WorldWideScience

Sample records for k1 flagellin induced

  1. Bacterial flagellin induces IL-6 expression in human basophils.

    Science.gov (United States)

    Jeon, Jun Ho; Ahn, Ki Bum; Kim, Sun Kyung; Im, Jintaek; Yun, Cheol-Heui; Han, Seung Hyun

    2015-05-01

    Binding of allergen to IgE on basophils positively affects allergic inflammation by releasing inflammatory mediators. Recently, basophils were shown to express pattern-recognition receptors, such as toll-like receptors (TLRs), for recognizing microbe-associated molecular patterns (MAMPs) that are independent of allergen-IgE binding. In this study, we investigated whether MAMP alone can induce IL-6 production in a human basophil cell line, KU812. Stimulation with flagellin in the absence of allergen-IgE association induced IL-6 expression in KU812 cells, while stimulation with lipoteichoic acid, peptidoglycan, or poly I:C did not under the same condition. Flagellin-induced IL-6 expression was also observed in human primary basophils. Flow cytometric analysis showed that KU812 cells expressed flagellin-recognizing TLR5 both on the cell surface and in the cytoplasm while TLR2 and TLR3 were observed only in the cytoplasm. We further demonstrated that although flagellin augmented the phosphorylation of mitogen-activated protein kinases including p38 kinase, ERK, and JNK, flagellin-induced IL-6 production was attenuated by inhibitors for p38 kinase and ERK, but not by JNK inhibitors. In addition, flagellin enhanced phosphorylation of signaling molecules including CREB, PKCδ, and AKT. The inhibitors for PKA and PKC also showed inhibitory effects. Interestingly, flagellin-induced IL-6 production was further enhanced by pretreatment with inhibitors for PI3K, implying that PI3K negatively affects the flagellin-induced IL-6 production. Furthermore, DNA binding activities of NF-κB, AP-1, and CREB, which play pivotal roles in the induction of IL-6 gene expression, were increased by flagellin. These results suggest that flagellin alone is sufficient to induce IL-6 gene expression via TLR5 signaling pathways in human basophils.

  2. Flagellin delivery by Pseudomonas aeruginosa rhamnolipids induces the antimicrobial protein psoriasin in human skin.

    Directory of Open Access Journals (Sweden)

    Ulf Meyer-Hoffert

    Full Text Available The opportunistic pathogen Pseudomonas aeruginosa can cause severe infections in patients suffering from disruption or disorder of the skin barrier as in burns, chronic wounds, and after surgery. On healthy skin P. aeruginosa causes rarely infections. To gain insight into the interaction of the ubiquitous bacterium P. aeruginosa and healthy human skin, the induction of the antimicrobial protein psoriasin by P. aeruginosa grown on an ex vivo skin model was analyzed. We show that presence of the P. aeruginosa derived biosurfactant rhamnolipid was indispensable for flagellin-induced psoriasin expression in human skin, contrary to in vitro conditions. The importance of the bacterial virulence factor flagellin as the major inducing factor of psoriasin expression in skin was demonstrated by use of a flagellin-deficient mutant. Rhamnolipid mediated shuttle across the outer skin barrier was not restricted to flagellin since rhamnolipids enable psoriasin expression by the cytokines IL-17 and IL-22 after topical application on human skin. Rhamnolipid production was detected for several clinical strains and the formation of vesicles was observed under skin physiological conditions. In conclusion we demonstrate herein that rhamnolipids enable the induction of the antimicrobial protein psoriasin by flagellin in human skin without direct contact of bacteria and responding cells. Hereby, human skin might control the microflora to prevent colonization of unwanted microbes in the earliest steps before potential pathogens can develop strategies to subvert the immune response.

  3. Sensitivity to Flg22 Is Modulated by Ligand-Induced Degradation and de Novo Synthesis of the Endogenous Flagellin-Receptor FLAGELLIN-SENSING2[W][OPEN

    Science.gov (United States)

    Smith, John M.; Salamango, Daniel J.; Leslie, Michelle E.; Collins, Carina A.; Heese, Antje

    2014-01-01

    FLAGELLIN-SENSING2 (FLS2) is the plant cell surface receptor that perceives bacterial flagellin or flg22 peptide, initiates flg22-signaling responses, and contributes to bacterial growth restriction. Flg22 elicitation also leads to ligand-induced endocytosis and degradation of FLS2 within 1 h. Why plant cells remove this receptor precisely at the time during which its function is required remains mainly unknown. Here, we assessed in planta flg22-signaling competency in the context of ligand-induced degradation of endogenous FLS2 and chemical interference known to impede flg22-dependent internalization of FLS2 into endocytic vesicles. Within 1 h after an initial flg22 treatment, Arabidopsis (Arabidopsis thaliana) leaf tissue was unable to reelicit flg22 signaling in a ligand-, time-, and dose-dependent manner. These results indicate that flg22-induced degradation of endogenous FLS2 may serve to desensitize cells to the same stimulus (homologous desensitization), likely to prevent continuous signal output upon repetitive flg22 stimulation. In addition to impeding ligand-induced FLS2 degradation, pretreatment with the vesicular trafficking inhibitors Wortmannin or Tyrphostin A23 impaired flg22-elicited reactive oxygen species production that was partially independent of BRASSINOSTEROID INSENSITIVE1-ASSOCIATED KINASE1. Interestingly, these inhibitors did not affect flg22-induced mitogen-activated protein kinase phosphorylation, indicating the ability to utilize vesicular trafficking inhibitors to target different flg22-signaling responses. For Tyrphostin A23, reduced flg22-induced reactive oxygen species could be separated from the defect in FLS2 degradation. At later times (>2 h) after the initial flg22 elicitation, recovery of FLS2 protein levels positively correlated with resensitization to flg22, indicating that flg22-induced new synthesis of FLS2 may prepare cells for a new round of monitoring the environment for flg22. PMID:24220680

  4. Flagellin of Pseudomonas aeruginosa induces transforming growth factor beta 1 expression in normal bronchial epithelial cells through mitogen activated protein kinase cascades

    Institute of Scientific and Technical Information of China (English)

    YANG Jing-jing; WANG Dan-dan; SUN Tie-ying

    2011-01-01

    flagellin vs. Control groups ((104.3±20.8) vs.(44.6±4.4) pg/ml (P <0.01)) and was ablated by either p38 or JNK inhibitors compared with flagellin treatment ((45.1±18.8)vs. (104.3±20.8) pg/ml and (48.1±20.8) vs. (104.3±20.8) pg/ml, respectively (P <0.05)). Flagellin also elevated the expression of IL-8 in BEAS-2B cells vs. The control groups ((554.9±57.7) vs. (51.4±2.2.9) pg/ml (P <0.01)), and p38 MAPK inhibitors weaken the expression by flagellin ((301.1 ±155.1) vs. (554.9±57.7) pg/ml (P <0.05)). Western blotting revealed that all three MAPK proteins, p38, JNK and ERK were activated by flagellin challenge in an early phase, respectively in 15 minutes (P <0.01), 30 minutes (P <0.01) and 15 minutes (P <0.01). TRAF6 siRNA which decreased expression of TRAF6, altered the activation of JNK, p38, and ERK following flagellin treatment, but its influence on the expression of TGF-β1 and IL-8 has no statistical significance.Conclusions Flagellin from P. Aeruginosa PAO1 induces TGF-β1 expression in normal bronchial epithelial cells,BEAS-2B, through the MAPK signal cascade in vitro. It suggests that the fibrogenesis/remodeling process may be initiated from an early stage of acute lung infection due to P. Aeruginosa.

  5. S6K1 controls autophagosome maturation in autophagy induced by sulforaphane or serum deprivation.

    Science.gov (United States)

    Hać, Aleksandra; Domachowska, Anna; Narajczyk, Magdalena; Cyske, Karolina; Pawlik, Anna; Herman-Antosiewicz, Anna

    2015-10-01

    It is well established that mTORC1 suppresses autophagy by phosphorylation and inactivation of proteins involved in autophagosome formation. However, the role of its substrate, p70S6 kinase1 (S6K1), in autophagy is quite controversial. In some models S6K1 activity correlates with autophagy suppression, however, some other studies show that S6K1 promotes rather than inhibits this process. Here, we investigated the role of S6K1 in prostate cancer cells (PC-3) and non-cancerous, mouse embryonic fibroblasts (MEF), either treated with autophagy inducer sulforaphane, an isothiocyanate derived from cruciferous plants, or deprived of serum. Our results indicate that constitutively active S6K1 decreases the level of LC3 processing and foci formation by autophagosomal vacuoles in cells treated with sulforaphane. On the other hand, presence of S6K1 is necessary for autophagosome maturation under conditions of autophagy induced by either sulforaphane or serum deprivation. Diminished level of S6K1 or lack of S6 kinases results in both, accumulation of autophagosomes and drop in the autophagolysosome number, and thus disturbs autophagy flux under stress conditions. Moreover, lack of S6 kinases reduces cell survival under stress conditions. Copyright © 2015 Elsevier GmbH. All rights reserved.

  6. The Legionella micdadei flagellin

    DEFF Research Database (Denmark)

    Bangsborg, Jette Marie; Hindersson, P; Shand, G;

    1995-01-01

    To study the structure and function of the Legionella flagellum, we screened a genomic L. micdadei library in Escherichia coli for expression of the flagellin (Fla) subunit. One recombinant clone, JM105 (pHI5588), producing a truncated Fla protein of 40.5 kDa was identified. The plasmid pHI5588...... shared a high degree of homology with other flagellin genes in the amino- and carboxy termini, whereas the central region was found to be nonconserved. The fla sequence will facilitate the cloning of Fla proteins from other Legionella species and the study of flagella in the pathogenesis of Legionnaires...

  7. Antibodies against Marinobacter algicola and Salmonella typhimurium flagellins do not cross-neutralize TLR5 activation.

    Directory of Open Access Journals (Sweden)

    Raul Terron-Exposito

    Full Text Available Flagellins evoke strong innate and adaptive immune responses. These proteins may play a key role as radioprotectors, exert antitumoral activity in certain types of tumor and reduce graft-versus-host disease in allogeneic hematopoietic stem cell transplant recipients. Notwithstanding, flagellins are highly immunogenic, and repeated use leads to their neutralization by systemic antibodies. This neutralization is not prevented by using functional deleted flagellins. These observations led us to explore the possibility of preventing initial neutralization by means of another functional flagellin that does not belong to common pathogenic bacteria but that has the capacity to activate TLR5. Here we characterized the functional capacity of the two-phase Marinobacter algicola (MA-derived flagellins (F and FR as systemic and mucosal adjuvants and compared their performance with that of Salmonella typhimurium (STF flagellins (FljB and FliC. We also report for the first time on the in vitro and in vivo capacity of various flagellins to trigger TLR5 activation in the presence of species-specific anti-flagellin antibodies, the cross-neutralization mediated by these antibodies, and the sequential use of these flagellins for TLR5 activation. Our results showed that MA flagellins behave in a similar way to STF ones, inducing pro-inflammatory cytokines (IL8, CCL20, CCL2 and evoking a strong in vivo antibody response against a model epitope. More importantly, MA flagellins were fully functional, in vitro or in vivo, in the presence of a high concentration of neutralizing anti-flagellin STF antibodies, and STF flagellin was not inhibited by neutralizing anti-flagellin MA antibodies. The use of active flagellins from distinct bacteria could be a useful approach to prevent systemic neutralization of this group of adjuvants and to facilitate the rational design of flagellin-based vaccines and/or other therapeutic treatments (against ischemia, acute renal failure

  8. Protection against multiple influenza A virus strains induced by candidate recombinant vaccine based on heterologous M2e peptides linked to flagellin.

    Directory of Open Access Journals (Sweden)

    Liudmila A Stepanova

    Full Text Available Matrix 2 protein ectodomain (M2e is considered a promising candidate for a broadly protective influenza vaccine. M2e-based vaccines against human influenza A provide only partial protection against avian influenza viruses because of differences in the M2e sequences. In this work, we evaluated the possibility of obtaining equal protection and immune response by using recombinant protein on the basis of flagellin as a carrier of the M2e peptides of human and avian influenza A viruses. Recombinant protein was generated by the fusion of two tandem copies of consensus M2e sequence from human influenza A and two copies of M2e from avian A/H5N1 viruses to flagellin (Flg-2M2eh2M2ek. Intranasal immunisation of Balb/c mice with recombinant protein significantly elicited anti-M2e IgG in serum, IgG and sIgA in BAL. Antibodies induced by the fusion protein Flg-2M2eh2M2ek bound efficiently to synthetic peptides corresponding to the human consensus M2e sequence as well as to the M2e sequence of A/Chicken/Kurgan/05/05 RG (H5N1 and recognised native M2e epitopes exposed on the surface of the MDCK cells infected with A/PR/8/34 (H1N1 and A/Chicken/Kurgan/05/05 RG (H5N1 to an equal degree. Immunisation led to both anti-M2e IgG1 and IgG2a response with IgG1 prevalence. We observed a significant intracellular production of IL-4, but not IFN-γ, by CD4+ T-cells in spleen of mice following immunisation with Flg-2M2eh2M2ek. Immunisation with the Flg-2M2eh2M2ek fusion protein provided similar protection from lethal challenge with human influenza A viruses (H1N1, H3N2 and avian influenza virus (H5N1. Immunised mice experienced significantly less weight loss and decreased lung viral titres compared to control mice. The data obtained show the potential for the development of an M2e-flagellin candidate influenza vaccine with broad spectrum protection against influenza A viruses of various origins.

  9. Protection against Multiple Influenza A Virus Strains Induced by Candidate Recombinant Vaccine Based on Heterologous M2e Peptides Linked to Flagellin

    Science.gov (United States)

    Kovaleva, Anna A.; Potapchuk, Marina V.; Korotkov, Alexandr V.; Sergeeva, Mariia V.; Kasianenko, Marina A.; Kuprianov, Victor V.; Ravin, Nikolai V.; Tsybalova, Liudmila M.; Skryabin, Konstantin G.; Kiselev, Oleg I.

    2015-01-01

    Matrix 2 protein ectodomain (M2e) is considered a promising candidate for a broadly protective influenza vaccine. M2e-based vaccines against human influenza A provide only partial protection against avian influenza viruses because of differences in the M2e sequences. In this work, we evaluated the possibility of obtaining equal protection and immune response by using recombinant protein on the basis of flagellin as a carrier of the M2e peptides of human and avian influenza A viruses. Recombinant protein was generated by the fusion of two tandem copies of consensus M2e sequence from human influenza A and two copies of M2e from avian A/H5N1 viruses to flagellin (Flg-2M2eh2M2ek). Intranasal immunisation of Balb/c mice with recombinant protein significantly elicited anti-M2e IgG in serum, IgG and sIgA in BAL. Antibodies induced by the fusion protein Flg-2M2eh2M2ek bound efficiently to synthetic peptides corresponding to the human consensus M2e sequence as well as to the M2e sequence of A/Chicken/Kurgan/05/05 RG (H5N1) and recognised native M2e epitopes exposed on the surface of the MDCK cells infected with A/PR/8/34 (H1N1) and A/Chicken/Kurgan/05/05 RG (H5N1) to an equal degree. Immunisation led to both anti-M2e IgG1 and IgG2a response with IgG1 prevalence. We observed a significant intracellular production of IL-4, but not IFN-γ, by CD4+ T-cells in spleen of mice following immunisation with Flg-2M2eh2M2ek. Immunisation with the Flg-2M2eh2M2ek fusion protein provided similar protection from lethal challenge with human influenza A viruses (H1N1, H3N2) and avian influenza virus (H5N1). Immunised mice experienced significantly less weight loss and decreased lung viral titres compared to control mice. The data obtained show the potential for the development of an M2e-flagellin candidate influenza vaccine with broad spectrum protection against influenza A viruses of various origins. PMID:25799221

  10. Global IP6K1 deletion enhances temperature modulated energy expenditure which reduces carbohydrate and fat induced weight gain

    OpenAIRE

    Zhu, Qingzhang; Ghoshal, Sarbani; Tyagi, Richa; Chakraborty, Anutosh

    2017-01-01

    Objective: IP6 kinases (IP6Ks) regulate cell metabolism and survival. Mice with global (IP6K1-KO) or adipocyte-specific (AdKO) deletion of IP6K1 are protected from diet induced obesity (DIO) at ambient (23 °C) temperature. AdKO mice are lean primarily due to increased AMPK mediated thermogenic energy expenditure (EE). Thus, at thermoneutral (30 °C) temperature, high fat diet (HFD)-fed AdKO mice expend energy and gain body weight, similar to control mice. IP6K1 is ubiquitously expressed; thus,...

  11. Flagellin stimulates protective lung mucosal immunity: role of cathelicidin-related antimicrobial peptide.

    Science.gov (United States)

    Yu, Fu-shin; Cornicelli, Matthew D; Kovach, Melissa A; Newstead, Michael W; Zeng, Xianying; Kumar, Ashok; Gao, Nan; Yoon, Sang Gi; Gallo, Richard L; Standiford, Theodore J

    2010-07-15

    TLRs are required for generation of protective lung mucosal immune responses against microbial pathogens. In this study, we evaluated the effect of the TLR5 ligand flagellin on stimulation of antibacterial mucosal immunity in a lethal murine Pseudomonas aeruginosa pneumonia model. The intranasal pretreatment of mice with purified P. aeruginosa flagellin induced strong protection against intratracheal P. aeruginosa-induced lethality, which was attributable to markedly improved bacterial clearance, reduced dissemination, and decreased alveolar permeability. The protective effects of flagellin on survival required TLR5 and were observed even in the absence of neutrophils. Flagellin induced strong induction of innate genes, most notably the antimicrobial peptide cathelicidin-related antimicrobial peptide. Finally, flagellin-induced protection was partially abrogated in cathelicidin-related antimicrobial peptide-deficient mice. Our findings illustrate the profound stimulatory effect of flagellin on lung mucosal innate immunity, a response that might be exploited therapeutically to prevent the development of gram-negative bacterial infection of the respiratory tract.

