschriftenkammer der Melker Stiftsbibliothek eröffnet sich uns eine neue Sichtweise auf das gegen Ende des Mittelalters wachsende Interesse an den Naturwissenschaften. Dies wurde durch die starke Aufwertung der Klosterbibliothek im Rahmen der 'Melker Reform' im 15. Jahrhundert noch weiter verstärkt. Diese Epoche fällt mit der Frühphase der Universität Wien und der 'ersten Wiener Schule der Astronomie' zusammen. Dieser Artikel beleuchtet ausgewählte astronomischen Werke in der Melker Stiftsbibliothek zwischen dem frühen 9 und dem 18. Jahrhundert. Einen Schwerpunkt stellt das Wirken der Wiener Schule der Astronomie dar, wobei wir u.a. die Melker Abschrift von Peuerbachs Gutachten über den Kometen von 1456 sowie die im Stift Melk durchgeführte Beobachtung der Mondfinsternis von 1457 durch Regiomontanus und Peuerbach beleuchten. Dieser Beitrag ist der einführende Übersichtsartikel zum Ausstellungsprojekt in der Melker Stiftsbibliothek im Rahmen des Internationalen Jahres der Astronomie 2009. The medieval period is commonly seen as a dark epoch for science in Europe. Especially monasteries were seen as institutions without interest in natural sciences except for medicine. Astronomy was allegedly only a tool to construct religious and civil calendars. The inventory of the medieval manuscript collection of the library of the Abbey of Melk allows a new view on the growing interest in the exact sciences towards the end of the medieval ages. This interest was intensified through the increased importance of the monastery library due to the monastery reform

  8. Protective effect of nuclear factor E2-related factor 2 on inflammatory cytokine response to brominated diphenyl ether-47 in the HTR-8/SVneo human first trimester extravillous trophoblast cell line.

    PubMed

    Park, Hae-Ryung; Loch-Caruso, Rita

    2014-11-15

    Polybrominated diphenyl ethers (PBDEs) are widely used flame retardants, and BDE-47 is a prevalent PBDE congener detected in human tissues. Exposure to PBDEs has been linked to adverse pregnancy outcomes in humans. Although the underlying mechanisms of adverse birth outcomes are poorly understood, critical roles for oxidative stress and inflammation are implicated. The present study investigated antioxidant responses in a human extravillous trophoblast cell line, HTR-8/SVneo, and examined the role of nuclear factor E2-related factor 2 (Nrf2), an antioxidative transcription factor, in BDE-47-induced inflammatory responses in the cells. Treatment of HTR-8/SVneo cells with 5, 10, 15, and 20μM BDE-47 for 24h increased intracellular glutathione (GSH) levels compared to solvent control. Treatment of HTR-8/SVneo cells with 20μM BDE-47 for 24h induced the antioxidant response element (ARE) activity, indicating Nrf2 transactivation by BDE-47 treatment, and resulted in differential expression of redox-sensitive genes compared to solvent control. Pretreatment with tert-butyl hydroquinone (tBHQ) or sulforaphane, known Nrf2 inducers, reduced BDE-47-stimulated IL-6 release with increased ARE reporter activity, reduced nuclear factor kappa B (NF-κB) reporter activity, increased GSH production, and stimulated expression of antioxidant genes compared to non-Nrf2 inducer pretreated groups, suggesting that Nrf2 may play a protective role against BDE-47-mediated inflammatory responses in HTR-8/SVneo cells. These results suggest that Nrf2 activation significantly attenuated BDE-47-induced IL-6 release by augmentation of cellular antioxidative system via upregulation of Nrf2 signaling pathways, and that Nrf2 induction may be a potential therapeutic target to reduce adverse pregnancy outcomes associated with toxicant-induced oxidative stress and inflammation.

  9. Investigation of gene expression pattern of 5HTR2a and MAO-A in PBMCs of individuals who had been exposed to air pollution in highly polluted area.

    PubMed

    Sheikhha, Hajar; Emadi, Somayeh; Jorbozedar, Saman; Deilami, Gholamreza D; Ahangari, Ghasem

    2014-01-01

    Exposure to air pollutants can lead to lung inflammatory responses such as allergic asthma. Serotonin elevation is shown to be present in peripheral blood of symptomatic asthmatic patients. In this study, two genes, 5HTR2a and MAO-A playing important roles in serotonin function, were analyzed in peripheral blood mononuclear cells (PBMCs) of individuals who had been exposed to air pollution and allergic asthmatic patients as well. Thus, cDNA was synthesized from PBMCs' mRNA of the subjects and then gene expression was measured by real-time PCR. 5HTR2A gene expression in PBMCs of both exposed individuals and allergic asthmatic patients was significantly increased compared with control group, whereas MAO-A gene expression showed no significant changes in neither groups. These results indicate that there may be association between increased 5HTR2a gene expression and exposure to air pollution as well as asthma incidence. Some relevant patents are also outlined in this article.

  10. MTA1 and MTA3 Regulate HIF1a Expression in Hypoxia-Treated Human Trophoblast Cell Line HTR8/Svneo

    PubMed Central

    Wang, Kai; Chen, Ying; Ferguson, Susan D.; Leach, Richard E.

    2015-01-01

    Hypoxia plays an important role in placental trophoblast differentiation and function during early pregnancy. Hypoxia-inducible factor 1 alpha (HIF1a) is known to regulate cellular adaption to hypoxic conditions. However, our current understanding of the role of HIF1a in trophoblast physiology is far from complete. Metastasis Associated Protein 1 and 3 (MTA1 and MTA3) are components of the Nucleosome Remodeling and Deacetylase (NuRD) complex, a chromatin remodeling complex, and are highly expressed in term placental trophoblasts. However, the role of MTA1 and MTA3 in the hypoxic placental environment of early pregnancy is unknown. In the present study, we examined the association among MTA1, MTA3 and HIF1a expression under hypoxic conditions in trophoblasts both in vivo and in vitro. We first investigated the localization of MTA1 and MTA3 with HIF1a expression in the placental trophoblast of 1st trimester placenta via immunohistochemistry. Our data reveals that under physiologically hypoxic environment, MTA1 and MTA3 along with HIF1a are highly expressed by villous trophoblasts. Next, we investigated the effect of hypoxia on these genes in vitro using the first trimester-derived HTR8/SVneo cell line and observed up-regulation of MTA1 and MTA3 as well as HIF1a protein following hypoxia treatment. To investigate the direct effect of MTA1 and MTA3 upon HIF1a, we over-expressed MTA1 and MTA3 genes in HTR8/SVneo cells respectively and examined protein levels of HIF1a via Western blot as well as HIF1a target gene expression using a luciferase assay driven by a hypoxia-response element promoter (HRE-luciferase). We found that over-expressions of MTA1 and MTA3 up-regulate both HIF1a protein level and HRE-luciferase activity under hypoxic condition. In summary, both MTA1 and MTA3 are induced by hypoxia and up-regulate HIF1a expression and HIF1a target gene expression in trophoblasts. These data suggest that MTA1 and MTA3 play critical roles in trophoblast function and

  11. Xanthohumol impairs glucose uptake by a human first-trimester extravillous trophoblast cell line (HTR-8/SVneo cells) and impacts the process of placentation.

    PubMed

    Correia-Branco, Ana; Azevedo, Cláudia F; Araújo, João R; Guimarães, João T; Faria, Ana; Keating, Elisa; Martel, Fátima

    2015-10-01

    In this study, we aimed to investigate modulation of glucose uptake by the HTR-8/SVneo human first-trimester extravillous trophoblast cell line by a series of compounds and to study its consequences upon cell proliferation, viability and migration. We observed that uptake of (3)H-deoxy-d-glucose ((3)H-DG; 10 nM) was time-dependent, saturable, inhibited by cytochalasin B (50 and 100 µM), phloretin (0.5 mM) and phloridzin (1 mM), insulin-insensitive and sodium-independent. In the short term (30 min), neither 5-HT (100-1000 µM), melatonin (10 nM) nor the drugs of abuse ethanol (100 mM), nicotine (100 µM), cocaine (25 µM), amphetamine (10-25 µM) and 3,4-methylenedioxy-N-methamphetamine (10 µM) affected (3)H-DG uptake, while dexamethasone (100-1000 µM), fluoxetine (100-300 µM), quercetin, epigallocatechin-3-gallate (30-1000 µM), xanthohumol (XH) and resveratrol (1-500 µM) decreased it. XH was the most potent inhibitor [IC50 = 3.55 (1.37-9.20) µM] of (3)H-DG uptake, behaving as a non-competitive inhibitor of (3)H-DG uptake, both after short- and long-term (24 h) treatment. The effect of XH (5 µM; 24 h) upon (3)H-DG uptake involved mammalian target of rapamycin, tyrosine kinases and c-Jun N-terminal kinases intracellular pathways. Moreover, XH appeared to decrease cellular uptake of lactate due to inhibition of the monocarboxylate transporter 1. Additionally, XH (24 h; 5 µM) decreased cell viability, proliferation, culture growth and migration. The effects of XH upon cell viability and culture growth, but not the antimigratory effect, were mimicked by low extracellular glucose conditions and reversed by high extracellular glucose conditions. We thus suggest that XH, by inhibiting glucose cellular uptake and impairing HTR-8/SVneo cell viability and proliferation, may have a deleterious impact in the process of placentation. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All

  12. Improved recovery of active GST-fusion proteins from insoluble aggregates: solubilization and purification conditions using PKM2 and HtrA2 as model proteins.

    PubMed

    Park, Dae-Wook; Kim, Sang-Soo; Nam, Min-Kyung; Kim, Goo-Young; Kim, Jungho; Rhim, Hyangshuk

    2011-04-01

    The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins. However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein, we used 1% sarkosyl for lysis and a 1:200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity, even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.

  13. Steric Constraint in the Primary Photoproduct of Sensory Rhodopsin II Is a Prerequisite for Light-Signal Transfer to HtrII†

    PubMed Central

    Ito, Motohiro; Sudo, Yuki; Furutani, Yuji; Okitsu, Takashi; Wada, Akimori; Homma, Michio; Spudich, John L.; Kandori, Hideki

    2013-01-01

    Sensory rhodopsin II (SRII, also called pharaonis phoborhodopsin, ppR) is responsible for negative phototaxis in Natronomonas pharaonis. Photoisomerization of the retinal chromophore from all-trans to 13-cis initiates conformational changes in the protein, leading to activation of the cognate transducer protein (HtrII). We previously observed enhancement of the C14–D stretching vibration of the retinal chromophore at 2244 cm−1 upon formation of the K state and interpreted that a steric constraint occurs at the C14D group in SRIIK. Here, we identify the counterpart of the C14D group as Thr204, because the C14–D stretching signal disappeared in T204A, T204S, and T204C mutants as well as a C14–HOOP (hydrogen out-of-plane) vibration at 864 cm−1. Although the K state of the wild-type bacteriorhodopsin (BR), a light-driven proton pump, possesses neither 2244 nor 864 cm−1 bands, both signals appeared for the K state of a triple mutant of BR that functions as a light sensor (P200T/V210Y/A215T). We found a positive correlation between these vibrational amplitudes of the C14 atom at 77 K and the physiological phototaxis response. These observations strongly suggest that the steric constraint between the C14 group of retinal and Thr204 of the protein is a prerequisite for light-signal transduction by SRII. PMID:18479149

  14. A Small Physiological Electric Field Mediated Responses of Extravillous Trophoblasts Derived from HTR8/SVneo Cells: Involvement of Activation of Focal Adhesion Kinase Signaling

    PubMed Central

    Zhang, Juan; Ren, Rongmei; Luo, Xuefeng; Fan, Ping; Liu, Xinghui; Liang, Shanshan; Ma, Lei; Yu, Ping; Bai, Huai

    2014-01-01

    Moderate invasion of trophoblast cells into endometrium is essential for the placental development and normal pregnancy. Electric field (EF)-induced effects on cellular behaviors have been observed in many cell types. This study was to investigate the effect of physiological direct current EF (dc EF) on cellular responses such as elongation, orientation and motility of trophoblast cells. Immortalized first trimester extravillous trophoblast cells (HTR-8/SVneo) were exposed to the dc EF at physiological magnitude. Cell images were recorded and analyzed by image analyzer. Cell lysates were used to detect protein expression by Western blot. Cultured in the dc EFs the cells showed elongation, orientation and enhanced migration rate compared with non-EF stimulated cells at field strengths of 100 mV/mm to 200 mV/mm. EF exposure increased focal adhesion kinase (FAK) phosphorylation in a time-dependent manner and increased expression levels of MMP-2. Pharmacological inhibition of FAK impaired the EF-induced responses including motility and abrogated the elevation of MMP-2 expression. However, the expression levels of integrins like integrin α1, α5, αV and β1 were not affected by EF stimulation. Our results demonstrate the importance of FAK activation in migration/motility of trophobalst cells driven by EFs. In addition, it raises the feasibility of using applied EFs to promote placentation through effects on trophoblast cells. PMID:24643246

  15. Mono-(2-ethylhexyl) phthalate induces apoptosis through miR-16 in human first trimester placental cell line HTR-8/SVneo.

    PubMed

    Meruvu, Sunitha; Zhang, Jian; Bedi, Yudhishtar Singh; Choudhury, Mahua

    2016-03-01

    Phthalates have been linked to adverse pregnancy complications. Mono-(2-ethylhexyl) phthalate, an active metabolite of di-(2-ethylhexyl) phthalate and an endocrine disruptor, has been shown to induce apoptosis in various cell types including placental cells. However, the mechanism of action of MEHP induced apoptosis is still unknown. We hypothesized that apoptosis may be mediated in part through altered microRNA(s) in placenta under MEHP exposure. In the present study, we report that MEHP increases miR-16 expression in a time- and dose-dependent manner (p<0.05), while inducing apoptosis in HTR-8/SVneo. Cells treated with MEHP showed a dose-dependent increase in cytotoxicity and reactive oxygen species along with decreased cell viability. Consistent with significant increase in apoptosis analyzed by flow cytometry, we detected decreased anti-apoptotic BCL-2 at transcriptional and translational levels with MEHP (p<0.05). Knockdown of miR-16 did not decrease the BCL-2/BAX protein expression ratio in the presence of MEHP when compared to negative control demonstrating that MEHP induces apoptosis directly through miR-16. In conclusion, our study demonstrates for the first time that MEHP induces miR-16, which in turn, alters BCL-2/BAX ratio leading to increased apoptosis. This study provides a novel insight into MEHP induced epigenetic regulation in placental apoptosis which may lead to pregnancy complications.

  16. Assessing the kinetics of high temperature oxidation of Inconel 617 in a dedicated HTR impure helium facility coupling thermogravimetry and gas phase chromatography

    NASA Astrophysics Data System (ADS)

    Chapovaloff, J.; Rouillard, F.; Combrade, P.; Pijolat, M.; Wolski, K.

    2013-10-01

    A new facility coupling thermogravimetric analysis (TGA) with gas phase chromatography (GPC) has been developed. This facility is dedicated for studying high temperature oxidation of Inconel 617 in impure helium environment containing H2O, H2 and CO at very low partial pressures (in the Pa range), which is representative of the high temperature reactor (HTR) concept developed within the Generation IV Forum. Simultaneous acquisition of mass gain and gas composition has allowed the influence of carbon monoxide and water vapour on the kinetics of oxidation to be studied. GPC measurements of gas consumption have allowed the plotting of individual mass gain curves for oxidation by H2O and CO. During isothermal exposure at 1123 K for 20 h, the oxidation was mainly due to water vapour with a minor contribution of carbon monoxide during the first hours. The contribution of water vapour to the oxidation kinetics was extracted. It was shown to obey a complete parabolic law and to be limited by an interfacial reaction during the first few hours of oxidation and to be controlled by a mixed interfacial and diffusion process, diffusion becoming the rate-determining step for long term oxidation. There was very good agreement between GPC measurements and the experimental TGA results.

  17. The HtrA/DegP family protease MamE is a bifunctional protein with roles in magnetosome protein localization and magnetite biomineralization

    PubMed Central

    Quinlan, Anna; Murat, Dorothée; Vali, Hojatollah; Komeili, Arash

    2011-01-01

    Summary Magnetotactic bacteria contain nanometer-sized, membrane-bound organelles, called magnetosomes, which are tasked with the biomineralization of small crystals of the iron oxide magnetite allowing the organism to use geomagnetic field lines for navigation. A key player in this process is the HtrA/DegP family protease MamE. In its absence, Magnetospirillum magneticum str AMB-1 is able to form magnetosome membranes but not magnetite crystals, a defect previously linked to the mislocalization of magnetosome proteins. In this work we use a directed genetic approach to find that MamE, and another predicted magnetosome-associated protease, MamO, likely function as proteases in vivo. However, as opposed to the complete loss of mamE where no biomineralization is observed, the protease-deficient variant of this protein still supports the initiation and formation of small, 20 nm-sized crystals of magnetite, too small to hold a permanent magnetic dipole moment. This analysis also reveals that MamE is a bifunctional protein with a protease-independent role in magnetosome protein localization and a protease-dependent role in maturation of small magnetite crystals. Together these results imply the existence of a previously unrecognized “checkpoint” in biomineralization where MamE moderates the completion of magnetite formation and thus committal to magneto-aerotaxis as the organism’s dominant mode of navigating the environment. PMID:21414040

  18. Magnetite Biomineralization in Magnetospirillum magneticum Is Regulated by a Switch-like Behavior in the HtrA Protease MamE.

    PubMed

    Hershey, David M; Browne, Patrick J; Iavarone, Anthony T; Teyra, Joan; Lee, Eun H; Sidhu, Sachdev S; Komeili, Arash

    2016-08-19

    Magnetotactic bacteria are aquatic organisms that produce subcellular magnetic particles in order to orient in the earth's geomagnetic field. MamE, a predicted HtrA protease required to produce magnetite crystals in the magnetotactic bacterium Magnetospirillum magneticum AMB-1, was recently shown to promote the proteolytic processing of itself and two other biomineralization factors in vivo Here, we have analyzed the in vivo processing patterns of three proteolytic targets and used this information to reconstitute proteolysis with a purified form of MamE. MamE cleaves a custom peptide substrate with positive cooperativity, and its autoproteolysis can be stimulated with exogenous substrates or peptides that bind to either of its PDZ domains. A misregulated form of the protease that circumvents specific genetic requirements for proteolysis causes biomineralization defects, showing that proper regulation of its activity is required during magnetite biosynthesis in vivo Our results represent the first reconstitution of the proteolytic activity of MamE and show that its behavior is consistent with the previously proposed checkpoint model for biomineralization. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Decommissioning of the Dragon High Temperature Reactor (HTR) Located at the Former United Kingdom Atomic Energy Authority (UKAEA) Research Site at Winfrith - 13180

    SciTech Connect

    Smith, Anthony A.

    2013-07-01

    The Dragon Reactor was constructed at the United Kingdom Atomic Energy Research Establishment at Winfrith in Dorset through the late 1950's and into the early 1960's. It was a High Temperature Gas Cooled Reactor (HTR) with helium gas coolant and graphite moderation. It operated as a fuel testing and demonstration reactor at up to 20 MW (Thermal) from 1964 until 1975, when international funding for this project was terminated. The fuel was removed from the core in 1976 and the reactor was put into Safestore. To meet the UK's Nuclear Decommissioning Authority (NDA) objective to 'drive hazard reduction' [1] it is necessary to decommission and remediate all the Research Sites Restoration Ltd (RSRL) facilities. This includes the Dragon Reactor where the activated core, pressure vessel and control rods and the contaminated primary circuit (including a {sup 90}Sr source) still remain. It is essential to remove these hazards at the appropriate time and return the area occupied by the reactor to a safe condition. (author)

  20. Magnetite Biomineralization in Magnetospirillum magneticum Is Regulated by a Switch-like Behavior in the HtrA Protease MamE*

    PubMed Central

    Hershey, David M.; Browne, Patrick J.; Iavarone, Anthony T.; Teyra, Joan; Lee, Eun H.; Sidhu, Sachdev S.; Komeili, Arash

    2016-01-01

    Magnetotactic bacteria are aquatic organisms that produce subcellular magnetic particles in order to orient in the earth's geomagnetic field. MamE, a predicted HtrA protease required to produce magnetite crystals in the magnetotactic bacterium Magnetospirillum magneticum AMB-1, was recently shown to promote the proteolytic processing of itself and two other biomineralization factors in vivo. Here, we have analyzed the in vivo processing patterns of three proteolytic targets and used this information to reconstitute proteolysis with a purified form of MamE. MamE cleaves a custom peptide substrate with positive cooperativity, and its autoproteolysis can be stimulated with exogenous substrates or peptides that bind to either of its PDZ domains. A misregulated form of the protease that circumvents specific genetic requirements for proteolysis causes biomineralization defects, showing that proper regulation of its activity is required during magnetite biosynthesis in vivo. Our results represent the first reconstitution of the proteolytic activity of MamE and show that its behavior is consistent with the previously proposed checkpoint model for biomineralization. PMID:27302060

  1. Results of a Neutronic Simulation of HTR-Proteus Core 4.2 using PEBBED and other INL Reactor Physics Tools: FY-09 Report

    SciTech Connect

    Hans D. Gougar

    2009-08-01

    The Idaho National Laboratory’s deterministic neutronics analysis codes and methods were applied to the computation of the core multiplication factor of the HTR-Proteus pebble bed reactor critical facility. A combination of unit cell calculations (COMBINE-PEBDAN), 1-D discrete ordinates transport (SCAMP), and nodal diffusion calculations (PEBBED) were employed to yield keff and flux profiles. Preliminary results indicate that these tools, as currently configured and used, do not yield satisfactory estimates of keff. If control rods are not modeled, these methods can deliver much better agreement with experimental core eigenvalues which suggests that development efforts should focus on modeling control rod and other absorber regions. Under some assumptions and in 1D subcore analyses, diffusion theory agrees well with transport. This suggests that developments in specific areas can produce a viable core simulation approach. Some corrections have been identified and can be further developed, specifically: treatment of the upper void region, treatment of inter-pebble streaming, and explicit (multiscale) transport modeling of TRISO fuel particles as a first step in cross section generation. Until corrections are made that yield better agreement with experiment, conclusions from core design and burnup analyses should be regarded as qualitative and not benchmark quality.

  2. Vom Erdmeridian zum Lichtzeitmeter.

    NASA Astrophysics Data System (ADS)

    Rotter, F.

    The metric unit of length, the metre, has a long history which begins in the year 1791. At the beginning, the metre was defined as one tenmillionth of the quarter of the meridian of the earth. This definition was often changed corresponding to the increasing precision of length-measurements. In 1983, a new definition was found which is based on the value of velocity of light in vacuo: c = 299792458 m/s.

  3. Zum Ursprung der Hominidae

    NASA Astrophysics Data System (ADS)

    Henke, Winfried

    1981-08-01

    A fundamental problem of hominisation is the branching of the human lineage leading to the genus Homo from other hominoids. At present discussed hypotheses of a Miocene separation of the pongid and hominid lineage are described under consideration of numerous new fossils from Europe, Asia and Africa. Of special interest is the possibility of an adhominisation of the genus Australopithecus (including A. afarensis).

  4. Vom Urknall zum Durchknall

    NASA Astrophysics Data System (ADS)

    Unzicker, Alexander

    Lautstarker Applaus erhob sich im Salon III/IV des Marriott-Hotels von Crystal City im amerikanischen Bundesstaat Virginia. In dem überfüllten Konferenzraum starrten alle wie gebannt auf die Leinwand, wo nicht mehr zu sehen war als ein nüchternes Diagramm aus zahlreichen Punkten und einer geschwungenen Kurve. Nureine eigenartige Personengruppe konnte sich davon zu Emotionen hinreißen lassen - Physiker auf der Jahrestagung der Astronomischen Gesellschaft, die ihren Begeisterungssturm noch minutenlang fortsetzten. Was war geschehen? Die im Diagramm aufgetragenen Daten bestätigten mit einer nie da gewesenen Genauigkeit ein fundamentales Naturgesetz zur Wärmeabstrahlung von heißen Körpern. 1900 von Max Planck entdeckt, leuchtete es nun in geradezu mathematischer Reinheit auf. Noch sensationeller war der Ursprung der Daten - Mikrowellensignale verschiedener Frequenzen, die nicht aus einem irdischen Labor stammten, sondern von einem heißen Urzustand des Universums! Ein Feuerball aus Wasserstoff und Helium, noch ohne jegliche Strukturen, die irgendwann Leben ermöglichen sollten, ließ damals seinem Licht freien Lauf. Mehr als zehn Milliarden Jahre war es bis zu den Detektoren des vom Menschen gebauten Satelliten COBE unterwegs, der wenige Tage zuvor die Daten übertragen hatte. Wenn ich das alles wie einen Film in meiner Vorstellung ablaufen lasse, bekomme ich immer eine Gänsehaut, als würde ich die inzwischen extrem abgekühlte Strahlung tatsächlich spüren. Ihre Gleichverteilung im Raum macht uns auch deutlich, dass wir uns nicht einbilden dürfen, an einem besonderen Ort im Universum zu leben - intelligente Aliens könnten sich seitdem überall entwickelt haben! Sollten sie - was nicht wahrscheinlich ist - uns wirklich von Zeit zu Zeit über die Schulter schauen, dann hätten sie an jenem Nachmittag des 13. Januar 1990, als der Vortrag stattfand, bestimmt anerkennend mit ihrem großen Kopf genickt.

  5. Multicopy suppressors of prc mutant Escherichia coli include two HtrA (DegP) protease homologs (HhoAB), DksA, and a truncated R1pA.

    PubMed Central

    Bass, S; Gu, Q; Christen, A

    1996-01-01

    We have isolated three multicopy suppressors of the conditional lethal phenotype of a prc (tsp) null strain of Escherichia coli. One of these suppressors included two novel putative protease genes in tandem that map to 3400 kb or 72.5 centisomes on the chromosome. We propose the names hhoA and hhoB, for htrA homolog, to denote that these genes encode proteins that are 58 and 35% identical, respectively, to the HtrA (DegP) serine protease and 36% identical to each other. The HhoA and HhoB proteins are predicted to be 455 and 355 amino acids, respectively, in length. The mature HhoA protein is periplasmic in location, and amino-terminal sequencing shows that it arises following cleavage of a 27-amino-acid signal peptide. Searches of the protein and DNA databases reveal a rapidly growing family of homologous genes in a variety of other bacteria, including several which are required for virulence in their host. Deletion of the hhoAB genes shows that they are not required for viability at high temperatures like the homologous htrA but grow more slowly than wild-type strains. A second multicopy prc suppressor is the dksA (dnaK suppressor) gene, which is also a multicopy suppressor of defects in the heat shock genes dnaK, dnaJ, and grpE. The dksA gene was independently isolated as a multicopy suppressor of a mukB mutation, which is required for chromosomal partitioning. A third dosage-dependent prc suppressor includes a truncated rare lipoprotein A (rlpA) gene. PMID:8576052

  6. Intranasal Vaccination in Mice with an Attenuated Salmonella enterica Serovar 908htr A Expressing Cp15 of Cryptosporidium: Impact of Malnutrition with Preservation of Cytokine Secretion

    PubMed Central

    Roche, James K.; Rojo, Ana Lara; Costa, Lourrany B.; Smeltz, Ronald; Manque, Patricio; Woehlbier, Ute; Bartelt, Luther; Galen, James; Buck, Gregory; Guerrant, Richard L.

    2013-01-01

    Cryptosporidium is a protozoan parasite associated with acute and persistent diarrhea that, even in asymptomatic persons, can impair normal growth and potentially cognitive and physical development in young children. The recent availability of the complete gene sequence for C. hominis antigen Cp15 allows examination of innovative vaccine regimens involving intra-nasal antigen priming with live bacterial vectors applicable to human populations. We used a recently described weaned mouse model of cryptosporidiosis, where nourished and malnourished vaccinated mice receive the Cp15 antigen recombinant with cytolysin A on a Salmonella serovar Typhi CVD 908-htrA vector, followed by parenteral exposure to antigen with adjuvant. After challenge with Cryptosporidium oocysts via gavage, parameters of infection and disease (stool shedding of parasites, growth rates) were quantified, and serum/lymphoid tissue harvested to elucidate the Cp15-specific adaptive immune response. In vaccinated nourished mice, the regimen was highly immunogenic, with strong antigen-specific IL-6 and IFN-γ secretion and robust Cp15-specific immunoglobulin titers. In vaccinated malnourished mice, secretion of cytokines, particularly IFN-γ, and antigen-specific humoral immunity were generally undiminished despite protein deprivation and stunted growth. In contrast, after natural (oral) challenge with an identical inoculum of Cryptosporidium oocysts, cytokine and humoral responses to Cp15 were less than one-fourth those in vaccinated mice. Nevertheless, vaccination resulted in only transient reduction in stool shedding of parasites and was not otherwise protective against disease. Overall, immunogenicity for a C. hominis antigen was documented in mice, even in the setting of prolonged malnutrition, using an innovative vaccine regimen involving intra-nasal antigen priming with a live enteric bacterial vector, that has potential applicability to vulnerable human populations irrespective of nutritional

  7. Serotonin, via HTR2 receptors, excites neurons in a cortical-like pre-motor nucleus necessary for song learning and production

    PubMed Central

    Wood, William E.; Lovell, Peter V.; Mello, Claudio V.; Perkel, David J.

    2011-01-01

    Serotonin (5-HT) is a neuromodulator that is important for neural development, learning and memory, mood, and perception. Dysfunction of the serotonin system is central to depression and other clinically important mood disorders and has been linked with learning deficits. In mammals, 5-HT release from the raphe nuclei in the brainstem can modulate the functional properties of cortical neurons, influencing sensory and motor processing. Birds also have serotonergic neurons in the dorsal raphe, suggesting that 5-HT plays similar roles in sensory and motor processing, perhaps modulating brain circuitry underlying birdsong. To investigate this possibility we measured the effects of 5-HT on spontaneous firing of projection neurons in the premotor robust nucleus of the arcopallium in brain slices from male zebra finches. These neurons are thought be akin to cortical layer V pyramidal neurons. 5-HT dramatically and reversibly enhanced the endogenous firing of RA neurons. Using pharmacological agonists and antagonists in vitro, we determined this action is mediated via HTR2 receptors, which we verified are expressed by in situ hybridization. Finally, focal administration of the serotonin selective reuptake inhibitor (SSRI) fluvoxamine revealed that endogenous 5-HT is sufficient to mediate this effect in vivo. These findings reveal a modulatory action of serotonin on the physiology of the song system circuitry and suggest a novel role of serotonin in regulating song production and/or learning; further understanding of the role of 5-HT in this system may help illuminate the complex role of this neuromodulator in social interactions and motor plasticity in humans. PMID:21957243

  8. Assignment of the gene encoding the 5-HT{sub 1E} serotonin receptor (S31) (locus HTR1E) to human chromosome 6q14-q15

    SciTech Connect

    Levy, F.O.; Tasken, K.; Solberg, R.

    1994-08-01

    The human gene for the 5-HT{sub 1E} serotonin receptor was recently cloned, but no chromosomal assignment has yet been given to this gene (locus HTR1E). In this work, we demonstrate by two independent polymerase chain reactions on a panel of human-hamster somatic cell hybrid genomic DNA that the 5-HT{sub 1E} serotonin receptor gene is localized on human chromosome 6. Furthermore, by means of in situ hybridization to human metaphase chromosomes, using the cloned 5-HT{sub 1E} receptor gene (phage clone {lambda}-S31) as a probe, we demonstrate that this gene is localized to the q14-q15 region on chromosome 6. Screening of genomic DNA from 15 unrelated Caucasian individuals, using as a probe the open reading frame of the cloned 5-HT{sub 1E} receptor gene, did not reveal any restriction fragment length polymorphisms with the enzymes BamHI, BanII, BglII, EcoRI, HincII, HindIII, HinfI, MspI, PstI, and PvuII. Since the 5-HT{sub 1E} receptor is found mainly in the cerebral cortex and abnormal function of the serotonergic system has been implicated in a variety of neurologic and psychiatric diseases, the precise chromosomal assignment of the 5-HT{sub 1E} receptor gene is the crucial first step toward the evaluation of this locus as a candidate for mutations in such syndromes. 28 refs., 2 figs., 2 tabs.

  9. Protective effect of nuclear factor E2-related factor 2 on inflammatory cytokine response to brominated diphenyl ether-47 in the HTR-8/SVneo human first trimester extravillous trophoblast cell line

    SciTech Connect

    Park, Hae-Ryung Loch-Caruso, Rita

    2014-11-15

    Polybrominated diphenyl ethers (PBDEs) are widely used flame retardants, and BDE-47 is a prevalent PBDE congener detected in human tissues. Exposure to PBDEs has been linked to adverse pregnancy outcomes in humans. Although the underlying mechanisms of adverse birth outcomes are poorly understood, critical roles for oxidative stress and inflammation are implicated. The present study investigated antioxidant responses in a human extravillous trophoblast cell line, HTR-8/SVneo, and examined the role of nuclear factor E2-related factor 2 (Nrf2), an antioxidative transcription factor, in BDE-47-induced inflammatory responses in the cells. Treatment of HTR-8/SVneo cells with 5, 10, 15, and 20 μM BDE-47 for 24 h increased intracellular glutathione (GSH) levels compared to solvent control. Treatment of HTR-8/SVneo cells with 20 μM BDE-47 for 24 h induced the antioxidant response element (ARE) activity, indicating Nrf2 transactivation by BDE-47 treatment, and resulted in differential expression of redox-sensitive genes compared to solvent control. Pretreatment with tert-butyl hydroquinone (tBHQ) or sulforaphane, known Nrf2 inducers, reduced BDE-47-stimulated IL-6 release with increased ARE reporter activity, reduced nuclear factor kappa B (NF-κB) reporter activity, increased GSH production, and stimulated expression of antioxidant genes compared to non-Nrf2 inducer pretreated groups, suggesting that Nrf2 may play a protective role against BDE-47-mediated inflammatory responses in HTR-8/SVneo cells. These results suggest that Nrf2 activation significantly attenuated BDE-47-induced IL-6 release by augmentation of cellular antioxidative system via upregulation of Nrf2 signaling pathways, and that Nrf2 induction may be a potential therapeutic target to reduce adverse pregnancy outcomes associated with toxicant-induced oxidative stress and inflammation. - Highlights: • BDE-47 stimulated ARE reporter activity and GSH production. • BDE-47 resulted in differential

  10. Modulation of FABP4 hypomethylation by DNMT1 and its inverse interaction with miR-148a/152 in the placenta of preeclamptic rats and HTR-8 cells.

    PubMed

    Yang, Anning; Zhang, Huiping; Sun, Yue; Wang, Yanhua; Yang, Xiaoming; Yang, Xiaoling; Zhang, Hui; Guo, Wei; Zhu, Guangrong; Tian, Jue; Jia, Yuexia; Jiang, Yideng

    2016-10-01

    Inflammation and dysregulated lipid metabolism are involved in the pathogenesis of preeclampsia, and fatty acid binding protein 4 (FABP4) is known to regulate both inflammation and lipid metabolism. In the present study, we elucidated the role of FABP4 using in vitro and in vivo models of preclampsia. We found increased expression of FABP4 in the placenta of preeclamptic rats, which was further confirmed in HTR-8 cells, an extravillous trophoblast cell line, treated with L-NAME. Overexpression of FABP4 in HTR-8 cells resulted in upregulated expression of pro-inflammatory cytokines IL-6 and TNF-α, and increased lipid accumulation, suggesting that FABP4 plays a role in preeclampsia. Furthermore, downregulation of methylation in the promotor resulted in increased FABP4 expression, which was mediated by downregulated DNA methyltransferase 1 (DNMT1). Bioinformatics analysis showed that miR-148a/152 regulated the expression of DNMT1, and additional in vitro studies revealed that miR-148a/152 inhibited DNMT1 expression by directly binding to its 3'-UTR. Interestingly, DNMT1 enhanced the expression of miR-148a/152 by downregulation of methylation in its promotor. Taken together, our results showed that FABP4 may be involved in the pathogenesis of preeclampsia, and the expression of FABP4 is enhanced by miR-148a/152 mediated inhibition of DNMT1 expression. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. A Single Dose of LSD Does Not Alter Gene Expression of the Serotonin 2A Receptor Gene (HTR2A) or Early Growth Response Genes (EGR1-3) in Healthy Subjects

    PubMed Central

    Dolder, Patrick C.; Grünblatt, Edna; Müller, Felix; Borgwardt, Stefan J.; Liechti, Matthias E.

    2017-01-01

    Rationale: Renewed interest has been seen in the use of lysergic acid diethylamide (LSD) in psychiatric research and practice. The repeated use of LSD leads to tolerance that is believed to result from serotonin (5-HT) 5-HT2A receptor downregulation. In rats, daily LSD administration for 4 days decreased frontal cortex 5-HT2A receptor binding. Additionally, a single dose of LSD acutely increased expression of the early growth response genes EGR1 and EGR2 in rat and mouse brains through 5-HT2A receptor stimulation. No human data on the effects of LSD on gene expression has been reported. Therefore, we investigated the effects of single-dose LSD administration on the expression of the 5-HT2A receptor gene (HTR2A) and EGR1-3 genes. Methods: mRNA expression levels were analyzed in whole blood as a peripheral biomarker in 15 healthy subjects before and 1.5 and 24 h after the administration of LSD (100 μg) and placebo in a randomized, double-blind, placebo-controlled, cross-over study. Results: LSD did not alter the expression of the HTR2A or EGR1-3 genes 1.5 and 24 h after administration compared with placebo. Conclusion: No changes were observed in the gene expression of LSD’s primary target receptor gene or genes that are implicated in its downstream effects. Remaining unclear is whether chronic LSD administration alters gene expression in humans. PMID:28701958

  12. Hax1 lacks BH modules and is peripherally associated to heavy membranes: implications for Omi/HtrA2 and PARL activity in the regulation of mitochondrial stress and apoptosis.

    PubMed

    Jeyaraju, D V; Cisbani, G; De Brito, O M; Koonin, E V; Pellegrini, L

    2009-12-01

    Hax1 has an important role in immunodeficiency syndromes and apoptosis. A recent report (Chao et al., Nature, 2008) proposed that the Bcl-2-family-related protein, Hax1, suppresses apoptosis in lymphocytes and neurons through a mechanism that involves its association to the inner mitochondrial membrane rhomboid protease PARL, to proteolytically activate the serine protease Omi/HtrA2 and eliminate active Bax. This model implies that the control of cell-type sensitivity to pro-apoptotic stimuli is governed by the PARL/Hax1 complex in the mitochondria intermembrane space and, more generally, that Bcl-2-family-related proteins can control mitochondrial outer-membrane permeabilization from inside the mitochondrion. Further, it defines a novel, anti-apoptotic Opa1-independent pathway for PARL. In this study, we present evidence that, in vivo, the activity of Hax1 cannot be mechanistically coupled to PARL because the two proteins are confined in distinct cellular compartments and their interaction in vitro is an artifact. We also show by sequence analysis and secondary structure prediction that Hax1 is extremely unlikely to be a Bcl-2-family-related protein because it lacks Bcl-2 homology modules. These results indicate a different function and mechanism of Hax1 in apoptosis and re-opens the question of whether mammalian PARL, in addition to apoptosis, regulates mitochondrial stress response through Omi/HtrA2 processing.

  13. Creation of a Full-Core HTR Benchmark with the Fort St. Vrain Initial Core and Assessment of Uncertainties in the FSV Fuel Composition and Geometry

    SciTech Connect

    Martin, William R.; Lee, John C.; baxter, Alan; Wemple, Chuck

    2012-03-31

    Information and measured data from the intial Fort St. Vrain (FSV) high temperature gas reactor core is used to develop a benchmark configuration to validate computational methods for analysis of a full-core, commercial HTR configuration. Large uncertainties in the geometry and composition data for the FSV fuel and core are identified, including: (1) the relative numbers of fuel particles for the four particle types, (2) the distribution of fuel kernel diameters for the four particle types, (3) the Th:U ratio in the initial FSV core, (4) and the buffer thickness for the fissile and fertile particles. Sensitivity studies were performed to assess each of these uncertainties. A number of methods were developed to assist in these studies, including: (1) the automation of MCNP5 input files for FSV using Python scripts, (2) a simple method to verify isotopic loadings in MCNP5 input files, (3) an automated procedure to conduct a coupled MCNP5-RELAP5 analysis for a full-core FSV configuration with thermal-hydraulic feedback, and (4) a methodology for sampling kernel diameters from arbitrary power law and Gaussian PDFs that preserved fuel loading and packing factor constraints. A reference FSV fuel configuration was developed based on having a single diameter kernel for each of the four particle types, preserving known uranium and thorium loadings and packing factor (58%). Three fuel models were developed, based on representing the fuel as a mixture of kernels with two diameters, four diameters, or a continuous range of diameters. The fuel particles were put into a fuel compact using either a lattice-bsed approach or a stochastic packing methodology from RPI, and simulated with MCNP5. The results of the sensitivity studies indicated that the uncertainties in the relative numbers and sizes of fissile and fertile kernels were not important nor were the distributions of kernel diameters within their diameter ranges. The uncertainty in the Th:U ratio in the intial FSV core was

  14. A genome-wide screen identifies Salmonella Enteritidis lipopolysaccharide biosynthesis and the HtrA heat shock protein as crucial factors involved in egg white persistence at chicken body temperature.

    PubMed

    Raspoet, R; Shearer, N; Appia-Ayme, C; Haesebrouck, F; Ducatelle, R; Thompson, A; Van Immerseel, F

    2014-05-01

    Eggs contaminated with Salmonella Enteritidis are an important source of human foodborne Salmonella infections. Salmonella Enteritidis is able to contaminate egg white during formation of the egg within the chicken oviduct, and it has developed strategies to withstand the antimicrobial properties of egg white to survive in this hostile environment. The mechanisms involved in the persistence of Salmonella Enteritidis in egg white are likely to be complex. To address this issue, a microarray-based transposon library screen was performed to identify genes necessary for survival of Salmonella Enteritidis in egg white at chicken body temperature. The majority of identified genes belonged to the lipopolysaccharide biosynthesis pathway. Additionally, we provide evidence that the serine protease/heat shock protein (HtrA) appears essential for the survival of Salmonella Enteritidis in egg white at chicken body temperature.

  15. Transient DNA damage induced by high-frequency electromagnetic fields (GSM 1.8 GHz) in the human trophoblast HTR-8/SVneo cell line evaluated with the alkaline comet assay.

    PubMed

    Franzellitti, Silvia; Valbonesi, Paola; Ciancaglini, Nicola; Biondi, Carla; Contin, Andrea; Bersani, Ferdinando; Fabbri, Elena

    2010-01-05

    One of the most controversial issue regarding high-frequency electromagnetic fields (HF-EMF) is their putative capacity to affect DNA integrity. This is of particular concern due to the increasing use of HF-EMF in communication technologies, including mobile phones. Although epidemiological studies report no detrimental effects on human health, the possible disturbance generated by HF-EMF on cell physiology remains controversial. In addition, the question remains as to whether cells are able to compensate their potential effects. We have previously reported that a 1-h exposure to amplitude-modulated 1.8 GHz sinusoidal waves (GSM-217 Hz, SAR=2 W/kg) largely used in mobile telephony did not cause increased levels of primary DNA damage in human trophoblast HTR-8/SVneo cells. Nevertheless, further investigations on trophoblast cell responses after exposure to GSM signals of different types and durations were considered of interest. In the present work, HTR-8/SVneo cells were exposed for 4, 16 or 24h to 1.8 GHz continuous wave (CW) and different GSM signals, namely GSM-217 Hz and GSM-Talk (intermittent exposure: 5 min field on, 10 min field off). The alkaline comet assay was used to evaluate primary DNA damages and/or strand breaks due to uncompleted repair processes in HF-EMF exposed samples. The amplitude-modulated signals GSM-217 Hz and GSM-Talk induced a significant increase in comet parameters in trophoblast cells after 16 and 24h of exposure, while the un-modulated CW was ineffective. However, alterations were rapidly recovered and the DNA integrity of HF-EMF exposed cells was similar to that of sham-exposed cells within 2h of recovery in the absence irradiation. Our data suggest that HF-EMF with a carrier frequency and modulation scheme typical of the GSM signal may affect the DNA integrity.

  16. The association of 5-HTR2A-1438A/G, COMTVal158Met, MAOA-LPR, DATVNTR and 5-HTTVNTR gene polymorphisms and borderline personality disorder in female heroin-dependent Chinese subjects.

    PubMed

    Yang, Mei; Mamy, Jules; Wang, Qiang; Liao, Yan-Hui; Seewoobudul, Vasish; Xiao, Shui-Yuan; Hao, Wei

    2014-04-03

    To explore the association between the 5-HTR2A-1438A/G, COMTVal158Met, MAOA-LPR, DATVNTR and 5-HTTVNTR polymorphisms with co-morbid borderline personality disorder (BPD) in female heroin-dependent patients. In a case control study, we compared the polymorphic distributions of 5-HTR2A-1438A/G, COMTVal158Met, MAOA-LPR, DATVNTR and 5-HTTVNTR in 296 female heroin-dependent patients (including 61 patients with BPD and 235 without BPD) and 101 normal females by genotypes, alleles, and interaction between genes. Female heroin-dependent subjects with BPD have lower frequency of the high activity allele (L: 4 repeats (4R)) of MAOA-LPR than those female heroin-dependent subjects without BPD, and have higher 5-HTTVNTR 10R/10R genotype frequency than normal female controls, with adjusted P-value<0.05 (after adjusted for multiple testing by 1000-fold permutation tests) respectively. By MDR (Multifactor Dimensionality Reduction) analyses, the interactive effects between MAOA-LPR and 5-HTTVNTR, and among MAOA-LPR, 5-HTTVNTR and rs6311 were close to the significance level (P=0.05) in predicting the risk of co-morbidity of BPD and heroin dependence relative to normal female controls, with 1000-fold permutation testing P-value<0.06 however >0.05 respectively. 5-HTTVNTR and MAOA-LPR may have independent predictive effects on co-morbid BPD in female heroin-dependent patients; the gene-gene interactions between MAOA-LPR and 5-HTTVNTR, and among MAOA-LPR, 5-HTTVNTR and rs6311 might also be involved in the etiology of this co-morbidity. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. Risperidone-Induced Adverse Drug Reactions and Role of DRD2 (–141 C Ins/Del) and 5HTR2C (–759 C>T) Genetic Polymorphisms in Patients with Schizophrenia

    PubMed Central

    Alladi, Charanraj Goud; Mohan, Anbarasan; Shewade, Deepak Gopal; Rajkumar, Ravi Philip; Adithan, Surendiran; Subramanian, Karthick

    2017-01-01

    Objective: To determine the adverse drug reaction (ADR) profile of risperidone and their association with dopamine (DRD2 − 141 C Ins/Del/rs1799732) and serotonin receptor (5HTR2C −759 C>T/rs3813929) gene polymorphisms in patients with schizophrenia. Materials and Methods: The study was conducted among 289 patients who were diagnosed with schizophrenia and were on treatment with risperidone (4–8 mg/day)-based therapy for a minimum of 4 weeks. Genotyping was carried by real-time quantitative polymerase chain reaction. All the patients were observed for the occurrences of ADRs during the study. Changes in prolactin levels and body weight were analyzed for a subgroup of 102 and 97 patients, respectively. Results: Risperidone-induced extrapyramidal symptoms (EPSs) were seen in 36.7% of patients. Among them, tremors were the most common symptom 31.8%. Risperidone-induced hyperprolactinemia and weight gain were seen in 87.2% and 53.6% in subgroup patients. Adverse effects such as sedation, gastrointestinal effects, and amenorrhea were seen in 9.7% (28/289), 5.1% (15/289), and 6.1% (7/114), respectively. Occurrence of DRD2 − 141 Ins/Del and Del/Del polymorphisms were significantly associated with increased prolactin levels in response to risperidone (odds ratio [OR] = 10.45; 95% confidence interval = 1.29–84.89, P = 0.004). No such association was observed with 5HTR2C (−759 C>T) polymorphism. Weight gain and EPS were not associated with the above genetic polymorphisms. Conclusion: Hyperprolactinemia, weight gain, and EPSs (>36.7%) were common adverse effects of risperidone. DRD2 – 141C Ins/Del and Del/Del polymorphisms were significantly associated with increased prolactin levels (OR = 10.45) in response to risperidone. PMID:28405133

  18. The association of 5-HTR2A-1438A/G, COMTVal158Met, MAOA-LPR, DATVNTR and 5-HTTVNTR gene polymorphisms and antisocial personality disorder in male heroin-dependent Chinese subjects.

    PubMed

    Yang, Mei; Kavi, Vasish; Wang, Wenfu; Wu, Zhimei; Hao, Wei

    2012-03-30

    To explore the association between the 5-HTR2A-1438A/G, COMTVal158Met, MAOA-LPR, DATVNTR and 5-HTTVNTR polymorphisms with comorbidity of antisocial personality disorder in male heroin-dependent patients. In case control study, we compared the polymorphic distributions of 5-HTR2A-1438A/G, COMTVal158Met, MAOA-LPR, DATVNTR and 5-HTTVNTR in 588 male heroin-dependent patients (including 311 patients with antisocial personality disorder and 277 patients without antisocial personality disorder) and 194 normal males by genotypes, alleles, and interaction between genes. Between male heroin-dependent patients with antisocial personality disorder and normal males, and between male heroin-dependent patients with and without antisocial personality disorder, the distributions of 5-HTTVNTR polymorphic genotypes and alleles were in statistical significance. Individuals carrying 10R allele were in higher risk of the comorbidity of antisocial personality disorder and heroin dependence. By MDR analyses, the interaction between 5-HTTVNTR and DATVNTR was close to statistical significance in predicting the risk of antisocial personality disorder in male heroin dependent patients. In male heroin dependent patients, individuals carrying 5-HTTVNTR 10R allele or/and DATVNTR 9R allele were in higher risks of co-occurring antisocial personality disorder, while individuals with 5-HTTVNTR 12R/12R and DATVNTR 10R/10R genotypes together were in lower risks of antisocial personality disorder. 5-HTTVNTR, and the interaction between 5-HTTVNTR and DATVNTR may be associated with the comorbidity of antisocial personality disorder in male heroin-dependent patients. Copyright © 2011 Elsevier Inc. All rights reserved.

  19. Bildung als "Kritische" Kategorie (Education as a "Critical" Category).

    ERIC Educational Resources Information Center

    Euler, Peter

    2003-01-01

    Argues that the demands on education are more pronounced than ever. Analyzes the relationship between education and critique. Reveals that the contradiction between the two is a decisive characteristic of education. Explains that educational theory is theory type and education is a necessary critical category of social development. (CAJ)

  20. Order Out of Chaos: A Case Study of the Application of Auftragstaktik by the 11th Panzer Division during the Chir River Battles 7-19 December 1942

    DTIC Science & Technology

    1989-03-01

    Jahrhundert als Geschichtsquelle, vol.2. ed. Dernot Bradley. (OsnabrUck: Biblio Verlag, 1981), 677. Hereafter cited as "Balck". All translations...die Lage so kritisch, daB ich cine Versamminung der Division nicht abwanten konnie. Zum Schutze 4cr 336. Div. hatte ich elnen Riegel aus Flak...demn Augenblick in den RUcken, als er zurn StoB nach Osten ansetzte. Zuerst mu~te cine lange Kolonne motonisicrwr Infutene dran glauben, darin die

  1. Johann Elert Bode's history of the Berlin Observatory until the year 1811. Edition of the manuscript. (German Title: Johann Elert Bodes Geschichte der Berliner Sternwarte bis zum Jahr 1811. Edition der Handschrift.)

    NASA Astrophysics Data System (ADS)

    Wielen, Roland; Wielen, Ute

    In this paper we present an edition of the manuscript entitled 'Entwurf einer litterarischen Geschichte der hiesigen Königl. Sternwarte bis zum Jahr 1811', written by Johann Elert Bode. The manuscript describes the history of the Berlin Observatory from its foundation in 1700 until 1811. The observatory belonged during this period to the Royal Prussian Academy of Sciences at Berlin. Bode served as director of the observatory from 1787 to 1825. We have transliterated and commentated Bode's manuscript. This manuscript of 14 pages in folio format is held for more than a century in the archives of the Astronomisches Rechen-Institut, which was founded in Berlin and has been moved to Heidelberg in 1945. In addition we edit the letter (Cabinet Order) of the Prussian King Friedrich Wilhelm III. to Bode, dated 6 November 1798, in which the king agrees to a modification of the observatory.

  2. Adaptation of the Endogenous Salmonella enterica Serovar Typhi clyA-Encoded Hemolysin for Antigen Export Enhances the Immunogenicity of Anthrax Protective Antigen Domain 4 Expressed by the Attenuated Live-Vector Vaccine Strain CVD 908-htrA

    PubMed Central

    Galen, James E.; Zhao, Licheng; Chinchilla, Magaly; Wang, Jin Yuan; Pasetti, Marcela F.; Green, Jeffrey; Levine, Myron M.

    2004-01-01

    Bacterial live-vector vaccines aim to deliver foreign antigens to the immune system and induce protective immune responses, and surface-expressed or secreted antigens are generally more immunogenic than cytoplasmic constructs. We hypothesize that an optimum expression system will use an endogenous export system to avoid the need for large amounts of heterologous DNA encoding additional proteins. Here we describe the cryptic chromosomally encoded 34-kDa cytolysin A hemolysin of Salmonella enterica serovar Typhi (ClyA) as a novel export system for the expression of heterologous antigens in the supernatant of attenuated Salmonella serovar Typhi live-vector vaccine strains. We constructed a genetic fusion of ClyA to the reporter green fluorescent protein and showed that in Salmonella serovar Typhi CVD 908-htrA, the fusion protein retains biological activity in both domains and is exported into the supernatant of an exponentially growing live vector in the absence of detectable bacterial lysis. The utility of ClyA for enhancing the immunogenicity of an otherwise problematic antigen was demonstrated by engineering ClyA fused to the domain 4 (D4) moiety of Bacillus anthracis protective antigen (PA). A total of 11 of 15 mice immunized intranasally with Salmonella serovar Typhi exporting the protein fusion manifested fourfold or greater rises in serum anti-PA immunoglobulin G, compared with only 1 of 16 mice immunized with the live vector expressing cytoplasmic D4 (P = 0.0002). In addition, the induction of PA-specific gamma interferon and interleukin 5 responses was observed in splenocytes. This technology offers exceptional versatility for enhancing the immunogenicity of bacterial live-vector vaccines. PMID:15557633

  3. Identification, expression, and pharmacology of a Cys{sub 23}-Ser{sub 23} substitution in the human 5-HT{sub 2C} receptor gene (HTR2C)

    SciTech Connect

    Lappalainen, J.; Ozaki, N.; Goldman, D.

    1995-05-20

    The function of brain serotonin-2C (5-HT{sub 2C}) receptors, including behavioral and neurochemical responses to 5-HT{sub 2C} agonist challenge, has been suggested to be abnormal in individuals with neuropsychiatric disorders. Thus, it is important to identify polymorphisms and functional variants within this gene. Using SSCP analysis, the authors identified a Cys{sub 23}-Ser{sub 23} substitution (designated 5-HT{sub 2Ccys} and 5-HT{sub 2Cser}) in the first hydrophobic region of the human 5-HT{sub 2C} receptor. Allele frequencies in unrelated Caucasians were 0.13 and 0.87 for 5-HT{sub 2Cser} and 5-HT{sub 2Ccys}, respectively. DNAs from informative CEPH families were typed for this polymorphism and analyzed with respect to 20 linked markers on the X chromosome. Linkage analysis placed the 5-HT{sub 2C} receptor gene (HTR2C) on Xq24. To evaluate whether this amino acid substitution causes a variant function of this receptor, recombinant human 5-HT{sub 2Ccys} and 5-HT{sub 2Cser} receptors were expressed in Xenopus oocytes and tested for responses to 5-HT using electrophysiological techniques. Concentration-response curves for 5-HT were not significantly different in oocytes expressing either form of the receptor, suggesting that the 5-HT{sub 2Ccys} and 5-HT{sub 2Cser} receptor proteins may not differ in their responses to serotonin under baseline physiological conditions. 43 refs., 3 figs., 1 tab.