  12. Adipocyte-specific deletion of Ip6k1 reduces diet-induced obesity by enhancing AMPK-mediated thermogenesis

    Science.gov (United States)

    Zhu, Qingzhang; Ghoshal, Sarbani; Rodrigues, Ana; Gao, Su; Asterian, Alice; Kamenecka, Theodore M.; Barrow, James C.

    2016-01-01

    Enhancing energy expenditure (EE) is an attractive strategy to combat obesity and diabetes. Global deletion of Ip6k1 protects mice from diet-induced obesity (DIO) and insulin resistance, but the tissue-specific mechanism by which IP6K1 regulates body weight is unknown. Here, we have demonstrated that IP6K1 regulates fat accumulation by modulating AMPK-mediated adipocyte energy metabolism. Cold exposure led to downregulation of Ip6k1 in murine inguinal and retroperitoneal white adipose tissue (IWAT and RWAT) depots. Adipocyte-specific deletion of Ip6k1 (AdKO) enhanced thermogenic EE, which protected mice from high-fat diet–induced weight gain at ambient temperature (23°C), but not at thermoneutral temperature (30°C). AdKO-induced increases in thermogenesis also protected mice from cold-induced decreases in body temperature. UCP1, PGC1α, and other markers of browning and thermogenesis were elevated in IWAT and RWAT of AdKO mice. Cold-induced activation of sympathetic signaling was unaltered, whereas AMPK was enhanced, in AdKO IWAT. Moreover, beige adipocytes from AdKO IWAT displayed enhanced browning, which was diminished by AMPK depletion. Furthermore, we determined that IP6 and IP6K1 differentially regulate upstream kinase-mediated AMPK stimulatory phosphorylation in vitro. Finally, treating mildly obese mice with the IP6K inhibitor TNP enhanced thermogenesis and inhibited progression of DIO. Thus, IP6K1 regulates energy metabolism via a mechanism that could potentially be targeted in obesity. PMID:27701146

  13. Outer membrane vesicles from flagellin-deficient Salmonella enterica serovar Typhimurium induce cross-reactive immunity and provide cross-protection against heterologous Salmonella challenge

    Science.gov (United States)

    Liu, Qiong; Liu, Qing; Yi, Jie; Liang, Kang; Hu, Bo; Zhang, Xiangmin; Curtiss, Roy; Kong, Qingke

    2016-01-01

    Outer membrane vesicles (OMVs) isolated from Salmonella Typhimurium are potentially useful for developing subunit vaccines because of high immunogenicity and protective efficacy. However, flagella might remain in OMV pellets following OMV purification, resulting in non-essential immune responses and counteraction of bacterial protective immune responses when developing a vaccine against infection of multiple serotypes Salmonella. In this study, a flagellin-deficient S. Typhimurium mutant was constructed. Lipopolysaccharide profiles, protein profiles and cryo-electron microscopy revealed that there were no significant differences between the wild-type and mutant OMVs, with the exception of a large amount of flagellin in the wild-type OMVs. Neither the wild-type OMVs nor the non-flagellin OMVs were toxic to macrophages. Mice immunized with the non-flagellin OMVs produced high concentrations of IgG. The non-flagellin OMVs elicited strong mucosal antibody responses in mice when administered via the intranasal route in addition to provoking higher cross-reactive immune responses against OMPs isolated from S. Choleraesuis and S. Enteritidis. Both intranasal and intraperitoneal immunization with the non-flagellin OMVs provided efficient protection against heterologous S. Choleraesuis and S. Enteritidis challenge. Our results indicate that the flagellin-deficient OMVs may represent a new vaccine platform that could be exploited to facilitate the production of a broadly protective vaccine. PMID:27698383

  14. Phenylpropanoid Defences in Nicotiana tabacum Cells: Overlapping Metabolomes Indicate Common Aspects to Priming Responses Induced by Lipopolysaccharides, Chitosan and Flagellin-22.

    Directory of Open Access Journals (Sweden)

    Msizi I Mhlongo

    Full Text Available Plants have evolved both constitutive and inducible defence strategies to cope with different biotic stimuli and stresses. Exposure of a plant to a challenging stress can lead to a primed state that allows it to launch a more rapid and stronger defence. Here we applied a metabolomic approach to study and compare the responses induced in Nicotiana tabacum cells by microbe-associated molecular pattern (MAMP molecules, namely lipopolysaccharides (LPS, chitosan (CHT and flagellin-22 (FLG22. Early response metabolites, extracted with methanol, were analysed by UHPLC-MS/MS. Using multivariate statistical tools the metabolic profiles induced by these elicitors were analysed. In the metabolic fingerprint of these agents a total of 19 cinnamic acid derivatives conjugated to quinic acids (chlorogenic acids, shikimic acid, tyramine, polyamines or glucose were found as discriminant biomarkers. In addition, treatment with the phytohormones salicylic acid (SA, methyljasmonic acid (MJ and abscisic acid (ABA resulted in differentially-induced phenylpropanoid pathway metabolites. The results indicate that the phenylpropanoid pathway is activated by these elicitors while hydroxycinnamic acid derivatives are commonly associated with the metabolic response to the MAMPs, and that the activated responses are modulated by both SA and MJ, with ABA not playing a role.

  15. Chimeric flagellin expressed by Salmonella typhimurium induces an ESAT-6-specific Th 1-type immune response and CTL effects following intranasal immunization

    Institute of Scientific and Technical Information of China (English)

    Hui Zhang; Liu Liu; Ke Wen; Jinlin Huang; Shizhong Geng; Junsong Shen; Zhiming Pan; Xinan Jiao

    2011-01-01

    The flagellin component FliC of Salmonella typhimurium is capable of activating the innate immune system via specific interactions with TLR5 and can also act as a carrier of foreign antigen to elicit antigen-specific immune responses.Thus,we constructed an attenuated Salmonella strain SL5928(fliC/esat) expressing chimeric flagellin that contained the ESAT-6 antigen coding sequence of Mycobacterium tuberculosis inserted into the highly variable region of the Salmonella flagellin coding gene fliCi.The chimeric flagellin functioned normally,as demonstrated using a flagella swarming assay and electron microscopy.To analyze the effects of chimeric flagellin,the cell-mediated immune response and cytotoxic T lymphocyte (CTL) effects specific for ESAT-6antigen were tested after intranasal immunization of mice with flagellated Salmonella SL5928(fliC/esat).The results showed that SL5928(fliC/esat) intranasal immunization can strongly elicit an ESAT-6-specific T helper (Th) 1-type immune response in mucosal lymphoid tissues,such as nasopharynx-associated lymph nodes,lung and Peyer's patches,and a Th 1/Th2 response was elicited in spleen and mesenteric lymph nodes.Furthermore,intranasal immunization of SL5928(fliC/esat) produced efficient CTL effects,as demonstrated using a 5-and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) assay.Thus,our study revealed that Salmonella flagellin acts as a carrier for foreign antigen and triggers strong Th 1 and CTL responses during intranasal immunization.Chimeric flagellin is potentially an effective strategy for the development of novel vaccines against tuberculosis in humans and animals.

  16. Inhibition of diabetic-cataract by vitamin K1 involves modulation of hyperglycemia-induced alterations to lens calcium homeostasis.

    Science.gov (United States)

    Sai Varsha, M K N; Raman, Thiagarajan; Manikandan, Ramar

    2014-11-01

    This study investigated the potential of vitamin K1 against streptozotocin-induced diabetic cataract in Wistar rats. A single, intraperitoneal injection of streptozotocin (STZ) (35 mg/kg) resulted in hyperglycemia, accumulation of sorbitol and formation of advanced glycation end product (AGE) in eye lens. Hyperglycemia in lens also resulted in superoxide anion and hydroxyl radical generation and less reduced glutathione suggesting oxidative stress in lens. Hyperglycemia also resulted in increase in lens Ca2+ and significant inhibition of lens Ca2+ ATPase activity. These changes were associated with cataract formation in diabetic animals. By contrast treatment of diabetic rats with vitamin K1 (5 mg/kg, sc, twice a week) resulted in animals with partially elevated blood glucose and with transparent lenses having normal levels of sorbitol, AGE, Ca2+ ATPase, Ca2+, and oxidative stress. Vitamin K 1 may function to protect against cataract formation in the STZ induced diabetic rat by affecting the homeostasis of blood glucose and minimizing subsequent oxidative and osmotic stress. Thus, these results show that Vitamin K1 inhibits diabetic-cataract by modulating lens Ca2+ homeostasis and its hypoglycemic effect through its direct action on the pancreas.

  17. Temporal changes of oxidative stress markers in Escherichia coli K1-induced experimental meningitis in a neonatal rat model.

    Science.gov (United States)

    Giridharan, Vijayasree V; Simões, Lutiana R; Dagostin, Valdemira S; Generoso, Jaqueline S; Rezin, Gislaine T; Florentino, Drielly; Muniz, Jhonata P; Collodel, Allan; Petronilho, Fabricia; Quevedo, Joao; Barichello, Tatiana

    2017-07-13

    Despite advances in antimicrobial therapy and advanced critical care neonatal bacterial meningitis has a mortality rate of over 10% and induces neurological sequelae in 20-50% of cases. Escherichia coli K1 (E. coli K1) is the most common gram-negative organism causing neonatal meningitis and is the second most common cause behind group B streptococcus. We previously reported that an E. coli K1 experimental meningitis infection in neonatal rats resulted in habituation and aversive memory impairment and a significant increase in cytokine levels in adulthood. In this present study, we investigated the oxidative stress profile including malondialdehyde (MDA) levels, carbonyl protein formation, myeloperoxidase activity (MPO) activity, superoxide dismutase (SOD) activity and catalase (CAT) activity 6, 12, 24, 48, 72 and 96h after E. coli K1 experimental meningitis infection. In addition, sulfhydryl groups, nitrite and nitrate levels and activity of the mitochondrial respiratory chain enzymes were also measured in the frontal cortex and hippocampus of neonatal rats. The results from this study demonstrated a significant increase in MDA, protein carbonyls and MPO activity and a simultaneous decrease in SOD activity in the hippocampus of the neonatal meningitis survivors but the same was not observed in frontal cortex. In addition, we also observed a significant increase in complex IV activity in the hippocampus and frontal cortex of meningitis survivor rats. Thus, the results from this study reaffirmed the possible role of oxidative stress, nitric oxide and its related compounds in the complex pathophysiology of E. coli K1-induced bacterial meningitis. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Protective effect of propolis on radiation-induced chromosomal damage on Chinese hamster ovary cells (CHO-K1)

    Energy Technology Data Exchange (ETDEWEB)

    Spigoti, Geyza; Bartolini, Paolo; Okazaki, Kayo [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)], e-mail: kokazaki@ipen.br; Tsutsumi, Shiguetoshi [Amazon Food Ltd., Tokyo (Japan)], e-mail: fwip5138@mb.infoweb.ne.jp

    2009-07-01

    In the last years, particular interest has been given to investigations concerning natural, effective and nontoxic compounds with radioprotective capacity in concert with increasing utilization of different types of ionizing radiation for various applications. Among them, propolis, a resinous mixture of substances collected by honey bees (Apis mellifera) has been considered promising since it presents several advantageous characteristics, i.e., antiinflammatory, anticarcinogenic, antimicrobial and free radical scavenging action. It is, therefore, a direct antioxidant that protects cells and organisms from the adverse effects of ionizing radiation. These relevant biological activities are mainly mediated by the flavonoids, present at relatively high concentrations in the propolis. Considering that the chemical composition and, consequently, the biological activity of propolis is variable according to the environmental plant ecology, the present study was conducted in order to evaluate the radioprotective capacity of Brazilian propolis, collected in the State of Rio Grande do Sul, against genotoxic damages induced by {sup 60}Co {gamma}-radiation in Chinese hamster ovary cells (CHO-K1). for this purpose, micronucleus induction was analyzed concerning irreparable damage, specifically related to DNA double-strand breaks, that are potentially carcinogenic. CHO-K1 cells were submitted to different concentrations of propolis (3 - 33 {mu}g/ml), 1 h before irradiation, with 1 Gy of {gamma} radiation (0.722 Gy/min). The data obtained showed a decreasing tendency in the quantity of radioinduced damage on cells previously treated with propolis. The radioprotective effect was more prominent at higher propolis concentration. The treatment with propolis alone did not induce genotoxic effects on CHO-K1 cells. Beside that, the treatment with propolis, associated or not with radiation, did not influence the kinetics of cellular proliferation. (author)

  19. Innate immune detection of flagellin positively and negatively regulates salmonella infection.

    Directory of Open Access Journals (Sweden)

    Marvin A Lai

    Full Text Available Salmonella enterica serovar Typhimurium is a flagellated bacterium and one of the leading causes of gastroenteritis in humans. Bacterial flagellin is required for motility and also a prime target of the innate immune system. Innate immune recognition of flagellin is mediated by at least two independent pathways, TLR5 and Naip5-Naip6/NlrC4/Caspase-1. The functional significance of each of the two independent flagellin recognition systems for host defense against wild type Salmonella infection is complex, and innate immune detection of flagellin contributes to both protection and susceptibility. We hypothesized that efficient modulation of flagellin expression in vivo permits Salmonella to evade innate immune detection and limit the functional role of flagellin-specific host innate defenses. To test this hypothesis, we used Salmonella deficient in the anti-sigma factor flgM, which overproduce flagella and are attenuated in vivo. In this study we demonstrate that flagellin recognition by the innate immune system is responsible for the attenuation of flgM(- S. Typhimurium, and dissect the contribution of each flagellin recognition pathway to bacterial clearance and inflammation. We demonstrate that caspase-1 controls mucosal and systemic infection of flgM(- S. Typhimurium, and also limits intestinal inflammation and injury. In contrast, TLR5 paradoxically promotes bacterial colonization in the cecum and systemic infection, but attenuates intestinal inflammation. Our results indicate that Salmonella evasion of caspase-1 dependent flagellin recognition is critical for establishing infection and that evasion of TLR5 and caspase-1 dependent flagellin recognition helps Salmonella induce intestinal inflammation and establish a niche in the inflamed gut.

  20. Innate immune detection of flagellin positively and negatively regulates salmonella infection.

    Science.gov (United States)

    Lai, Marvin A; Quarles, Ellen K; López-Yglesias, Américo H; Zhao, Xiaodan; Hajjar, Adeline M; Smith, Kelly D

    2013-01-01

    Salmonella enterica serovar Typhimurium is a flagellated bacterium and one of the leading causes of gastroenteritis in humans. Bacterial flagellin is required for motility and also a prime target of the innate immune system. Innate immune recognition of flagellin is mediated by at least two independent pathways, TLR5 and Naip5-Naip6/NlrC4/Caspase-1. The functional significance of each of the two independent flagellin recognition systems for host defense against wild type Salmonella infection is complex, and innate immune detection of flagellin contributes to both protection and susceptibility. We hypothesized that efficient modulation of flagellin expression in vivo permits Salmonella to evade innate immune detection and limit the functional role of flagellin-specific host innate defenses. To test this hypothesis, we used Salmonella deficient in the anti-sigma factor flgM, which overproduce flagella and are attenuated in vivo. In this study we demonstrate that flagellin recognition by the innate immune system is responsible for the attenuation of flgM(-) S. Typhimurium, and dissect the contribution of each flagellin recognition pathway to bacterial clearance and inflammation. We demonstrate that caspase-1 controls mucosal and systemic infection of flgM(-) S. Typhimurium, and also limits intestinal inflammation and injury. In contrast, TLR5 paradoxically promotes bacterial colonization in the cecum and systemic infection, but attenuates intestinal inflammation. Our results indicate that Salmonella evasion of caspase-1 dependent flagellin recognition is critical for establishing infection and that evasion of TLR5 and caspase-1 dependent flagellin recognition helps Salmonella induce intestinal inflammation and establish a niche in the inflamed gut.

  1. Activation of SphK1 by K6PC-5 Inhibits Oxygen-Glucose Deprivation/Reoxygenation-Induced Myocardial Cell Death.

    Science.gov (United States)

    Shao, Jun-jie; Peng, Yi; Wang, Li-ming; Wang, Jian-kai; Chen, Xin

    2015-11-01

    In the current study, we evaluated the potential effect of a novel sphingosine kinase 1 (SphK1) activator, K6PC-5, on oxygen-glucose deprivation (OGD)/reoxygenation-induced damages to myocardial cells. We demonstrated that K6PC-5 increased intracellular sphingosine-1-phosphate (S1P) content and remarkably inhibited OGD/reoxygenation-induced death of myocardial cells (H9c2/HL-1 lines and primary murine myocardiocytes). SphK1 inhibitors, B-5354c and SKI-II, or SphK1-siRNA knockdown not only aggregated OGD/reoxygenation-induced cytotoxicity but also nullified the cytoprotection by K6PC-5. On the other hand, overexpression of SphK1 alleviated H9c2 cell death by OGD/reoxygenation, and K6PC-5-mediated cytoprotection was also enhanced in SphK1 overexpressed cells. Molecularly, OGD/reoxygenation activated the mitochondrial death pathway, evidenced by reactive oxygen species (ROS) production, mitochondrial membrane potential reduction, and p53-cyclophilin D (Cyp-D) association, which were all alleviated by K6PC-5 or overexpression of SphK1, but exacerbated by SphK1 knockdown. Furthermore, OGD/reoxygenation induced prodeath ceramide production in myocardial cells, which was largely suppressed by K6PC-5. In the meantime, adding a cell-permeable short-chain ceramide (C6) mimicked OGD/reoxygenation actions and induced ROS production and the mitochondrial death pathway in myocardial cells. Together, we conclude that K6PC-5 inhibits OGD/reoxygenation-induced myocardial cell death probably through activating SphK1. The results of the study indicate a potential benefit of K6PC-5 on ischemic heart disease.