  4. Ein Framework für echtzeitfähige Ethernet-Netzwerke

    NASA Astrophysics Data System (ADS)

    Dopatka, Frank

    Im Rahmen dieser Arbeit wird die Entwicklung eines formalen Framewoks vorgestellt, mit dessen Hilfe zwischen der Kompatibilität zum verbreiteten baumförmigen Standard-Ethernet und der Einhaltung von Echtzeitanforderungen einer automatisierten Anlage variiert werden kann. Damit kann bereits vor der Auswahl einer bestimmten Technologie eine Schedule der Echtzeit-Übertragungen offline kalkuliert und simuliert werden, sobald die Anforderungen der Geräte im industriellen Echtzeit-Netzwerk bekannt sind.

  5. Amplituden der Kernphasen im Bereich der Kaustik B und Untersuchung der Struktur der Übergangszone zum inneren Erdkern mit spektralen Amplituden der diffraktierten Phase PKP(BC)

    NASA Astrophysics Data System (ADS)

    Wolf, Michael D. C.

    2002-04-01

    unterschiedliche Weise. Eine Deutung des Verhaltens erfordert die Berechnung von Abklingspektren. Dabei wird die Abschwächung des PKP(BC)diff Signals für acht Frequenzen zwischen 6.4 s und 1.25 Hz ermittelt und als Spektrum dargestellt. Die Form des Abklingspektrums ist charakteristisch für die Beschaffenheit der Geschwindigkeitsstruktur direkt oberhalb der Grenze zum inneren Erdkern (GIK). Die Beben, deren Kernphasen im Regionalnetz als diffraktierte Kernphasen BCdiff registriert werden, liegen in einem Entfernungsbereich jenseits von 150 °. In dieser Distanz befinden sich die Erdbebenherde der Tonga-Fidschi-Subduktionszone, deren Breitbandaufzeichnungen verwendet werden. Die Auswertung unkorrigierter Wellenformen ergibt Abklingspektren, die mit plausiblen Erdmodellen nicht in Einklang zu bringen sind. Aus diesem Grund werden die Daten einer spektralen Stationskorrektur unterzogen, die eigens zu diesem Zweck ermittelt wird. Am Beginn der Auswertung steht eine Prüfung bekannter Erdmodelle mit unterschiedlichen Geschwindigkeitsstrukturen oberhalb der GIK. Zu den untersuchten Modellen zählen PREM, IASP91, AK135Q, PREM2, SP6, OICM2 und eine Variante des PREM. Die Untersuchung ergibt, da Modelle, die einen verringerten Gradienten oberhalb der GIK aufweisen, eine bessere Übereinstimmung mit den gemessenen Daten zeigen als Modelle ohne diese Übergangszone. Zur Verifikation dieser These wird ein Erdmodell, das keinen verringerten Gradienten oberhalb der GIK besitzt (PREM), durch eine Reihe unterschiedlicher Geschwindigkeitsverläufe in diesem Bereich ergänzt und deren synthetische Seismogramme berechnet. Das Resultat der Untersuchung sind zwei Varianten des PREM, deren Frequenzanalyse eine gute Übereinstimmung mit den Daten zeigt. Das Abklingspektrum des Erdmodells PD47, das in einer 380 km mächtigen Schicht einen negativen Gradienten besitzt, zeigt eine groe Ähnlichkeit mit den gemessenen Spektren. Dennoch kann es nicht als realistisches Modell angesehen werden, da der Punkt

  6. Safe Hydrogen Generation by Nuclear HTR

    SciTech Connect

    Sochet, Isabelle; Viossat, Anne-Laure; Rouyer, Jean-Loup

    2004-07-01

    Several concepts of new high temperature nuclear reactors are designed to generate electricity and hydrogen. Hydrogen processes envisaged here are sulfur iodine thermo-chemical process and high temperature electrolysis. Proximity of hydrogen generation is a safety challenge for nuclear reactor. This paper describes prevention and protection against hydrogen hazards as a function of inventories and type of operation of the processes. This study is important for the designers because long distance between reactor and hydrogen facility induces difficult technological equipment. (authors)

  7. Serotonin skews human macrophage polarization through HTR2B and HTR7.

    PubMed

    de las Casas-Engel, Mateo; Domínguez-Soto, Angeles; Sierra-Filardi, Elena; Bragado, Rafael; Nieto, Concha; Puig-Kroger, Amaya; Samaniego, Rafael; Loza, Mabel; Corcuera, María Teresa; Gómez-Aguado, Fernando; Bustos, Matilde; Sánchez-Mateos, Paloma; Corbí, Angel L

    2013-03-01

    Besides its role as a neurotransmitter, serotonin (5-hydroxytryptamine, 5HT) regulates inflammation and tissue repair via a set of receptors (5HT(1-7)) whose pattern of expression varies among cell lineages. Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair, we evaluated whether 5HT modulates human macrophage polarization. 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting IL-10 production, upregulated the expression of M2 polarization-associated genes (SERPINB2, THBS1, STAB1, COL23A1), and reduced the expression of M1-associated genes (INHBA, CCR2, MMP12, SERPINE1, CD1B, ALDH1A2). Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines, both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT. In fact, blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers. 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2(M-CSF) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages. Therefore, 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7), whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies.

  8. Auf dem Weg zum universellen Quantencomputer

    NASA Astrophysics Data System (ADS)

    Jaksch, Dieter; Calarco, Tommaso; Zoller, Peter

    2000-11-01

    Die Quantenmechanik eröffnet faszinierende Perspektiven für die Kommunikation und die Informationsverarbeitung. Um universell programmierbare Quantenrechner realisieren zu können bedarf es der Implementierung von Konzepten zur Quanteninformationsverarbeitung die sich auf eine große Anzahl von Qubits anwenden lassen.

  9. Longitude - critical examination of a bestselling book (German Title: Längengrad - Kritische Betrachtung eines Bestsellers)

    NASA Astrophysics Data System (ADS)

    Lühning, Felix

    The history of longitude determination at sea in connection with John Harrison's clock constructions was widely disseminated by Dana Sobel's novel. It is shown that this novel, however, is very inaccurate and even flawed in its basic concept and in many details. This contribution traces the true historical courses and yields distinct insights in the history of the longitude problem.

  10. Leonard Cohen: Suzanne. Kritische Anmerkungen zur Interpretation (Critical Comments on Interpretation)

    ERIC Educational Resources Information Center

    Faulstich, Werner

    1977-01-01

    Criticizes a critique by J. Marsden of Leonard Cohen's pop song "Suzanne" as material for high school foreign language teaching. States that Marsden's article is concerned only with the text. The music agrees with the theme (loneliness) and with the "social milieu." (Text is in German.) (IFS/WGA)

  11. Wie kritisch sind unsere Englischlehrwerke? (How Critical are Our English Teaching Texts?)

    ERIC Educational Resources Information Center

    Mueller, Heinrich

    1974-01-01

    Using selected lessons from the textbooks "Learning English" (Klett) and "English for Today" (Schrodel-Lensing) for the beginner's level, it is shown that modern-oriented texts, critical of content and having an emancipated outlook, are not presented at all, or only in the beginning. (Text is in German.) (IFS/WGA)

  12. An Unusual Variant of Anlage Tumor of Pineal Region in an Infant

    PubMed Central

    Kothari, Kanchan; Goel, Naina; Mahore, Amit

    2015-01-01

    A 9-month-old male child was brought with complaints of increasing head size for 2 months, increasing lethargy and vomiting for the last 2 days. Radiology revealed a heterogeneously enhancing, globular lesion in the pineal region with hydrocephalus. Near total excision of the tumor was carried out. The histopathological examination of the lesion showed heterogenous elements in the form of mature neuroepithelial and ectomesenchymal tissue. The pathology and radiology of this unusual lesion is discussed with relevant review of literature. PMID:25977909

  13. Differentiation of cartilaginous anlage in entire embryonic mouse limbs cultured in a rotating bioreactor.

    NASA Astrophysics Data System (ADS)

    Duke, P.; Oakley, C.; Montufar-Solis, D.

    The embryonic mammalian limb is sensitive both in vivo and in vitro to changes in gravitational force. Hypergravity of centrifugation and microgravity of space decreased size of elements due to precocious or delayed chondrogenesis respectively. In recapitulating spaceflight experiments, premetatarsals were cultured in suspension in a low stress, low sheer rotating bioreactor, and found to be shorter than those cultured in standard culture dishes, and cartilage development was delayed. This study only measured length of the metatarsals, and did not account for possible changes in width and/or in form of the skeletal elements. Shorter cartilage elements in limbbuds cultured in the bioreactor may be due to the ability of the system to reproduce a more in vivo 3D shape than traditional organ cultures. Tissues subjected to traditional organ cultures become flattened by their own weight, attachment to the filter, and restrictions imposed by nutrient diffusion. The purpose of the current experiment was to determine if entire limb buds could be successfully cultured in the bioreactor, and to compare the effects on 3D shape with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were placed either in the bioreactor, in Trowell culture, or fixed as controls. Limbbuds were cultured for six days, fixed, and processed either as whole mounts or embedded for histology. Qualitative analysis revealed that the Trowell culture specimens were flattened, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections of limbbuds from both types of cultures had excellent cartilage differentiation, with apparently more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Morphometric quantitation of the cartilaginous elements for comparisons of the two culture systems was complicated due to some limb buds fusing together during culture. This problem was especially noticeable in the younger limbs, and may be rectified by embedding the limbs in a matrix (e.g. alginate) to maintain integrity of the tissue while in culture in the bioreactor. The bioreactor supported differentiation of skeletal elements in entire limbs, and maintained better external limb morphology than did the Trowell system. Supported by NIH/NIDCR Training Grant T358DE07252-08.

  14. Molekulare Methoden zum Nachweis, zur Quantifizierung und zum Monitoring der Mykotoxinbildung lebensmittelrelevanter Pilze

    NASA Astrophysics Data System (ADS)

    Geisen, Rolf

    Schimmelpilze kommen ubiquitär vor und spielen besonders bei pflanzlichen Lebensmitteln und Rohprodukten eine besondere Rolle als Verderbsorganismen. Es wird geschätzt, dass 20-25 % der jährlichen Produktion an pflanzlichen Produkten durch Schimmelpilze verdorben werden (Smith et al., 1994). Viele der lebensmittelrelevanten Schimmelpilze sind zudem in der Lage, Mykotoxine, toxische Sekundärmetabolite, zu bilden, was das Ausmaß des Problems deutlich macht. Die wichtigsten mykotoxinbildenden Spezies gehören zu den Fusarien (Trichothecene, Fumonisine, Zearalenon), Aspergillen (Aflatoxin, Ochratoxin, Cyclopiazonsäure) und Penicillien (Patulin, Ochratoxin). Für viele Mykotoxine, wie die Aflatoxine, Ochratoxin, Fumonisine und Trichothecene sind Grenzwerte erlassen worden, die die Verkehrsfähigkeit betroffener Produkte regeln. Die Einhaltung der Grenzwerte kann sehr genau durch offizielle chemisch-analytische Methoden, wie HPLC, GC-MS etc. kontrolliert werden. Diese analytischen Methoden sind aber für die Anwendung eines HACCP-Ansatzes zur Kontrolle der Mykotoxinbildung nur bedingt geeignet, da sie Endpunktkontrollen darstellen und nur das über eine längere Zeit gebildete Mykotoxin bestimmen. Sie sagen daher nichts über die biologischen Bedingungen zur Zeit der Bildung durch den Pilz aus.

  15. Supplement to Johann Elert Bode's history of the Berlin Observatory until the year 1811. Scans of the manuscript and of related documents. (German Title: Supplement zu Johann Elert Bodes Geschichte der Berliner Sternwarte bis zum Jahr 1811. Scans der Handschrift und zugehöriger Dokumente.)

    NASA Astrophysics Data System (ADS)

    Wielen, Roland; Wielen, Ute

    As a supplement to our edition (Wielen R. and Wielen U. 2010a) of the hand-written manuscript entitled 'Entwurf einer litterarischen Geschichte der hiesigen Königl. Sternwarte bis zum Jahr 1811', authored by Johann Elert Bode, we give in this paper colour scans of the manuscript. The manuscript describes the history of the Berlin Observatory from its foundation in 1700 until 1811. This manuscript of 14 pages in folio format is held for more than a century in the archives of the Astronomisches Rechen-Institut, which was founded in Berlin and has been moved to Heidelberg in 1945. In addition we give the scan of a copy of the hand-written cabinet order of the Prussian King Friedrich Wilhelm III., dated 6 November 1798, concerning the modification of the observatory. Both manuscripts have been transliterated and commentated by us in the publication cited above (Wielen R. and Wielen U. 2010a). For an easy comparison with Bode's manuscript, we give here also scans of the three printed versions of a shorter history of the Berlin Observatory published by him earlier.

  16. Evaluation of the Serotonergic Genes htr1A, htr1B, htr2A, and slc6A4 in Aggressive Behavior of Golden Retriever Dogs

    PubMed Central

    Vos-Loohuis, M.; Schilder, M. B. H.; van Oost, B. A.; Hazewinkel, H. A. W.; Wade, C. M.; Karlsson, E. K.; Lindblad-Toh, K.; Liinamo, A. E.; Leegwater, P. A. J.

    2007-01-01

    Aggressive behavior displays a high heritability in our study group of Golden Retriever dogs. Alterations in brain serotonin metabolism have been described in aggressive dogs before. Here, we evaluate whether four genes of the canine serotonergic system, coding for the serotonin receptors 1A, 1B, and 2A, and the serotonin transporter, could play a major role in aggression in Golden Retrievers. We performed mutation screens, linkage analysis, an association study, and a quantitative genetic analysis. There was no systematic difference between the coding DNA sequence of the candidate genes in aggressive and non-aggressive Golden Retrievers. An affecteds-only parametric linkage analysis revealed no strong major locus effect on human-directed aggression related to the candidate genes. An analysis of 41 single nucleotide polymorphisms (SNPs) in the 1 Mb regions flanking the genes in 49 unrelated human-directed aggressive and 49 unrelated non-aggressive dogs did not show association of SNP alleles, genotypes, or haplotypes with aggression at the candidate loci. We completed our analyses with a study of the effect of variation in the candidate genes on a collection of aggression-related phenotypic measures. The effects of the candidate gene haplotypes were estimated using the Restricted Maximum Likelihood method, with the haplotypes included as fixed effects in a linear animal model. We observed no effect of the candidate gene haplotypes on a range of aggression-related phenotypes, thus extending our conclusions to several types of aggressive behavior. We conclude that it is unlikely that these genes play a major role in the variation in aggression in the Golden Retrievers that we studied. Smaller phenotypic effects of these loci could not be ruled out with our sample size. Electronic supplementary material The online version of this article (doi:10.1007/s10519-007-9179-7) contains supplementary material, which is available to authorized users. PMID:18066658

  17. FABRICATION OF URANIUM OXYCARBIDE KERNELS AND COMPACTS FOR HTR FUEL

    SciTech Connect

    Dr. Jeffrey A. Phillips; Eric L. Shaber; Scott G. Nagley

    2012-10-01

    As part of the program to demonstrate tristructural isotropic (TRISO)-coated fuel for the Next Generation Nuclear Plant (NGNP), Advanced Gas Reactor (AGR) fuel is being irradiation tested in the Advanced Test Reactor (ATR) at Idaho National Laboratory (INL). This testing has led to improved kernel fabrication techniques, the formation of TRISO fuel particles, and upgrades to the overcoating, compaction, and heat treatment processes. Combined, these improvements provide a fuel manufacturing process that meets the stringent requirements associated with testing in the AGR experimentation program. Researchers at Idaho National Laboratory (INL) are working in conjunction with a team from Babcock and Wilcox (B&W) and Oak Ridge National Laboratory (ORNL) to (a) improve the quality of uranium oxycarbide (UCO) fuel kernels, (b) deposit TRISO layers to produce a fuel that meets or exceeds the standard developed by German researches in the 1980s, and (c) develop a process to overcoat TRISO particles with the same matrix material, but applies it with water using equipment previously and successfully employed in the pharmaceutical industry. A primary goal of this work is to simplify the process, making it more robust and repeatable while relying less on operator technique than prior overcoating efforts. A secondary goal is to improve first-pass yields to greater than 95% through the use of established technology and equipment. In the first test, called “AGR-1,” graphite compacts containing approximately 300,000 coated particles were irradiated from December 2006 to November 2009. The AGR-1 fuel was designed to closely replicate many of the properties of German TRISO-coated particles, thought to be important for good fuel performance. No release of gaseous fission product, indicative of particle coating failure, was detected in the nearly 3-year irradiation to a peak burn up of 19.6% at a time-average temperature of 1038–1121°C. Before fabricating AGR-2 fuel, each fabrication process was improved and changed. Changes to the kernel fabrication process included replacing the carbon black powder feed with a surface-modified carbon slurry and shortening the sintering schedule. AGR-2 TRISO particles were produced in a 6-inch diameter coater using a charge size about 21-times that of the 2-inch diameter coater used to coat AGR-1 particles. The compacting process was changed to increase matrix density and throughput by increasing the temperature and pressure of pressing and using a different type of press. AGR-2 fuel began irradiation in the ATR in late spring 2010.

  18. Fabrication of Uranium Oxycarbide Kernels for HTR Fuel

    SciTech Connect

    Charles Barnes; CLay Richardson; Scott Nagley; John Hunn; Eric Shaber

    2010-10-01

    Babcock and Wilcox (B&W) has been producing high quality uranium oxycarbide (UCO) kernels for Advanced Gas Reactor (AGR) fuel tests at the Idaho National Laboratory. In 2005, 350-µm, 19.7% 235U-enriched UCO kernels were produced for the AGR-1 test fuel. Following coating of these kernels and forming the coated-particles into compacts, this fuel was irradiated in the Advanced Test Reactor (ATR) from December 2006 until November 2009. B&W produced 425-µm, 14% enriched UCO kernels in 2008, and these kernels were used to produce fuel for the AGR-2 experiment that was inserted in ATR in 2010. B&W also produced 500-µm, 9.6% enriched UO2 kernels for the AGR-2 experiments. Kernels of the same size and enrichment as AGR-1 were also produced for the AGR-3/4 experiment. In addition to fabricating enriched UCO and UO2 kernels, B&W has produced more than 100 kg of natural uranium UCO kernels which are being used in coating development tests. Successive lots of kernels have demonstrated consistent high quality and also allowed for fabrication process improvements. Improvements in kernel forming were made subsequent to AGR-1 kernel production. Following fabrication of AGR-2 kernels, incremental increases in sintering furnace charge size have been demonstrated. Recently small scale sintering tests using a small development furnace equipped with a residual gas analyzer (RGA) has increased understanding of how kernel sintering parameters affect sintered kernel properties. The steps taken to increase throughput and process knowledge have reduced kernel production costs. Studies have been performed of additional modifications toward the goal of increasing capacity of the current fabrication line to use for production of first core fuel for the Next Generation Nuclear Plant (NGNP) and providing a basis for the design of a full scale fuel fabrication facility.

  19. Transport Corrections in Nodal Diffusion Codes for HTR Modeling

    SciTech Connect

    Abderrafi M. Ougouag; Frederick N. Gleicher

    2010-08-01

    The cores and reflectors of High Temperature Reactors (HTRs) of the Next Generation Nuclear Plant (NGNP) type are dominantly diffusive media from the point of view of behavior of the neutrons and their migration between the various structures of the reactor. This means that neutron diffusion theory is sufficient for modeling most features of such reactors and transport theory may not be needed for most applications. Of course, the above statement assumes the availability of homogenized diffusion theory data. The statement is true for most situations but not all. Two features of NGNP-type HTRs require that the diffusion theory-based solution be corrected for local transport effects. These two cases are the treatment of burnable poisons (BP) in the case of the prismatic block reactors and, for both pebble bed reactor (PBR) and prismatic block reactor (PMR) designs, that of control rods (CR) embedded in non-multiplying regions near the interface between fueled zones and said non-multiplying zones. The need for transport correction arises because diffusion theory-based solutions appear not to provide sufficient fidelity in these situations.

  20. Examination and improvement of the SHEM energy group structure for HTR and deep burn HTR design and analysis

    NASA Astrophysics Data System (ADS)

    Ngeleka, Tholakele Prisca

    The purpose of this study was to study and improve the SHEM energy group structures (281 and 361) and General Atomics-193 energy group structure utilizing the more systematic, consistent, and sophisticated energy group selection method referred to as contribution and point-wise cross-section driven (CPXSD) method. The SHEM-281 and -361 energy group structures were developed for LWR and General Atomics energy group structure was developed for the fast reactors. Pebble bed and Prismatic hexagonal block type fuel are used for cell analysis. DRAGON transport code was used for this task taking advantage of its capability to compute adjoint fluxes for reactor analysis. MCNP5 was used for generation of the reference solution selected due to its accuracy of neutron transport calculations. Comparisons with DRAGON calculations are presented. Pebble fuel element and Prismatic hexagonal block models were created for both codes. In the DRAGON code, analysis are conducted for the starting energy group structure by computing both forward and adjoint fluxes as well as the reaction rates and k-effective. Then forward and adjoint fluxes were used in computing the importance function of the groups, and the groups with high importance function are subdivided accordingly. The whole energy group interval of interest was divided into fast, epithermal and thermal regions. Firstly, the improvement was done for fast region and a new library was created and applied in the fuel cell analysis until the selected target criteria’s were met (10 pcm relative deviation of Δk/k and 1 percent deviation of reaction rate of interest). Then similar procedure was repeated for epithermal and thermal regions. The dominant parameters for each energy region were considered as required such as the fission cross section for fast region, absorption and scattering cross sections for epithermal region and absorption cross section for thermal region and k-effective applied for all energy regions. Pebble fuel element and the Prismatic hexagonal block were analyzed for depletion based on the improved energy group structure SHEM_TPN-531.

  1. Zum Glück gibt es Garantie!

    NASA Astrophysics Data System (ADS)

    Mehrmann, Volker

    Die numerische Lösung von Gleichungssystemen ist eine zentrale Aufgabe bei der Untersuchung fast jeden wissenschaftlichen Problems. Die Beschreibung physikalischer, chemischer oder biologischer Prozesse erfolgt im Wesentlichen immer durch Systeme von Gleichungen. Diese können aus Differentialgleichungen oder auch algebraischen Gleichungen bestehen. Lösungsverfahren dieser Systeme führen jedoch meist auf das Lösen von sehr großen linearen Gleichungssystemen. Daher ist es notwendig Methoden zu entwickeln um diese linearen Gleichungssysteme effizient zu lösen.

  2. Zum Wissenschaftsverständnis der modernen Evolutionsbiologie

    NASA Astrophysics Data System (ADS)

    Sommer, Ralf J.

    Die moderne Evolutionsbiologie hat ihren Ursprung in den Arbeiten von Charles Darwin und Alfred Wallace (Darwin 1963). Der gemeinsame Ausgangspunkt des Evolutionsgedanken ist dabei die Beobachtung, dass die biologische Welt nicht konstant ist. Biologische Systeme und alle darin lebenden Organismen unterliegen über längere Zeiträume hinweg einer stetigen Veränderung. Diese grundlegende Eigenschaft biologischer Systeme macht die Biologie zu einer historischen Wissenschaft und stellt einen wichtigen Gegensatz zu großen Teilen der Physik dar. Obwohl die Aussage von der Veränderlichkeit der Arten heute trivial klingt, war sie im 19. Jahrhundert eine Revolution, da die Konstanz der Arten und der Welt eine vorherrschende Stellung im damaligen Weltbild hatte (Amundson 2005).

  3. GLATT Wirbelschichttechnologie zum Coating von Pulvern, Pellets und Mikropellets

    NASA Astrophysics Data System (ADS)

    Grave, Annette; Pöllinger, Norbert

    Wirbelschichtverfahren wurden ursprünglich in der chemischen Verfahrenstechnik angewandt. Ende der 1950er Jahre fand die Wirbelschichttrocknung Eingang in die pharmazeutische Industrie, da eine verbesserte Trocknungseffizienz im Vergleich zu bestehenden Verfahren erzielt werden konnte. Viele Granulationsprozesse wurden durch Feuchtgranulation in einem Zwangsmischer durchgeführt, worauf ein Trocknungsschritt in einem Hordentrockner folgte. Je nach Produktqualität kann eine Hordentrocknung allerdings mehrere Tage dauern. Derart lange Trocknungszeiten können bei Anwendung der Wirbelschichttrocknung häufig auf weniger als eine Stunde verkürzt werden. Die Wirbelschichttrocknung ist eine besonders effektive und schonende Art der Trocknung, da die gesamte Oberfläche der einzelnen Partikel für den Wärme- und Feuchteübergang zur Verfügung steht.

  4. Werner Heisenberg zum 100. Geburtstag: Pionier der Quantenmechanik

    NASA Astrophysics Data System (ADS)

    Jacobi, Manfred

    2001-11-01

    Werner Heisenberg war eine der prägendsten Gestalten der Physik des 20. Jahrhunderts. Zu seinen wichtigsten Verdiensten gehören die Grundlegung der Quantenmechanik, die Formulierung der Unschärferelationen sowie die Beteiligung an der Ausarbeitung der Kopenhagener Deutung der Quantenmechanik. Darüber hinaus lieferte er Arbeiten von fundamentalem Charakter zur Theorie des Atomkerns, zur kosmischen Strahlung und zur Quantenfeldtheorie. Während des Krieges war er an den Arbeiten des Uranvereins beteiligt, der die Möglichkeit einer Entwicklung von Kernwaffen untersuchte, jedoch über Vorarbeiten zur Reaktorphysik nicht hinauskam. Wegen dieser Tätigkeit wurde er bei Kriegsende für einige Monate in England interniert. Nach seiner Rückkehr widmete er sich vor allem dem Aufbau der Physik in Deutschland, die während der NS-Zeit nahezu ihrer gesamten Substanz beraubt worden war.

  5. Zum Problem der Hochschulreform in Spanien: Einige ausgewahlte Daten.

    ERIC Educational Resources Information Center

    Val, Jose Cajide; Philipp, Rita Radl; Castro, Ana Porto

    1998-01-01

    Investigates the teaching, research, and management entailed in four new degree programs--physics, agricultural engineering, agricultural food-processing technology, and pharmacy courses--at Spain's University of Santiago de Compostela. Reports students' opinions of reforms in these courses, revealing dissatisfaction with facilities for practical…

  6. Untersuchungen zum Harnsäuremetabolismus von Littorina littorea (Gastropoda)

    NASA Astrophysics Data System (ADS)

    Heil, K. P.; Eichelberg, D.

    1983-12-01

    Periwinkles, as typical inhabitants of sea-shores, are subjected to extreme changes of environmental conditions, which affect their excretion. In Littorina littorea uric acid, urea and ammonium were detected particularly in the kidney, but the only metabolite excreted was ammonium. Only the concentration of uric acid was dependent on the availability of water; decreasing periods of submersion during low tide and raised salinities caused a higher concentration of uric acid, while increasing periods of submersion and lowered salinities effected the opposite. Transfer of periwinkles within their intertidal habitat and laboratory experiments to test the effect of salinity showed that the concentration of uric acid in the kidney is adaptable. The dependence of uric acid concentration in the kidney on environmental conditions and the ammoniotelic excretion of L. littorea are discussed with regard to its particular living conditions. It is suggested that uric acid serves as nitrogen depot and has a particular function in osmoregulation.

  7. Zum Uebersetzen fachlicher Texte (On the Translation of Technical Texts)

    ERIC Educational Resources Information Center

    Friederich, Wolf

    1975-01-01

    Reviews a 1974 East German publication on translation of scientific literature from Russian to German. Considers terminology, different standard levels of translation in East Germany, and other matters related to translation. (Text is in German.) (DH)

  8. Vom Big Business zum Smart Business in der Energiewirtschaft

    NASA Astrophysics Data System (ADS)

    Klaus, Jürgen; Anthonijsz, Jos

    Kaum eine Branche hat in den letzten zehn Jahren einen tiefgreifenderen Wandel erfahren als die Energiewirtschaft. Einstmals geprägt durch Großkonzerne, die flächendeckend alle Formen von Energiedienstleistungen erbracht haben, stellt sich heute eine gänzlich veränderte Landschaft dar. Nicht nur das es heute eine Vielzahl von Erzeugern gibt, die zunehmend dezentral aufgestellt sind, auch die klassischen Marktrollen wechseln: Erzeuger werden zu Händlern, Verbraucher werden zu Erzeugern. Darüber hinaus drängen heute neue, vormals branchenfremde, Marktteilnehmer in die Energiewirtschaft. Leicht nachzuvollziehen, dass es eine neue Art der Kommunikation braucht. Ziel ist alle Akteure zu vernetzen, um so neuartige Geschäftsmodelle zu ermöglichen. Das Internet of Things (IoT) und die Serviceplattformen bieten hierzu die geeignete Grundlage und Lösungen für neue und zukünftige Prozesse in der Energiebranche. Hierbei stehen auch Themen, wie Big Data, Datenformate, Datennutzung und -sicherheit im Fokus.

  9. Lothar Jaenicke (1923-2015) zum Gedächtnis.

    PubMed

    Hummel, Hans E; Langner, Simone S

    2017-07-26

    Among the post-war European biochemists, L. Jaenicke (1923-2015) stands out as a distinctive character with bright ideas and professional elegance, yet with personal modesty. His contributions to diverse fields such as C1 metabolic pathways, signal compounds (aquatic sex attractants) of brown algae and chemotaxis appear as distinctive features not covered by his peers. His rich scientific œuvre is documented in 291 journal articles, nine books, and a huge number of almost 570 book reviews, recensions, biographies and commentaries on the politics of science and the history of biochemistry. A quantitative approach is attempted here to make his diversified life's work more transparent through graphical analysis by the novel method of ergography. At a glance, his different strands of activities stand out clearly and speak for themselves, without the need for excessive verbal interpretation.

  10. Hermann Wilhelm Abich im Kaukasus: Zum zweihundertsten Geburtstag

    NASA Astrophysics Data System (ADS)

    Seibold, Ilse; Seibold, Eugen

    2006-11-01

    Hermann Abich was born in 1806 in Berlin and died in 1886 in Graz. He grew up in a wealthy family which had friendly relations with famous scientists like Alexander von Humboldt, Leopold von Buch or Carl Ritter. After his studies in Heidelberg and Berlin he turned to extended fieldwork at the volcanoes of Italy. In 1833 1834 he published excellent petrological/chemical results and got soon a good scientific reputation. Thus he was nominated as Professor for Geology and Mineralogy of the prestigious Russian University in Dorpat (now Tartu, Esthonia) in 1842. In 1844 he was sent to Armenia by the Russian authorities. For the next three decades his fieldwork with about 190 publications was concentrated on the Great and Lesser Caucasus. This was a period of Russian expansion to the South with long-lasting regional fights. But he enjoyed the support of powerful governors. He was an indefatigable and enthusiastic explorer and a precise observer and designer. His interests covered many fields: morphology, glaciology, structural geology, volcanology with Thermal Springs, mineral resources from hydrocarbons, coal, salt to ores, stratigraphy and paleontology as a base for geological maps. But he also gave advice for practical problems, and he was active in meteorology, botany and archaeology. Alltogether he became “the Father of Caucasus Geology”. The following sketch stresses only on three aspects of his activities. He was one of the first pioneers in hydrocarbon exploration, especially around the anticlines with the mud volcanoes near Baku. In many respects, however, his fundamental ideas were erronous. He explained the structure of the Great Caucasus by the traditional theories of Leopold von Buch and Elie de Beaumont. The Caucasus anticline “was elevated by forces acting from beneath”. Following them he tried to discover regularities in the strike of mountain chains. Similarily he treated volcanism like Alexander von Humboldt and Leopold von Buch with their two groups of phenomena: voluminous, mostly basaltic “elevation craters” versus isolated, mostly trachytic and relatively small cones of “true volcanoes”. In spite of the isolation of the Caucasus region he had cultivated continuously contacts with leading geologists in Europe and was honoured by many institutions. He left Russia in 1876 for Vienna planning to write there the final monograph volumes about his investigations but he died before he could complete them.

  11. Zum Problem der Hochschulreform in Spanien: Einige ausgewahlte Daten.

    ERIC Educational Resources Information Center

    Val, Jose Cajide; Philipp, Rita Radl; Castro, Ana Porto

    1998-01-01

    Investigates the teaching, research, and management entailed in four new degree programs--physics, agricultural engineering, agricultural food-processing technology, and pharmacy courses--at Spain's University of Santiago de Compostela. Reports students' opinions of reforms in these courses, revealing dissatisfaction with facilities for practical…

  12. Kommunikationsanforderungen an verteilte Echtzeitsysteme in der Fertigungsautomatisierung

    NASA Astrophysics Data System (ADS)

    Just, Roman; Trsek, Henning

    Kommunikationssysteme der Automatisierung müssen hohe zeitliche Anforderungen erfüllen, damit die entsprechenden industriellen Anwendungen realisiert werden können. Im Gegensatz zum IT-Bereich sind diese Anforderungen jedoch häufig nicht genau bekannt, was insbesondere beim Einsatz von drahtlosen Technologien Probleme bereiten kann1. In dieser Arbeit werden Verkehrsmuster einer realen Anlage aus dem Bereich der Fertigungsautomatisierung bestimmt. Die Zwischenankunfts- und Latenzzeiten einzelner Sensorund Aktorsignale ermöglichen Rückschlüsse auf zeitliche Anforderungen und Charakteristiken der untersuchten Anwendung. Im Anschluss werden die erzielten Erkenntnisse hinsichtlich ausgewählter Kommunikationsanforderungen analysiert und aktuell gültige Anforderungen von realen Automatisierungsanlagen abgeleitet. Weiterhin werden sie zukünftig zur Entwicklung realitätsnaher Simulationsmodelle genutzt.

  13. Mecp2 deficiency leads to altered Htr2c pre-mRNA editing and HTR2C isoform distribution in mouse hippocampus and cerebellum

    USDA-ARS?s Scientific Manuscript database

    Rett Syndrome (RTT) is a neurodevelopmental disorder caused by mutations in MECP2, a methyl-CpG binding protein and transcriptional repressor. CpG methylation plays an important role in genomic imprinting since imprinted genes are regulated by regions of differentially methylated CpGs (or ICs). A ...

  14. Fracture of a HTR-PMI cranioplastic implant after severe TBI.

    PubMed

    López González, Antonio; Pérez Borredá, Pedro; Conde Sardón, Rebeca

    2015-02-01

    A 13-year-old girl with a large left fronto-parietal hard-tissue replacement patient-matched implant (HTR®-PMI) cranioplasty-since she suffered from a traumatic brain injury (TBI) 6 years ago-had a new severe TBI that detached and fractured the implant as well as caused a left subdural hematoma and a large frontal contusion. The hematoma and contusion were removed and the implant was substituted by a provisional titanium mesh. To the best of our knowledge, this is the first case reported about an HTR®-PMI fracture. It is theorized that the bone ingrowth into the macroporous implants, like those of hydroxyapatite, gives strength and resistance to the implant. But in the case we describe, no macroscopic bone ingrowth was detected 6 years after implantation and the traumatic force that impacted over the cranioplasty exceeded its properties.

  15. Carbon monoxide formation in UO2 kerneled HTR fuel particles containing oxygen getters

    NASA Astrophysics Data System (ADS)

    Proksch, E.; Strigl, A.; Nabielek, H.

    1986-01-01

    Mass spectrometric measurements of CO in irradiated UO2 fuel particles containing oxygen getters are summarized. Uranium carbide addition in the 3% to 15% range reduces the CO release by factors between 25 and 80, up to burn-up levels as high as 70% FIMA. Unintentional gettering by SiC in TRISO coated particles with failed inner pyrocarbon layers results in CO reduction factors between 15 and 110. For ZrC, ambiguous results are obtained; ZrC probably results in CO reduction by a factor of 40; Ce2O3 and La2O3 seem less effective than the carbides; for Ce2O3, reduction factors between 3 and 15 are found. However, the results are possibly incorrect due to premature oxidation of the getter already during fabrication. Addition of SiO2 + Al2O3 has no influence on CO release.

  16. Graphite irradiation testing for HTR technology at the High Flux Reactor in Petten

    NASA Astrophysics Data System (ADS)

    Vreeling, J. A.; Wouters, O.; Laan, J. G. van der

    2008-10-01

    In 2001 the Nuclear Research and Consultancy Group started a large graphite irradiation program for the development of High Temperature Reactor technology in the European framework. The irradiation experiments, containing present day available graphite grades, are performed at the High Flux Reactor in Petten. The grades are NBG-10, NBG-17, NBG-18, NBG-20, NBG-25, PCEA, PPEA, PCIB, LPEB, IG-110 and IG-430. In the fifth framework programme (2001-2004) and sixth framework programme (2005-2009) four irradiation experiments are foreseen, resulting in design curves at irradiation temperatures between 650 °C and 950 °C. The post-irradiation testing is focused on dimensional changes, dynamic Young's modulus, coefficient of thermal expansion and coefficient of thermal conductivity. The irradiation programme and preliminary results from the first irradiation experiment at 750 °C to 8 dpa will be discussed in this paper.

  17. Modulating telomerase activity in tumor patients by targeting dyskerin binding site for hTR.

    PubMed

    Katunaric, M; Zamolo, G

    2012-09-01

    Telomeres shortening, which leads to apoptosis, is prevented by telomerase adding small repeated segments of DNA to the telomeres. The telomerase level has been correlated with progression of several cancer types, including acute leukemia, breast, prostate, lung cancer and melanoma. Suppression of telomerase activity was found to reduce metastatic potential but could have serious side effects in normal proliferative cells. One of the proteins stabilizing the telomerase complex called dyskerin reduces the maximum telomerase activity. We suggest a possible therapeutic agent which would disable the interaction of dyskerin and telomerase, but would not completely inhibit telomerase activity.

  18. Evaluation of silver transport through SiC during the German HTR fuel program

    NASA Astrophysics Data System (ADS)

    van der Merwe, J. J.

    2009-12-01

    For direct cycle gas cooled high-temperature reactor designs operating conditions may be limited as a result of excessive maintenance dose rates caused by the 110mAg source term on the turbine. The accurate prediction of silver fission and activation products' release during normal operation is required to ensure regulatory compliance and economic viability of planned power plants. Fuel qualification programs should provide satisfactory results to ensure correct analyses, but will not be available for many years. In the meantime data from the German fuel development program are utilized to develop and validate analyses methods. Previously, from primarily material and fuel component test results, transport models and parameters through all the fuel materials and components were derived. Good verification of silver transport in fuel materials and components were achieved with the exception of SiC. An earlier study of all German fuel irradiation tests identified a set of applicable tests to be used to evaluate silver transport through high-quality SiC. Simplified first estimate analyses were performed on different transport models. This was followed by an in depth study with a more complete diffusion model on all applicable irradiation test results to derive new transport parameters for silver in SiC. These new parameters were applied in an evaluation of some heat-up tests of irradiated fuel spheres to assess the range for which they are valid.

  19. Opposite displacement of helix F in attractant and repellent signaling by sensory rhodopsin-Htr complexes.

    PubMed

    Sasaki, Jun; Tsai, Ah-lim; Spudich, John L

    2011-05-27

    Two forms of the phototaxis receptor sensory rhodopsin I distinguished by differences in its photoactive site have been shown to be directly correlated with attractant and repellent signaling by the dual-signaling protein. In prior studies, differences in the photoactive site defined the two forms, namely the direction of light-induced proton transfer from the chromophore and the pK(a) of an Asp counterion to the protonated chromophore. Here, we show by both in vivo and in vitro measurements that the two forms are distinct protein conformers with structural similarities to two conformers seen in the light-driven proton transport cycle of the related protein bacteriorhodopsin. Measurements of spontaneous cell motility reversal frequencies, an in vivo measure of histidine kinase activity in the phototaxis system, indicate that the two forms are a photointerconvertible pair, with one conformer activating and the other inhibiting the kinase. Protein conformational changes in these photoconversions monitored by site-directed spin labeling show that opposite structural changes in helix F, distant from the photoactive site, correspond to the opposite phototaxis signals. The results provide the first direct evidence that displacements of helix F are directly correlated with signaling and impact our understanding of the sensory rhodopsin I signaling mechanism and the evolution of diverse functionality in this protein family.

  20. Immature teratoma of the tectum mesencephali with histopathological detection of rudimentary eye anlage in a 3-year-old boy: Report of a rare case.

    PubMed

    Dubinski, Daniel; Mittelbronn, Michel; Marquardt, Gerhard; Tews, Dominique S; Noack, Anika; Behmanesh, Bedjan; Seifert, Volker; Freiman, Thomas M

    2016-12-01

    Intracranial teratoma is a rare neoplasm derived from omnipotent germinal cells that can contain mesoderm, endoderm and/or ectoderm layer tissue. Histologically teratomas are characterized by abnormal structures like teeth or bone that can be further subdivided into mature and immature according to the presence of incompletely differentiated tissue. Characteristic intracranial teratomas are space-occupying lesions in the pineal region and often present with hydrocephalic symptoms due to aqueduct stenosis. A 3-year-old boy presented with a peracute hemiparesis, fatigue and speech deficit. MRI diagnostics showed a cystic, partially solid, inhomogeneous contrast-enhancing formation at the top of the tectum mesencephali with consecutive aqueduct compression. The patient underwent a sub-occipital craniotomy via a supracerebellar approach and complete resection was achieved. The histopathological examination mainly showed mature tissue of ectodermal, mesodermal and endodermal origin. However, small areas of undifferentiated neuroectodermal tissue within an optic vesicle formation were detected, leading to the diagnosis of an immature teratoma. In due course, the patient was discharged in good health without neurological deficits. To our knowledge, optic vesicle-containing intracranial germ cell tumors are extremely rare. Here we report a unique case with immature neuroectodermal tissue within an optic vesicle formation in an otherwise mature teratoma.

  1. [The role of the family in childhood and adolescent binge eating - a systematic review].

    PubMed

    Tetzlaff, Anne; Hilbert, Anja

    2014-01-01

    Fragestellung: Während der Einfluss der Familie bei Anorexia Nervosa und Bulimia Nervosa im Kindes- und Jugendalter belegt ist und bereits in Übersichtsarbeiten zusammengefasst wurde, liegen derzeit wenige Befunde zum Zusammenhang zu Essanfällen ohne kompensatorische Verhaltensweisen vor. Ziel dieser systematischen Übersichtsarbeit ist es daher, familiäre Einflussfaktoren auf die Entstehung und Aufrechterhaltung von Essanfällen zu beschreiben. Methodik: Eine systematische Datenbanksuche für Studien zum Zusammenhang von familiären Faktoren und Essanfällen wurde durchgeführt. Ergebnisse: Die eingeschlossenen Studien zeigten einheitlich, dass eine unsichere Bindung des Kindes, eine geringere Familienfunktionalität und geringere emotionale Unterstützung mit Essanfällen querschnittlich assoziiert sind, elterliche Arbeitslosigkeit sowie elterliche Depressionen retrospektive Korrelate darstellen und weniger Familienmahlzeiten und häufige kritische Kommentare über Figur und Gewicht innerhalb der Familie variable Risikofaktoren für Essanfälle sind. Inkonsistente Befunde fanden sich hingegen bezüglich der Familienstrukturen, dem Vorliegen elterlicher Essstörungen und Diäthalten sowie dem Erkennen von Essanfällen beim eigenen Kind. Geschlechterunterschiede hinsichtlich familiärer Beziehungen und gewichtsbezogener Stigmatisierung wurden identifiziert. Schlussfolgerungen: Ebenso wie bei anderen Essstörungen im Kindes- und Jugendalter spielen familiäre Einflussfaktoren auch bei Essanfällen eine wichtige Rolle. Daher könnten eine Diagnostik familiärer Einflüsse und familientherapeutische Interventionen in der Behandlung von Essanfällen im Kindes- und Jugendalter hilfreich sein. Mithilfe von prospektiven Studiendesigns könnten die divergierenden Ergebnisse aufgeklärt werden.

  2. Retrospective analysis of a VACM (vacuum-assisted closure and mesh-mediated fascial traction) treatment manual for temporary abdominal wall closure - results of 58 consecutive patients.

    PubMed

    Beltzer, Christian; Eisenächer, Alexander; Badendieck, Steffen; Doll, Dietrich; Küper, Markus; Lenz, Stefan; Krapohl, Björn Dirk

    2016-01-01

    Einleitung: Das optimale Behandlungskonzept eines temporären Bauchdeckenverschlusses (temporary abdominal closure, TAC) bei kritisch kranken viszeralchirurgischen Patienten mit offenem Abdomen („open abdomen“, OA) ist weiterhin unklar. Durch eine VACM-Therapie (vacuum-assisted closure and mesh-mediated fascial traction) scheinen sich gegenüber anderen Verfahren des TAC höhere Faszienverschlussraten (delayed primary fascial closure rate, FCR) realisieren zu lassen. Material und Methoden: Patienten unserer Klinik (n=58), welche in den Jahren 2005 bis 2008 mittels eines VAC/VACM-Behandlungsmanuals behandelt wurden, wurden retrospektiv analysiert. Ergebnisse: Die FCR aller Patienten betrug insgesamt 48,3% (95%-Konfidenzintervall: 34,95–61,78). Bei Patienten, bei denen im Verlauf ein Vicryl-Netz auf Faszienebene implantiert wurde (VACM-Therapie), konnte eine FCR von 61,3% realisiert werden. Die Letalität der mittels VACM therapierten Patienten betrug 45,2% (95%-KI: 27,32–63,97).Schlussfolgerung: Die Ergebnisse der eigenen Untersuchung bestätigen bisherige Studienergebnisse, die eine akzeptable FCR bei non-trauma-Patienten durch Anwendung der VACM-Therapie zeigen konnten. Die VACM-Therapie scheint aktuell Therapiekonzept der ersten Wahl bei Patienten mit OA und Indikation zum TAC zu sein.

  3. Qualitätszentrierte Softwareentwicklung in wissenschaftlichen Arbeitsgruppen Auf dem Weg vom Prototypen zum Produkt

    NASA Astrophysics Data System (ADS)

    Maleike, D.; Neuhaus, J.; Heimann, T.; Nolden, M.; Poxleitner, J.; Schöbinger, M.; Schwarz, T.; Seitel, M.; Wegner, I.; Wolber, P.; Meinzer, H.-P.; Wolf, I.

    Neue Bildverarbeitungsmethoden werden häufig hauptsächlich für Publikationszwecke entwickelt. DieWeiterentwicklung bis zur Verwendbarkeit für klinische Anwender erfordert zusätzlichen Aufwand und wird daher oft nicht weiter verfolgt. In dieser Arbeit wurden mehrere Bildanalyseverfahren unserer Abteilung konsequent zu einem stabilen und benutzbaren Softwareprodukt weiter entwickelt, so dass es für den Einsatz bei klinischen Partnern und als Demonstrationssoftware auf Messen verwendbar ist. Wir beschreiben das Vorgehen bei der Weiterentwicklung, welche Erfahrungen gemacht wurden und welcher zusätzliche Aufwand damit verbunden ist.

  4. Jahre Entwicklung der Instandhaltung - von der ausfallorientierten Instandhaltung zum gemeinsamen TPM und RCM

    NASA Astrophysics Data System (ADS)

    Iske, Friedhelm

    Zur Einleitung meines Beitrages möchte ich von einem Gespräch mit einem Mitarbeiter berichten, das ich als junger Vorgesetzter einer Instandhaltungsgruppe 1988 führte. Der engagierte Mitarbeiter feierte damals sein vierzigjähriges Dienstjubiläum und war stolz auf das von ihm Geleistete sowie auf den besonderen Einsatz seiner Altersgruppe, die nach dem Zweiten Weltkrieg das Werk wieder aufgebaut hatte. Auf meine Frage, was denn damals die erste Aufgabe in der Firma war, bekam ich kurz und knapp und mit einer Selbstverständlichkeit die selbstbewusste Antwort: "Unser Pferd füttern und mit dem Pferd die innerbetrieblichen Transporte erledigen“. Als junger, technisch orientierter Vorgesetzter war ich über diese Antwort sehr überrascht. Gedanklich weit entfernt war die Vorstellung, dass in der Vergangenheit Transporte mit einem Pferd erledigt wurden.

  5. Zum Gebrauch von Key Words im Englischunterricht (On the Use of Key Words in Teaching English)

    ERIC Educational Resources Information Center

    Bliemel, Willibald

    1975-01-01

    Discusses using key words as carriers of meaning for sentences, offering sample sentences in the fields of (1) dictation and guided narration, (2) structural pattern drills, (3) spoken dialog, and (4) silent reading. Here key words are important in deducing the meaning of new words. (Text is in German.) (IFS/WGA)

  6. Ein Erfahrungsbericht zum Thema Interaktion (A Report of Experience on the Theme of Interaction)

    ERIC Educational Resources Information Center

    Schroedter-Albers, Henning

    1977-01-01

    Describes work at a Goethe Institute branch with two groups of foreign students learning German, in which radio news, after preparatory work by the teacher, was used to induce question-and-answer dialogue. Many types of teaching aids and exercises used are described, including three-way conversation. (Text is in German.) (IFS/WGA)

  7. Neuerscheinungen zum Fremdsprachenunterricht 1: Englisch (Recent Publications in the Teaching of Foreign Languages 1: English)

    ERIC Educational Resources Information Center

    Neusprachliche Mitteilungen, 1976

    1976-01-01

    Reprints reviews of 20 recent (1973-74) books on English teaching, including: works on English at the primary level, communicative competence, teaching literature; three grammar systems and some text collections; modern English drama, English short stories; American novels and lyric poetry; aspects of linguistics; four lexicons. (Text is in…

  8. Das Studium: Vom Start zum Ziel: Lei(d)tfaden für Studierende

    NASA Astrophysics Data System (ADS)

    Messing, Barbara

    Wer studieren will, hat viele Fragen und muss viele Entscheidungen treffen. Dieser Leitfaden hilft, eine Wahl zutreffen, sich an der Hochschule zurecht zu finden und Hürden rechtzeitig zu erkennen und zu bewältigen. Es geht um Fragen wie: Wer unterstützt mich finanziell? Studentenwohnheim oder WG? Was bringt eine Vorlesung? Wie komme ich an eine Abschlussarbeit - und wie schreibe ich sie? Weitere Themen sind: Struktur der Hochschule, Lebensabschnitt Studium, Motivation beim Lernen, Zeitmanagement im Studium, Recherchieren und Lesen, Verstehen und Entwickeln, Mathematik im Studium, Vorträge halten und Prüfungen bestehen.

  9. Neue Erkenntnisse zum wenig bekannten RRab-Veraenderlichen SY Canis Minoris

    NASA Astrophysics Data System (ADS)

    Wunder, Edgar

    2013-02-01

    948 CCD measurements with V filter in 2012/13 enable an accurate determination of the period of the RRab variable SY CMi. At present (and probably since JD 2444000) the following elements are valid: HJD (Max) = 24 55943.5136 + 0.5658182 * E Before JD 2444000 the pulsation of SY CMi can be described by: HJD (Max) = 24 25234.682 + 0.5658236 * E The V magnitude range of SY CMi is V=14.8 in maximum to V=16.1 in minimum. However, the heights and depths of the maxima / minima vary, indicating a probable Blazhko effect. The rise duration from minimum to maximum is 13 % of the period.

  10. Japans Hauptstadt in Deutschland -. wie Düsseldorf sich zum wichtigsten Ziel japanischer Investitionen machte

    NASA Astrophysics Data System (ADS)

    Jäger, Kristina

    2017-04-01

    Local actors, such as municipal authorities, can promote and shape the settlement of foreign companies in a particular manner. In many instances, actions of economic promotion can lead to several conflicts; firstly between foreign investors and their national representatives, and secondly with (supra)regional industries or business circles. Here, flexible and long-term settlement projects, the permission of migrant infrastructure, and moreover, their high-profile marketing may attract foreign companies and long-term investors.

  11. Mit Training vom Tragen Wissen zum Kompetenten Handeln? (With Training from Inert Knowledge to Competent Acting?).

    ERIC Educational Resources Information Center

    Wahl, Diethelm

    2002-01-01

    Argues that all forms of education have difficulty moving from knowledge to competent acting. Discusses practical exercise as an alternative to bridge the gap. Recognizes that exercises have to be embedded in trans-situational aims and planning and must include inert emotions. Proposes different training schemes for methodological-didactic and…

  12. Noch einmal: Zum phonologischen Aspekt in der Lautschulung (Once Again: The Phonological Aspect of Teaching Pronunciation)

    ERIC Educational Resources Information Center

    Germer, Erich

    1973-01-01

    Argues against the dominance of applied phonology in the teaching of English pronunciation. A follow-up to the author's Einige Probleme der Aussprache im Englischunterricht der Hauptschule'' (Some Problems of Pronunciation in English Instruction in the Elementary School), Englisch, Mar 1972. (RS)

  13. Zum Problem der Hochschulreform in Spanien: Einige Ausgewählte Daten

    NASA Astrophysics Data System (ADS)

    Val, José Cajide; Philipp, Rita Radl; Castro, Ana Porto

    1998-09-01

    Over the past ten years there has been a sweeping reform of the courses taught and qualifications awarded by Spanish universities. For that reason, an investigation was launched in 1996-97 at the University of Santiago de Compostela (in Northwest Spain) into the teaching, research and management entailed in four new degree courses: physics (years one to four), agricultural engineering (first and second years), agricultural food-processing technology (years one to three) and pharmacy. This article reports on students' opinions of the reforms in these courses, revealing dissatisfaction with facilities for practical course work, lack of connection with prospective employment, and a call for more information about courses.

  14. Anmerkungen zum Normenkatalog Englisch (Notes on the List of Norms for English)

    ERIC Educational Resources Information Center

    Kann, Hans-Joachim

    1976-01-01

    Criticizes the acceptance of the error quotient in evaluating linguistic achievement in the current "uniform" graduation examination in English. Offers examples of the unsuitability of this requirement and calls for a new, more balanced solution. (Text is in German.) (IFS/WGA)

  15. Ein Erfahrungsbericht zum Thema Interaktion (A Report of Experience on the Theme of Interaction)

    ERIC Educational Resources Information Center

    Schroedter-Albers, Henning

    1977-01-01

    Describes work at a Goethe Institute branch with two groups of foreign students learning German, in which radio news, after preparatory work by the teacher, was used to induce question-and-answer dialogue. Many types of teaching aids and exercises used are described, including three-way conversation. (Text is in German.) (IFS/WGA)

  16. Zum Auf und Ab des Meeresspiegels in Skandinavien: Langer Streit um Eustasie oder Isostasie

    NASA Astrophysics Data System (ADS)

    Seibold, Eugen; Seibold, Ilse

    2012-03-01

    The phenomenon of the rise of the Scandinavian shield during the Holocene and the concomitant fall in level of the Baltic Sea has been investigated for centuries. Already in medieval times, there were reports about the coastlines of the Gulf of Bothnia that are full of relevant observations. During the eighteenth century, scientists such as Celsius and Linnaeus collected observations such as these. The result was that the search for the possible explanations of this rise-and-fall phenomenon intensified. The generally favoured explanation was that there was an active sinking of sea level in the Baltic rather than an active rising of the land surface in Fennoscandia. This was because water was seen as mobile, in contrast to a „terra firma". The relevant discussion was often emotional, and here, we try to illustrate it using material from the Geologenarchiv Freiburg (von Hoff, von Buch and Goethe). No more than a few decades later, it became obvious by the theory of Ice Age that both the sea level and the land could be mobile (eustatic sea level changes—glacial isostasy). Additionally, of course, plate tectonics had some influence: Norway is situated at the western end of the Eurasian plate and is part of a passive continental margin. There are still open research problems, many of which can be addressed using modern methods of satellite-based geophysics and geodesy. Some other aspects as the permanent uplift trend of Scandinavia since the Cambrium or the rhythmic to and fro of magma in the upper mantle during the Pleistocene are mentioned.