  2. Role of hepatocyte S6K1 in palmitic acid-induced endoplasmic reticulum stress, lipotoxicity, insulin resistance and in oleic acid-induced protection.

    Science.gov (United States)

    Pardo, Virginia; González-Rodríguez, Águeda; Muntané, Jordi; Kozma, Sara C; Valverde, Ángela M

    2015-06-01

    The excess of saturated free fatty acids, such as palmitic acid, that induces lipotoxicity in hepatocytes, has been implicated in the development of non-alcoholic fatty liver disease also associated with insulin resistance. By contrast, oleic acid, a monounsaturated fatty acid, attenuates the effects of palmitic acid. We evaluated whether palmitic acid is directly associated with both insulin resistance and lipoapoptosis in mouse and human hepatocytes and the impact of oleic acid in the molecular mechanisms that mediate both processes. In human and mouse hepatocytes palmitic acid at a lipotoxic concentration triggered early activation of endoplasmic reticulum (ER) stress-related kinases, induced the apoptotic transcription factor CHOP, activated caspase 3 and increased the percentage of apoptotic cells. These effects concurred with decreased IR/IRS1/Akt insulin pathway. Oleic acid suppressed the toxic effects of palmitic acid on ER stress activation, lipoapoptosis and insulin resistance. Besides, oleic acid suppressed palmitic acid-induced activation of S6K1. This protection was mimicked by pharmacological or genetic inhibition of S6K1 in hepatocytes. In conclusion, this is the first study highlighting the activation of S6K1 by palmitic acid as a common and novel mechanism by which its inhibition by oleic acid prevents ER stress, lipoapoptosis and insulin resistance in hepatocytes.

  3. Flagellin Encoded in Gene-Based Vector Vaccines Is a Route-Dependent Immune Adjuvant.

    Directory of Open Access Journals (Sweden)

    Hamada F Rady

    Full Text Available Flagellin has been tested as a protein-based vaccine adjuvant, with the majority of studies focused on antibody responses. Here, we evaluated the adjuvant activity of flagellin for both cellular and humoral immune responses in BALB/c mice in the setting of gene-based immunization, and have made several novel observations. DNA vaccines and adenovirus (Ad vectors were engineered to encode mycobacterial protein Ag85B, with or without flagellin of Salmonella typhimurium (FliC. DNA-encoded flagellin given IM enhanced splenic CD4+ and CD8+ T cell responses to co-expressed vaccine antigen, including memory responses. Boosting either IM or intranasally with Ad vectors expressing Ag85B without flagellin led to durable enhancement of Ag85B-specific antibody and CD4+ and CD8+ T cell responses in both spleen and pulmonary tissues, correlating with significantly improved protection against challenge with pathogenic aerosolized M. tuberculosis. However, inclusion of flagellin in both DNA prime and Ad booster vaccines induced localized pulmonary inflammation and transient weight loss, with route-dependent effects on vaccine-induced T cell immunity. The latter included marked reductions in levels of mucosal CD4+ and CD8+ T cell responses following IM DNA/IN Ad mucosal prime-boosting, although antibody responses were not diminished. These findings indicate that flagellin has differential and route-dependent adjuvant activity when included as a component of systemic or mucosally-delivered gene-based prime-boost immunization. Clear adjuvant activity for both T and B cell responses was observed when flagellin was included in the DNA priming vaccine, but side effects occurred when given in an Ad boosting vector, particularly via the pulmonary route.

  4. Flagellin Encoded in Gene-Based Vector Vaccines Is a Route-Dependent Immune Adjuvant.

    Science.gov (United States)

    Rady, Hamada F; Dai, Guixiang; Huang, Weitao; Shellito, Judd E; Ramsay, Alistair J

    2016-01-01

    Flagellin has been tested as a protein-based vaccine adjuvant, with the majority of studies focused on antibody responses. Here, we evaluated the adjuvant activity of flagellin for both cellular and humoral immune responses in BALB/c mice in the setting of gene-based immunization, and have made several novel observations. DNA vaccines and adenovirus (Ad) vectors were engineered to encode mycobacterial protein Ag85B, with or without flagellin of Salmonella typhimurium (FliC). DNA-encoded flagellin given IM enhanced splenic CD4+ and CD8+ T cell responses to co-expressed vaccine antigen, including memory responses. Boosting either IM or intranasally with Ad vectors expressing Ag85B without flagellin led to durable enhancement of Ag85B-specific antibody and CD4+ and CD8+ T cell responses in both spleen and pulmonary tissues, correlating with significantly improved protection against challenge with pathogenic aerosolized M. tuberculosis. However, inclusion of flagellin in both DNA prime and Ad booster vaccines induced localized pulmonary inflammation and transient weight loss, with route-dependent effects on vaccine-induced T cell immunity. The latter included marked reductions in levels of mucosal CD4+ and CD8+ T cell responses following IM DNA/IN Ad mucosal prime-boosting, although antibody responses were not diminished. These findings indicate that flagellin has differential and route-dependent adjuvant activity when included as a component of systemic or mucosally-delivered gene-based prime-boost immunization. Clear adjuvant activity for both T and B cell responses was observed when flagellin was included in the DNA priming vaccine, but side effects occurred when given in an Ad boosting vector, particularly via the pulmonary route.

  5. The peptide derived from the Ig-like domain of human herpesvirus 8 K1 protein induces death in hematological cancer cells

    Directory of Open Access Journals (Sweden)

    Daniluk Urszula

    2012-08-01

    Full Text Available Abstract Background Although significant progress has been made in the treatment of lymphomas, many lymphomas exhibit resistance to cell death, suggesting a defective Fas signaling, which remains poorly understood. We previously reported that cells expressing the K1 protein of human herpesvirus 8 (HHV-8 resist death through the complex formation of the Ig-like domain of K1 with Fas. Recently, we investigated whether peptides derived from the Ig-like domain of the K1 protein may affect cell death. Methods K1 positive and negative cell lines were incubated with the K1-derived peptides, and cell death (apoptotic and necrotic was assessed by flow cytometry and LDH assay. Activation of caspases was assessed by fluorometric assay and flow cytometry. Fas receptor-independent, peptide-mediated cell killing was tested in the Fas-resistant Daudi cell line and Jurkat cell clones deficient in caspase-8 and FADD functionality. Activation of TNF receptors I and II was blocked by pre-incubation with corresponding blocking antibodies. The effect of the K1 peptide in vivo was tested in a mouse xenograft model. Results We observed that the peptide S20-3 enhanced cell death in K1-positive BJAB cells and HHV-8 positive primary effusion lymphoma (PEL cell lines. Similar effects of this peptide were observed in B-cell lymphoma and T-lymphoblastic leukemia cells without K1 expression but not in normal human peripheral blood mononuclear cells. A single intratumoral injection of the S20-3 peptide decreased the growth of Jurkat xenografts in SCID mice. The mechanism of tumor cell death induced by the S20-3 peptide was associated with activation of caspases, but this activity was only partially inhibited by the pan-caspase inhibitor z-VAD. Furthermore, the K1 peptide also killed Fas-resistant Daudi cells, and this killing effect was inhibited by pre-incubation of cells with antibodies blocking TNFRI. Conclusion Taken together, these findings indicate that the S20

  6. DNA-DSB in CHO-K1 cells induced by heavy-ions: Break rejoining and residual damage (GSI)

    Science.gov (United States)

    Taucher-Scholz, G.; Heilmann, J.; Becher, G.; Kraft, G.

    1994-01-01

    DNA double strand breaks (DSB's) are the critical lesions involved in cellular effects of ionizing radiation. Therefore, the evaluation of DSB induction in mammalian cells after heavy ion irradiation is an essential task for the assessment of high-LET radiation risk in space. Of particular interest has been the question of how the biological efficiency for the cellular inactivation endpoint relates to the initial lesions (DSBs) at varying LETs. For cell killing, an increased Relative Biological Efficiency (RBE) has been determined for highLET radiation around 100-200 keV/mu m. At higher LET, the RBE's decrease again to values below one for the very heavy particles. At GSI, DSB-induction was measured in CHO-K1 cells following irradiation with accelerated particles covering a wide LET range. The electrophoretic elution of fragmented DNA out of agarose plugs in a constant electrical field was applied for the detection of DSB's. The fraction of DNA retained was determined considering the relative intensities of ethidium bromide fluorescence in the well and in the gel lane. Dose-effect curves were established, from which the RBE for DSB induction was calculated at a fraction of 0.7 of DNA retained In summary, these rejoining studies are in line with an enhanced severity of the DNA DSB's at higher LET's, resulting in a decreased repairability of the induced lesions. However, no information concerning the fidelity of strand breaks rejoining is provided in these studies. To assess correct rejoining of DNA fragments an experimental system involving individual DNA hybridization bands has been set up. In preliminary experiments Sal I generated DNA fragments of 0.9 Mbp were irradiated with xrays and incubated for repair However, restitution of the original signals was not observed, probably due to the high radiation dose necessary for breakage of a fragment of this size. A banding pattern with NotI hybridization signals in a higher MW range (3Mbp) has been obtained by varying

  7. Bacterial flagellin triggers cardiac innate immune responses and acute contractile dysfunction.

    Directory of Open Access Journals (Sweden)

    Joelle Rolli

    Full Text Available BACKGROUND: Myocardial contractile failure in septic shock may develop following direct interactions, within the heart itself, between molecular motifs released by pathogens and their specific receptors, notably those belonging to the toll-like receptor (TLR family. Here, we determined the ability of bacterial flagellin, the ligand of mammalian TLR5, to trigger myocardial inflammation and contractile dysfunction. METHODOLOGY/PRINCIPAL FINDINGS: TLR5 expression was determined in H9c2 cardiac myoblasts, in primary rat cardiomyocytes, and in whole heart extracts from rodents and humans. The ability of flagellin to activate pro-inflammatory signaling pathways (NF-kappaB and MAP kinases and the expression of inflammatory cytokines was investigated in H9c2 cells, and, in part, in primary cardiomyocytes, as well as in the mouse myocardium in vivo. The influence of flagellin on left ventricular function was evaluated in mice by a conductance pressure-volume catheter. Cardiomyocytes and intact myocardium disclosed significant TLR5 expression. In vitro, flagellin activated NF-kappaB, MAP kinases, and the transcription of inflammatory genes. In vivo, flagellin induced cardiac activation of NF-kappaB, expression of inflammatory cytokines (TNF alpha, IL-1 beta, IL-6, MIP-2 and MCP-1, and provoked a state of reversible myocardial dysfunction, characterized by cardiac dilation, reduced ejection fraction, and decreased end-systolic elastance. CONCLUSION/SIGNIFICANCE: These results are the first to indicate that flagellin has the ability to trigger cardiac innate immune responses and to acutely depress myocardial contractility.

  8. Effects of inhibition of serine palmitoyltransferase (SPT and sphingosine kinase 1 (SphK1 on palmitate induced insulin resistance in L6 myotubes.

    Directory of Open Access Journals (Sweden)

    Agnieszka Mikłosz

    Full Text Available BACKGROUND: The objective of this study was to examine the effects of short (2 h and prolonged (18 h inhibition of serine palmitoyltransferase (SPT and sphingosine kinase 1 (SphK1 on palmitate (PA induced insulin resistance in L6 myotubes. METHODS: L6 myotubes were treated simultaneously with either PA and myriocin (SPT inhibitor or PA and Ski II (SphK1inhibitor for different time periods (2 h and 18 h. Insulin stimulated glucose uptake was measured using radioactive isotope. Expression of insulin signaling proteins was determined using Western blot analyses. Intracellular sphingolipids content [sphinganine (SFA, ceramide (CER, sphingosine (SFO, sphingosine-1-phosphate (S1P] were estimated by HPLC. RESULTS: Our results revealed that both short and prolonged time of inhibition of SPT by myriocin was sufficient to prevent ceramide accumulation and simultaneously reverse palmitate induced inhibition of insulin-stimulated glucose transport. In contrast, prolonged inhibition of SphK1 intensified the effect of PA on insulin-stimulated glucose uptake and attenuated further the activity of insulin signaling proteins (pGSK3β/GSK3β ratio in L6 myotubes. These effects were related to the accumulation of sphingosine in palmitate treated myotubes. CONCLUSION: Myriocin is more effective in restoration of palmitate induced insulin resistance in L6 myocytes, despite of the time of SPT inhibition, comparing to SKII (a specific SphK1 inhibitor. Observed changes in insulin signaling proteins were related to the content of specific sphingolipids, namely to the reduction of ceramide. Interestingly, inactivation of SphK1 augmented the effect of PA induced insulin resistance in L6 myotubes, which was associated with further inhibition of insulin stimulated PKB and GSK3β phosphorylation, glucose uptake and the accumulation of sphingosine.

  9. Rapamycin-enhanced mitomycin C-induced apoptotic death is mediated through the S6K1-Bad-Bak pathway in peritoneal carcinomatosis.

    Science.gov (United States)

    Song, X; Dilly, A-K; Kim, S-Y; Choudry, H A; Lee, Y J

    2014-06-05

    Peritoneal carcinomatosis (PC) is the most common secondary cancerous disease, and more effective novel regimens are needed. In this study, we identified a novel combination treatment for PC, chemotherapeutic agent mitomycin C in combination with mTOR (mammalian target of rapamycin) inhibitor rapamycin. We observed that the combination of mitomycin C and rapamycin induced synergistic cytotoxicity and apoptosis, which was mediated through an increase in caspase activation. The combination of mitomycin C and rapamycin inactivated p70 S6 ribosomal kinase (S6K1) and dephosphorylated Bad, leading to dissociation of Bcl-xL from Bak, which resulted in Bak oligomerization, mitochondria dysfunction and cytochrome c release. PF-4708671, a S6K1-specific inhibitor, enhanced the combination treatment-induced apoptosis, whereas S6K1 E389 DeltaCT-HA (S6K1 active form) dramatically decreased the induction of apoptosis. In addition, the combination treatment significantly inhibited LS174T intraperitoneal tumor growth in vivo. This study provides a preclinical rationale for apoptosis induction linked with the mTOR pathway through a combination of chemotherapeutic agents and mTOR inhibitor, and will support this combinatorial strategy to PC patients.

  10. Functional Activity of Antibodies Directed towards Flagellin Proteins of Non-Typhoidal Salmonella.

    Directory of Open Access Journals (Sweden)

    Girish Ramachandran

    Full Text Available Non-typhoidal Salmonella (NTS serovars Typhimurium and Enteritidis are major causes of invasive bacterial infections in children under 5 years old in sub-Saharan Africa, with case fatality rates of ~20%. There are no licensed NTS vaccines for humans. Vaccines that induce antibodies against a Salmonella Typhi surface antigen, Vi polysaccharide, significantly protect humans against typhoid fever, establishing that immune responses to Salmonella surface antigens can be protective. Flagella proteins, abundant surface antigens in Salmonella serovars that cause human disease, are also powerful immunogens, but the functional capacity of elicited anti-flagellar antibodies and their role in facilitating bacterial clearance has been unclear. We examined the ability of anti-flagellar antibodies to mediate microbial killing by immune system components in-vitro and assessed their role in protecting mice against invasive Salmonella infection. Polyclonal (hyperimmune sera and monoclonal antibodies raised against phase 1 flagellin proteins of S. Enteritidis and S. Typhimurium facilitated bacterial uptake and killing of the homologous serovar pathogen by phagocytes. Polyclonal anti-flagellar antibodies accompanied by complement also achieved direct bacterial killing. Serum bactericidal activity was restricted to Salmonella serovars expressing the same flagellin used as immunogen. Notably, individual anti-flagellin monoclonal antibodies with complement were not bactericidal, but this biological activity was restored when different monoclonal anti-flagellin antibodies were combined. Passive transfer immunization with a monoclonal IgG antibody specific for phase 1 flagellin from S. Typhimurium protected mice against lethal challenge with a representative African invasive S. Typhimurium strain. These findings have relevance for the use of flagellin proteins in NTS vaccines, and confirm the role of anti-flagellin antibodies as mediators of protective immunity.

  11. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus.

    Science.gov (United States)

    Zhao, Yue; Yang, Jieling; Shi, Jianjin; Gong, Yi-Nan; Lu, Qiuhe; Xu, Hao; Liu, Liping; Shao, Feng

    2011-09-14

    Inflammasomes are large cytoplasmic complexes that sense microbial infections/danger molecules and induce caspase-1 activation-dependent cytokine production and macrophage inflammatory death. The inflammasome assembled by the NOD-like receptor (NLR) protein NLRC4 responds to bacterial flagellin and a conserved type III secretion system (TTSS) rod component. How the NLRC4 inflammasome detects the two bacterial products and the molecular mechanism of NLRC4 inflammasome activation are not understood. Here we show that NAIP5, a BIR-domain NLR protein required for Legionella pneumophila replication in mouse macrophages, is a universal component of the flagellin-NLRC4 pathway. NAIP5 directly and specifically interacted with flagellin, which determined the inflammasome-stimulation activities of different bacterial flagellins. NAIP5 engagement by flagellin promoted a physical NAIP5-NLRC4 association, rendering full reconstitution of a flagellin-responsive NLRC4 inflammasome in non-macrophage cells. The related NAIP2 functioned analogously to NAIP5, serving as a specific inflammasome receptor for TTSS rod proteins such as Salmonella PrgJ and Burkholderia BsaK. Genetic analysis of Chromobacterium violaceum infection revealed that the TTSS needle protein CprI can stimulate NLRC4 inflammasome activation in human macrophages. Similarly, CprI is specifically recognized by human NAIP, the sole NAIP family member in human. The finding that NAIP proteins are inflammasome receptors for bacterial flagellin and TTSS apparatus components further predicts that the remaining NAIP family members may recognize other unidentified microbial products to activate NLRC4 inflammasome-mediated innate immunity. © 2011 Macmillan Publishers Limited. All rights reserved

  12. Functional Activity of Antibodies Directed towards Flagellin Proteins of Non-Typhoidal Salmonella.

    Science.gov (United States)

    Ramachandran, Girish; Tennant, Sharon M; Boyd, Mary A; Wang, Jin Y; Tulapurkar, Mohan E; Pasetti, Marcela F; Levine, Myron M; Simon, Raphael

    2016-01-01

    Non-typhoidal Salmonella (NTS) serovars Typhimurium and Enteritidis are major causes of invasive bacterial infections in children under 5 years old in sub-Saharan Africa, with case fatality rates of ~20%. There are no licensed NTS vaccines for humans. Vaccines that induce antibodies against a Salmonella Typhi surface antigen, Vi polysaccharide, significantly protect humans against typhoid fever, establishing that immune responses to Salmonella surface antigens can be protective. Flagella proteins, abundant surface antigens in Salmonella serovars that cause human disease, are also powerful immunogens, but the functional capacity of elicited anti-flagellar antibodies and their role in facilitating bacterial clearance has been unclear. We examined the ability of anti-flagellar antibodies to mediate microbial killing by immune system components in-vitro and assessed their role in protecting mice against invasive Salmonella infection. Polyclonal (hyperimmune sera) and monoclonal antibodies raised against phase 1 flagellin proteins of S. Enteritidis and S. Typhimurium facilitated bacterial uptake and killing of the homologous serovar pathogen by phagocytes. Polyclonal anti-flagellar antibodies accompanied by complement also achieved direct bacterial killing. Serum bactericidal activity was restricted to Salmonella serovars expressing the same flagellin used as immunogen. Notably, individual anti-flagellin monoclonal antibodies with complement were not bactericidal, but this biological activity was restored when different monoclonal anti-flagellin antibodies were combined. Passive transfer immunization with a monoclonal IgG antibody specific for phase 1 flagellin from S. Typhimurium protected mice against lethal challenge with a representative African invasive S. Typhimurium strain. These findings have relevance for the use of flagellin proteins in NTS vaccines, and confirm the role of anti-flagellin antibodies as mediators of protective immunity.

  13. Malaria Vaccine Development: Are Bacterial Flagellin Fusion Proteins the Bridge between Mouse and Humans?

    Directory of Open Access Journals (Sweden)

    Daniel Y. Bargieri

    2011-01-01

    Full Text Available In the past 25 years, the development of an effective malaria vaccine has become one of the biggest riddles in the biomedical sciences. Experimental data using animal infection models demonstrated that it is possible to induce protective immunity against different stages of malaria parasites. Nonetheless, the vast body of knowledge has generated disappointments when submitted to clinical conditions and presently a single antigen formulation has progressed to the point where it may be translated into a human vaccine. In parallel, new means to increase the protective effects of antigens in general have been pursued and depicted, such as the use of bacterial flagellins as carriers/adjuvants. Flagellins activate pathways in the innate immune system of both mice and humans. The recent report of the first Phase I clinical trial of a vaccine containing a Salmonella flagellin as carrier/adjuvant may fuel the use of these proteins in vaccine formulations. Herein, we review the studies on the use of recombinant flagellins as vaccine adjuvants with malarial antigens in the light of the current state of the art of malaria vaccine development. The available information indicates that bacterial flagellins should be seriously considered for malaria vaccine formulations to the development of effective human vaccines.

  14. Allergic Reactions Induced by Intravenous Vitamin K1 Infusion and Pharmaceutical Intervention%静脉滴注维生素K1致过敏反应3例及药学干预

    Institute of Scientific and Technical Information of China (English)

    傅征然; 王晓玲

    2013-01-01

    Three children who underwent renal biopsy received an infusion of vitamin K1 10 mg once daily to prevent puncture site bleeding.They developed allergic reactions such as chest tightness,shortness of breath,dyspnea,and skin rash one to five minutes after starting therapy.The medication was stopped immediately.Ten to twenty minutes after drug withdrawal,the allergic symptoms subsided.Clinical pharmacists analyzed the cause of allergic reaction,and implemented pharmaceutical intervention effectively.%3例肾内科患儿拟行肾穿刺术接受维生素K110 mg静脉滴注预防穿刺部位出血,用药后1~5 min出现胸闷、喘憋、呼吸困难、皮疹等过敏反应.立即停用维生素K1,过敏反应相关症状于停药后10~20 min消失.本文分析了过敏反应发生的原因,并实施了有效的药学干预.

  15. Flagellins of Salmonella Typhi and nonpathogenic Escherichia coli are differentially recognized through the NLRC4 pathway in macrophages.

    Science.gov (United States)

    Yang, Jingyi; Zhang, Ejuan; Liu, Fang; Zhang, Yan; Zhong, Maohua; Li, Yaoming; Zhou, Dihan; Chen, Yaoqing; Cao, Yuan; Xiao, Yang; He, Benxia; Yang, Yi; Sun, Ying; Lu, Mengji; Yan, Huimin

    2014-01-01

    Flagellin is recognized by both Toll-like receptor (TLR)5 and NAIP5/NLRC4 inflammasome receptors. We hypothesized that the flagellins derived from different bacteria might differentially activate TLR5 and/or NAIP5/NLRC4 signal pathways. To test this, the immune recognition of recombinant flagellins derived from pathogenic Salmonella Typhi (SF) and the nonpathogenic Escherichia coli K12 strain MG1655 (KF) were examined by the activation of TLR5 and NLRC4 pathways in various cell types. While flagellins SF and KF were not distinguishable in activating the TLR5 pathway, KF induced significantly less interleukin-1β production and pyroptotic cell death in peritoneal macrophages than SF, and showed markedly lower efficiency in activating caspase-1 through the NLRC4 pathway than SF. Macrophages may differentially recognize flagellins by intracellular sensors and thereby initiate the immune response to invading pathogenic bacteria. Our findings suggest an active role of flagellin as an important determinant in host differential immune recognition and for the control of bacteria infection.

  16. Antiproliferative effects on colon adenocarcinoma cells induced by co-administration of vitamin K1 and Lactobacillus rhamnosus GG.

    Science.gov (United States)

    Orlando, Antonella; Linsalata, Michele; Russo, Francesco

    2016-06-01

    Vitamin K (VK), an essential nutrient associated with the clotting cascade, has also been demonstrated to have anticancer properties in various cancer cells including colon cancer cells. Also probiotics have gained interest as potential anticancer agents. Among them, Lactobacillus rhamnosus GG (L.GG) has been shown to inhibit cell proliferation and polyamine biosynthesis as well as to induce apoptosis in different human gastrointestinal cancer cells. Nevertheless, the exact mechanisms involved in these actions are not completely elucidated. Therefore, the aims of the present study were to evaluate in three differently graded human colon cancer cells (namely Caco-2, HT-29 and SW480) the effects of increasing VK1 concentrations, administered alone or in combination with viable L.GG, on the cell proliferation evaluated by MTT test, apoptosis investigated by Bax/Bcl-2 ratio and the percentage of the apoptotic cells, and the cell cycle evaluated by MUSE cell analyzer. Both VK1 and L.GG administered alone up to 72 h, caused inhibition of proliferation, induction of apoptosis and the cell cycle arrest in all the tested colon cancer cells. When VK1 and L.GG were co-administered, the addition of increasing VK1 concentrations potentiated the probiotic antiproliferative effect in a dose-dependent manner, being also related to the individual features of each cell line. The effect was more evident in Caco-2 and HT-29 cells compared to the less differentiated SW480. The enhanced antiproliferative efficacy due to co-administration of L.GG and VK1 could represent a suitable option in a functional food strategy for cancer growth inhibition and chemoprevention.

  17. Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition.

    Directory of Open Access Journals (Sweden)

    Laura Helft

    Full Text Available Microbe-associated molecular patterns (MAMPs are molecules, or domains within molecules, that are conserved across microbial taxa and can be recognized by a plant or animal immune system. Although MAMP receptors have evolved to recognize conserved epitopes, the MAMPs in some microbial species or strains have diverged sufficiently to render them unrecognizable by some host immune systems. In this study, we carried out in vitro evolution of the Arabidopsis thaliana flagellin receptor FLAGELLIN-SENSING 2 (FLS2 to isolate derivatives that recognize one or more flagellin peptides from bacteria for which the wild-type Arabidopsis FLS2 confers little or no response. A targeted approach generated amino acid variation at FLS2 residues in a region previously implicated in flagellin recognition. The primary screen tested for elevated response to the canonical flagellin peptide from Pseudomonas aeruginosa, flg22. From this pool, we then identified five alleles of FLS2 that confer modest (quantitatively partial recognition of an Erwinia amylovora flagellin peptide. Use of this Erwinia-based flagellin peptide to stimulate Arabidopsis plants expressing the resulting FLS2 alleles did not lead to a detectable reduction of virulent P. syringae pv. tomato growth. However, combination of two identified mutations into a single allele further increased FLS2-mediated responses to the E. amylovora flagellin peptide. These studies demonstrate the potential to raise the sensitivity of MAMP receptors toward particular targets.

  18. Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition

    Science.gov (United States)

    Helft, Laura; Thompson, Mikayla

    2016-01-01

    Microbe-associated molecular patterns (MAMPs) are molecules, or domains within molecules, that are conserved across microbial taxa and can be recognized by a plant or animal immune system. Although MAMP receptors have evolved to recognize conserved epitopes, the MAMPs in some microbial species or strains have diverged sufficiently to render them unrecognizable by some host immune systems. In this study, we carried out in vitro evolution of the Arabidopsis thaliana flagellin receptor FLAGELLIN-SENSING 2 (FLS2) to isolate derivatives that recognize one or more flagellin peptides from bacteria for which the wild-type Arabidopsis FLS2 confers little or no response. A targeted approach generated amino acid variation at FLS2 residues in a region previously implicated in flagellin recognition. The primary screen tested for elevated response to the canonical flagellin peptide from Pseudomonas aeruginosa, flg22. From this pool, we then identified five alleles of FLS2 that confer modest (quantitatively partial) recognition of an Erwinia amylovora flagellin peptide. Use of this Erwinia-based flagellin peptide to stimulate Arabidopsis plants expressing the resulting FLS2 alleles did not lead to a detectable reduction of virulent P. syringae pv. tomato growth. However, combination of two identified mutations into a single allele further increased FLS2-mediated responses to the E. amylovora flagellin peptide. These studies demonstrate the potential to raise the sensitivity of MAMP receptors toward particular targets. PMID:27270917

  19. Identification of IRS-1 Ser-1101 as a target of S6K1 in nutrient- and obesity-induced insulin resistance.

    Science.gov (United States)

    Tremblay, Frédéric; Brûlé, Sophie; Hee Um, Sung; Li, Yu; Masuda, Kohei; Roden, Michael; Sun, Xiao Jian; Krebs, Michael; Polakiewicz, Roberto D; Thomas, George; Marette, André

    2007-08-28

    S6K1 has emerged as a critical signaling component in the development of insulin resistance through phosphorylation and inhibition of IRS-1 function. This effect can be triggered directly by nutrients such as amino acids or by insulin through a homeostatic negative-feedback loop. However, the role of S6K1 in mediating IRS-1 phosphorylation in a physiological setting of nutrient overload is unresolved. Here we show that S6K1 directly phosphorylates IRS-1 Ser-1101 in vitro in the C-terminal domain of the protein and that mutation of this site largely blocks the ability of amino acids to suppress IRS-1 tyrosine and Akt phosphorylation. Consistent with this finding, phosphorylation of IRS-1 Ser-1101 is increased in the liver of obese db/db and wild-type, but not S6K1(-/-), mice maintained on a high-fat diet and is blocked by siRNA knockdown of S6K1 protein. Finally, infusion of amino acids in humans leads to the concomitant activation of S6K1, phosphorylation of IRS-1 Ser-1101, a reduction in IRS-1 function, and insulin resistance in skeletal muscle. These findings indicate that nutrient- and hormonal-dependent activation of S6K1 causes insulin resistance in mice and humans, in part, by mediating IRS-1 Ser-1101 phosphorylation.

  20. Inhibition of p70 S6 kinase (S6K1) activity by A77 1726, the active metabolite of leflunomide, induces autophagy through TAK1-mediated AMPK and JNK activation.

    Science.gov (United States)

    Xu, Xiulong; Sun, Jing; Song, Ruilong; Doscas, Michelle E; Williamson, Ashley J; Zhou, Jingsong; Sun, Jun; Jiao, Xinan; Liu, Xiufan; Li, Yi

    2017-03-31

    mTOR activation suppresses autophagy by phosphorylating ULK1 at S757 and suppressing its enzymatic activity. Here we report that feedback activation of mTOR in the PI-3 kinase pathway by two p70 S6 kinase (S6K1) inhibitors (PF-4708671 and A77 1726, the active metabolite of an immunosuppressive drug leflunomide) or by S6K1 knockdown did not suppress but rather induced autophagy. Suppression of S6K1 activity led to the phosphorylation and activation of AMPK, which then phosphorylated ULK1 at S555. While mTOR feedback activation led to increased phosphorylation of ULK1 at S757, this modification did not the disrupt ULK1-AMPK interaction nor dampen ULK1 S555 phosphorylation and the induction of autophagy. In addition, inhibition of S6K1 activity led to JNK activation, which also contributed to autophagy. 5Z-7-oxozeaenol, a specific inhibitor of TAK1, or TAK1 siRNA blocked A77 1726-induced activation of AMPK and JNK, and LC3 lipidation. Taken together, our study establishes S6K1 as a key player in the PI-3 kinase pathway to suppress autophagy through inhibiting AMPK and JNK in a TAK1-dependent manner.

  1. Differential regulation of epidermal growth factor receptor by hydrogen peroxide and flagellin in cultured lung alveolar epithelial cells.

    Science.gov (United States)

    Nishi, Hiroyuki; Maeda, Noriko; Izumi, Shunsuke; Higa-Nakamine, Sayomi; Toku, Seikichi; Kakinohana, Manabu; Sugahara, Kazuhiro; Yamamoto, Hideyuki

    2015-02-05

    In previous studies, we found that stimulation of Toll-like receptor 5 (TLR5) by flagellin induced the activation of mitogen-activated protein kinase (MAPK)-activated protein kinase-2 (MAPKAPK-2) through activation of the p38 MAPK pathway in cultured alveolar epithelial A549 cells. Our studies strongly suggested that MAPKAPK-2 phosphorylated epidermal growth factor receptor (EGFR) at Ser1047. It has been reported that phosphorylation of Ser1047 after treatment with tumor necrosis factor α (TNFα) induced the internalization of EGFR. In the present study, we first found that treatment of A549 cells with hydrogen peroxide induced the activation of MAPKAPK-2 and phosphorylation of EGFR at Ser1047 within 30 min. This was different from flagellin treatment because hydrogen peroxide treatment induced the phosphorylation of EGFR at Tyr1173 as well as Ser1047, indicating the activation of EGFR. We also found that KN93, an inhibitor of CaM kinase II, inhibited the hydrogen peroxide-induced phosphorylation of EGFR at Ser1047 through inhibition of the activation of the p38 MAPK pathway. Furthermore, we examined the internalization of EGFR by three different methods. Flow cytometry with an antibody against the extracellular domain of EGFR and biotinylation of cell surface proteins revealed that flagellin, but not hydrogen peroxide, decreased the amount of cell-surface EGFR. In addition, activation of extracellular signal-regulated kinase by EGF treatment was reduced by flagellin pre-treatment. These results strongly suggested that hydrogen peroxide activated the p38 MAPK pathway via activation of CaM kinase II and that flagellin and hydrogen peroxide regulate the functions of EGFR by different mechanisms.

  2. Amplification and flagellin typing of pseudomonas aeruginosa by molecular method

    Directory of Open Access Journals (Sweden)

    gholamreza goudarzi

    2011-08-01

    Conclusion: This method could be utilized to determine flagellated (Motile and non-flagellated strains of P. aeruginosa, genotyping, amplification of full coding sequence of fliC gene in order to clone and express recombinant flagellin protein.

  3. GD3 expression in CHO-K1 cells increases growth rate, induces morphological changes, and affects cell-substrate interactions.