  17. Untersuchungen zum hydraulischen Grundbruch in Baugruben in homogenen Böden

    NASA Astrophysics Data System (ADS)

    Koltuk, Serdar; Fernandez-Steeger, Tomas M.; Azzam, Rafig

    2016-09-01

    In this study, two problems are discussed with respect to hydraulic heave. Firstly, the verification of safety against hydraulic heave according to DIN EN 1997-1 (2014-03) yields different results depending on the considered limit state conditions. It is demonstrated that this problem can be solved by using hydraulic gradients as a parameter for the limit state condition. Secondly, the form of a potential failure body appearing in three-dimensional groundwater models is unknown. Based on numerical analyses, it has been shown that an average hydraulic gradient that is determined by using an assumed three-dimensional failure body with the width suggested by Terzaghi for two-dimensional cases leads to a quicksand condition in an infinitesimal soil column directly adjacent to the wall, when its value is equal to the critical hydraulic gradient of the soil. The model tests show that Terzaghi's method or quicksand condition refers to certain deformations in an excavation base. The hydraulic gradient required for the first sign of a quicksand condition in loose sands may be lower than the theoretically required hydraulic gradient, whereas it is higher in dense sands.

  18. Japans Hauptstadt in Deutschland - - wie Düsseldorf sich zum wichtigsten Ziel japanischer Investitionen machte

    NASA Astrophysics Data System (ADS)

    Jäger, Kristina

    2017-02-01

    Local actors, such as municipal authorities, can promote and shape the settlement of foreign companies in a particular manner. In many instances, actions of economic promotion can lead to several conflicts; firstly between foreign investors and their national representatives, and secondly with (supra)regional industries or business circles. Here, flexible and long-term settlement projects, the permission of migrant infrastructure, and moreover, their high-profile marketing may attract foreign companies and long-term investors.

  19. Leistungsmessungen zum Einsatz der J2EE Technologie für Feldbussimulationen

    NASA Astrophysics Data System (ADS)

    Garreis, Thorsten; Jürgens, Hendrik; Schmidtmann, Uwe; von Cölin, Gerd

    Die Flexibilisierung der Fertigung und kurze Produktzyklen setzen einen beschleunigten, möglichst fehlerfreien Entwicklungsprozess voraus. Dieses Ziel kann jedoch nur erreicht werden, wenn bereits parallel zur Hardwareentwicklung die Software mit Hilfe einer Simulation der Hardware entwickelt und getestet werden kann. Darüber hinaus können mit Simulationen alternative Lösungen studiert werden, die zu optimierten Verfahren führen. In dieser Arbeit werden Leistungstests an der Simulationsumgebung drd-Simulator präsentiert, die auf der J2EE-Technologie1 basiert. Für die Messungen wird eine Soft-SPS eingesetzt, deren Feldbus vom Typ ModbusTCP simuliert wird. Erste Messungen zeigen, dass J2EE eine leistungsstarke und skalierbare Plattform für die Realisierung von Feldbussimulationen ist. In einem Zyklusbereich ab 100 ms ist das E/A-Verhalten echtzeitfähig, wobei meherere Steuerungen auf den Bus zugreifen können. Die J2EE-Technologie zeichnet sich zudem durch eine gute Ausfallsicherheit und dynamische Rekonfigurier-barkeit aus, so dass Tests dynamisch zur Laufzeit ergänzt oder verändert werden können.

  20. Nochmals: Zum franzoesischen Chanson im Unterricht (Once More: On the French Chanson in Teaching)

    ERIC Educational Resources Information Center

    Scheel, Hans-Ludwig

    1976-01-01

    Examines two recent books on the French chanson, and finds that the chanson is a desirable item in teaching French, specifically at Secondary Grade 2, for various reasons, including the students' interest in the subject. (Text is in German.) (IFS/WGA)

  1. Neuerscheinungen zum Fremdsprachenunterricht 2: Franzoesisch (Recent Publications in Foreign Language Teaching 2: French)

    ERIC Educational Resources Information Center

    Neusprachliche Mitteilungen, 1976

    1976-01-01

    Reviews 15 publications on teaching French (1972-1975). Encluded are books on: spoken vs. written French, linguistic grammar and teaching grammar, minimum French grammar, error linguistics, text collections for Secondary Grade 2, a stylistic exercise book, a collection for dictation, and literature. (Text is in German.) (IFS/WGA)

  2. Nochmals: Zum franzoesischen Chanson im Unterricht (Once More: On the French Chanson in Teaching)

    ERIC Educational Resources Information Center

    Scheel, Hans-Ludwig

    1976-01-01

    Examines two recent books on the French chanson, and finds that the chanson is a desirable item in teaching French, specifically at Secondary Grade 2, for various reasons, including the students' interest in the subject. (Text is in German.) (IFS/WGA)

  3. Smart Meter Rollout: Intelligente Messsysteme als Schnittstelle zum Kunden im Smart Grid und Smart Market

    NASA Astrophysics Data System (ADS)

    Vortanz, Karsten; Zayer, Peter

    Das Gesetz zur Digitalisierung der Energiewende ist verabschiedet. Ab 2017 sind moderne Messeinrichtungen (mME) und intelligente Messsysteme (iMSys) zu verbauen und zu betreiben. Der "deutsche Weg" für die Einführung von Smart Metern sieht einen stufenweisen Rollout sowie ein Höchstmaß an Informations- und Datensicherheit vor. Dabei spielen iMSys und mME eine wichtige Rolle bei der Neugestaltung der intelligenten Netze (Smart Grids) und des neuen Marktmodells (Smart Market). Dieser Beitrag beschäftigt sich mit den neuen Gesetzen, den Marktrollen und ihren Aufgaben, Datenschutz und Datensicherheit, dem iMSys als sichere Lösung, dem sicheren Betrieb von Smart Meter Gateways, Smart Grid - Smart Market, dem Zusammenspiel zwischen reguliertem Bereich und Markt, den Einsatzbereichen der iMSys sowie den Auswirkungen auf Prozesse und Systeme und gibt Handlungsempfehlungen.

  4. Die Schönbuchbahn. Von der Nebenbahn zum S-Bahn-Standard

    NASA Astrophysics Data System (ADS)

    Brauer, Tobias

    2017-09-01

    Since the successful reactivation of the schönbuch railway between Böblingen and Dettenhausen the number of passengers has increased continuously. Due to this high demand of over 8000 people per day and the increase of popularity the current expansion projects include the electrification of the 17 km long track and the installation of double-track sections for providing a 15-minute-service between Böblingen and Holzgerlingen. To realize the new operational concept, Zweckverband Schönbuchbahn (ZVS), the public transport authority, has ordered nine light rail-styled vehicles (with costs around 51,3 mio. euro for developing and construction). In this way the schönbuch railway is an important factor to guarantee a sustainable and forward-looking mobility in the metropolitan area of Stuttgart and for restricting car traffic.

  5. Lenin und Kuhn zum Verhältnis von Krise und Revolution

    NASA Astrophysics Data System (ADS)

    Engler, Fynn Ole

    In seinem Exil im schweizerischen Gebirgsdörfchen Sörenberg verfasste Lenin, der intellektuelle Kopf der Sozialdemokratischen Arbeiterpartei Russlands (Bolschewiki), im Mai/Juni 1915 seine wohl umfangreichste theoretische Abhandlung über die Ursachen des Zusammenbruchs der Zweiten Internationale. Den konkreten Anlass dafür lieferten die Ansichten führender sozialdemokratischer Arbeiterparteien seit dem Ausbruch des Ersten Weltkriegs im August 1914.

  6. Anmerkungen zum Studienabbruch (Some Remarks about Students' Dropout). ZIFF Papiere 73.

    ERIC Educational Resources Information Center

    Peters, Otto

    This article presents reflections on the high dropout rate at distance education universities in general and at the FernUniversitat in particular. The introduction outlines four reasons why the dropout rate should not be as high as it is. The first chapter--on general aspects--addresses problems of definition of dropout, the astounding differences…

  7. Fission product retention in TRISCO coated UO sub 2 particle fuels subjected to HTR simulated core heating tests

    SciTech Connect

    Baldwin, C.A.; Kania, M.J.

    1990-11-01

    Results of the examination and analysis of 25,730 individual microspheres from spherical fuel elements HFR-K3/1 and HFR-K3/3 are reported. The parent spheres were irradiated in excess of end-of-life exposure and subsequently subjected to simulated core heating tests in a special high-temperature furnace at Forschungszentrum, Juelich, GmbH (KFA). Following the heating tests, the spheres were electrolytically deconsolidated to obtain unbonded fuel particles for Irradiated Microsphere Gamma Analyzer (IMGA) analysis. For sphere HFR-K3/1, which was heated for 500 h at 1600{degree}C, only four particles were identified as having released fission products. The remaining particles from the sphere showed no statistical evidence of fission product release. Scanning Electron Microscopy (SEM) examination showed that three of the defect particles had large sections of the TRISO coating missing, while the fourth appeared normal. For sphere HFR-K3/3, which was heated for 100 h at 1800{degree}C, the IMGA data revealed that fission product release (cesium) from individual particles was significant and that there was large particle-to-particle variation in retention capabilities. Individual particle release (cesium) averaged ten times the KFA-measured integral spherical fuel element release value. In addition, the bimodal distribution of the individual particle data indicated that two distinct modes of failure at fuel temperatures of 1800{degree}C and above may exist. 6 refs., 6 figs., 4 tabs.

  8. Association of the 5HTR2A gene with suicidal behavior: CASE-control study and updated meta-analysis

    PubMed Central

    2013-01-01

    Background The polymorphism rs6313 (T102C) has been associated with suicidal behavior in case–control and meta-analysis studies, but results and conclusions remain controversial. The aim of the present study was to examine the association between T102C with suicidal behavior in a case–control study and, to assess the combined evidence – this case–control study and available data from other related studies – we carried out a meta-analysis. Methods We conducted a case–control study that included 161 patients with suicide attempts and 244 controls; we then performed a meta-analysis. The following models were evaluated in the meta-analysis: A) C allele vs T allele; B) T allele vs C allele; C) Caucasian population, D) Asian population, and E) suicide attempters with schizophrenia. Results We found an association between attempted suicide and control participants for genotype (χ2=6.28, p=0.04, df=2) and allele (χ2=6.17, p=0.01, df=1, OR 1.48 95% IC: 1.08-2.03) frequencies in the case–control study. The meta-analysis, comprising 23 association studies (including the present one), showed that the rs6313 polymorphism is not associated with suicidal behavior for the following comparisons:T allele vs C allele (OR: 1.03; 95% CI 0.93-1.13; p(Z)=0.44); C allele vs T allele: (OR:0.99; 95% CI: 0.90-1.08; p(Z)=0.22); Caucasians (OR:1.09; 95% CI: 0.96-1.23), and Asians (OR:0.96; 95% CI: 0.84-1.09). Conclusion Our results showed association between the rs6313 (T102C) polymorphism and suicidal behavior in the case–control study. However, the meta-analysis showed no evidence of association. Therefore, more studies are necessary to determine conclusively an association between T102C and suicidal behavior. PMID:23311440

  9. Effects of thermal treatment on the mechanical integrity of silicon carbide in HTR fuel up to 2200 °C

    NASA Astrophysics Data System (ADS)

    Rohbeck, Nadia; Xiao, Ping

    2014-08-01

    Achieving inherent safety of the High Temperature Reactor relies on the exceptional performance of its fuel. The design foresees complete encapsulation of all fissionable material by layers of carbon and silicon carbide (SiC) forming the tristructural-isotropic fuel particle. Its mechanical integrity and ability to fully retain fission products even in the event of an accident is a vital safety concern. The present study investigates the effect of post-deposition annealing on the SiC coating at design-based accident temperatures and beyond. Therefore, samples of simulated fuel have been fabricated by fluidized bed chemical vapour deposition and thermally treated in inert atmosphere up to 2200 °C. Nanoindentation and crush test measurements showed only minor reductions of elastic modulus and fracture strength up to 2000 °C. Substantial weight loss and crystal growth were observed at annealing temperatures of 2100 °C and above. Raman spectroscopy identified the formation of a multi-layered graphene film covering the SiC grains after annealing and scanning electron microscopy revealed significant porosity formation within the coating from 1800 °C onwards. These observations were attributed towards an evaporation-precipitation mechanism of SiC at very elevated temperatures that only slightly diminishes the hardness, elastic modulus or fracture strength, but might still be problematic in respect to fission product retention of the SiC layer.

  10. Fuzzy logic applications for failure analysis and diagnosis of a primary circuit of the HTR nuclear power plant

    NASA Astrophysics Data System (ADS)

    Kitowski, J.; Ksiażek, E.

    1985-10-01

    The paper presents some preliminary results of diagnostic analysis of the stimulated primary circuit of the THTR-300 nuclear power plant, based on the fuzzy sets theory. Two methods are used: the solution of the fuzzy relational equations and its extension to fuzzy logic. The first stage of the diagnostic system is the identification of the plant in the sense of fuzzy relations and the second stage is the failure or disturbance recognition. According to the fuzzy logic, lower and upper bounds of the failure vectors are obtained. Such an expert system can supply the operator in a correct recognition of the current state of the complex plant.

  11. Neuroprotective effects of HTR1A antagonist WAY-100635 on scopolamine-induced delirium in rats and underlying molecular mechanisms.

    PubMed

    Qiu, Yimin; Chen, Dongmei; Huang, Xiaojing; Huang, Lina; Tang, Liang; Jiang, Jihong; Chen, Lianhua; Li, Shitong

    2016-10-19

    Limited surveys have assessed the performance of 5-hydroxytreptamine receptor 1A and its antagonist WAY-100635 in pharmacological manipulations targeting delirium therapies. The purpose of this paper was to assess the central pharmacological activity of WAY-100635 in a rat model of scopolamine-induced delirium and its underlying mechanism. A delirium rat model was established by intraperitoneal injection of scopolamine and behavioral changes evaluated through open field and elevated plus maze experiments. Concentrations of monoamines in the hippocampus and amygdalae were detected by high performance liquid chromatography. The effect of WAY-100635 on the recovery of rats from delirium was assessed by stereotactic injection of WAY-100635 and its mechanism of action determined by measuring mRNA and protein expression via real time PCR and western blotting methods. The total distance and the number of crossing and rearing in the elevated plus maze test and the time spent in the light compartment in the dark/light test of scopolamine-treated rats were significantly increased while the percentage of time spent in the open arms was decreased, showing the validity of the established delirium rat model. The measurement of the concentrations of noradrenaline, 3,4-dihydroxyphenylacetic acid, the homovanillic acid, 5-hydroxy-3-indoleacetic acid and serotonin concentrations in the cerebrospinal fluid (CSF) of scopolamine-induced delirium rats were significantly increased. The intra-hippocampus and intra-BLA injections of WAY-100635 improved the delirium-like behavior of rats by significantly reducing the expression of NLRP3 inflammasome and the release of IL1-β and IL8 into CSF. Taken together, these findings indicate that WAY-100635 may exert a therapeutic effect on post-operative delirium by controlling neurotransmission as well as suppressing neuroinflammation in the central nervous system.

  12. Final Report on Utilization of TRU TRISO Fuel as Applied to HTR Systems Part II: Prismatic Reactor Cross Section Generation

    SciTech Connect

    Vincent Descotes

    2011-03-01

    The deep-burn prismatic high temperature reactor is made up of an annular core loaded with transuranic isotopes and surrounded in the center and in the periphery by reflector blocks in graphite. This disposition creates challenges for the neutronics compared to usual light water reactor calculation schemes. The longer mean free path of neutrons in graphite affects the neutron spectrum deep inside the blocks located next to the reflector. The neutron thermalisation in the graphite leads to two characteristic fission peaks at the inner and outer interfaces as a result of the increased thermal flux seen in those assemblies. Spectral changes are seen at least on half of the fuel blocks adjacent to the reflector. This spectral effect of the reflector may prevent us from successfully using the two step scheme -lattice then core calculation- typically used for light water reactors. We have been studying the core without control mechanisms to provide input for the development of a complete calculation scheme. To correct the spectrum at the lattice level, we have tried to generate cross-sections from supercell calculations at the lattice level, thus taking into account part of the graphite surrounding the blocks of interest for generating the homogenised cross-sections for the full-core calculation. This one has been done with 2 to 295 groups to assess if increasing the number of groups leads to more accurate results. A comparison with a classical single block model has been done. Both paths were compared to a reference calculation done with MCNP. It is concluded that the agreement with MCNP is better with supercells, but that the single block model remains quite close if enough groups are kept for the core calculation. 26 groups seems to be a good compromise between time and accu- racy. However, some trials with depletion have shown huge variations of the isotopic composition across a block next to the reflector. It may imply that at least an in- core depletion for the number density calculation may be necessary in the complete calculation scheme.

  13. Lehrer in der Bundesrepublik Deutschland. Eine Kritische Analyse Statistischer Daten uber das Lehrpersonal an Allgemeinbildenden Schulen. (Education in the Federal Republic of Germany. A Statistical Study of Teachers in Schools of General Education.)

    ERIC Educational Resources Information Center

    Kohler, Helmut

    The purpose of this study was to analyze the available statistics concerning teachers in schools of general education in the Federal Republic of Germany. An analysis of the demographic structure of the pool of full-time teachers showed that in 1971 30 percent of the teachers were under age 30, and 50 percent were under age 35. It was expected that…

  14. Lernen und Lehren von Mathematik-Analysen zum Unterrichtshandeln II - Band 6, IDM-Reihe, Untersuchungen zum Mathematikunterricht. (Learning and Teaching of Mathematics - Analysis of Instructional Actions II - Volume 6, IDM Series, Inquiries into Mathematics Instruction.)

    ERIC Educational Resources Information Center

    Bauersfeld, Heinrich; And Others

    This set of five papers, written in German with abstracts in English, was collected by the Institut fuer Didaktit der Mathematik (Institute for the Teaching of Mathematics) at the University of Bielefeld in West Germany. In the first paper, Bauersfeld considers domains of subjective experiences as the best issue for an interactive theory of…

  15. A study on effects of and stance over tuition fees.

    PubMed

    Karay, Yassin; Matthes, Jan

    2016-01-01

    Zielsetzung: Durch die in Deutschland zurzeit wieder abgeschafften Studiengebühren wurden Nachteile wie z.B. eine verlängerte Studiendauer befürchtet, während der Nachweis eines Nutzens bislang aussteht. Wir haben den Einfluss von Gebühren in Höhe von € 500 pro Semester auf den Studienverlauf Kölner Medizinstudierender sowie deren Einstellung gegenüber diesen Abgaben untersucht.Methodik: Für 1.324 Studierende, die im vorklinischen Studienabschnitt durchweg, zeitweise oder nie Studiengebühren zu zahlen hatten, wurde die Einhaltung der Mindeststudienzeit und die Studienabbrecherquote bis zum ersten Abschnitt der ärztlichen Prüfung analysiert. Mithilfe einer Regressionsanalyse wurden etwaige Effekte durch Studiengebühren und demografische Faktoren untersucht. Zusätzlich beantworteten 400 Studierende in einer online-Befragung Fragen zu ihrer Belastung durch und ihrer Einstellung zu Studiengebühren.Ergebnisse: Studiengebühren hatten keinen erkennbaren Einfluss auf Studiendauer oder Abbrecherquote. Laut Angaben in der online-Befragung wichen zuzeiten der Studiengebühren signifikant mehr Studierende vom vorgeschlagenen Stundenplan ab und sie verwendeten signifikant mehr Zeit darauf, nebenher Geld zu verdienen. 51% der Studierenden, die Gebühren gezahlt hatten und sogar 71% derer, die dies nie mussten, fanden Studiengebühren nicht oder eher nicht gerechtfertigt. Mehr als zwei Drittel der Studierenden konnten keine anhaltende Verbesserung durch Studiengebühren erkennen.Schlussfolgerung: Studiengebühren hatten an der Medizinischen Fakultät zu Köln keinen Einfluss auf Abbrecherquote und Studiendauer. Sie veränderten aber offenbar das Studierverhalten durch die vermehrte Notwendigkeit, neben dem Studium Geld zu verdienen. Die Kölner Medizinstudierenden sahen die Studiengebühren mehrheitlich kritisch und nahmen keinen nachhaltigen Nutzen im Sinne einer Verbesserung der Ausbildungsqualität wahr.

  16. Englisch an Gesamtschulen: Handbuch zum Fremdsprachenunterricht, 7. Schuljahr (English in Comprehensive Schools: A Handbook for Foreign Language Instruction, Grade 7).

    ERIC Educational Resources Information Center

    Freudenstein, Reinhold; And Others

    This guide for teaching English at the seventh grade level in German comprehensive schools contains the following major headings and subtopics: (1) Fundamentals and Postulates: student grouping problems, recommendations for a plan of instruction and use of materials, and learning goals in secondary education; (2) the Audiolingual Conception: a…

  17. Zytoprotektion mit Amifostin (Ethyol(®)) in der Chemotherapie: Meta-Analyse zum pharmakokinetischen Interaktionspotential mit Zytostatika.

    PubMed

    Czejka, Martin; Schüller, Johannes; Kletzl, Heidemarie

    2017-08-25

    The cytoprotective agent amifostine (AMI) is capable to protect healthy cells (contrary to tumor cells) due to higher activity of alkaline phosphatase at the membrane site of normal cells. In seven clinical trials the influence of AMI on the pharmacokinetics of different cytostatics was investigated. Preadministration of AMI increased Cmax of doxorubicin (+ 44 %, p < 0.06), epirubicin (+ 31 %, P < 0.08), mitomycin C (+ 41 %, p < 0.01) and docetaxel (+ 31 % and + 17 %, not significant). In contrary, the peak concentration of pirarubicin , the tetrahydropyranyl-prodrug of doxorubicin was decreased (- 50 %, P < 0.03), leading to an equal higher concentrationof doxorubicin in the blood . In accordance to the peak concentrations, the AUC'ast was increased by chemoprotection: doxorubicin + 53 % (p < 0.01) and epirubicin + 23 % (not significant), docetaxel + 25 % and + 31 % (not significant). AUC'ast of mitomycin C and paclitaxel seemed to be unaffected by preadministered AMI. A particular inhibition of the protein binding by AMI has been identified as one reason for higher serum concentrations of anthracycline drugs. After cytoprotection, a possible increase of the cytostatic's Serum concentrations should be taken into account for optimal dosage schedules.

  18. Einige linguistische und methodisch-didaktische Anmerkungen zum "potentiellen Wortschatz" (Some Linguistic and Methodological Comments on Passive Vocabulary)

    ERIC Educational Resources Information Center

    Lehmann, Volkmar

    1976-01-01

    Helpful factors in acquiring passive vocabulary are: word-building laws, international words, and genetic similarities. Another is "meaning variants" of the foreign words, derived by processes analogous to those of the native tongue. This supports the validity of reading knowledge as an early goal. (Text is in German.) (IFS/WGA)

  19. Zum Ausgleich von generativer und energetischer Sprachbetrachtung (A Comparison of the "Generative" and "Energetic" Views of Language)

    ERIC Educational Resources Information Center

    Weisgerber, Leo

    1972-01-01

    Discussion of two basic conceptions: Wilhelm von Humboldt's idea of language as energeia'' existing within and without man, and Noam Chomsky's idea of language generated by the speaker according to an innate apparatus. Revised version of lectures presented at the University of Bonn, West Germany in August 1971. (RS)

  20. Zum Ausgleich von generativer und energetischer Sprachbetrachtung (A Comparison of the "Generative" and "Energetic" Views of Language)

    ERIC Educational Resources Information Center

    Weisgerber, Leo

    1972-01-01

    Discussion of two basic conceptions: Wilhelm von Humboldt's idea of language as energeia'' existing within and without man, and Noam Chomsky's idea of language generated by the speaker according to an innate apparatus. Revised version of lectures presented at the University of Bonn, West Germany in August 1971. (RS)

  1. Stellungnahme zum Mannheimer Gutachten aus fremdsprachendidaktischer Sicht (Critique of the Mannheim Rating from the Standpoint of Foreign Language Teaching).

    ERIC Educational Resources Information Center

    Heuer, Helmut

    1978-01-01

    Criticizes the goals stated in the Mannheim Rating (of teaching texts for German as a foreign language) as being vaguely defined, and the criteria as being faultily formulated. Not considered are: text science, learning psychology, and psychology of motivation. Actual classroom teaching experience is insufficiently considered. (IFS/WGA)

  2. Untersuchungen an Plattfischen und Nordseegarnelen (Crangon crangon) im Eulitoral des Wattenmeeres nach dem Übergang zum Bodenleben

    NASA Astrophysics Data System (ADS)

    Berghahn, R.

    1983-06-01

    In spring and early summer, Pleuronectes platessa, Platichthys flesus, Solea solea, and Crangon crangon transform to benthic postlarvae in the shallow water region of the North Frisian Wadden Sea (eastern North Sea). During the first weeks after metamorphosis, they remain in the puddles and drainage gulleys on the tidal flats that are exposed during low tide. Positive rheotaxis prompting the juveniles to swim against the ebb tide currents is an important mechanism for maintaining their position on the flats. When sunlight intensity increases, they continue their locomotory and feeding activity, even during daytime, in the pools still present at low tide. On warm, cloudless days, temperature levels in these pools may increase to lethal thresholds. Consequently, the animals attempt to escape from higher tidal flats near the shore through the drainage gulleys. Such migrations bring about changes in their distribution patterns. Plaice can scarcely be found in near-shore areas, and they are absent at both high and low tide. The responses observed were correlated to physical and chemical data recorded during the observation period. The ecological significance of tidal pools as habitats is discussed.

  3. Zum Problem der Fehlergraduierung in deutsch-franzoesischen Uebersetzungen (On the Problem of Grading Errors in German-French Translations)

    ERIC Educational Resources Information Center

    Kielhoefer, Bernd

    1975-01-01

    Discusses the correction system for German-French translations used in German universities. The following are proposed: (1) Differentiation of, and a grading scale for, errors; (2) Distinguishing interstructural vs. intrastructural errors, (with suggestion for grading); (3) Localizing errors in grammar or vocabulary, with consequences for grading.…

  4. Zum Einsatz von Bildergeschichten bei der Weiterentwicklung der Gespraechsfaehigkeit (Using Picture Stories in the Further Development of Ability in Conversation)

    ERIC Educational Resources Information Center

    Lusensky, Helmuth

    1975-01-01

    In continuation courses for teachers, discussion of a picture course was found valuable, providing: nearness to actual teaching practice, expansion and solidifying of a topical vocabulary, possibility of guiding conversation, a favorable climate for conversation, and little financial (and technological) outlay. (Text is in German.) (IFS/WGA)

  5. Mapping of the serotonin 5-HT{sub 1D{alpha}} autoreceptor gene (HTR1D) on chromosome 1 using a silent polymorphism in the coding region

    SciTech Connect

    Ozaki, N.; Lappalainen, J.; Linnoila, M.

    1995-04-24

    Serotonin (5-HT){sub ID} receptors are 5-HT release-regulating autoreceptors in the human brain. Abnormalities in brain 5-HT function have been hypothesized in the pathophysiology of various psychiatric disorders, including obsessive-compulsive disorder, autism, mood disorders, eating disorders, impulsive violent behavior, and alcoholism. Thus, mutations occurring in 5-HT autoreceptors may cause or increase the vulnerability to any of these conditions. 5-HT{sub 1D{alpha}} and 5-HT{sub 1D{Beta}} subtypes have been previously localized to chromosomes 1p36.3-p34.3 and 6q13, respectively, using rodent-human hybrids and in situ localization. In this communication, we report the detection of a 5-HT{sub 1D{alpha}} receptor gene polymorphism by single strand conformation polymorphism (SSCP) analysis of the coding sequence. The polymorphism was used for fine scale linkage mapping of 5-HT{sub 1D{alpha}} on chromosome 1. This polymorphism should also be useful for linkage studies in populations and in families. Our analysis also demonstrates that functionally significant coding sequence variants of the 5-HT{sub 1D{alpha}} are probably not abundant either among alcoholics or in the general population. 14 refs., 1 fig., 1 tab.

  6. Endocrine disruptors, polychlorinated biphenyls-induced gC1qR-dependent apoptosis in human trophoblast cell line HTR-8/SVneo.

    PubMed

    Gu, Ping-Qing; Gao, Ling-Juan; Li, Li; Liu, Zhu; Luan, Fu-qi; Peng, Yu-Zhu; Guo, Xi-Rong

    2012-02-01

    Although an association exists between exposure to polychlorinated biphenyls (PCBs) and spontaneous miscarriage, the mechanisms underlying this phenomenon remain unclear. In this study, PCBs content in plasma was detected by gas chromatography-mass spectrometry (GC-MS) and decidua tissues were examined for the expression of globular heads of C1q receptor (gC1qR) using Western blot in patients who underwent induced abortion and spontaneous abortion. Results showed increased PCBs content and gC1qR expression in patients who experienced spontaneous abortion. In vitro, Western blot analysis demonstrated significantly higher caspase 3 expression and apoptotic cell counts in green fluorescent protein (GFP)-gC1qR vector group. Additionally, gC1qR and caspase 3 showed decreased expression following PCBs plus gC1qR small interfering RNA (siRNA) treatment. The percentage of apoptotic cells increased in cells treated with PCBs alone or PCB plus negative siRNA. These data suggest that maternal exposure to PCBs is associated with adverse pregnancy outcomes and that upregulation of gC1qR is important for PCBs-mediated trophoblast cell apoptosis.

  7. miR-155* mediates suppressive effect of PTEN 3'-untranslated region on AP-1/NF-κB pathway in HTR-8/SVneo cells.

    PubMed

    Xue, P; Zheng, M; Diao, Z; Shen, L; Liu, M; Gong, P; Sun, H; Hu, Y

    2013-08-01

    Among miRNAs, miR-155 is a known regulator of immune system. Accumulating studies have revealed the connections between miR-155 and activator protein 1 (AP-1)/nuclear factor (NF)-κB. However, miR-155*, a miR-155 paralog, has so far been less studied. Here we demonstrated that miR-155*, induced by lipopolysaccharide (LPS) in an AP-1/NF-κB dependent manner, played a positive feedback role in AP-1/NF-κB pathway via targeting interleukin-1 receptor-associated kinase M (IRAKM) and NF-κB inhibitor interacting Ras-like 1 (NKIRAS1) in trophoblasts. Our study further proved that miR-155*-targeted PTEN 3'-untranslated region (3'UTR) increased IRAKM and NKIRAS1 expression by competing for miR-155* binding, thereby suppressing AP-1/NF-κB activation induced by LPS.

  8. Identification of a cys-ser substitution in the 5-HT{sub 2C} (HTR2C) receptor gene and allelic association to violent behavior and alcoholism

    SciTech Connect

    Lappalainen, J.; Ozaki, N.; Goldman, D.

    1994-09-01

    Several lines of evidence suggest that brain serotonergic functions, including behavioral and neurochemical responses to 5-HT{sub 2C} agonist, are abnormal in some individuals with alcoholism and aggressive behaviors. The aim of the present study was to identify coding sequence variants in the human 5-HT{sub 2C} receptor gene which may cause abnormal or variant function of this receptor. Using SSCP analysis, a non-conservative cys-ser substitution was found in the 5-HT{sub 2C} receptor (designated 5-HT{sub 2Ccys} and 5-HT{sub 2Cser}). The polymorphism was typed in CEPH families to genetically map the gene. To test for association of the variant to alcoholism, violent behavior and serotonin function, the 5-HT{sub 2C} genotypes of 151 non-related Finnish male alcoholic violent offenders and impulsive fire setters and 127 Finnish psychiatrically interviewed healthy male volunteers were determined. CSF 5-HIAA concentrations were available for 74 alcoholic violent offenders and 25 healthy volunteers. Linkage analysis placed the 5-HT{sub 2C} gene on Xq21, a region that has been previously shown to contain genes for several mental retardation syndromes. The 5-HT{sub 2Ccys}/5-HT{sub 2Cser} genotype frequencies in alcoholic violent offenders and controls differed significantly (0.90/0.10 and 0.82/0.18, respectively, P=0.048). The association was found to be strongest in the violent offenders who did not fulfill the criteria for antisocial personality disorder (5-HT{sub 2Ccys}/5-HT{sub 2Cser} 0.93/0.07, p=0.021). No association was found between CSF 5-HIAA concentrations and 5-HT{sub 2C} genotype. These results implicate a 5-HT{sub 2C} receptor amino acid substitution in predisposition to alcohol abuse and violent behavior in a subgroup of alcoholics.

  9. PRELIMINARY RESULTS OF THE AGC-4 IRRADIATION IN THE ADVANCED TEST REACTOR AND DESIGN OF AGC-5 (HTR16-18469)

    SciTech Connect

    Davenport, Michael; Petti, D. A.

    2016-11-01

    The United States Department of Energy’s Advanced Reactor Technologies (ART) Program will irradiate up to six nuclear graphite creep experiments in the Advanced Test Reactor (ATR) located at the Idaho National Laboratory (INL). The graphite experiments are being irradiated over an approximate eight year period to support development of a graphite irradiation performance data base on the new nuclear grade graphites now available for use in high temperature gas reactors. The goals of the irradiation experiments are to obtain irradiation performance data, including irradiation creep, at different temperatures and loading conditions to support design of the Very High Temperature Gas Reactor (VHTR), as well as other future gas reactors. The experiments each consist of a single capsule that contain six stacks of graphite specimens, with half of the graphite specimens in each stack under a compressive load, while the other half of the specimens are not be subjected to a compressive load during irradiation. The six stacks have differing compressive loads applied to the top half of diametrically opposite pairs of specimen stacks. A seventh specimen stack in the center of the capsule does not have a compressive load. The specimens are being irradiated in an inert sweep gas atmosphere with on-line temperature and compressive load monitoring and control. There are also samples taken of the sweep gas effluent to measure any oxidation or off-gassing of the specimens that may occur during initial start-up of the experiment. The first experiment, AGC-1, started its irradiation in September 2009, and the irradiation was completed in January 2011. The second experiment, AGC-2, started its irradiation in April 2011 and completed its irradiation in May 2012. The third experiment, AGC-3, started its irradiation in late November 2012 and completed in the April of 2014. AGC-4 is currently being irradiated in the ATR. This paper will briefly discuss the preliminary irradiation results of the AGC-4 experiment, as well as the design of AGC-5.

  10. Color-specific conditioning effects due to both orange and blue stimuli are observed in a Halobacterium salinarum strain devoid of putative methylatable sites on HtrI.

    PubMed

    Lucia, S; Cercignani, G; Frediani, A; Petracchi, D

    2003-01-01

    Behavioral responses of Halobacterium salinarum appear as changes in the frequency of motion reversals. Turning on orange light decreases the reversal frequency, whereas blue light induces reversals. Light pulses normally induce the same response as step-up stimuli. However, anomalous behavioral reactions, including inverse responses, are seen when stimuli are applied in sequence. The occurrence of a prior stimulus is conditioning for successive stimulation on a time scale of the same order of adaptational processes. These prolonged conditioning effects are color-specific. The only adaptation process identified so far is methylation of the transducers, and this could be somehow color-specific. Therefore we tested for the behavioral anomalies in a mutant in which all methylation sites on the transducer have been eliminated. The results show that behavioral anomalies are unaffected by the absence of methylation processes on the transducer.

  11. Olanzapine-induced weight gain is associated with the -759C/T and -697G/C polymorphisms of the HTR2C gene.

    PubMed

    Godlewska, B R; Olajossy-Hilkesberger, L; Ciwoniuk, M; Olajossy, M; Marmurowska-Michałowska, H; Limon, J; Landowski, J

    2009-08-01

    Weight gain, a serious problem associated with some antipsychotic drugs, notably olanzapine and clozapine, was suggested to be associated with -759C/T polymorphism of the 5-HT2C receptor gene. This study aimed to examine a potential association of two functional polymorphisms of the promoter region of this gene: -759C/T (rs3813929) and -697G/C (rs518147), with weight gain after 6 weeks of olanzapine monotherapy. It included 107 patients with schizophrenia; among them 36 are first-episode drug-naive patients. Analysis was carried out by PCR-restriction fragment length polymorphism. A protective effect of -759T and -697C alleles was found: significantly less patients with -697C (3/51) and no patient with -759T (0/28) alleles experienced body mass index increase >or=10% (P=0.0006 and 0.002, respectively). The same was true for drug-naive patients possessing any of the variant alleles. There was a significant association of haplotypes with a >or=10% body mass index increase (P=0.001). On the basis of the additional statistical analysis, the more important role of -697C allele was suggested.

  12. Serotonin 1B Receptor Gene (HTR1B) Methylation as a Risk Factor for Callous-Unemotional Traits in Antisocial Boys.

    PubMed

    Moul, Caroline; Dobson-Stone, Carol; Brennan, John; Hawes, David J; Dadds, Mark R

    2015-01-01

    The serotonin system is thought to play a role in the aetiology of callous-unemotional (CU) traits in children. Previous research identified a functional single nucleotide polymorphism (SNP) from the promoter region of the serotonin 1B receptor gene as being associated with CU traits in boys with antisocial behaviour problems. This research tested the hypothesis that CU traits are associated with reduced methylation of the promoter region of the serotonin 1B receptor gene due to the influence of methylation on gene expression. Participants (N = 117) were boys with antisocial behaviour problems aged 3-16 years referred to University of New South Wales Child Behaviour Research Clinics. Participants volunteered a saliva sample from which the genotype of a SNP from the promoter region of the serotonin 1B receptor gene and the methylation levels of 30 CpG sites from 3 CpG regions surrounding the location of this polymorphism were assayed. Lower levels of serotonin 1B receptor gene methylation were associated with higher levels of CU traits. This relationship, however, was found to be moderated by genotype and carried exclusively by two CpG sites for which levels of methylation were negatively associated with overall methylation levels in this region of the gene. Results provide support to the emerging literature that argues for a genetically-driven system-wide alteration in serotonin function in the aetiology of CU traits. Furthermore, the results suggest that there may be two pathways to CU traits that involve methylation of the serotonin 1B receptor gene; one that is driven by a genotypic risk and another that is associated with risk for generally increased levels of methylation. Future research that aims to replicate and further investigate these results is required.

  13. High-temperature gas-cooled reactor technology development program. Annual progress report for period ending December 31, 1980

    SciTech Connect

    Not Available

    1981-08-01

    Research activities are described concerning HTGR chemistry; fueled graphite development; prestressed concrete pressure vessel development; structural materials; HTGR graphite studies; HTR core evaluation; reactor physics; shielding; application and project assessments; and HTR Core Flow Test Loop studies.

  14. Neue Lautzeichen im Advanced Learners Dictionary (ALD). Stellungnahmen zum Pro und Kontra (New Sound Symbols in the Advanced Learners Dictionary [ALD]. Considerations Pro and Con)

    ERIC Educational Resources Information Center

    Zielsprache Englisch, 1976

    1976-01-01

    The phonetic symbols in the "Advanced Learners Dictionary" (Oxford University Press, London) are discussed critically in articles by L. Alfes, H. Arndt, E. Bauch, G. Dahlmann-Resing, W. Friedrich, E. Germer, B. Haycraft, H. P. Kelz. Reference is made to an earlier article "Neue Zeichen", by H. G. Hoffmann. (Text is in German.)…

  15. Komm, Lass dir was erzahlen. Versuch einer Methodologie zum Geschichtenerzahlen fur Deutsch (A Methodology for Storytelling in the German Language Classroom).

    ERIC Educational Resources Information Center

    Wicke, Rainer E.

    A methodology is presented for storytelling in the elementary school German language classroom. Intended for the instructor in Alberta, this guide is illustrated with examples of types of activities and techniques useful for teaching German to the elementary school student. (JC)

  16. Philippus Feselius - Biographical notes on the unknown medicus of Kepler's Tertius Interveniens. (German Title: Philippus Feselius - Biographische Notizen zum unbekannten Medicus aus Keplers Tertius Interveniens)

    NASA Astrophysics Data System (ADS)

    Lenke, Nils; Roudet, Nicolas

    Until now, Philipp Feselius has been perceived only indirectly as Kepler's antagonist. Not much is known about his life besides his work as Baden private physician and his book against astrology which was cited intensely in Kepler's «Tertius Interveniens». This paper traces the stations of his career as a physician, about his presumable provenance and education in Strasbourg, his academic career in Tübingen, Strasbourg, Rostock and Padua, the doctorate in Basel in 1592, up to his employment, in 1599, as a court physician in Sulzburg and later in Durlach. Further hand-written and printed traces of Feselius are presented, and his social environment is investigated so that his personality becomes clearer, and relations can be established between his education and his writing against astrology.

  17. Zum Problem der Differenzierung von praepositionalem Objekt und praepositionaler Adverbailbestimmung in der deutschen Gegenwartssprache (On the Problem of Distinguishing Prepositional Object from Prepositional Adverbial in Contemporary German)

    ERIC Educational Resources Information Center

    Hambarzumjan, Rusanna

    1974-01-01

    Making this distinction is difficult because German here deals in prepositional phrases. More precise criteria for distinguishing may be expected from dependence grammar, and from application of the concepts of governance ("Rektion") and valence. Reference is made to experiments by J. D. Apresjan. (Text is in German.) (IFS/WGA)

  18. Fremdsprachen im Fernsehen. Einige Bemerkungen zum Franzoesischkurs des 3. Fernsehprogramms "Les Gammas" (Foreign Languages on Television. Some Observations on the French Course "Les Gammas" on Television Program 3)

    ERIC Educational Resources Information Center

    Laitenberger-Wegener, Heide

    1976-01-01

    Criticizes the television French course "Les Gammas" for the usual TV faults: the "student" is 80 to 90 percent onlooker-listener, and 10 to 20 percent participant. TV can teach essentially only auditory understanding. The course is described in some detail; it is found to have high motivational value. (Text is in German.) (IFS/WGA)

  19. Neue Lautzeichen im Advanced Learners Dictionary (ALD). Stellungnahmen zum Pro und Kontra (New Sound Symbols in the Advanced Learners Dictionary [ALD]. Considerations Pro and Con)

    ERIC Educational Resources Information Center

    Zielsprache Englisch, 1976

    1976-01-01

    The phonetic symbols in the "Advanced Learners Dictionary" (Oxford University Press, London) are discussed critically in articles by L. Alfes, H. Arndt, E. Bauch, G. Dahlmann-Resing, W. Friedrich, E. Germer, B. Haycraft, H. P. Kelz. Reference is made to an earlier article "Neue Zeichen", by H. G. Hoffmann. (Text is in German.)…

  20. On the 300th anniversary of the death of Gottfried Kirch, astronomer and calendar-maker (German Title: Zum 300. Todestag des Astronomen und Kalendermachers Gottfried Kirch)

    NASA Astrophysics Data System (ADS)

    Herbst, Klaus-Dieter

    2010-12-01

    Almost ten years ago, an article on Gottfried Kirch written by the present author appeared in Vol. 8 of this series, in which new biographical material drawn mainly from unpublished sources (church and burgher registers, letters) was presented. In the meantime, Kirch's correspondence appeared in print. The life of this scientist of early modern times can be traced in more detail from the correspondence and from newly found sources - numerous large writing calendars written by Kirch, previously thought to be lost. Kirch's 300th year of death offers an apt opportunity to prepare a sketch from this rich fund to recall his personality not only as an astronomer and calendar maker, but also as someone who was inclined to the spirit of the early Enlightenment and grounded in the Christianity movement of the Pietists.

  1. Zum Problem der Assimilation im Franzoesischen (On the Problem of Assimilation in French. A Contribution to Presentation of Facts in Handbooks of Phonetics)

    ERIC Educational Resources Information Center

    Bufe, Wolfgang

    1976-01-01

    Handbooks of French phonetics disagree with the findings of phonetic research. Discussed are: differentiating between teaching pronunciation and auditory perception: the difference between acoustic and semantic hearing; and the importance of nasal regressive and progressive assimilation in contemporary spoken French. (Text is in German.) (IFS/WGA)

  2. Die Indirekte Rede als Diskursstrategie: Innovative Lehrmethoden zum Konjunktiv I (Indirect Speech as a Discourse Strategy: Innovative Teaching Methods for the Subjunctive).

    ERIC Educational Resources Information Center

    Gramberg, Anne-Kathrin; Heinze, Karin U.

    1993-01-01

    This article talks about the subjunctive of indirect speech, in which its important functions and meanings are depicted. An analysis of the instructional materials used in the first and second years of language study, followed by practical curriculum recommendations, demonstrates how this grammatical phenomenon can be established in an advanced…

  3. Friedrich Möglich - sein Beitrag zum Aufbau der Physik in der DDR. Friedrich Möglich - langjähriger Mitherausgeber und Chefredakteur der Annalen der Physik

    NASA Astrophysics Data System (ADS)

    Rompe, Robert

    Friedrich Möglich, ein Schüler von Max von Laue und Erwin Schrödinger, übernahm 1947 die Chefredaktion der Annalen der Physik.Translated AbstractFriedrich Möglich - His Contributions to the Formation of Physics in GDRFriedrich Möglich a student of Max von Laue and Erwin Schrödinger took over as editor in chief of Annalen der Physik in 1947.

  4. Komm, Lass dir was erzahlen. Versuch einer Methodologie zum Geschichtenerzahlen fur Deutsch (A Methodology for Storytelling in the German Language Classroom).

    ERIC Educational Resources Information Center

    Wicke, Rainer E.

    A methodology is presented for storytelling in the elementary school German language classroom. Intended for the instructor in Alberta, this guide is illustrated with examples of types of activities and techniques useful for teaching German to the elementary school student. (JC)

  5. Erwaegungen zum Begriffsfeld Methodik aus fremdsprachendidaktischer Sicht (Considerations on the Concept "Methodik," from the Point of View of Foreign Language Teaching)

    ERIC Educational Resources Information Center

    Schroeder, Konrad; Weller, Franz-Rudolf

    1975-01-01

    The German term "Methodik" is compared with three other terms: "Methodologie,""Methode," and "Didaktik," as well as with the French and English equivalents of these three. Criteria are developed for a methodology of foreign-language teaching, and problem areas for such a discipline are sketched. (Text is in German.) (IFS/WGA)

  6. Zum Problem der Korrektur und Bewertung des commentaire de texte als Arbeits - und Pruefungsform (On the Problem of Correcting and Grading Text Commentary in Graduation Exams in French)

    ERIC Educational Resources Information Center

    Hornung, Walter

    1976-01-01

    Discusses two conflicting concepts of correcting and grading high school graduation examinations in French: "mathematical" grading of complex exercises versus objective grading of reduced segments. Recommends higher valuation on goal-orientation as against extreme objectivity. Students' performance must be evaluated individually. Various concrete…

  7. Zum Problem des Tests, insbesondere des Einstufungstests, im Deutschunterricht fuer Auslaender (On the Problem of Tests, Particularly Placement Tests, in Teaching German to Foreigners)

    ERIC Educational Resources Information Center

    Breitung, H. A.; And Others

    1974-01-01

    New placement procedure at Humboldt University includes interviews and placement tests. Interviews reveal response ability, tempo, pronunciation, comprehension, etc. The 60-minute test that follows is described and results discussed, as well as difficulty level and grading. Results: better grouping of students, less shifting, better work. (Text is…

  8. Die Indirekte Rede als Diskursstrategie: Innovative Lehrmethoden zum Konjunktiv I (Indirect Speech as a Discourse Strategy: Innovative Teaching Methods for the Subjunctive).

    ERIC Educational Resources Information Center

    Gramberg, Anne-Kathrin; Heinze, Karin U.

    1993-01-01

    This article talks about the subjunctive of indirect speech, in which its important functions and meanings are depicted. An analysis of the instructional materials used in the first and second years of language study, followed by practical curriculum recommendations, demonstrates how this grammatical phenomenon can be established in an advanced…

  9. Entwicklung und lmplementierung von Analysemethoden zum Erfassen vonGeschwindigkeitsfeldem mit dem PIV Verfahren (Development and Implementation of Analytical Methods for Detecting Velocity Fields using PIV- Method)

    DTIC Science & Technology

    2016-04-26

    Particle-Image-Velocimetry 1 064nm beam dump 532nm output / Dielectric polarizer Doubling crystal Laser mirrors and plates Output couple Nd:YAG Rod...zuriickgesetzt. Der Anwender befindet sic :1 wieder an dem Funkt im Programm, nachdem er die Bildpaare eingelesen hatte. 5.2.3. Auswertung der Daten und

  10. The time resolution domain of stellar radio astronomy

    NASA Technical Reports Server (NTRS)

    Bookbinder, J.

    1985-01-01

    The high time resolution (HTR) radio observation of late-type stars and RS CVn systems is discussed. Some examples of these sources are addressed, identifying what information HTR observations can provide. HTR can provide important information on flares in late-type stars, and can be used to study coronal structure and the particle acceleration mechanism in these stars. The possible use of HTR to establish the nature of quiescent emission form RS CVn systems is discussed.

  11. The time resolution domain of stellar radio astronomy

    NASA Technical Reports Server (NTRS)

    Bookbinder, J.

    1985-01-01

    The high time resolution (HTR) radio observation of late-type stars and RS CVn systems is discussed. Some examples of these sources are addressed, identifying what information HTR observations can provide. HTR can provide important information on flares in late-type stars, and can be used to study coronal structure and the particle acceleration mechanism in these stars. The possible use of HTR to establish the nature of quiescent emission form RS CVn systems is discussed.

  12. Linguistic Challenges in Mendelian Genetics: Teachers' Talk in Action

    ERIC Educational Resources Information Center

    Thörne, Karin; Gericke, Niklas M.; Hagberg, Mariana

    2013-01-01

    This study investigates Swedish teachers' use of language when teaching Mendelian genetics in compulsory school. The primary objective of the study is to explore how teachers use the related concepts "gene," "allele," and "anlag" (a Swedish variant of the German word "anlage") and how these are related to…

  13. Linguistic Challenges in Mendelian Genetics: Teachers' Talk in Action

    ERIC Educational Resources Information Center

    Thörne, Karin; Gericke, Niklas M.; Hagberg, Mariana

    2013-01-01

    This study investigates Swedish teachers' use of language when teaching Mendelian genetics in compulsory school. The primary objective of the study is to explore how teachers use the related concepts "gene," "allele," and "anlag" (a Swedish variant of the German word "anlage") and how these are related to…

  14. Taxonomy and phylogeny of two species of the genus Deviata (Protista, Ciliophora) from China, with description of a new soil form, Deviata parabacilliformis sp. nov.

    PubMed

    Li, Fengchao; Lv, Zhao; Yi, Zhenzhen; Al-Farraj, Saleh A; Al-Rasheid, Khaled A S; Shao, Chen

    2014-11-01

    The morphology and morphogenesis of a soil hypotrichous ciliate, Deviata parabacilliformis sp. nov., isolated from northern China, were investigated. D. parabacilliformis measures about 75-210 × 25-60 µm in vivo, with an elongate and flexible body. It possesses one right marginal row, two to four left marginal rows and three dorsal kineties. The main morphogenetic features of D. parabacilliformis are: (i) the oral primordium originates de novo; (ii) anlage IV of the opisthe originates from parental frontoventral row V, anlage V originates de novo, and anlage VI forms from frontoventral row VI; and (iii) anlage I of the proter originates from the anterior portion of the parental paroral, anlage II originates from the buccal cirrus, anlage III originates from the parabuccal cirri, anlage IV originates from parental frontoventral row IV and anlage V forms from the anterior of parental frontoventral row VI. The morphology of an edaphic population of another species of the genus Deviata, Deviata bacilliformis (Gelei 1954) Eigner 1995, was also investigated. This work also provides the first record of SSU rRNA gene sequences for species of the genus Deviata. Molecular phylogenetic analysis suggests that Deviata is not monophyletic, and its position is poorly resolved due to weak phylogenetic signal of the 18S marker in the Stichotrichida. © 2014 IUMS.

  15. Metabolism

    MedlinePlus

    ... symptoms. Metabolic diseases and conditions include: Hyperthyroidism (pronounced: hi-per-THIGH-roy-dih-zum). Hyperthyroidism is caused ... or through surgery or radiation treatments. Hypothyroidism (pronounced: hi-po-THIGH-roy-dih-zum). Hypothyroidism is caused ...

  16. RNA splicing and editing modulation of 5-HT(2C) receptor function: relevance to anxiety and aggression in VGV mice.

    PubMed

    Martin, C B P; Ramond, F; Farrington, D T; Aguiar, A S; Chevarin, C; Berthiau, A-S; Caussanel, S; Lanfumey, L; Herrick-Davis, K; Hamon, M; Madjar, J J; Mongeau, R

    2013-06-01

    Changes in serotonin(2C) receptor (5-HTR2c) editing, splicing and density were found in conditions such as depression and suicide, but mechanisms explaining the changes in 5-HTR2c function are unknown. Thus, mice expressing only the fully edited VGV isoform of 5-HTR2c, in which clinically relevant behavioral changes are associated with alterations in splicing and receptor density, were studied. VGV mice displayed enhanced anxiety-like behavior in response to a preferential 5-HTR2c agonist in the social interaction test. Nearly half of interactions between pairs of VGV congeners consisted of fighting behaviors, whereas no fighting occurred in wild-type (WT) mice. VGV mice also exhibited a striking increase in freezing behaviors in reaction to an innately aversive ultrasonic stimulus. This behavioral phenotype occurred in conjunction with decreased brain 5-HT turnover during stress. These functional data were put in relation with the 5-HTR2c mRNA splicing process generating a truncated protein (5-HTR2c-Tr) in addition to the full-length receptor (5-HTR2c-Fl). 5-HTR2c-Tr mRNA was less abundant in many brain regions of VGV mice, which concomitantly had more 5-HTR2c than WT mice. Fluorescence resonance energy transfer and bioluminescence resonance energy transfer studies in transfected living HEK293T cells showed that 5-HTR2c-Tr interacts with 5-HTR2c-Fl. The 5-HTR2c-Tr was localized in the endoplasmic reticulum where it retained 5-HTR2c-Fl, preventing the latter to reach the plasma membrane. Consequently, 5-HTR2c-Tr decreased (3)H-mesulergine binding to 5-HTR2c-Fl at the plasma membrane in a concentration-dependent manner and more strongly with edited 5-HTR2c-Fl. These results suggest that 5-HTR2c pre-mRNA editing and splicing are entwined processes determining increased 5-HTR2c levels in pathological conditions through a deficit in 5-HTR2c-Tr.

  17. Development of the ciliary body: morphological changes in the distal portion of the optic cup in the human.

    PubMed

    Peces-Peña, M D; de la Cuadra-Blanco, C; Vicente, A; Mérida-Velasco, J R

    2013-01-01

    This study seeks to determine the main events that occur in the development of the ciliary body (CB) in the 5-14th week of development. The CB develops from the distal portion of the optic cup (OC) and the neighboring mesenchyme. During the 5th week of development, 4 zones were observed in the distal portion of the OC: in zone 1, the epithelia of the outer and inner layers of the OC came into contact. This contact coincided with the appearance of mainly apical granule pigments. This zone corresponded to the anlage of the epithelial layers of the CB. In zone 2, the cells surrounded the marginal sinus and contained scarce pigment granules and nuclei in the basal position. This zone corresponded to the anlage of the iris. Zone 3 was triangular in shape and its vertex ran towards the marginal sinus and corresponded to common cell progenitors. Zone 4 corresponded to the retinal pigment epithelium anlage and the neural retina anlage. We determined the onset of the stroma and the ciliary muscle anlage at the end of the 7th week. In the 13-14th week, we observed the anlage of the orbicularis ciliaris (pars plana of the CB) and corona ciliaris (pars plicata of the CB), in addition to the anlage of the ciliary muscle. Our study, therefore, establishes a precise timetable of the development of the CB. © 2013 S. Karger AG, Basel.