    Science.gov (United States)

    Daniotti, Jose L; Zurita, Adolfo R; Trindade, Vera M T; Maccioni, Hugo J F

    2002-11-01

    We have generated a panel of CHO-K1 cell clones with different glycolipid compositions by stable transfection of appropriate glycosyltransferases and studied the morphological and growth phenotype of a clone stably expressing Sial-T2. Compared with the GM3 expressing parental cells, Sial-T2 transfectants show low expression of GM3 and neo expression of GD3 and GT3. These cells show about 60% reduction of the mean cell area, and about 2-fold increase of the mean colony area and growth rate. Cells over expressing Sial-T2 showed a flattened appearance, and with time in culture they detached from the substrate leaving adhered material that was GD3 immunoreactive. No apoptotic or proteome differences could be detected in the Sial-T2 transfectants. Thus, increased expression of GD3 and GT3 influence parameters of growth and social behavior of CHO-K1 cells. However, the molecular and cellular basis underlying these influences requires further investigation.

  4. Relationship between genotype and phenotype of flagellin C in Salmonella

    Institute of Scientific and Technical Information of China (English)

    Wan-Sheng Ji; Jia-Lu Hu; Jun-Wen Qiu; Bo-Rong Pan; Dao-Rong Peng; Bing-Long Shi; Shao-Juan Zhou; Kai-Chun Wu; Dai-Ming Fan

    2001-01-01

    AIM: To discover the relationship between the genotype and antigen serotype of flagellin C among Salmonella strains. METHODS: Fragment of Salmonella flagellin C in plasmid pLS408 was cloned, sequenced and compared with the corresponding sequence in other strains. Salmonella strains including two typhi strains, one paratyphoid strain, one enteritidis and one typhimurium strain were isolated from outpatients. Genome DNA was purified respectively from these clinical isolstes, then the corresponding flagellin C fragment was amplified by polymerase chain reaction, and the amplification products were analyzed by agarose gel electrophoreeis. RESULTS: The cloned fragment includes 582 nucleotides encoding the variable region and partial conservative region of Salmonella flagellin C in plasmid pLS408. With comparison to the corresponding sequences reported previously, there is only a little difference from other strains with the same flagellar serotype in both nucleotide and amino acid level. Specific PCR products were amplified in Salmonella strains with flagellar eerotype H-1-d including S. Muenchen, typhi and typhimurium, but not in S.paratyphoid C or S. Enteritidis strains. CONCLUSION: In this experiment, the specificity of nucleotide sequence could be found in flagellin C central variable regions as it exists in flagellar serotypes in Salmonella. It may be helpful to developing a rapid,sensitive, accurate and PCR-based method to detect Salmonella strains with serotype H-1-d.

  5. Arcobacter spp. possess two very short flagellins of which FlaA is essential for motility

    OpenAIRE

    Ho, H.; Lipman, L.J.A.; Wösten, M.M.S.M.; van Asten, A.J.; Gaastra, W

    2008-01-01

    Like Campylobacter and Helicobacter spp., Arcobacter spp. possess two flagellin genes (flaA and flaB) located adjacent to each other. The aim of this study was to characterize the flagellin proteins of Arcobacter spp., because these proteins are known virulence factors in the Epsilonproteobacteria, to which these three species belong. With the exception of Arcobacter nitrofigilis, Arcobacter flagellins are almost half the size of those in other Epsilonproteobacteria. Arcobacter flagellin prot...

  6. Self-assembly of flagellin on Au(111) surfaces.

    Science.gov (United States)

    González Orive, Alejandro; Pissinis, Diego E; Diaz, Carolina; Miñán, Alejandro; Benítez, Guillermo A; Rubert, Aldo; Daza Millone, Antonieta; Rumbo, Martin; Hernández Creus, Alberto; Salvarezza, Roberto C; Schilardi, Patricia L

    2014-11-01

    The adsorption of flagellin monomers from Pseudomonas fluorescens on Au(111) has been studied by Atomic Force Microscopy (AFM), Scanning Tunneling Microscopy (STM), X-ray Photoelectron Spectroscopy (XPS), Surface Plasmon Resonance (SPR), and electrochemical techniques. Results show that flagellin monomers spontaneously self-assemble forming a monolayer thick protein film bounded to the Au surface by the more hydrophobic subunit and exposed to the environment the hydrophilic subunit. The films are conductive and allow allocation of electrochemically active cytochrome C. The self-assembled films could be used as biological platforms to build 3D complex molecular structures on planar metal surfaces and to functionalize metal nanoparticles.

  7. Protein glycosylation in Helicobacter pylori: beyond the flagellins?

    Directory of Open Access Journals (Sweden)

    Patrick S Hopf

    Full Text Available Glycosylation of flagellins by pseudaminic acid is required for virulence in Helicobacter pylori. We demonstrate that, in H. pylori, glycosylation extends to proteins other than flagellins and to sugars other than pseudaminic acid. Several candidate glycoproteins distinct from the flagellins were detected via ProQ-emerald staining and DIG- or biotin- hydrazide labeling of the soluble and outer membrane fractions of wild-type H. pylori, suggesting that protein glycosylation is not limited to the flagellins. DIG-hydrazide labeling of proteins from pseudaminic acid biosynthesis pathway mutants showed that the glycosylation of some glycoproteins is not dependent on the pseudaminic acid glycosylation pathway, indicating the existence of a novel glycosylation pathway. Fractions enriched in glycoprotein candidates by ion exchange chromatography were used to extract the sugars by acid hydrolysis. High performance anion exchange chromatography with pulsed amperometric detection revealed characteristic monosaccharide peaks in these extracts. The monosaccharides were then identified by LC-ESI-MS/MS. The spectra are consistent with sugars such as 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (Pse5Ac7Ac previously described on flagellins, 5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (Pse5Am7Ac, bacillosamine derivatives and a potential legionaminic acid derivative (Leg5AmNMe7Ac which were not previously identified in H. pylori. These data open the way to the study of the mechanism and role of protein glycosylation on protein function and virulence in H. pylori.

  8. Characterization and functional analysis of seven flagellin genes in Rhizobium leguminosarum bv. viciae. Characterization of R. leguminosarum flagellins

    Directory of Open Access Journals (Sweden)

    Tambalo Dinah D

    2010-08-01

    Full Text Available Abstract Background Rhizobium leguminosarum bv. viciae establishes symbiotic nitrogen fixing partnerships with plant species belonging to the Tribe Vicieae, which includes the genera Vicia, Lathyrus, Pisum and Lens. Motility and chemotaxis are important in the ecology of R. leguminosarum to provide a competitive advantage during the early steps of nodulation, but the mechanisms of motility and flagellar assembly remain poorly studied. This paper addresses the role of the seven flagellin genes in producing a functional flagellum. Results R. leguminosarum strains 3841 and VF39SM have seven flagellin genes (flaA, flaB, flaC, flaD, flaE, flaH, and flaG, which are transcribed separately. The predicted flagellins of 3841 are highly similar or identical to the corresponding flagellins in VF39SM. flaA, flaB, flaC, and flaD are in tandem array and are located in the main flagellar gene cluster. flaH and flaG are located outside of the flagellar/motility region while flaE is plasmid-borne. Five flagellin subunits (FlaA, FlaB, FlaC, FlaE, and FlaG are highly similar to each other, whereas FlaD and FlaH are more distantly related. All flagellins exhibit conserved amino acid residues at the N- and C-terminal ends and are variable in the central regions. Strain 3841 has 1-3 plain subpolar flagella while strain VF39SM exhibits 4-7 plain peritrichous flagella. Three flagellins (FlaA/B/C and five flagellins (FlaA/B/C/E/G were detected by mass spectrometry in the flagellar filaments of strains 3841 and VF39SM, respectively. Mutation of flaA resulted in non-motile VF39SM and extremely reduced motility in 3841. Individual mutations of flaB and flaC resulted in shorter flagellar filaments and consequently reduced swimming and swarming motility for both strains. Mutant VF39SM strains carrying individual mutations in flaD, flaE, flaH, and flaG were not significantly affected in motility and filament morphology. The flagellar filament and the motility of 3841 strains

  9. Resistance exercise-induced S6K1 kinase activity is not inhibited in human skeletal muscle despite prior activation of AMPK by high-intensity interval cycling

    DEFF Research Database (Denmark)

    Apró, William; Moberg, Marcus; Hamilton, D. Lee

    2015-01-01

    .05) and eukaryotic elongation factor 2 phosphorylation was reduced (∼55%, P influenced by the two modes of exercise; ER induced a significant increase in gene and protein expression of MuRF1 (P ...Combining endurance and strength training in the same session has been reported to reduce the anabolic response to the latter form of exercise. The underlying mechanism, based primarily on results from rodent muscle, is proposed to involve AMPK-dependent inhibition of mTORC1 signaling....... This hypothesis was tested in eight trained male subjects who in randomized order performed either resistance exercise only (R) or interval cycling followed by resistance exercise (ER). Biopsies taken from the vastus lateralis before and after endurance exercise and repeatedly after resistance exercise were...

  10. The adjuvant activity of alphavirus replicons is enhanced by incorporating the microbial molecule flagellin into the replicon.

    Directory of Open Access Journals (Sweden)

    Maria L Knudsen

    Full Text Available Ligands of pattern recognition receptors (PRRs including Toll-like receptors (TLRs stimulate innate and adaptive immune responses and are considered as potent adjuvants. Combinations of ligands might act in synergy to induce stronger and broader immune responses compared to stand-alone ligands. Alphaviruses stimulate endosomal TLRs 3, 7 and 8 as well as the cytoplasmic PRR MDA-5, resulting in induction of a strong type I interferon (IFN response. Bacterial flagellin stimulates TLR5 and when delivered intracellularly the cytosolic PRR NLRC4, leading to secretion of proinflammatory cytokines. Both alphaviruses and flagellin have independently been shown to act as adjuvants for antigen-specific antibody responses. Here, we hypothesized that alphavirus and flagellin would act in synergy when combined. We therefore cloned the Salmonella Typhimurium flagellin (FliC gene into an alphavirus replicon and assessed its adjuvant activity on the antibody response against co-administered antigen. In mice immunized with recombinant alphavirus, antibody responses were greatly enhanced compared to soluble FliC or control alphavirus. Both IgG1 and IgG2a/c responses were increased, indicating an enhancement of both Th1 and Th2 type responses. The adjuvant activity of FliC-expressing alphavirus was diminished but not abolished in the absence of TLR5 or type I IFN signaling, suggesting the contribution of several signaling pathways and some synergistic and redundant activity of its components. Thus, we have created a recombinant adjuvant that stimulates multiple signaling pathways of innate immunity resulting in a strong and broad antibody response.

  11. A conserved TLR5 binding and activation hot spot on flagellin

    Science.gov (United States)

    Song, Wan Seok; Jeon, Ye Ji; Namgung, Byeol; Hong, Minsun; Yoon, Sung-il

    2017-01-01

    Flagellin is a bacterial protein that polymerizes into the flagellar filament and is essential for bacterial motility. When flagellated bacteria invade the host, flagellin is recognized by Toll-like receptor 5 (TLR5) as a pathogen invasion signal and eventually evokes the innate immune response. Here, we provide a conserved structural mechanism by which flagellins from Gram-negative γ-proteobacteria and Gram-positive Firmicutes bacteria bind and activate TLR5. The comparative structural analysis using our crystal structure of a complex between Bacillus subtilis flagellin (bsflagellin) and TLR5 at 2.1 Å resolution, combined with the alanine scanning analysis of the binding interface, reveals a common hot spot in flagellin for TLR5 activation. An arginine residue (bsflagellin R89) of the flagellin D1 domain and its adjacent residues (bsflagellin E114 and L93) constitute a hot spot that provides shape and chemical complementarity to a cavity generated by the loop of leucine-rich repeat 9 in TLR5. In addition to the flagellin D1 domain, the D0 domain also contributes to TLR5 activity through structurally dispersed regions, but not a single focal area. These results establish the groundwork for the future design of flagellin-based therapeutics. PMID:28106112

  12. New Flagellin Gene for Salmonella enterica serovar Typhi from the East Indonesian Archipelago

    NARCIS (Netherlands)

    M. Hatta; A.R. Sultan; R. Pastoor; H.L. Smits

    2011-01-01

    Phase variation is a property unique of some Salmonella enterica serovar Typhi strains from Indonesia. Salmonella Typhi isolates from Indonesia have been described that in addition to the phase 1 Hd flagellin gene contain a second flagellin gene named z66. S. Typhi isolates from Indonesia with a mut

  13. Arcobacter spp. possess two very short flagellins of which FlaA is essential for motility

    NARCIS (Netherlands)

    Ho, H.; Lipman, L.J.A.; Wösten, M.M.S.M.; van Asten, A.J.; Gaastra, W.

    2008-01-01

    Like Campylobacter and Helicobacter spp., Arcobacter spp. possess two flagellin genes (flaA and flaB) located adjacent to each other. The aim of this study was to characterize the flagellin proteins of Arcobacter spp., because these proteins are known virulence factors in the Epsilonproteobacteria,

  14. Evidence for an heterogeneous glycosylation of the Clostridium tyrobutyricum ATCC 25755 flagellin.

    Science.gov (United States)

    Bédouet, L; Arnold, F; Robreau, G; Batina, P; Talbot, F; Binet, A

    1998-01-01

    Glycosylation analysis of the flagellin from the Gram-positive species Clostridium tyrobutyricum has been supplemented. Amino acid analysis of the glycopeptides obtained after pronase digestion of flagellin indicated that O-glycosylation which was previously demonstrated after nonreductive beta-elimination, probably occurred via the hydroxyl group of serine. Otherwise, beta-elimination partly deglycosylated flagellin. After this treatment carbohydrates were still linked to protein as shown by a digoxigenin-hydrazide labelling. Therefore, in addition to linkages via serine, alkaline resistant linkages exist on the flagellin and some glycans may be linked to the protein core via the amide nitrogen of asparagine or via the hydroxyl group of tyrosine. Furthermore, according to an immunological analysis, glycans attached to flagellin via alkaline sensitive linkages may be different from those attached via alkaline resistant linkages.

  15. Contribution of six flagellin genes to the flagellum biogenesis of Vibrio vulnificus and in vivo invasion.

    Science.gov (United States)

    Kim, Soo Young; Thanh, Xuan Tran Thi; Jeong, Kwangjoon; Kim, Seong Bin; Pan, Sang O; Jung, Che Hun; Hong, Seol Hee; Lee, Shee Eun; Rhee, Joon Haeng

    2014-01-01

    Vibrio vulnificus is a halophilic pathogenic bacterium that is motile due to the presence of a single polar flagellum. V. vulnificus possesses a total of six flagellin genes organized into two loci (flaFBA and flaCDE). We proved that all six of the flagellin genes were transcribed, whereas only five (FlaA, -B, -C, -D, and -F) of the six flagellin proteins were detected. To understand roles of the six V. vulnificus flagellins in motility and virulence, mutants with single and multiple flagellin deletions were constructed. Mutations in flaB or flaC or the flaCDE locus resulted in a significant decrease in motility, adhesion, and cytotoxicity, whereas single mutations in the other flagellin genes or the flaFBA locus showed little or no effect. The motility was completely abolished only in the mutant lacking all six flagellin genes (flaFBA flaCDE). Surprisingly, a double mutation of flaB and flaD, a gene sharing 99% identity with the flaB at the amino acid level, resulted in the largest decrease in motility, adhesion, and cytotoxicity except for the mutant in which all six genes were deleted (the hexa mutant). Additionally, the 50% lethal doses (LD50s) of the flaB flaD and the flaFBA flaCDE mutants increased 23- and 91-fold in a mouse model, respectively, and the in vitro and in vivo invasiveness of the mutants was significantly decreased compared to that of the wild type. Taken together, the multiple flagellin subunits differentially contribute to the flagellum biogenesis and the pathogenesis of V. vulnificus, and among the six flagellin genes, flaB, flaD, and flaC were the most influential components.

  16. Fusion proteins of flagellin and the major birch pollen allergen Bet v 1 show enhanced immunogenicity, reduced allergenicity, and intrinsic adjuvanticity.