  18. Expression of messenger RNA coding for 5-HT receptor, alpha and beta adrenoreceptor (subtypes) during oestrus and dioestrus in the bovine uterus.

    PubMed

    Ontsouka, E C; Reist, M; Graber, H; Blum, J W; Steiner, A; Hirsbrunner, G

    2004-12-01

    Serotoninergic and adrenergic receptors (5-HTR and AR) are involved in the regulation of uterine contractility. The objective of this study was to compare mRNA levels of 5-HTR(1A), 5-HTR(1B), 5-HTR(1D), 5-HTR(1F), 5-HTR(2A), 5-HTR(2B), 5-HTR(2C), 5-HTR(4) and alpha(1A), alpha(1B), alpha(1D), alpha(2AD), alpha(2B), alpha(2C), and beta(1), beta(2), beta(3)-AR in oestrus and dioestrus, and at three uterine locations (tip, middle and base) using quantitative real-time polymerase chain reaction. Uterine specimens consisting of endometrium and myometrium including vessels and serosa were collected from cows in oestrus (n = 10) and dioestrus (n = 15) respectively. Levels of 5-HTR and AR mRNA were expressed relative to the geometric mean of ribosomal RNA (18S), ubiquitin and glyceraldehyde phosphate dehydrogenase by the mean values of geNorm algorithm. 5-HTR(1A), 5-HTR(2C) and beta(3)-AR mRNA could not be detected in uterine tissues. The mRNA levels of 5-HTR(1F) and 5-HTR(2B) were lower (P < 0.05), but of 5-HTR(4) were higher (P < 0.05) in oestrus than in dioestrus. The mRNA levels of alpha(1A)-AR, alpha(2AD)-AR, alpha(2B)-AR were lower (P < 0.05), but of alpha(2C)-AR and beta(2)-AR were higher (P < 0.05) in oestrus than dioestrus. The mRNA levels of 5-HTR(1B) and 5-HTR(1D) (oestrus) and of alpha(2AD)-AR (dioestrus) differed among uterine locations (base > middle > tip; P < 0.05). The mRNA expression of 5-HTR and AR (subtypes) in bovine uterus was associated with cycle activity and varied according to uterine location. Additional studies on protein level will be carried out in order to elucidate the role of these receptor families on uterine contractility, which may then help to clarify clinical relevance.

  19. Structure and Function of the Virulence-Associated High-Temperature Requirement A of Mycobacterium tuberculosis

    SciTech Connect

    MohamedMohaideen, Nilofar N.; Palaninathan, Satheesh K.; Morin, Paul M.; Williams, Brad J.; Braunstein, Miriam; Tichy, Shane E.; Locker, Joseph; Russell, David H.; Jacobs, Jr., William R.; Sacchettini, James C.

    2008-06-13

    The high-temperature requirement A (HtrA) family of serine proteases has been shown to play an important role in the environmental and cellular stress damage control system in Escherichia coli. Mycobacterium tuberculosis (Mtb) has three putative HtrA-like proteases, HtrA1, HtrA2, and HtrA3. The deletion of htrA2 gives attenuated virulence in a mouse model of TB. Biochemical analysis reveals that HtrA2 can function both as a protease and as a chaperone. The three-dimensional structure of HtrA2 determined at 2.0 {angstrom} resolution shows that the protease domains form the central core of the trimer and the PDZ domains extend to the periphery. Unlike E. coli DegS and DegP, the protease is naturally active due to the formation of the serine protease-like catalytic triad and its uniquely designed oxyanion hole. Both protease and PDZ binding pockets of each HtrA2 molecule are occupied by autoproteolytic peptide products and reveal clues for a novel autoregulatory mechanism that might have significant importance in HtrA-associated virulence of Mtb.

  20. HTR1B, ADIPOR1, PPARGC1A, and CYP19A1 and Obesity in a Cohort of Caucasians and African Americans: An Evaluation of Gene-Environment Interactions and Candidate Genes

    PubMed Central

    Edwards, Todd L.; Velez Edwards, Digna R.; Villegas, Raquel; Cohen, Sarah S.; Buchowski, Maciej S.; Fowke, Jay H.; Schlundt, David; Long, Ji Rong; Cai, Qiuyin; Zheng, Wei; Shu, Xiao-Ou; Hargreaves, Margaret K.; Jeffrey, Smith; Williams, Scott M.; Signorello, Lisa B.; Blot, William J.; Matthews, Charles E.

    2012-01-01

    The World Health Organization estimates that the number of obese and overweight adults has increased to 1.6 billion, with concomitant increases in comorbidity. While genetic factors for obesity have been extensively studied in Caucasians, fewer studies have investigated genetic determinants of body mass index (BMI; weight (kg)/height (m)2) in African Americans. A total of 38 genes and 1,086 single nucleotide polymorphisms (SNPs) in African Americans (n = 1,173) and 897 SNPs in Caucasians (n = 1,165) were examined in the Southern Community Cohort Study (2002–2009) for associations with BMI and gene × environment interactions. A statistically significant association with BMI survived correction for multiple testing at rs4140535 (β = −0.04, 95% confidence interval: −0.06, −0.02; P = 5.76 × 10−5) in African Americans but not in Caucasians. Gene-environment interactions were observed with cigarette smoking and a SNP in ADIPOR1 in African Americans, as well as between a different SNP in ADIPOR1 and physical activity in Caucasians. A SNP in PPARGC1A interacted with alcohol consumption in African Americans, and a different SNP in PPARGC1A was nominally associated in Caucasians. A SNP in CYP19A1 interacted with dietary energy intake in African Americans, and another SNP in CYP191A had an independent association with BMI in Caucasians. PMID:22106445

  1. AXIAL SKELETAL AND HOX EXPRESSION DOMAIN ALTERATIONS INDUCED BY RETINOIC ACID, VALPROIC ACID AND BROMOXYNIL DURING MURINE DEVELOPMENT

    EPA Science Inventory

    ABSTRACT

    Retinoic acid (RA) alters the developmental fate of the axial skeletal anlage. "Anteriorizations" or "posteriorizations", the assumption of characteristics of embryonic areas normally anterior or posterior to the affected tissues, are correlated with altered emb...

  2. AXIAL SKELETAL AND HOX EXPRESSION DOMAIN ALTERATIONS INDUCED BY RETINOIC ACID, VALPROIC ACID AND BROMOXYNIL DURING MURINE DEVELOPMENT

    EPA Science Inventory

    ABSTRACT

    Retinoic acid (RA) alters the developmental fate of the axial skeletal anlage. "Anteriorizations" or "posteriorizations", the assumption of characteristics of embryonic areas normally anterior or posterior to the affected tissues, are correlated with altered emb...

  3. The role of the serotonergic system in suicidal behavior

    PubMed Central

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

    2013-01-01

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

  4. An in situ hybridization study of Runx2, Osterix, and Sox9 in the anlagen of mouse mandibular condylar cartilage in the early stages of embryogenesis.

    PubMed

    Shibata, Shunichi; Yokohama-Tamaki, Tamaki

    2008-09-01

    Mandibular condylar cartilage is the best-studied mammalian secondary cartilage, differing from primary cartilage in that it originates from alkaline phosphatase-positive progenitor cells. We previously demonstrated that three transcription factors related to bone and cartilage formation, namely Runx2, Osterix and Sox9, are simultaneously expressed in the anlage of mandibular condylar cartilage (condylar anlage) at embryonic day (E)14. In this study, expression of these transcription factors was investigated in the anlagen of mandibular bone (mandibular anlagen) from E11.0 to 14.0. Runx2 mRNA was first expressed in the mandibular anlage at E11.5. Osterix mRNA was first expressed at E12.0, and showed a different expression pattern from that of Runx2 from E12.5 to E14.0, confirming that Osterix acts downstream of Runx2. Sox9 mRNA was expressed in Meckel's cartilage and its anlagen throughout the experimental period, but not clearly in the mandibular anlagen until E13.0. At E13.5, the condylar anlage was morphologically identified at the posterior end of the mandibular anlage, and enhanced Sox9 mRNA expression was detected here. At this stage, Runx2 and Osterix mRNA were simultaneously detected in the condylar anlage. These results indicate that the Sox9 mRNA-expressing condylar anlage is derived from Runx2/Osterix mRNA-expressing mandibular anlage, and that upregulation of Sox9 in this region acts as a trigger for subsequent condylar cartilage formation.

  5. Solvent-Responsive Molecularly Imprinted Nanogels for Targeted Protein Analysis in MALDI-TOF Mass Spectrometry.

    PubMed

    Bertolla, Maddalena; Cenci, Lucia; Anesi, Andrea; Ambrosi, Emmanuele; Tagliaro, Franco; Vanzetti, Lia; Guella, Graziano; Bossi, Alessandra Maria

    2017-03-01

    Molecular imprinted poly(acrylamido)-derivative nanogels have shown their selectivity to bind the protein human serum transferrin (HTR) and also showed their capability for instantaneous solvent-induced modification upon the addition of acetonitrile. Integrated to matrix-assisted laser desorption/ionization time-of-flight mass analysis the HTR-imprinted solvent-responsive nanogels permitted the determination of HTR straight from serum and offered novel perspectives in targeted protein analysis.

  6. Serotonin as a homeostatic regulator of lactation.

    PubMed

    Collier, R J; Hernandez, L L; Horseman, N D

    2012-08-01

    Serotonin (5-HT), a neurotransmitter produced in mammary epithelial cells (MECs), acts via autocrine-paracrine mechanisms on MECs to regulate milk secretion in a variety of species. Recent studies in dairy cows reported that 5-HT ligands affect milk yield and composition. We determined the mRNA expression of bovine 5-HT receptor (5-HTR) subtypes in bovine mammary tissue (BMT) and cultured bovine MECs. We then used pharmacologic agents to evaluate functional activities of 5-HTR subtypes. The mRNAs for five receptor isoforms (5-HTR1B, 5-HTR2A, 5-HTR2B, 5-HTR4, and 5-HTR7) were identified by conventional reverse transcription PCR, real-time PCR, and in situ hybridization in BMT. In addition to luminal MEC expression, 5-HTR4 was expressed in myoepithelium, and 5-HTR1B, HTR2A, and HTR2B were expressed in small mammary blood vessels. Studies to date report that there are multiple 5-HTR isoforms in mammary tissue of rodents, humans, and cattle. Inhibition of the 5-HT reuptake transporter with selective 5-HT reuptake inhibitors (SSRIs) disrupted tight junctions and decreased milk protein mRNA expression in mouse, human, and bovine mammary cells. Selective 5-HT reuptake inhibitors act to increase the cellular exposure to 5-HT by preventing reuptake of 5-HT by the cell and eventual degradation. Increasing 5-HT concentration in milk via inhibiting its reuptake (SSRI), or by increasing the precursor for 5-HT synthesis 5-hydroxytryptophan, accelerated decline in milk synthesis at dry-off. We conclude that the 5-HT system in mammary tissue acts as a homeostatic regulator of lactation.

  7. Deletion of the Serotonin Receptor Type 3A in Mice Leads to Sudden Cardiac Death During Pregnancy.

    PubMed

    Park, Hyewon; Oh, Chang-Myung; Park, Junbeom; Park, Hyelim; Cui, Shanyu; Kim, Hyung Suk; Namkung, Jun; Park, Sang-kyu; Pak, Hui-Nam; Lee, Moon-Hyoung; Kim, Hail; Joung, Boyoung

    2015-01-01

    The serotonin receptor type 3 (Htr3) blocker is associated with QT prolongation and torsades de pointes. However, little is known about effects of Htr3 on the heart arrhythmia. An electrophysiological study Involving knock-out (KO) female mice lacking functional Htr3a (Htr3a(-/-)) and their wild-type littermates during non-pregancy (NP) and late pregnancy (LP) was performed. Htr3a mRNA was present in the wild-type, but not in the Htr3a(-/-)mouse hearts. Serotonin and tryptophan hydroxylase 1 (Tph1), a rate-limiting enzyme of serotonin synthesis in hearts, is increased during pregnancy. The heart weight and size were increased in the pregnant mice regardless of a mutation. The QTc intervals were prolonged after pregnancy in both the wild (NP: 171.2±16.8 vs. LP: 247.7±14.3 ms; P<0.001) and Htr3a(-/-)mice (NP: 187.9±18.7 vs. LP: 275.6±11.0 ms, P<0.001). Compared with wild-type LP mice, Htr3a(-/-)LP mice had increased spontaneous ventricle tarchycardia (VT; 56% vs. 0%, P=0.002), VT inducibility (66% vs. 25%, P=0.002) and mortality (56% vs. 0%, P=0.002). Pharmacologic administration of serotonin and Htr3 agonists (m-CPBG) decreased the QT interval in wild mice, but not in Htr3a(-/-)mice. Htr3a is present in mouse hearts. Serotonin and Tph1 were increased during pregnancy. The deletion of Htr3a was related to fatal arrhythmias and sudden cardiac death during pregnancy, and its activation reversed the QT prolongation.

  8. Morphology, morphogenesis and small subunit rRNA gene sequence of a soil hypotrichous ciliate, Perisincirra paucicirrata (Ciliophora, Kahliellidae), from the shoreline of the Yellow River, North China.

    PubMed

    Li, Fengchao; Xing, Yi; Li, Jiamei; Al-Rasheid, Khaled A S; He, Songke; Shao, Chen

    2013-01-01

    The morphology, morphogenesis, and 18S rRNA gene sequence of a soil hypotrichous ciliate Perisincirra paucicirrata, isolated from north China, were investigated. Perisincirra paucicirrata differs from its congeners in: (1) having a body length to width ratio in vivo of 4:1, (2) its adoral zone occupying between 15% and 25% of the total body length, and (3) the presence of two parabuccal cirri, three left (with 10-16 cirri each) and two right marginal rows (with 14-24 cirri each), and three dorsal kineties. Our study offers a first attempt to begin to map the morphogenetic processes of the genus, which are mainly characterised by the following: the formation of four frontal ventral transverse anlagens for each daughter cell, with the proter's anlage I originating from the reorganised anterior part of the parental paroral; the paroral and endoral anlage developed from the reorganised old endoral and do not contribute the first frontal cirrus; the frontoventral transverse anlage I contributing the left frontal cirrus; anlage II generating the middle frontal and the buccal cirri; anlage III developing the right frontal cirrus and the anterior parabuccal cirrus; and anlage IV contributing the posterior parabuccal cirrus. As an additional contribution, we judge that the inner one or the two right rows of P. kahli and P. longicirrata are marginal rows. Phylogenetic analysis based on SSU rDNA sequences suggests that Perisincirra is related to sporadotrichids, but provides no credible evidence for its taxonomic position.

  9. Polymorphisms of the serotonin transporter and receptor genes: susceptibility to substance abuse.

    PubMed

    Herman, Aryeh I; Balogh, Kornelia N

    2012-06-01

    Serotonin (5-hydroxytryptamine [5-HT]) is an important neurotransmitter implicated in regulating substance-use disorder (SUD) acquisition, maintenance, and recovery. During the past several years, an abundance of research has begun discovering and describing specific 5-HT genetic polymorphisms associated with SUDs. Genetic variations in the 5-HT system, such as SLC6A4, HTR1B, HTR2A, HTR2C, HTR3 (HTR3A, HTR3B, HTR3C, HTR3D, and HTR3E), likely play a role contributing to SUD patient heterogeneity. The 5-HT transporter-linked polymorphic region S allele, located in SLC6A4, has now been modestly associated with alcohol dependence in two large meta-analyses. Additional 5-HT genes may also play a role but have not been extensively investigated. A limited number of SUD treatment studies have included 5-HT gene variation as moderating treatment outcomes, but the results have been equivocal. Future research on 5-HT addiction genetics should adopt whole-genome sequencing technology, utilize large study samples, and collect data from multiple ethnic groups. Together, these methods will build on the work already conducted with the aim of utilizing 5-HT genetics in SUD treatment settings.

  10. Inhibition of telomerase RNA decay rescues telomerase deficiency caused by dyskerin or PARN defects

    PubMed Central

    Shukla, Siddharth; Schmidt, Jens C.; Goldfarb, Katherine C.; Cech, Thomas R.; Parker, Roy

    2016-01-01

    Mutations in the human telomerase RNA (hTR), the telomerase RNP component dyskerin (DKC1), and the poly(A) ribonuclease (PARN) can lead to reduced levels of hTR and dyskeratosis congenita (DC). However, the enzymes and mechanisms responsible for hTR degradation are unknown. We demonstrate that defects in dyskerin binding lead to hTR degradation by PAPD5-mediated oligoadenylation promoting 3’ to 5’ degradation by EXOSC10, as well as decapping and 5’ to 3’ decay by the cytoplasmic DCP2 and XRN1 enzymes. PARN increases hTR levels by deadenylating hTR, thereby limiting its degradation by EXOSC10. Telomerase activity and proper hTR localization in dyskerin- or PARN-deficient cells can be rescued by knockdown of DCP2 and/or EXOSC10. Prevention of hTR RNA decay also leads to a rescue of localization of DC-associated hTR mutants. These results suggest that inhibition of RNA decay pathways might be a useful therapy for some telomere pathologies. PMID:26950371

  11. Telomerase Reverse Transcriptase Is Required for the Localization of Telomerase RNA to Cajal Bodies and Telomeres in Human Cancer Cells

    PubMed Central

    Tomlinson, Rebecca L.; Abreu, Eladio B.; Ziegler, Tania; Ly, Hinh; Counter, Christopher M.

    2008-01-01

    Telomere maintenance by telomerase is critical for the unlimited division potential of most human cancer cells. The two essential components of human telomerase, telomerase RNA (hTR) and telomerase reverse transcriptase (hTERT), are recruited from distinct subnuclear sites to telomeres during S phase. Throughout the remainder of the cell cycle hTR is found primarily in Cajal bodies. The localization of hTR to Cajal bodies and telomeres is specific to cancer cells where telomerase is active and is not observed in primary cells. Here we show that the trafficking of hTR to both telomeres and Cajal bodies depends on hTERT. RNA interference–mediated depletion of hTERT in cancer cells leads to loss of hTR from both Cajal bodies and telomeres without affecting hTR levels. In addition, expression of hTERT in telomerase-negative cells (including primary and ALT cancer cell lines) induces hTR to localize to both sites. Factors that did not stimulate hTR localization in our experiments include increased hTR RNA levels and Cajal body numbers, and expression of SV40 large T antigen and oncogenic Ras. Our findings suggest that the trafficking of telomerase to Cajal bodies and telomeres in cancer cells correlates with and depends on the assembly of the enzyme. PMID:18562689

  12. Deep Burn: Development of Transuranic Fuel for High-Temperature Helium-Cooled Reactors- Monthly Highlights October 2010

    SciTech Connect

    Snead, Lance Lewis; Besmann, Theodore M; Collins, Emory D; Bell, Gary L

    2010-11-01

    The DB Program monthly highlights report for September 2010, ORNL/TM-2010/252, was distributed to program participants by email on October 26. This report discusses: (1) Core and Fuel Analysis; (2) Spent Fuel Management; (3) Fuel Cycle Integration of the HTR (high temperature helium-cooled reactor); (4) TRU (transuranic elements) HTR Fuel Qualification; (5) HTR Spent Fuel Recycle - (a) TRU Kernel Development (ORNL), (b) Coating Development (ORNL), (c) Characterization Development and Support, (d) ZrC Properties and Handbook; and (6) HTR Fuel Recycle.

  13. Telomerase reverse transcriptase is required for the localization of telomerase RNA to cajal bodies and telomeres in human cancer cells.

    PubMed

    Tomlinson, Rebecca L; Abreu, Eladio B; Ziegler, Tania; Ly, Hinh; Counter, Christopher M; Terns, Rebecca M; Terns, Michael P

    2008-09-01

    Telomere maintenance by telomerase is critical for the unlimited division potential of most human cancer cells. The two essential components of human telomerase, telomerase RNA (hTR) and telomerase reverse transcriptase (hTERT), are recruited from distinct subnuclear sites to telomeres during S phase. Throughout the remainder of the cell cycle hTR is found primarily in Cajal bodies. The localization of hTR to Cajal bodies and telomeres is specific to cancer cells where telomerase is active and is not observed in primary cells. Here we show that the trafficking of hTR to both telomeres and Cajal bodies depends on hTERT. RNA interference-mediated depletion of hTERT in cancer cells leads to loss of hTR from both Cajal bodies and telomeres without affecting hTR levels. In addition, expression of hTERT in telomerase-negative cells (including primary and ALT cancer cell lines) induces hTR to localize to both sites. Factors that did not stimulate hTR localization in our experiments include increased hTR RNA levels and Cajal body numbers, and expression of SV40 large T antigen and oncogenic Ras. Our findings suggest that the trafficking of telomerase to Cajal bodies and telomeres in cancer cells correlates with and depends on the assembly of the enzyme.

  14. "Land der Berge, Land der Seen": Zum Oesterreichbild in amerikanischen Deutsch-Lehrbuechern ("Land of Mountains, Land of Lakes": On the Picture of Austria Presented by German Textbooks in the USA).

    ERIC Educational Resources Information Center

    Koppensteiner, Juergen

    1978-01-01

    Nearly all German textbooks currently used in the USA either entirely fail to mention Austria, or else portray it by means of myths, cliches (Apfelstrudel, Sachertorte, Gemuetlichkeit), and often with outright untruths. The American student gets a picture of Austria as an "underdeveloped Disneyland of Europe". (WGA)

  15. "Zielsprache Englisch" interviewt Hans-Eberhard Piepho zum Thema "Neues" Volkshochschul-Zertifikat Englisch 1 ("Zielsprache Englisch" Interviews Hans-Eberhard Piepho on the New People's University Certificate 1 in English).

    ERIC Educational Resources Information Center

    Zielsprache Englisch, 1979

    1979-01-01

    Discusses aspects of the revised People's University Certificate English 1, including: (1) function of catalogs in setting forth learning goals (topics, situations, intentions) for textbook writers and course directors; (2) advantages of authentic exercise texts; (3) weighting linguistic skills; (4) vocabulary work; and (5) developing grammatical…

  16. Literature Research on the Mechanical Properties of Fibre Composise Materials Analysis of the State of the Art. Volume I. (Schrifttumsrecherche zum Festigkeitsverhalten von Faserverbundwerkstoffen-Analyse des Standes der Technik-),

    DTIC Science & Technology

    1980-08-11

    only one resin system. Surface treatment with poly- amide primer and polyurethane enamel proved to be advantageous (see also section 2). The following...coat of polyurethane paint there was no Stion in life under the same test conditions. The environmental stresses ts were environmental changes lasting...drilled), - simultaneous shaping and curing (’ pultrusion ’ process), - spar flange embedded in the skin to reduce the number of fasteners, - increased

  17. Ergebnisse einer Lehrerbefragung zum Hintergrund des Englischunterrichts in der Hauptschuloberstufe (Results of a Teacher Questionnaire on the Background of Instruction in English in the Upper Grades of the Hauptschule [9-grade "terminal" school]).

    ERIC Educational Resources Information Center

    Hellwig, Karlheinz

    1980-01-01

    The questionnaire dealt with: earmarks of weaker students, parents' attitude, the teachers' training and working conditions, problems of individual instruction, time-frame, existence of professional associations, evaluation of professional conferences, and the use of media. Some evaluations are appended. (IFS/WGA)

  18. Astronomical dilettante or misunderstood genius? On Johann Hieronymus Schroeter's image in the history of science. (German Title: Astronomischer Dilettant oder verkanntes Genie? Zum Bild Johann Hieronymus Schroeters in der Wissenschaftsgeschichte)

    NASA Astrophysics Data System (ADS)

    Oestmann, Günther

    The paper deals with contemporary assessments of Johann Hieronymus Schroeter's (1745-1816) astronomical works - especially by Wilhelm Olbers and Carl Friedrich Gauß - and also later judgements of the scientific importance and significance of his observations voiced by astronomers and historians.

  19. Ergebnisse einer Lehrerbefragung zum Hintergrund des Englischunterrichts in der Hauptschuloberstufe (Results of a Teacher Questionnaire on the Background of Instruction in English in the Upper Grades of the Hauptschule [9-grade "terminal" school]).

    ERIC Educational Resources Information Center

    Hellwig, Karlheinz

    1980-01-01

    The questionnaire dealt with: earmarks of weaker students, parents' attitude, the teachers' training and working conditions, problems of individual instruction, time-frame, existence of professional associations, evaluation of professional conferences, and the use of media. Some evaluations are appended. (IFS/WGA)

  20. Auf dem Weg zu einem neuen Weltcurriculum? Zum Grundbildungskonzept von PISA und der Aufgabenzuweisung an die Schule (Towards a World Curriculum? -- The Concept of Basic Education (Literacy) underlying PISA and the tasks allocated to Schooling).

    ERIC Educational Resources Information Center

    Fuchs, Hans-Werner

    2003-01-01

    Recognizes that the debate on the Program for International Student Assessment 2000 (PISA) has been dominated by comparisons of the ranks achieved and by the question of the consequences to be drawn from the results. Discusses the educational concept forming the basis for the investigation and aims of PISA. (CAJ)

  1. Partielle Geschlechtertrennung--Enttauschte Hoffnungen? Monoedukative Lernumgebungen zum Chancenausgleich im Unterricht auf dem Prufstand (Partial Separation of the Sexes--Disappointed Hopes? An Assessment of Non-Educative Learning Environments for Equalizing Educational Opportunities).

    ERIC Educational Resources Information Center

    Ludwig, Peter H.

    2003-01-01

    Argues that the thesis of discrimination against girls in coeducation schools has been replaced by a belief that, during certain phases or in specific subjects, abandonment of coeducation would promote equal opportunities. Questions whether classic or recent surveys provide empirical evidence for this moderate skeptical attitude towards…

  2. James Baldwin und das Rassenproblem in den USA. Unterrichtsprojekt zum Komplex "Texte und ihre gesellschaftliche Bedingtheit" (James Baldwin and the Race Problem in the USA. Teaching Project in the Series "Texts and their Social Context")

    ERIC Educational Resources Information Center

    Kerschgens, Edda

    1977-01-01

    Reports on an 18-hour teaching project with students at a teachers' college. Four Baldwin texts were used. Questions considered included whether Baldwin's treatment of the race problem reveals any changes or shifts of emphasis. Suggestions are made for adaptation to teaching in grades 11-13. (Text is in German.) (IFS/WGA)

  3. Partielle Geschlechtertrennung--Enttauschte Hoffnungen? Monoedukative Lernumgebungen zum Chancenausgleich im Unterricht auf dem Prufstand (Partial Separation of the Sexes--Disappointed Hopes? An Assessment of Non-Educative Learning Environments for Equalizing Educational Opportunities).

    ERIC Educational Resources Information Center

    Ludwig, Peter H.

    2003-01-01

    Argues that the thesis of discrimination against girls in coeducation schools has been replaced by a belief that, during certain phases or in specific subjects, abandonment of coeducation would promote equal opportunities. Questions whether classic or recent surveys provide empirical evidence for this moderate skeptical attitude towards…

  4. The archivalia of the Astronomisches Rechen-Institut on the calendar in Prussia. Edition of the documents. (German Title: Die Archivalien des Astronomischen Rechen-Instituts zum Kalender in Preußen. Edition der Dokumente.)