    Science.gov (United States)

    Kitzmüller, Claudia; Kalser, Julia; Mutschlechner, Sonja; Hauser, Michael; Zlabinger, Gerhard J; Ferreira, Fatima; Bohle, Barbara

    2017-04-26

    Recombinant fusion proteins of flagellin and antigens have been demonstrated to induce strong innate and adaptive immune responses. Such fusion proteins can enhance the efficacy of allergen-specific immunotherapy. We sought to characterize different fusion proteins of flagellin and the major birch pollen allergen Bet v 1 for suitability as allergy vaccines. A truncated version of flagellin (NtCFlg) was genetically fused to the N- or C-terminus of Bet v 1. Toll-like receptor (TLR) 5 binding was assessed with HEK293 cells expressing TLR5. Upregulation of CD40, CD80, CD83, and CD86 on monocyte-derived dendritic cells from allergic patients was analyzed by using flow cytometry. The T cell-stimulatory capacity of the fusion proteins was assessed with naive and Bet v 1-specific T cells. IgE binding was tested in inhibition ELISAs and basophil activation tests. Mice were immunized with the fusion proteins in the absence and presence of aluminum hydroxide. Cellular and antibody responses were monitored. Murine antibodies were tested for blocking capacity in basophil activation tests. Both fusion proteins matured monocyte-derived dendritic cells through TLR5. Compared with Bet v 1, the fusion proteins showed stronger T cell-stimulatory and reduced IgE-binding capacity and induced murine Bet v 1-specific antibodies in the absence of aluminum hydroxide. However, only antibodies induced by means of immunization with NtCFlg fused to the C-terminus of Bet v 1 inhibited binding of patients' IgE antibodies to Bet v 1. Bet v 1-flagellin fusion proteins show enhanced immunogenicity, reduced allergenicity, and intrinsic adjuvanticity and thus represent promising vaccines for birch pollen allergen-specific immunotherapy. However, the sequential order of allergen and adjuvant within a fusion protein determines its immunologic characteristics. Copyright © 2017 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

  17. The effect of oral vitamin K1 on protein induced by vitamin K absence Ⅱ in newborn infants%口服维生素K1对新生儿维生素K缺乏诱导蛋白的影响

    Institute of Scientific and Technical Information of China (English)

    梁友玲; 林超琼; 鲁杰

    2002-01-01

    目的探讨口服维生素K1(VK1)对新生儿VK缺乏症诱导蛋白Ⅱ(PIVKA-Ⅱ)的影响及口服VK1的最佳剂量.方法随机将101例足月新生儿分为口服VK1 2.5 mg组、5 mg组及对照组三组,出生时均采集脐血,用酶联免疫吸附法测定PIVKA-Ⅱ含量,3 d后复查三组新生儿血PIVKA-Ⅱ含量.结果PIVKA-Ⅱ≥2 mg/ml人数2.5 mg组由8例降至4例,5 mg组由11例降至2例,对照组由11例降至8例.经统计学处理,2.5 mg组与5 mg组比较P>0.05;口服VK1组与对照组比较P<0.05.结论PIVKA-Ⅱ的变化与口服VK1剂量的差异无显著性.

  18. Flagellin Diversity in Clostridium botulinum Groups I and II: a New Strategy for Strain Identification▿

    Science.gov (United States)

    Paul, Catherine J.; Twine, Susan M.; Tam, Kevin J.; Mullen, James A.; Kelly, John F.; Austin, John W.; Logan, Susan M.

    2007-01-01

    Strains of Clostridium botulinum are traditionally identified by botulinum neurotoxin type; however, identification of an additional target for typing would improve differentiation. Isolation of flagellar filaments and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that C. botulinum produced multiple flagellin proteins. Nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) analysis of in-gel tryptic digests identified peptides in all flagellin bands that matched two homologous tandem flagellin genes identified in the C. botulinum Hall A genome. Designated flaA1 and flaA2, these open reading frames encode the major structural flagellins of C. botulinum. Colony PCR and sequencing of flaA1/A2 variable regions classified 80 environmental and clinical strains into group I or group II and clustered isolates into 12 flagellar types. Flagellar type was distinct from neurotoxin type, and epidemiologically related isolates clustered together. Sequencing a larger PCR product, obtained during amplification of flaA1/A2 from type E strain Bennett identified a second flagellin gene, flaB. LC-MS analysis confirmed that flaB encoded a large type E-specific flagellin protein, and the predicted molecular mass for FlaB matched that observed by SDS-PAGE. In contrast, the molecular mass of FlaA was 2 to 12 kDa larger than the mass predicted by the flaA1/A2 sequence of a given strain, suggesting that FlaA is posttranslationally modified. While identification of FlaB, and the observation by SDS-PAGE of different masses of the FlaA proteins, showed the flagellin proteins of C. botulinum to be diverse, the presence of the flaA1/A2 gene in all strains examined facilitates single locus sequence typing of C. botulinum using the flagellin variable region. PMID:17351097

  19. Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells

    Science.gov (United States)

    Starost, Laura Julia; Karassek, Sascha; Sano, Yasuteru; Kanda, Takashi; Kim, Kwang Sik; Dobrindt, Ulrich; Rüter, Christian; Schmidt, Marcus Alexander

    2016-01-01

    Pertussis toxin (PTx), the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood–brain barrier (BBB) in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218’s effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB. PMID:27754355

  20. Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells.

    Science.gov (United States)

    Starost, Laura Julia; Karassek, Sascha; Sano, Yasuteru; Kanda, Takashi; Kim, Kwang Sik; Dobrindt, Ulrich; Rüter, Christian; Schmidt, Marcus Alexander

    2016-10-13

    Pertussis toxin (PTx), the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood-brain barrier (BBB) in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218's effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB.

  1. Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Laura Julia Starost

    2016-10-01

    Full Text Available Pertussis toxin (PTx, the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood–brain barrier (BBB in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218’s effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB.

  2. Sublingual flagellin protects against acute pneumococcal pneumonia in a TLR5-dependent and NLRC4-independent fashion.

    Science.gov (United States)

    Muñoz-Wolf, Natalia; Rial, Analía; Fougeron, Delphine; Tabareau, Julien; Sirard, Jean-Claude; Chabalgoity, José A

    2016-09-01

    To evaluate efficacy of sublingual flagellin to treat acute pneumonia. Mice were treated sublingually with flagellin and challenged intranasally with a lethal dose of pneumococcus. Flagellins lacking TLR5 or NLRC4 activation domains were used to assess their contribution to protection. Sublingual flagellin protected mice in a TLR5-dependent, NLRC4-independent fashion. Neutrophils were required for protection. Flagellin-stimulated lung epithelial cells recapitulated the lung's transcriptional profile suggesting they could be targeted by flagellin in vivo. Ligation of TLR5, a pathogen recognition receptor not naturally engaged by pneumococcus, protects mice from invasive pneumonia when administered via sublingual route. This can be a highly cost-effective alternative therapy against pneumonia.

  3. Expression, purification, and functional characterisation of Flagellin, a TLR5-ligand

    Directory of Open Access Journals (Sweden)

    Irshad Ahmed Hajam

    2013-06-01

    Full Text Available Flagellin, a Toll-like receptor 5 (TLR5-ligand, is known for its activities like adjuvant, induction of pro-inflammatory cytokines and innate immunity. In this context, fliC gene of Salmonella Typhimurium was cloned into pET32a expression plasmid using in-house designed gene specific primers. The frame and orientation of the inserted fliC gene was confirmed upon colony PCR, restriction enzyme analysis and sequencing. Sequence analysis of fliC revealed proper orientation of the gene and had 1,485 nucleotides. Following transformation of pET-fliC plasmid into Escherichia coli BL21 (DE3 cells, the gene was expressed after inducing with IPTG (Isopropylβ-D-1-thiogalactopyranoside. The polyHis-tag-fliC was ~70kDa as confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. The identity/authenticity of the recombinant-fliC was confirmed by its specific reactivity with commercial anti-fliC MAb of S. Typhimurium. Further, the antigenic and functional properties of recombinant-fliC were determined espousing its ability to induce antigen specific antibodies in G pigs and increased m-RNA expression of certain pro-inflammatory mediators like TNF-α and GM-CSF in vitro.

  4. Determination of the physiological 2:2 TLR5:flagellin activation stoichiometry revealed by the activity of a fusion receptor

    Energy Technology Data Exchange (ETDEWEB)

    Ivičak-Kocjan, Karolina; Panter, Gabriela; Benčina, Mojca [Laboratory of Biotechnology, National Institute of Chemistry, 1000 Ljubljana (Slovenia); The Centre of Excellence EN-FIST, 1000 Ljubljana (Slovenia); Jerala, Roman, E-mail: roman.jerala@ki.si [Laboratory of Biotechnology, National Institute of Chemistry, 1000 Ljubljana (Slovenia); The Centre of Excellence EN-FIST, 1000 Ljubljana (Slovenia); The Faculty of Chemistry and Chemical Technology, University of Ljubljana, 1000 Ljubljana (Slovenia)

    2013-05-24

    Highlights: •The chimeric protein fusing flagellin to the TLR5 ectodomain is constitutively active. •Mutation P736H within the BB-loop of TLR5 TIR domain renders the receptor inactive. •The R90D mutation in flagellin inactivated autoactivation of the chimeric protein. •The 2:2 stoichiometry of the TLR5:flagellin complex is physiologically relevant. -- Abstract: Toll-like receptor 5 (TLR5) recognizes flagellin of most flagellated bacteria, enabling activation of the MyD88-dependent signaling pathway. The recently published crystal structure of a truncated zebrafish TLR5 ectodomain in complex with an inactive flagellin fragment indicated binding of two flagellin molecules to a TLR5 homodimer, however this complex did not dimerize in solution. In the present study, we aimed to determine the physiological stoichiometry of TLR5:flagellin activation by the use of a chimeric protein composed of an active flagellin fragment linked to the N-terminus of human TLR5 (SF-TLR5). This construct was constitutively active. Inactivation by the R90D mutation within flagellin demonstrated that autoactivation of the chimeric protein depended solely on the specific interaction between TLR5 and flagellin. Addition of wild-type hTLR5 substantially lowered autoactivation of SF-TLR5 in a concentration dependent manner, an effect which was reversible by the addition of exogenous Salmonella typhimurium flagellin, indicating the biological activity of a TLR5:flagellin complex with a 2:2 stoichiometry. These results, in addition to the combinations of inactive P736H mutation within the BB-loop of the TIR domain of TLR5 and SF-TLR5, further confirm the mechanism of TLR5 activation.

  5. Consequences of flagellin export through the type III secretion system of Pseudomonas syringae reveal a major difference in the innate immune systems of mammals and the model plant Nicotiana benthamiana.

    Science.gov (United States)

    Wei, Hai-Lei; Chakravarthy, Suma; Worley, Jay N; Collmer, Alan

    2013-04-01

    Bacterial flagellin is perceived as a microbe (or pathogen)-associated molecular pattern (MAMP or PAMP) by the extracellular pattern recognition receptors, FLS2 and TLR5, of plants and mammals respectively. Flagellin accidently translocated into mammalian cells by pathogen type III secretion systems (T3SSs) is recognized by nucleotide-binding leucine-rich repeat receptor NLRC4 as a pattern of pathogenesis and induces a death-associated immune response. The non-pathogen Pseudomonas fluorescens Pf0-1, expressing a Pseudomonas syringae T3SS, and the plant pathogen P. syringae pv. tomato DC3000 were used to seek evidence of an analogous cytoplasmic recognition system for flagellin in the model plant Nicotiana benthamiana. Flagellin (FliC) was secreted in culture and translocated into plant cells by the T3SS expressed in Pf0-1 and DC3000 and in their ΔflgGHI flagellar pathway mutants. ΔfliC and ΔflgGHI mutants of Pf0-1 and DC3000 were strongly reduced in elicitation of reactive oxygen species production and in immunity induction as indicated by the ability of challenge bacteria inoculated 6 h later to translocate a type III effector-reporter and to elicit effector-triggered cell death. Agrobacterium-mediated transient expression in N. benthamiana of FliC with or without a eukaryotic export signal peptide, coupled with virus-induced gene silencing of FLS2, revealed no immune response that was not FLS2 dependent. Transiently expressed FliC from DC3000 and Pectobacterium carotovorum did notinduce cell death in N. benthamiana, tobacco or tomato leaves. Flagellin is the major Pseudomonas MAMP perceived by N. benthamiana, and although flagellin secretion through the plant cell wall by the T3SS may partially contribute to FLS2-dependent immunity, flagellin in the cytosol does not elicit immune-associated cell death. We postulate that a death response to translocated MAMPs would produce vulnerability to the many necrotrophic pathogens of plants, such as P

  6. Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion

    Science.gov (United States)

    Radomska, Katarzyna A.; Wösten, Marc M. S. M.; Ordoñez, Soledad R.; Wagenaar, Jaap A.; van Putten, Jos P. M.

    2017-01-01

    Flagella-driven motility enables bacteria to reach their favorable niche within the host. The human foodborne pathogen Campylobacter jejuni produces two heavily glycosylated structural flagellins (FlaA and FlaB) that form the flagellar filament. It also encodes the non-structural FlaC flagellin which is secreted through the flagellum and has been implicated in host cell invasion. The mechanisms that regulate C. jejuni flagellin biogenesis and guide the proteins to the export apparatus are different from those in most other enteropathogens and are not fully understood. This work demonstrates the importance of the putative flagellar protein FliS in C. jejuni flagella assembly. A constructed fliS knockout strain was non-motile, displayed reduced levels of FlaA/B and FlaC flagellin, and carried severely truncated flagella. Pull-down and Far Western blot assays showed direct interaction of FliS with all three C. jejuni flagellins (FlaA, FlaB, and FlaC). This is in contrast to, the sensor and regulator of intracellular flagellin levels, FliW, which bound to FlaA and FlaB but not to FlaC. The FliS protein but not FliW preferred binding to glycosylated C. jejuni flagellins rather than to their non-glycosylated recombinant counterparts. Mapping of the binding region of FliS and FliW using a set of flagellin fragments showed that the C-terminal subdomain of the flagellin was required for FliS binding, whereas the N-terminal subdomain was essential for FliW binding. The separate binding subdomains required for FliS and FliW, the different substrate specificity, and the differential preference for binding of glycosylated flagellins ensure optimal processing and assembly of the C. jejuni flagellins. PMID:28659885

  7. Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion

    Directory of Open Access Journals (Sweden)

    Katarzyna A. Radomska

    2017-06-01

    Full Text Available Flagella-driven motility enables bacteria to reach their favorable niche within the host. The human foodborne pathogen Campylobacter jejuni produces two heavily glycosylated structural flagellins (FlaA and FlaB that form the flagellar filament. It also encodes the non-structural FlaC flagellin which is secreted through the flagellum and has been implicated in host cell invasion. The mechanisms that regulate C. jejuni flagellin biogenesis and guide the proteins to the export apparatus are different from those in most other enteropathogens and are not fully understood. This work demonstrates the importance of the putative flagellar protein FliS in C. jejuni flagella assembly. A constructed fliS knockout strain was non-motile, displayed reduced levels of FlaA/B and FlaC flagellin, and carried severely truncated flagella. Pull-down and Far Western blot assays showed direct interaction of FliS with all three C. jejuni flagellins (FlaA, FlaB, and FlaC. This is in contrast to, the sensor and regulator of intracellular flagellin levels, FliW, which bound to FlaA and FlaB but not to FlaC. The FliS protein but not FliW preferred binding to glycosylated C. jejuni flagellins rather than to their non-glycosylated recombinant counterparts. Mapping of the binding region of FliS and FliW using a set of flagellin fragments showed that the C-terminal subdomain of the flagellin was required for FliS binding, whereas the N-terminal subdomain was essential for FliW binding. The separate binding subdomains required for FliS and FliW, the different substrate specificity, and the differential preference for binding of glycosylated flagellins ensure optimal processing and assembly of the C. jejuni flagellins.

  8. Adjuvant role of Pseudomonas flagellin for Acinetobacter baumannii biofilm associated protein

    Science.gov (United States)

    Sefidi, Mozhgan Derakhshan; Rasooli, Iraj; Owlia, Parviz; Talei, Daryush; Astaneh, Shakiba Darvish Alipour; Nazarian, Shahram

    2016-01-01

    AIM To study immunogenicity of Pseudomonas N terminal flagellin as an adjuvant for Acinetobacter baumannii (A. baumannii) biofilm associated protein (Bap). METHODS The N terminal flagellin gene was amplified. The pET28a (+) and polymerase chain reaction products were digested with HindIII and EcoR I. The ligation of N terminal flagellin into pET28a (‏+) was performed using T4 DNA ligase and was then transformed into Escherichia coli BL21 (DE3) as a suitable expression host. pET28a (‏+) vector harboring a conserved region of Bap from our previous work was used. The recombinant proteins were expressed, analyzed by SDS-PAGE method and was purified by affinity chromatography with His-Tag residues followed by confirmation with western blotting. Mice were immunized with recombinant N terminal flagellin and Bap subunits. The immunized animals were intranasally (i.n) challenged with A. baumannii and Pseudomonas aeruginosa (P. aeruginosa). RESULTS The flagellin enhanced the immunogenicity of Bap causing an increase in specific IgG titers in serum (P Bap-Flagellin immunized group challenged with A. baumannii showed significantly lower bacterial load compared to the control group. The bacterial loads were studied in internal organs. A. baumannii infected immunized animals with Bap-Flagellin exhibited internal organs with minor bacterial load while P. aeruginosa PAO1 infected group showed heavy bacterial load of (4.3 ± 0.12) × 106, (1.1 ± 0.01) × 106 and (2.2 ± 0.22) × 106 per gram of lungs, liver and spleen respectively. Bacterial loads were detected per gram of lungs, liver and spleen of the mice group immunized with Bap were (1.2 ± 0.06) × 107, (11.1 ± 0.041) × 105 and (3.6 ± 0.42) × 106 respectively. In vivo neutralization assay indicated that all experimental mice groups, except for Flagellin administered group was significantly (P < 0.05) protected against A. baumannii. CONCLUSION These results demonstrate that P. aeruginosa Flagellin as an adjuvant for

  9. DNA-Encoded Flagellin Activates Toll-Like Receptor 5 (TLR5, Nod-like Receptor Family CARD Domain-Containing Protein 4 (NRLC4, and Acts as an Epidermal, Systemic, and Mucosal-Adjuvant

    Directory of Open Access Journals (Sweden)

    Steven E. Applequist

    2013-09-01

    Full Text Available Eliciting effective immune responses using non-living/replicating DNA vaccines is a significant challenge. We have previously shown that ballistic dermal plasmid DNA-encoded flagellin (FliC promotes humoral as well as cellular immunity to co-delivered antigens. Here, we observe that a plasmid encoding secreted FliC (pFliC(-gly produces flagellin capable of activating two innate immune receptors known to detect flagellin; Toll-like Receptor 5 (TLR5 and Nod-like Receptor family CARD domain-containing protein 4 (NRLC4. To test the ability of pFliC(-gly to act as an adjuvant we immunized mice with plasmid encoding secreted FliC (pFliC(-gly and plasmid encoding a model antigen (ovalbumin by three different immunization routes representative of dermal, systemic, and mucosal tissues. By all three routes we observed increases in antigen-specific antibodies in serum as well as MHC Class I-dependent cellular immune responses when pFliC(-gly adjuvant was added. Additionally, we were able to induce mucosal antibody responses and Class II-dependent cellular immune responses after mucosal vaccination with pFliC(-gly. Humoral immune responses elicited by heterologus prime-boost immunization with a plasmid encoding HIV-1 from gp160 followed by protein boosting could be enhanced by use of pFliC(-gly. We also observed enhancement of cross-clade reactive IgA as well as a broadening of B cell epitope reactivity. These observations indicate that plasmid-encoded secreted flagellin can activate multiple innate immune responses and function as an adjuvant to non-living/replicating DNA immunizations. Moreover, the capacity to elicit mucosal immune responses, in addition to dermal and systemic properties, demonstrates the potential of flagellin to be used with vaccines designed to be delivered by various routes.

  10. Flagellin peptide flg22 gains access to long-distance trafficking in Arabidopsis via its receptor, FLS2

    Energy Technology Data Exchange (ETDEWEB)

    Jelenska, Joanna [University of Chicago; Greenberg, Jean T. [University of Chicago; Mirzadeh, Saed [ORNL; Davern, Sandra M. [ORNL; Standaert, Robert F. [ORNL

    2017-03-01

    Diverse pathogen-derived molecules, such as bacterial flagellin and its conserved peptide flg22, are recognized in plants via plasma membrane receptors and induce both local and systemic immune responses. The fate of such ligands was unknown: whether and by what mechanism(s) they enter plant cells and whether they are transported to distal tissues. We used biologically active fluorophore and radiolabeled peptides to establish that flg22 moves to distal organs with the closest vascular connections. Remarkably, entry into the plant cell via endocytosis together with the FLS2 receptor is needed for delivery to vascular tissue and long-distance transport of flg22. This contrasts with known routes of long distance transport of other non-cell-permeant molecules in plants, which require membrane-localized transporters for entry to vascular tissue. Thus, a plasma membrane receptor acts as a transporter to enable access of its ligand to distal trafficking routes.

  11. Hamiltonian[k,k+1]-因子%Hamiltonian [k, k + 1]-Factor

    Institute of Scientific and Technical Information of China (English)

    蔡茂诚; 方奇志; 李延军

    2003-01-01

    A Hamiltonian [k, k + 1]-factor is a [k, k + 1]-factor containing a Hamiltonian cycle. A simple graph G of order n is n/2-critical if δ(G) ≥ n/2 but δ(G - e) < n/2 for any edge e ∈ E(G). Let k ≥ 2 be an integer and G be an n/2-critical graph with n ≥ 4k - 6 and n ≥ 7. In this paper it is proved that for any given Hamiltonian cycle C of G, G has a [k, k + 1]-factor containing C. This result is an improvement on some recent results about the existence of Hamiltonian [k, k + 1]-factor.%本文考虑n/2-临界图中Hamiltonian[k,k+1]-因子的存在性.Hamiltonian[k,k+1]-因子是指包含Hamiltonian圈的[k,k+1]-因子;给定阶数为n的简单图G,若δ(G)≥n/2而δ(G\\e)<n/2(对任意的e∈E(G)),则称G为n/2-临界图.设k为大于等于2的整数,G为n/2-临界图(其中n≥4k-6且n≥7),我们证明了对于G的任何Hamiltonian圈C,G中必存在包含C的[k,k+1]-因子.该结果改进了现有的一些有关Hamiltonian[k,k+1]-因子存在性的结果.

  12. Biochemical and immunological analyses of the flagellin of Clostridium tyrobutyricum ATCC 25755.

    Science.gov (United States)

    Arnold, F; Bédouet, L; Batina, P; Robreau, G; Talbot, F; Lécher, P; Malcoste, R

    1998-01-01

    The monoclonal antibody 21E7-B12 (IgG3) can be used in a direct method of Clostridium tyrobutyricum detection based on an immunoenzymatic assay. Immunoelectron microscopy demonstrated that the 21E7-B12 antibody recognized the surface-exposed epitopes on the flagellar filaments of C. tyrobutyricum. After flagellar extraction, the purified flagellin showed an apparent molecular mass of 46 kDa with an isoelectric point of 3.6. Sugar staining, mild periodate oxidation and beta-elimination experiments showed that the flagellin was glycosylated and that the 21E7-B12 epitope was located in the sugar moiety. Amino acid composition showed that the flagellar filament protein contained a high percentage of serine and threonine, while proline was absent. The first 23 residues of the N-terminal were determined and sequence homology with other flagellins was found.

  13. Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus

    Science.gov (United States)

    Yuan, Ye; Wang, Xiuli; Guo, Sheping; Liu, Yang; Ge, Hui; Qiu, Xuemei

    2010-11-01

    The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. Bacteria flagellins play an important role during infection and induction of the host immune response. Thus, flagellin proteins are an ideal target for vaccines. We amplified the complete flagellin subunit gene ( flaA) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 62.78 kDa. We purified and characterized the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for further studies into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus. In addition, the purified FlaA protein can be used for further functional and structural studies.

  14. P70S6K 1 regulation of angiogenesis through VEGF and HIF-1{alpha} expression

    Energy Technology Data Exchange (ETDEWEB)

    Bian, Chuan-Xiu; Shi, Zhumei [Department of Pathology, Cancer Center, Nanjing Medical University, Nanjing 210029 (China); Meng, Qiao; Jiang, Yue; Liu, Ling-Zhi [Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107 (United States); Jiang, Bing-Hua, E-mail: binghjiang@yahoo.com [Department of Pathology, Cancer Center, Nanjing Medical University, Nanjing 210029 (China); Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107 (United States)

    2010-07-30

    Research highlights: {yields} P70S6K1 regulates VEGF expression; {yields} P70S6K1 induces transcriptional activation through HIF-1{alpha} binding site; {yields} P70S6K1 regulates HIF-1{alpha}, but not HIF-1{beta} protein expression; {yields} P70S6K1 mediates tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression. -- Abstract: The 70 kDa ribosomal S6 kinase 1 (p70S6K1), a downstream target of phosphoinositide 3-kinase (PI3K) and ERK mitogen-activated protein kinase (MAPK), is an important regulator of cell cycle progression, and cell proliferation. Recent studies indicated an important role of p70S6K1 in PTEN-negative and AKT-overexpressing tumors. However, the mechanism of p70S6K1 in tumor angiogenesis remains to be elucidated. In this study, we specifically inhibited p70S6K1 activity in ovarian cancer cells using vector-based small interfering RNA (siRNA) against p70S6K1. We found that knockdown of p70S6K1 significantly decreased VEGF protein expression and VEGF transcriptional activation through the HIF-1{alpha} binding site at its enhancer region. The expression of p70S6K1 siRNA specifically inhibited HIF-1{alpha}, but not HIF-1{beta} protein expression. We also found that p70S6K1 down-regulation inhibited ovarian tumor growth and angiogenesis, and decreased cell proliferation and levels of VEGF and HIF-1{alpha} expression in tumor tissues. Our results suggest that p70S6K1 is required for tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression, providing a molecular mechanism of human ovarian cancer mediated by p70S6K1 signaling.

  15. Hyperactive S6K1 mediates oxidative stress and endothelial dysfunction in aging: inhibition by resveratrol.

    Science.gov (United States)

    Rajapakse, Angana G; Yepuri, Gautham; Carvas, João M; Stein, Sokrates; Matter, Christian M; Scerri, Isabelle; Ruffieux, Jean; Montani, Jean-Pierre; Ming, Xiu-Fen; Yang, Zhihong

    2011-04-22

    Mammalian target of rapamycin (mTOR)/S6K1 signalling emerges as a critical regulator of aging. Yet, a role of mTOR/S6K1 in aging-associated vascular endothelial dysfunction remains unknown. In this study, we investigated the role of S6K1 in aging-associated endothelial dysfunction and effects of the polyphenol resveratrol on S6K1 in aging endothelial cells. We show here that senescent endothelial cells displayed higher S6K1 activity, increased superoxide production and decreased bioactive nitric oxide (NO) levels than young endothelial cells, which is contributed by eNOS uncoupling. Silencing S6K1 in senescent cells reduced superoxide generation and enhanced NO production. Conversely, over-expression of a constitutively active S6K1 mutant in young endothelial cells mimicked endothelial dysfunction of the senescent cells through eNOS uncoupling and induced premature cellular senescence. Like the mTOR/S6K1 inhibitor rapamycin, resveratrol inhibited S6K1 signalling, resulting in decreased superoxide generation and enhanced NO levels in the senescent cells. Consistent with the data from cultured cells, an enhanced S6K1 activity, increased superoxide generation, and decreased bioactive NO levels associated with eNOS uncoupling were also detected in aortas of old WKY rats (aged 20-24 months) as compared to the young animals (1-3 months). Treatment of aortas of old rats with resveratrol or rapamycin inhibited S6K1 activity, oxidative stress, and improved endothelial NO production. Our data demonstrate a causal role of the hyperactive S6K1 in eNOS uncoupling leading to endothelial dysfunction and vascular aging. Resveratrol improves endothelial function in aging, at least in part, through inhibition of S6K1. Targeting S6K1 may thus represent a novel therapeutic approach for aging-associated vascular disease.

  16. Immunoglobulin A antibodies directed against Campylobacter jejuni flagellin present in breast-milk.

    OpenAIRE

    Nachamkin, I; Fischer, S H; Yang, X. H.; Benitez, O.; Cravioto, A.

    1994-01-01

    We studied the relationship between IgA anti-campylobacter flagellin antibodies in breast milk samples and protection of breastfed infants living in a rural Mexican village from campylobacter infection. There were fewer episodes of campylobacter infection (symptomatic and asymptomatic combined) in infants breastfed with milk containing specific anti-flagellin antibodies (1.2/child/year, 95% CI 0.6-1.8) versus non-breastfed children (3.3/child/year, 95% CI 1.8-4.8; P < 0.01). Infants breastfed...

  17. Expression of recombinant Newcastle disease virus F protein in Pichia pastoris and its immunogenicity using flagellin as the adjuvant.

    Science.gov (United States)

    Kang, Xilong; Wang, Jing; Jiao, Yang; Tang, Peipei; Song, Li; Xiong, Dan; Yin, Yuelan; Pan, Zhiming; Jiao, Xinan

    2016-12-01

    Newcastle disease (ND), a highly contagious, acute, and potent infectious disease caused by Newcastle disease virus (NDV), has a considerable impact on the global poultry industry. Although both live attenuated and inactivated vaccines are used to prevent and control the spread of ND among chickens, the increasing number of ND outbreaks in commercial poultry flocks worldwide indicates that routine vaccinations are insufficient to control ND. Hence, efforts are being invested into developing alternative and more effective vaccination strategies. In this study, we focus on F protein, the neutralizing and protective antigen of NDV, and flagellin (FliC), a toll-like receptor 5 (TLR5) agonist that is an effective inducer of innate immune responses. We amplified F gene from velogenic NDV strain F48E8. The recombinant histidine (His)-tagged F protein was efficiently expressed in a Pichia pastoris (P. pastoris) eukaryotic system and verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. The conditions for F protein expression in P. pastoris were optimal. The immunogenicity of F protein with FliC as the adjuvant was evaluated in a C3H/HeJ mouse model. FliC was found to enhance both F-specific and NDV-specific IgG responses and F-specific cellular immune responses following intraperitoneal co-administration with F protein. Thus, the recombinant F protein expressed by P. pastoris when used with flagellin as the adjuvant has potential as a subunit vaccine candidate. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Reptile Toll-like receptor 5 unveils adaptive evolution of bacterial flagellin recognition.

    Science.gov (United States)

    Voogdt, Carlos G P; Bouwman, Lieneke I; Kik, Marja J L; Wagenaar, Jaap A; van Putten, Jos P M

    2016-01-07

    Toll-like receptors (TLR) are ancient innate immune receptors crucial for immune homeostasis and protection against infection. TLRs are present in mammals, birds, amphibians and fish but have not been functionally characterized in reptiles despite the central position of this animal class in vertebrate evolution. Here we report the cloning, characterization, and function of TLR5 of the reptile Anolis carolinensis (Green Anole lizard). The receptor (acTLR5) displays the typical TLR protein architecture with 22 extracellular leucine rich repeats flanked by a N- and C-terminal leucine rich repeat domain, a membrane-spanning region, and an intracellular TIR domain. The receptor is phylogenetically most similar to TLR5 of birds and most distant to fish TLR5. Transcript analysis revealed acTLR5 expression in multiple lizard tissues. Stimulation of acTLR5 with TLR ligands demonstrated unique responsiveness towards bacterial flagellin in both reptile and human cells. Comparison of acTLR5 and human TLR5 using purified flagellins revealed differential sensitivity to Pseudomonas but not Salmonella flagellin, indicating development of species-specific flagellin recognition during the divergent evolution of mammals and reptiles. Our discovery of reptile TLR5 fills the evolutionary gap regarding TLR conservation across vertebrates and provides novel insights in functional evolution of host-microbe interactions.

  19. Effects of recombinant flagellin B and its ND1 domain from Vibrio anguillarum on macrophages from gilthead seabream (Sparus aurata L.) and rainbow trout (Oncorhynchus mykiss, W.).

    Science.gov (United States)

    González-Stegmaier, Roxana; Romero, Alex; Estepa, Amparo; Montero, Jana; Mulero, Victoriano; Mercado, Luis

    2015-01-01

    Flagellin is the principal component of flagellum in Gram negative and positive bacteria, and it is also the ligand that activates the Toll-like receptor 5 (TLR5) in mammals and fish. In higher vertebrates, flagellin induces the activation of the membrane-bound TLR5 (TLR5M), which promotes the expression of proinflammatory cytokines and chemokines and the co-stimulatory molecules present in antigen-presenting cells needed for the activation of T cells. In the present study, we report the production of two recombinant proteins of Vibrio anguillarum: i) a full length flagellin B (FlaB) (rFla) and ii) the amino-terminus of the D1 domain (rND1) of the same protein, the region mainly responsible for binding to TLR5 and for the immunostimulatory activity of flagellin. The effects of these recombinant proteins were assessed in vitro using head kidney macrophages of gilthead seabream (Sparus aurata L., Perciformes, Sparidae) and rainbow trout (Oncorhynchus mykiss W., Salmoniformes, Salmonidae). In both species, 3 h of stimulation with rFla and rND1 induced expression of the proinflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), and of the chemokine IL-8. In gilthead seabream macrophages stimulated with rFla and rND1, a 900- and 6-fold increase were observed for IL-1β transcription, while a 900- and 3-fold increase were recorded for IL-8 transcription, respectively, as compared to non-stimulated macrophages. In rainbow trout, rFla increased expression of IL-8 40-fold in macrophages, whereas rND1 increased expression of the chemokine 3-fold, as compared to non-stimulated cells. The results obtained for rFla and rND1 demonstrate their modulatory capabilities in vitro, suggesting that rFla and rND1 could be evaluated as immunostimulatory candidates for use in farmed fish. However, further in vivo studies are needed to confirm and expand on the present results.

  20. Role of GluK1 kainate receptors in seizures, epileptic discharges, and epileptogenesis.

    Science.gov (United States)

    Fritsch, Brita; Reis, Janine; Gasior, Maciej; Kaminski, Rafal M; Rogawski, Michael A

    2014-04-23

    Kainate receptors containing the GluK1 subunit have an impact on excitatory and inhibitory neurotransmission in brain regions, such as the amygdala and hippocampus, which are relevant to seizures and epilepsy. Here we used 2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA), a potent and selective agonist of kainate receptors that include the GluK1 subunit, in conjunction with mice deficient in GluK1 and GluK2 kainate receptor subunits to assess the role of GluK1 kainate receptors in provoking seizures and in kindling epileptogenesis. We found that systemic ATPA, acting specifically via GluK1 kainate receptors, causes locomotor arrest and forelimb extension (a unique behavioral characteristic of GluK1 activation) and induces myoclonic behavioral seizures and electrographic seizure discharges in the BLA and hippocampus. In contrast, the proconvulsant activity of systemic AMPA, kainate, and pentylenetetrazol is not mediated by GluK1 kainate receptors, and deletion of these receptors does not elevate the threshold for seizures in the 6 Hz model. ATPA also specifically activates epileptiform discharges in BLA slices in vitro via GluK1 kainate receptors. Olfactory bulb kindling developed similarly in wild-type, GluK1, and GluK2 knock-out mice, demonstrating that GluK1 kainate receptors are not required for epileptogenesis or seizure expression in this model. We conclude that selective activation of kainate receptors containing the GluK1 subunit can trigger seizures, but these receptors are not necessary for seizure generation in models commonly used to identify therapeutic agents for the treatment of epilepsy.