    NASA Astrophysics Data System (ADS)

    Wielen, Roland; Wielen, Ute

    In the archives of the Astronomisches Rechen-Institut at Heidelberg, there is an old set of 31 documents which are related to the calendar used in Prussia and which originated in the period from 1700 to 1854. The oldest document is an original print of the 'Calendar Edict' issued on 10 May 1700. In this edict, Friedrich III., Elector of Brandenburg, gave a monopoly for issuing calendars in his country to an academy which was founded slightly later. He founded at the same time an observatory in Berlin. The main task of the employed astronomers was to edit the 'Improved Calendar' which was newly introduced in his Protestant country. The Astronomisches Rechen-Institut, which was founded in Berlin and was moved to Heidelberg in 1945, considers this Calendar Edict as his foundation document too. All the other documents are handwritten, mainly letters, but also a detailed expose 'On the Calendar Issues in the Prussian State' from 1843. Two of the scripts stem from the 18th century. The remaining documents are related to the work of the Royal Prussian Calendar Deputation and were written between 1816 and 1854. In this paper we describe, commentate, and transliterate all the documents of this 'Kalender-Konvolut'.

  5. Video: einige Bemerkungen zum medien-spezifischen Ort und zur didaktischen Basis (Video: Some Notes on Its Place among Teaching Media and on Its Theoretical Basis in FL Teaching)

    ERIC Educational Resources Information Center

    Bauer, Hans-Ludwig

    1976-01-01

    Reports on the introduction of video recorders at various branches of the Goethe Institute. The characteristics of video are compared with those of films, tapes and printed matter. Video's advantage is that through it authentic, unprepared material can be presented, and it provides strong motivation. (Text is in German.) (IFS/WGA)

  6. "Land der Berge, Land der Seen": Zum Oesterreichbild in amerikanischen Deutsch-Lehrbuechern ("Land of Mountains, Land of Lakes": On the Picture of Austria Presented by German Textbooks in the USA).

    ERIC Educational Resources Information Center

    Koppensteiner, Juergen

    1978-01-01

    Nearly all German textbooks currently used in the USA either entirely fail to mention Austria, or else portray it by means of myths, cliches (Apfelstrudel, Sachertorte, Gemuetlichkeit), and often with outright untruths. The American student gets a picture of Austria as an "underdeveloped Disneyland of Europe". (WGA)

  7. James Baldwin und das Rassenproblem in den USA. Unterrichtsprojekt zum Komplex "Texte und ihre gesellschaftliche Bedingtheit" (James Baldwin and the Race Problem in the USA. Teaching Project in the Series "Texts and their Social Context")

    ERIC Educational Resources Information Center

    Kerschgens, Edda

    1977-01-01

    Reports on an 18-hour teaching project with students at a teachers' college. Four Baldwin texts were used. Questions considered included whether Baldwin's treatment of the race problem reveals any changes or shifts of emphasis. Suggestions are made for adaptation to teaching in grades 11-13. (Text is in German.) (IFS/WGA)

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

    PubMed

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

    2017-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  10. The optimum number and types of plasma samples necessary for an accurate activated partial thromboplastin time-based heparin therapeutic range.

    PubMed

    Marlar, Richard A; Gausman, Jana

    2013-01-01

    Monitoring of unfractionated heparin therapy by activated partial thromboplastin time (aPTT) using the ex vivo method for determining the aPTT-based heparin therapeutic range (HTR) is the standard of practice. Many intrinsic and extrinsic factors influence its accuracy. To investigate the optimum number and types of samples acceptable for an accurate ex vivo HTR determination. Values from patients receiving unfractionated heparin are used to determine the HTR by published guidelines. The number and types of samples are changed to investigate the effect on HTR parameters. Absolute minimum number of samples for an accurate HTR is 20, with fewer than 10% of the samples from the same patient or 50% of the samples with international normalized ratio of 1.3 to 1.5. The ex vivo HTR method is the best protocol currently available; however, the number of samples used affects its accuracy. The optimum number of samples is 30 or more but the absolute minimum number is 20. In addition, limitation of specific sample types also affects the HTR parameters. An inaccurate HTR may be calculated if inappropriate sample number or types of samples are used.

  11. Hardware Model Of A Shipboard Zonal Electrical Distribution System (ZEDS): Alternating Current/Direct Current (AC/DC)

    DTIC Science & Technology

    2010-06-01

    STC-5 Generator Characteristics from Testing [ 10 ] ....................................................... 54 Table 5: LA 55-P and HTR 50-SB...200 kHz DC ... 10 kHz The results of the testing, shown in Figure 29 and Figure 30, indicated that the HTR 50-SB has sufficient fidelity for this...53 10 5.0 THE ZEDS HARDWARE MODEL ARCHITECTURE

  12. 75 FR 81246 - Privacy Act of 1974; System of Records

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-27

    ... maintenance of the system: 10 U.S.C. 138, Assistant Secretaries of Defense; DoD Directive 5125.01, Assistant... Individual Ready Reserve (IRR) member's health treatment record (HTR) from the Department of Veterans Affairs (VA) upon mobilization of the individual. The VA is required to maintain the HTR on members of the...

  13. How to Conceive of Critical Educational Theory Today?

    ERIC Educational Resources Information Center

    Masschelein, Jan

    2004-01-01

    This paper starts from a brief sketch of the classical figure of critical educational theory or science (Kritische Erziehungswissenshaft). Critical educational theory presents itself as the privileged guardian of the critical principle of education (Bildung) and its emancipatory promise. It involves the possibility of saying "I" in order to speak…

  14. Responsie van Verschillende Munitieartikelen bij Opwarming en Brand (Response of Munitions Items Due to Heating or a Fire)

    DTIC Science & Technology

    2007-09-01

    werkzaamheden thermische responsie van artikelen. (met KS32) valt binnen deze normen net als Het project is gestart met bet ontwikkelen De kritische temperatuur...RDX, HMX of TNT en bindersystemnen als HTPB. Ook bijvoorbeeld de moderne 155 munitie van Rheinmetall (met KS32) valt binnen deze normen , net als

  15. Modulation of anxiety by cortical serotonin 1A receptors

    PubMed Central

    Piszczek, Lukasz; Piszczek, Agnieszka; Kuczmanska, Joanna; Audero, Enrica; Gross, Cornelius T.

    2015-01-01

    Serotonin (5-HT) plays an important role in the modulation of behavior across animal species. The serotonin 1A receptor (Htr1a) is an inhibitory G-protein coupled receptor that is expressed both on serotonin and non-serotonin neurons in mammals. Mice lacking Htr1a show increased anxiety behavior suggesting that its activation by serotonin has an anxiolytic effect. This outcome can be mediated by either Htr1a population present on serotonin (auto-receptor) or non-serotonin neurons (hetero-receptor), or both. In addition, both transgenic and pharmacological studies have shown that serotonin acts on Htr1a during development to modulate anxiety in adulthood, demonstrating a function for this receptor in the maturation of anxiety circuits in the brain. However, previous studies have been equivocal about which Htr1a population modulates anxiety behavior, with some studies showing a role of Htr1a hetero-receptor and others implicating the auto-receptor. In particular, cell-type specific rescue and suppression of Htr1a expression in either forebrain principal neurons or brainstem serotonin neurons reached opposite conclusions about the role of the two populations in the anxiety phenotype of the knockout. One interpretation of these apparently contradictory findings is that the modulating role of these two populations depends on each other. Here we use a novel Cre-dependent inducible allele of Htr1a in mice to show that expression of Htr1a in cortical principal neurons is sufficient to modulate anxiety. Together with previous findings, these results support a hetero/auto-receptor interaction model for Htr1a function in anxiety. PMID:25759645

  16. 5-HT3 receptor influences the washing phenotype and visual organization in obsessive-compulsive disorder supporting 5-HT3 receptor antagonists as novel treatment option.

    PubMed

    Lennertz, Leonhard; Wagner, Michael; Grabe, Hans Jörgen; Franke, Petra E; Guttenthaler, Vera; Rampacher, Friederike; Schulze-Rauschenbach, Svenja; Vogeley, Andrea; Benninghoff, Jens; Ruhrmann, Stephan; Pukrop, Ralf; Klosterkötter, Joachim; Falkai, Peter; Maier, Wolfgang; Mössner, Rainald

    2014-01-01

    A role of the HTR3A-E genes in obsessive-compulsive disorder (OCD) can be expected based on promising effects of 5-HT3 receptor antagonists as adjunctive treatment of OCD. We therefore genotyped six common coding or promoter variants within the HTR3A-E genes in a case-control-sample consisting of N=236 OCD patients and N=310 control subjects and in N=58 parent-child-trios. Given the heterogeneous OCD phenotype, we also investigated OCD symptom dimensions and cognitive endophenotypes in subsamples. OCD patients scoring high for the washing subtype were significantly more likely to carry the c.256G-allele of the HTR3E variant rs7627615 (p=0.0001) as compared to OCD patients low for this symptom dimension. Visual organization was impaired in OCD patients and unaffected relatives as compared to healthy control subjects and carriers of the HTR3E c.256G/c.256G-genotype performed significantly worse (p=0.007). The case-control analyses revealed a nominal significant association of the HTR3D variant rs1000592 (p.H52R) with OCD (p=0.029) which was also evident after combination of the case-control and the trio-results (p=0.024). In male subjects, the variant rs6766410 (p.N163K) located in the HTR3C was significantly associated with OCD (p=0.007). The association findings of the HTR3C and the HTR3E remained significant after correction for the number of variants investigated. These findings indicate a role of common variants of the HTR3A-E genes in OCD and OCD-related phenotypes and further support the use of 5-HT3 receptor antagonists as novel treatment options. The HTR3E gene is a novel candidate gene impacting on the individual expression of OC symptoms and OCD-related cognitive dysfunction.

  17. Ontogeny of the Saccus Vasculosus, a Seasonal Sensor in Fish.

    PubMed

    Maeda, Ryosuke; Shimo, Takayuki; Nakane, Yusuke; Nakao, Nobuhiro; Yoshimura, Takashi

    2015-11-01

    TSH secreted from the pars distalis (PD) of the pituitary gland stimulates the thyroid gland. In contrast, TSH secreted from the pars tuberalis (PT) of the pituitary gland regulates seasonal reproduction. The ontogeny of thyrotrophs and the regulatory mechanisms of TSH are apparently different between the PD and the PT. Interestingly, fish do not have an anatomically distinct PT, and the saccus vasculosus (SV) of fish is suggested to act as a seasonal sensor. Thus, it is possible that the SV is analogous to the PT. Here we examined the ontogeny of the pituitary gland and SV using rainbow trout. A histological analysis demonstrated the development of the pituitary anlage followed by that of the SV. Lhx3 and Pit-1, which are required for the development of PD thyrotrophs, clearly labeled the pituitary anlage. The common glycoprotein α-subunit (CGA) and TSH β-subunit (TSHB) genes were also detected in the pituitary anlage. In contrast, none of these genes were detected in the SV anlage. We then performed a microarray analysis and identified parvalbumin (Pvalb) as a marker for SV development. Because Pvalb expression was not detected in the pituitary anlage, no relationship was observed between the development of the SV and the pituitary gland. In contrast to embryos, Lhx3, Pit-1, CGA, and TSHB were all expressed in the adult SV. These results suggest that the morphological differentiation of SV occurs during the embryonic stage but that the functional differentiation into a seasonal sensor occurs in a later developmental stage.

  18. Identification of Methylation Sites and Effects of Phototaxis Stimuli on Transducer Methylation in Halobacterium salinarum

    PubMed Central

    Perazzona, Bastianella; Spudich, John L.

    1999-01-01

    The two transducers in the phototaxis system of the archaeon Halobacterium salinarum, HtrI and HtrII, are methyl-accepting proteins homologous to the chemotaxis transducers in eubacteria. Consensus sequences predict three glutamate pairs containing potential methylation sites in HtrI and one in HtrII. Mutagenic substitution of an alanine pair for one of these, Glu265-Glu266, in HtrI and for the homologous Glu513-Glu514 in HtrII eliminated methylation of these two transducers, as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis autofluorography. Photostimulation of the repellent receptor sensory rhodopsin II (SRII) induced reversible demethylation of HtrII, while no detectable change in the extent of methylation of HtrI was observed in response to stimulation of its cognate sensory rhodopsin, the attractant receptor SRI. Cells containing HtrI or HtrII with all consensus sites replaced by alanine still exhibited phototaxis responses and behavioral adaptation, and methanol release assays showed that methyl group turnover was still induced in response to photostimulation of SRI or SRII. By pulse-chase experiments with in vivo l-[methyl-3H]methionine-labeled cells, we found that repetitive photostimulation of SRI complexed with wild-type (or nonmethylatable) HtrI induced methyl group turnover in transducers other than HtrI to the same extent as in wild-type HtrI. Both attractant and repellent stimuli cause a transient increase in the turnover rate of methyl groups in wild-type H. salinarum cells. This result is unlike that obtained with Escherichia coli, in which attractant stimuli decrease and repellent stimuli increase turnover rate, and is similar to that obtained with Bacillus subtilis, which also shows turnover rate increases regardless of the nature of the stimulus. We found that a CheY deletion mutant of H. salinarum exhibited the E. coli-like asymmetric pattern, as has recently also been observed in B. subtilis. Further, we demonstrate

  19. The primary structures of the Archaeon Halobacterium salinarium blue light receptor sensory rhodopsin II and its transducer, a methyl-accepting protein.

    PubMed

    Zhang, W; Brooun, A; Mueller, M M; Alam, M

    1996-08-06

    Recently, a large family of transducer proteins in the Archaeon Halobacterium salinarium was identified. On the basis of the comparison of the predicted structural domains of these transducers, three distinct subfamilies of transducers were proposed. Here we report isolation, complete gene sequences, and analysis of the encoded primary structures of transducer gene htrII, a member of family B, and its blue light receptor gene (sopII) of sensory rhodopsin II (SRII). The start codon ATG of the 714-bp sopII gene is one nucleotide beyond the termination codon TGA of the 2298-bp htrII gene. The deduced protein sequence of HtrII predicts a eubacterial chemotaxis transducer type with two hydrophobic membrane-spanning segments connecting sizable domains in the periplasm and cytoplasm. HtrII has a common feature with HtrI, the sensory rhodopsin I transducer; like HtrI, HtrII possesses a hydrophilic loop structure just after the second transmembrane segment. The C-terminal 299 residues (765 amino acid residues total) of HtrII show strong homology to the signaling and methylation domain of eubacterial transducer Tsr. The hydropathy plot of the primary structure of SRII indicates seven membrane-spanning alpha-helical segments, a characteristic feature of retinylidene proteins ("rhodopsins") from a widespread family of photoactive pigments. SRII shows high identity with SRI (42%), bacteriorhodopsin (BR) (32%), and halorhodopsin (24%). The crucial positions for retinal binding sites in these proteins are nearly identical, with the exception of Met-118 (numbering according to the mature BR sequence), which is replaced by Val in SRII. In BR, residues Asp-85 and Asp-96 are crucial in proton pumping. In SRII, the position corresponding to Asp-85 in BR is conserved, but the corresponding position of Asp-96 is replaced by an aromatic Tyr. Coexpression of the htrII and sopII genes restores SRII phototaxis to a mutant (Pho81) that contains a deletion in the htrI/sopI and insertion in

  20. Development of the stapes and associated structures in human embryos.

    PubMed

    Rodríguez-Vázquez, J F

    2005-08-01

    The objective of this study was to clarify the development of the stapes in humans and its relationship with the cartilage of the second branchial arch. The study was carried out in 25 human embryos between 6 and 28 mm crown-rump length. The stapes develops at the cranial end of the second branchial arch through an independent anlage of the cartilage of this arch. Between the stapedial anlage and the cranial end of the Reichert's cartilage there is a formation called the interhyale, the internal segment of which gives rise to the tendon of the stapedial muscle. The stapedial anlage is a unique formation with two distinct parts: the superior part that will comprise the base and the inferior part that will be crossed by the stapedial artery during embryonic development and will constitute the limbs and the head of the stapes. According to the results, the otic capsule is not involved in formation of the base of the stapes.

  1. Mol-Gastronomie Käsefondue: eine kolloidale Schmelze

    NASA Astrophysics Data System (ADS)

    Vilgis, Thomas A.

    2004-11-01

    An Weihnachten oder zum Jahreswechsel kommt gern das Fonduegeschirr zum Einsatz, weil sich damit viele hungrige Gäste unterhaltsam sättigen lassen. Käsefondues sind allerdings eine physikalisch anspruchsvolle Angelegenheit. Die Kunst besteht darin, die viskoelastische Käseschmelze während des Genusses homogen zu halten.

  2. Einleitung

    NASA Astrophysics Data System (ADS)

    Zahoransky, Richard; Allelein, Hans-Josef; Bollin, Elmar; Oehler, Helmut; Schelling, Udo

    Mit der Beherrschung der Energie, zunächst des Feuers, schaffte sich die Menschheit die Basis zur technischen und wirtschaftlichen Entwicklung. Energie in ihren verschiedenen Erscheinungsformen ist zur Nahrungszubereitung, zum Wohnen, zum Transport, zur Kommunikation, in der Technik, Industrie und in der Freizeit unverzichtbar. Energie gehört zu den Grundbedürfnissen.

  3. Ton- und Bildübertragung

    NASA Astrophysics Data System (ADS)

    Plaßmann, Wilfried

    Stichworte zum Kapitel: Analoge Rundfunk-Stereoübertragung; Digitale Farbfernsehtechnik: Zielsetzung, Vorgaben, DVB-Projekt, MPEG-2 Standard, Redundanz- und Irrelevanzreduktion, Differenz-Pulsmodulation, Diskrete Kosinus-Transformation, Fehler-Erkennung und -Korrektur, Energieverwischung, 16-QAPSK-Modulation, Gleichwellennetz, Statistischer Multiplex, Daten zum DVB-T.

  4. Deep Burn: Development of Transuranic Fuel for High-Temperature Helium-Cooled Reactors- Monthly Highlights September 2010

    SciTech Connect

    Snead, Lance Lewis; Besmann, Theodore M; Collins, Emory D; Bell, Gary L

    2010-10-01

    The DB Program monthly highlights report for August 2010, ORNL/TM-2010/184, was distributed to program participants by email on September 17. This report discusses: (1) Core and Fuel Analysis - (a) Core Design Optimization in the HTR (high temperature helium-cooled reactor) Prismatic Design (Logos), (b) Core Design Optimization in the HTR Pebble Bed Design (INL), (c) Microfuel analysis for the DB HTR (INL, GA, Logos); (2) Spent Fuel Management - (a) TRISO (tri-structural isotropic) repository behavior (UNLV), (b) Repository performance of TRISO fuel (UCB); (3) Fuel Cycle Integration of the HTR (high temperature helium-cooled reactor) - Synergy with other reactor fuel cycles (GA, Logos); (4) TRU (transuranic elements) HTR Fuel Qualification - (a) Thermochemical Modeling, (b) Actinide and Fission Product Transport, (c) Radiation Damage and Properties; (5) HTR Spent Fuel Recycle - (a) TRU Kernel Development (ORNL), (b) Coating Development (ORNL), (c) Characterization Development and Support, (d) ZrC Properties and Handbook; and (6) HTR Fuel Recycle - (a) Graphite Recycle (ORNL), (b) Aqueous Reprocessing, (c) Pyrochemical Reprocessing METROX (metal recovery from oxide fuel) Process Development (ANL).

  5. Antagonism of serotonin receptor 1B decreases viability and promotes apoptosis in the COS canine osteosarcoma cell line.

    PubMed

    Viall, A K; Goodall, C P; Stang, B; Marley, K; Chappell, P E; Bracha, S

    2016-06-01

    Serotonin receptor 1B (5HTR1B) traditionally exhibits anti-proliferative activity in osteoblasts. We examined the expression and function of 5HTR1B in the COS canine osteosarcoma cell line and normal canine osteoblasts. Equal levels of 5HTR1B gene and protein expression were found between normal and malignant osteoblasts. Treatment with serotonin enhanced viability of osteosarcoma cells but not normal osteoblasts. Challenge with the 5HTR1B agonist anpirtoline caused no change in cell viability. Rather incubation with the specific receptor antagonist SB224289 caused reduction in osteoblast viability, with this effect more substantial in osteosarcoma cells. Investigation of this inhibitory activity showed 5HTR1B antagonism induces apoptosis in malignant cells. Evaluation of phosphorylated levels of CREB and ERK, transcriptional regulators associated with serotonin receptor signalling in osteoblasts, revealed aberrant 5HTR1B signalling in COS. Our results confirm the presence of 5HTR1B in a canine osteosarcoma cell line and highlight this receptor as a possible novel therapeutic target.

  6. Serotonin receptor expression along the dorsal–ventral axis of mouse hippocampus

    PubMed Central

    Tanaka, Kenji F.; Samuels, Benjamin Adam; Hen, René

    2012-01-01

    Using in situ hybridization, we describe, for the first time, the profiles of expression of serotonin receptors (Htr/5-HTR) along the dorsal–ventral axis of mouse hippocampus. cRNA probes for most Htrs, excluding Htr6, were used. All hippocampal subregions and the entorhinal cortex cells providing input into the hippocampus were examined. The study shows that some, but not all, Htrs are expressed in the cells of the hippocampal circuitry. At both the subfield and the cell type levels, a somewhat overlapping pattern is observed. Four serotonin receptors, Htr1a, Htr2a, Htr2c and Htr7, display an expression pattern that changes along the dorsal–ventral axis of the hippocampus. Given the proposed functional differentiation of the hippocampus along its long axis, with the dorsal pole more involved in cognitive functions and the ventral pole more involved in mood and anxiety, our results suggest that serotonin receptors enriched in the ventral pole probably contribute to mood- and anxiety-related behaviours. PMID:22826340

  7. Transducer-binding and transducer-mutations modulate photoactive-site-deprotonation in sensory rhodopsin I.

    PubMed

    Jung, K H; Spudich, E N; Dag, P; Spudich, J L

    1999-10-05

    Sensory rhodopsin I (SRI) is a seven-transmembrane helix retinylidene protein that mediates color-sensitive phototaxis responses through its bound transducer HtrI in the archaeon Halobacterium salinarum. Deprotonation of the Schiff base attachment site of the chromophore accompanies formation of the SRI signaling state, S(373). We measured the rate of laser flash-induced S(373) formation in the presence and absence of HtrI, and the effects of mutations in SRI or HtrI on the kinetics of this process. In the absence of HtrI, deprotonation occurs rapidly (halftime 10 micros) if the proton acceptor Asp76 is ionized (pK(a) = approximately 7), and only very slowly (halftime > 10 ms) when Asp76 is protonated. Transducer-binding, although it increases the pK(a) of Asp76 so that it is protonated throughout the range of pH studied, results in a first order, pH-independent rate of S(373) formation of approximately 300 micros. Therefore, the complexation of HtrI facilitates the proton-transfer reaction, increasing the rate approximately 50-fold at pH6. Arrhenius analysis shows that HtrI-binding accelerates the reaction primarily by an entropic effect, suggesting HtrI constrains the SRI molecule in the complex. Function-perturbing mutations in SRI and HtrI also alter the rate of S(373) formation and the lambda(max) of the parent state as assessed by laser flash-induced kinetic difference spectroscopy, and shifts to longer wavelength are correlated with slower deprotonation. The data indicate that HtrI affects electrostatic interactions of the protonated Schiff base and not only receives the signal from SRI but also optimizes the photochemical reaction process for SRI signaling.

  8. The nucleus accumbens 5-HTR₄-CART pathway ties anorexia to hyperactivity.

    PubMed

    Jean, A; Laurent, L; Bockaert, J; Charnay, Y; Dusticier, N; Nieoullon, A; Barrot, M; Neve, R; Compan, V

    2012-12-11

    In mental diseases, the brain does not systematically adjust motor activity to feeding. Probably, the most outlined example is the association between hyperactivity and anorexia in Anorexia nervosa. The neural underpinnings of this 'paradox', however, are poorly elucidated. Although anorexia and hyperactivity prevail over self-preservation, both symptoms rarely exist independently, suggesting commonalities in neural pathways, most likely in the reward system. We previously discovered an addictive molecular facet of anorexia, involving production, in the nucleus accumbens (NAc), of the same transcripts stimulated in response to cocaine and amphetamine (CART) upon stimulation of the 5-HT(4) receptors (5-HTR(4)) or MDMA (ecstasy). Here, we tested whether this pathway predisposes not only to anorexia but also to hyperactivity. Following food restriction, mice are expected to overeat. However, selecting hyperactive and addiction-related animal models, we observed that mice lacking 5-HTR(1B) self-imposed food restriction after deprivation and still displayed anorexia and hyperactivity after ecstasy. Decryption of the mechanisms showed a gain-of-function of 5-HTR(4) in the absence of 5-HTR(1B), associated with CART surplus in the NAc and not in other brain areas. NAc-5-HTR(4) overexpression upregulated NAc-CART, provoked anorexia and hyperactivity. NAc-5-HTR(4) knockdown or blockade reduced ecstasy-induced hyperactivity. Finally, NAc-CART knockdown suppressed hyperactivity upon stimulation of the NAc-5-HTR(4). Additionally, inactivating NAc-5-HTR(4) suppressed ecstasy's preference, strengthening the rewarding facet of anorexia. In conclusion, the NAc-5-HTR(4)/CART pathway establishes a 'tight-junction' between anorexia and hyperactivity, suggesting the existence of a primary functional unit susceptible to limit overeating associated with resting following homeostasis rules.

  9. Conformational states of HAMP domains interacting with sensory rhodopsin membrane systems: an integrated all-atom and coarse-grained molecular dynamics simulation approach.

    PubMed

    Sahoo, Bikash Ranjan; Fujiwara, Toshimichi

    2016-12-20

    Understanding the downstream signaling mechanism of sensory rhodopsin and its cognate transducer complex (srII-htrII) has long been a challenge in the field of photoreceptor research. Here, an integration of all-atom and coarse-grained (CG) molecular dynamics (MD) simulations in different srII-htrII complex states is carried out. It is shown that the cytoplasmic four-helix HAMP dimer gives rise to a gear-box model interaction with discrete hydrophobic packing in Natronomonas pharaonis (Np). Structural analysis in all-atom and CG-MD reveals a stable conformational state in the physiological environment (323 K and 1.15 M salt). Comparative analysis in the ground and intermediate state conformations reveals substantial inter-HAMP interactions in the intermediate state with uniform clockwise (+10° to +30°) and counterclockwise (-20° to -40°) rotations in the α1 helix and the α2 helix of the monomer, respectively. Low temperature and low salt environments (283 K and 0.15 M) significantly affect srII-htrII binding affinity in both states with unusual helix bending. The distinguished control cable, knob-into-holes packing and piston-like movements in HAMP helices are found in the intermediate state complex. The N-terminal htrII (159 residues) coupled with srII yields a binding energy (ΔGbind) of -309.22, -436.53 and -331.11 kJ mol(-1) in the MM/PBSA calculation for the NphtrII homodimer, the NpsrII-htrII ground state conformation and the NpsrII-htrII intermediate state conformation, respectively. Only the HAMP1 domain shows a very low ΔGbind value (-21.03 kJ mol(-1)) for the ground state in comparison to that for the intermediate state (-54.68 kJ mol(-1)). The structural analysis highlights the key residues that include Y199(srII), T189(srII), E43(htrII), T86(htrII), M100(htrII), E116(htrII), E126(htrII) and S130(htrII) for complex stabilization and signal transduction.

  10. Pathophysiology of hemolytic transfusion reactions.

    PubMed

    Davenport, Robertson D

    2005-07-01

    Hemolytic transfusion reactions (HTR) are systemic reactions provoked by immunologic red blood cell (RBC) incompatibility. Clinical and experimental observations of such reactions indicate that they proceed through phases of humoral immune reaction, activation of phagocytes, productions of cytokine mediators, and wide-ranging cellular responses. HTR have many features in common with the systemic inflammatory response syndrome (SIRS). Knowledge of the pathophysiologic mechanisms in HTR suggest that newer biological agents that target complement intermediates or proinflammatory cytokines may be effective agents in the treatment of severe HTRs.

  11. Deep Burn Develpment of Transuranic Fuel for High-Temperature Helium-Cooled Reactors - July 2010