  1. Plasmid Transfer of Plasminogen K1-5 Reduces Subcutaneous Hepatoma Growth by Affecting Inflammatory Factors

    Directory of Open Access Journals (Sweden)

    Lea A. Koch

    2014-01-01

    Full Text Available There is evidence that plasminogen K1-5 (PlgK1-5 directly affects tumour cells and inflammation. Therefore, we analysed if PlgK1-5 has immediate effects on hepatoma cells and inflammatory factors in vitro and in vivo. In vitro, effects of plasmid encoding PlgK1-5 (pK1-5 on Hepa129, Hepa1-6, and HuH7 cell viability, apoptosis, and proliferation as well as VEGF and TNF-alpha expression and STAT3-phosphorylation were investigated. In vivo, tumour growth, proliferation, vessel density, and effects on vascular endothelial growth factor (VEGF and tumour necrosis factor alpha (TNF-alpha expression were examined following treatment with pK1-5. In vivo, pK1-5 halved cell viability; cell death was increased by up to 15% compared to the corresponding controls. Proliferation was not affected. VEGF, TNF-alpha, and STAT3-phosphorylation were affected following treatment with pK1-5. In vivo, ten days after treatment initiation, pK1-5 reduced subcutaneous tumour growth by 32% and mitosis by up to 77% compared to the controls. Vessel density was reduced by 50%. TNF-alpha levels in tumour and liver tissue were increased, whereas VEGF levels in tumours and livers were reduced after pK1-5 treatment. Taken together, plasmid gene transfer of PlgK1-5 inhibits hepatoma (cell growth not only by reducing vessel density but also by inducing apoptosis, inhibiting proliferation, and triggering inflammation.

  2. The destructive citrus pathogen, 'Candidatus Liberibacter asiaticus' encodes a functional flagellin characteristic of a pathogen-associated molecular pattern.

    Directory of Open Access Journals (Sweden)

    Huasong Zou

    Full Text Available Huanglongbing (HLB is presently the most devastating citrus disease worldwide. As an intracellular plant pathogen and insect symbiont, the HLB bacterium, 'Candidatus Liberibacter asiaticus' (Las, retains the entire flagellum-encoding gene cluster in its significantly reduced genome. Las encodes a flagellin and hook-associated protein (Fla of 452 amino acids that contains a conserved 22 amino acid domain (flg22 at positions 29 to 50 in the N-terminus. The phenotypic alteration in motility of a Sinorhizobium meliloti mutant lacking the fla genes was partially restored by constitutive expression of Fla(Las. Agrobacterium-mediated transient expression in planta revealed that Fla(Las induced cell death and callose deposition in Nicotiana benthamiana, and that the transcription of BAK1 and SGT1, which are associated with plant innate immunity, was upregulated. Amino acid substitution experiments revealed that residues 38 (serine and 39 (aspartate of Fla(Las were essential for callose induction. The synthetic flg22(Las peptide could not induce plant cell death but retained the ability to induce callose deposition at a concentration of 20 µM or above. This demonstrated that the pathogen-associated molecular pattern (PAMP activity of flg22 in Las was weaker than those in other well-studied plant pathogenic bacteria. These results indicate that Fla(Las acts as a PAMP and may play an important role in triggering host plant resistance to the HLB bacteria.

  3. Flagellin and outer surface proteins from Borrelia burgdorferi are not glycosylated

    OpenAIRE

    Štěrba, Ján

    2012-01-01

    Glycosylation of four proteins from Borrelia burgdorferi s.s. was investigated ? flagellins FlaA, FlaB, and outer surface proteins OspA and OspB. Glycosylation of these four proteins was not proved by any of the used techniques. However, other glycan-staining positive proteins were present in the borrelia samples. These proteins were suggested to originate in the culture medium.

  4. Distribution and Polymorphism of the Flagellin Genes from Isolates of Campylobacter coli and Campylobacter jejuni

    Science.gov (United States)

    1993-05-01

    American Society for Microbioloc% Distribution and Polymorphism of the Flagellin Genes from Isolates of Campylobacter coli and Campylobacter jejuni RICHARD...in Campylobacter jejuni . serogroups both the flaA and flaB genes are extremely Mol. M;crobiol. 5:1151-1158. z homologous. Within most LIO heat-labile...irllwn hungatei. J1. Bacteriol. 123:-28 proteins of Campylobacter jejuni 81116. Infect. Immun. 59: 42. Thomashow, L S., and S. C. Rittenberg. 198

  5. Residue Specific and Chirality Dependent Interactions between Carbon Nanotubes and Flagellin.

    Science.gov (United States)

    Macwan, Isaac G; Zhao, Zihe; Sobh, Omar T; Mukerji, Ishita; Dharmadhikari, Bhushan; Patra, Prabir K

    2016-01-01

    Flagellum is a lash-like cellular appendage found in many single-celled living organisms. The flagellin protofilaments contain 11-helix dual turn structure in a single flagellum. Each flagellin consists of four sub-domains - two inner domains (D0, D1) and two outer domains (D2, D3). While inner domains predominantly consist of α-helices, the outer domains are primarily beta sheets with D3. In flagellum, the outermost sub-domain is the only one that is exposed to the native environment. This study focuses on the interactions of the residues of D3 of an R-type flagellin with 5nm long chiral (5,15) and arm-chair (12,12) single-walled carbon nanotubes (SWNT) using molecular dynamics simulation. It presents the interactive forces between the SWNT and the residues of D3 from the perspectives of size and chirality of the SWNT. It is found that the metallic (arm-chair) SWNT interacts the most with glycine and threonine residues through van der Waals and hydrophobic interactions, whereas the semiconducting (chiral) SWNT interacts largely with the area of protein devoid of glycine by van der Waals, hydrophobic interactions, and hydrogen bonding. This indicates a crucial role that glycine plays in distinguishing metallic from semiconducting SWNTs.

  6. Exposure to bacterial products lipopolysaccharide and flagellin and hepatocellular carcinoma: a nested case-control study.

    Science.gov (United States)

    Fedirko, Veronika; Tran, Hao Quang; Gewirtz, Andrew T; Stepien, Magdalena; Trichopoulou, Antonia; Aleksandrova, Krasimira; Olsen, Anja; Tjønneland, Anne; Overvad, Kim; Carbonnel, Franck; Boutron-Ruault, Marie-Christine; Severi, Gianluca; Kühn, Tilman; Kaaks, Rudolf; Boeing, Heiner; Bamia, Christina; Lagiou, Pagona; Grioni, Sara; Panico, Salvatore; Palli, Domenico; Tumino, Rosario; Naccarati, Alessio; Peeters, Petra H; Bueno-de-Mesquita, H B; Weiderpass, Elisabete; Castaño, José María Huerta; Barricarte, Aurelio; Sánchez, María-José; Dorronsoro, Miren; Quirós, J Ramón; Agudo, Antonio; Sjöberg, Klas; Ohlsson, Bodil; Hemmingsson, Oskar; Werner, Mårten; Bradbury, Kathryn E; Khaw, Kay-Tee; Wareham, Nick; Tsilidis, Konstantinos K; Aune, Dagfinn; Scalbert, Augustin; Romieu, Isabelle; Riboli, Elio; Jenab, Mazda

    2017-04-04

    Leakage of bacterial products across the gut barrier may play a role in liver diseases which often precede the development of liver cancer. However, human studies, particularly from prospective settings, are lacking. We used a case-control study design nested within a large prospective cohort to assess the association between circulating levels of anti-lipopolysaccharide (LPS) and anti-flagellin immunoglobulin A (IgA) and G (IgG) (reflecting long-term exposures to LPS and flagellin, respectively) and risk of hepatocellular carcinoma. A total of 139 men and women diagnosed with hepatocellular carcinoma between 1992 and 2010 were matched to 139 control subjects. Multivariable rate ratios (RRs), including adjustment for potential confounders, hepatitis B/C positivity, and degree of liver dysfunction, were calculated with conditional logistic regression. Antibody response to LPS and flagellin was associated with a statistically significant increase in the risk of hepatocellular carcinoma (highest vs. lowest quartile: RR = 11.76, 95% confidence interval = 1.70-81.40; P trend = 0.021). This finding did not vary substantially by time from enrollment to diagnosis, and did not change after adjustment for chronic infection with hepatitis B and C viruses. These novel findings, based on exposures up to several years prior to diagnosis, support a role for gut-derived bacterial products in hepatocellular carcinoma development. Further study into the role of gut barrier failure and exposure to bacterial products in liver diseases is warranted.

  7. Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

    Directory of Open Access Journals (Sweden)

    Qing Zeng

    2017-09-01

    Full Text Available Escherichia coli (E. coli K1 sepsis and meningitis is a severe infection characterized by high mortality in neonates. Successful colonization and translocation across the intestinal mucosa have been regarded as the critical steps for E. coli K1 sepsis and meningitis. We recently reported that the probiotic mixture, Golden Bifido (containing live Lactobacillus bulgaricus, Bifidobacterium, and Streptococcus thermophilus, LBS has a preventive role against neonatal E. coli K1 bacteremia and meningitis. However, the interaction between the neonatal gut barrier, probiotics and E. coli K1 is still not elucidated. The present study aims to investigate how LBS exerts its protective effects on neonatal gut barrier during E. coli K1 infection. The beneficial effects of LBS were explored in vitro and in vivo using human colon carcinoma cell lines HT-29 and rat model of neonatal E. coli K1 infection, respectively. Our results showed that stimulation with E. coli K1 was able to cause intestinal barrier dysfunction, which were reflected by E. coli K1-induced intestinal damage and apoptosis of intestinal epithelial cells, reduction of mucin, immunoglobulin A (IgA and tight junction proteins expression, as well as increase in intestinal permeability, all these changes facilitate E. coli K1 intestinal translocation. However, these changes were alleviated when HT-29 cells were treated with LBS before E. coli K1 infection. Furthermore, we found that LBS-treated neonatal rats (without E. coli K1 infection have showed higher production of mucin, ZO-1, IgA, Ki67 in intestinal mucosa as well as lower intestinal permeability than that of non-treated rats, indicating that LBS could accelerate the development of neonatal intestinal defense. Taken together, our results suggest that enhancement of the neonatal intestinal defense to fight against E. coli K1 translocation could be the potential mechanism to elucidate how LBS confers a protective effect against neonatal E

  8. The bacterial elicitor flagellin activates its receptor in tomato cells according to the address-message concept.

    Science.gov (United States)

    Meindl, T; Boller, T; Felix, G

    2000-09-01

    flg22, a peptide corresponding to the most conserved domain of bacterial flagellin, acts as a potent elicitor in plants. Here, we have used an iodinated derivative of flg22 ((125)I-labeled Tyr-flg22) as a molecular probe for the flagellin receptor in tomato cells. This radioligand showed rapid binding to a single class of specific, saturable, high-affinity receptor sites in intact cells and membrane preparations. Binding, although essentially nonreversible under physiological conditions, was not covalent, and chemical cross-linking was required to specifically label a single polypeptide of 115 kD. Intact flagellin and elicitor-active flagellin peptides but not biologically inactive analogs efficiently competed for binding of radioligand. Peptides lacking the C terminus of the conserved domain, previously found to act as competitive antagonists of elicitor action in tomato cells, also competed for binding of radioligand. Thus, this novel, high-affinity binding site exhibited all the characteristics expected of a functional receptor of bacterial flagellin. For a model of receptor activation, we propose a two-step mechanism according to the address-message concept, in which binding of the N terminus (address) is the first step and activation of responses with the C terminus (message) is the second step.

  9. Characterization of pro-inflammatory flagellin proteins produced by Lactobacillus ruminis and related motile Lactobacilli.

    Directory of Open Access Journals (Sweden)

    B Anne Neville

    Full Text Available Lactobacillus ruminis is one of at least twelve motile but poorly characterized species found in the genus Lactobacillus. Of these, only L. ruminis has been isolated from mammals, and this species may be considered as an autochthonous member of the gastrointestinal microbiota of humans, pigs and cows. Nine L. ruminis strains were investigated here to elucidate the biochemistry and genetics of Lactobacillus motility. Six strains isolated from humans were non-motile while three bovine isolates were motile. A complete set of flagellum biogenesis genes was annotated in the sequenced genomes of two strains, ATCC25644 (human isolate and ATCC27782 (bovine isolate, but only the latter strain produced flagella. Comparison of the L. ruminis and L. mali DSM20444(T motility loci showed that their genetic content and gene-order were broadly similar, although the L. mali motility locus was interrupted by an 11.8 Kb region encoding rhamnose utilization genes that is absent from the L. ruminis motility locus. Phylogenetic analysis of 39 motile bacteria indicated that Lactobacillus motility genes were most closely related to those of motile carnobacteria and enterococci. Transcriptome analysis revealed that motility genes were transcribed at a significantly higher level in motile L. ruminis ATCC27782 than in non-motile ATCC25644. Flagellin proteins were isolated from L. ruminis ATCC27782 and from three other Lactobacillus species, while recombinant flagellin of aflagellate L. ruminis ATCC25644 was expressed and purified from E. coli. These native and recombinant Lactobacillus flagellins, and also flagellate L. ruminis cells, triggered interleukin-8 production in cultured human intestinal epithelial cells in a manner suppressed by short interfering RNA directed against Toll-Like Receptor 5. This study provides genetic, transcriptomic, phylogenetic and immunological insights into the trait of flagellum-mediated motility in the lactobacilli.

  10. Combining Flagellin and human β-defensin-3 to combat bacterial Infections

    Directory of Open Access Journals (Sweden)

    Ofra eSabag

    2014-12-01

    Full Text Available The discovery and therapeutic use of antibiotics made a major contribution to the reduction of human morbidity and mortality. However, the growing resistance to antibiotics has become a matter of huge concern. In this study we aimed to develop an innovative approach to treat bacterial infections utilizing two components: the human antibacterial peptide β-defensin-3 (BD3 and the bacterial protein flagellin (F. This combination was designed to provide an efficient weapon against bacterial infections with the peptide killing the bacteria directly, while the flagellin protein triggers the immune system and acts against bacteria escaping from the peptide’s action. We designed, expressed and purified the fusion protein FBD3 and its two components, the F protein and the native BD3 peptide. FBD3 fusion protein and native BD3 peptide had antibacterial activity in vitro against various bacterial strains. FBD3 and F proteins could also recognize their receptor expressed on target cells and stimulated secretion of IL-8. In addition, F and FBD3 proteins had a partial protective effect in mice infected by pathogenic E. coli bacteria that cause a lethal disease. Moreover, we were able to show partial protection of mice infected with E. coli using a flagellin sequence from Salmonella.We also explored flagellin’s basic mechanisms of action, focusing on its effects on CD4+ T cells from healthy donors. We found that F stimulation caused an increase in the mRNA levels of the Th1 response cytokines IL12A and IFNγ. In addition, F stimulation affected its own receptor.

  11. Immunogenicity and efficacy of flagellin-envelope fusion dengue vaccines in mice and monkeys.

    Science.gov (United States)

    Liu, Ge; Song, Langzhou; Beasley, David W C; Putnak, Robert; Parent, Jason; Misczak, John; Li, Hong; Reiserova, Lucia; Liu, Xiangyu; Tian, Haijun; Liu, Wenzhe; Labonte, Darlene; Duan, Lihua; Kim, Youngsun; Travalent, Linda; Wigington, Devin; Weaver, Bruce; Tussey, Lynda

    2015-05-01

    The envelope (E) protein of flaviviruses includes three domains, EI, EII, and EIII, and is the major protective antigen. Because EIII is rich in type-specific and subcomplex-specific neutralizing epitopes and is easy to express, it is particularly attractive as a recombinant vaccine antigen. VaxInnate has developed a vaccine platform that genetically links vaccine antigens to bacterial flagellin, a Toll-like receptor 5 ligand. Here we report that tetravalent dengue vaccines (TDVs) consisting of four constructs, each containing two copies of EIII fused to flagellin (R3.2x format), elicited robust and long-lived neutralizing antibodies (geometric mean titers of 200 to 3,000), as measured with a 50% focus reduction neutralization test (FRNT50). In an immunogenicity study, rhesus macaques (n = 2) immunized subcutaneously with 10 μg or 90 μg of TDV three or four times, at 4- to 6-week intervals, developed neutralizing antibodies to four dengue virus (DENV) serotypes (mean post-dose 3 FRNT50 titers of 102 to 601). In an efficacy study, rhesus macaques (n = 4) were immunized intramuscularly with 16 μg or 48 μg of TDV or a placebo control three times, at 1-month intervals. The animals that received 48-μg doses of TDV developed neutralizing antibodies against the four serotypes (geometric mean titers of 49 to 258) and exhibited reduced viremia after DENV-2 challenge, with a group mean viremia duration of 1.25 days and 2 of 4 animals being completely protected, compared to the placebo-treated animals, which all developed viremia, with a mean duration of 4 days. In conclusion, flagellin-EIII fusion vaccines are immunogenic and partially protective in a nonhuman primate model.

  12. The flagellin hypervariable region is a potential flagella display domain in probiotic Escherichia coli strain Nissle 1917.

    Science.gov (United States)

    Yang, Ying; Yang, Yi; Ou, Bingming; Xia, Pengpeng; Zhou, Mingxu; Li, Luan; Zhu, Guoqiang

    2016-09-01

    The most studied probiotic, Escherichia coli strain Nissle 1917 (EcN) possesses flagella of serotype H1. To explore the potential to use EcN flagellin in flagella display applications, we investigated the effect of deleting amino acids in the hypervariable region of flagellin on EcNc (EcN cured of its two cryptic plasmids pMUT1 and pMUT2). Two EcNc flagellin isogenic mutants with deletions of amino acid residual from 277 to 286 and from 287 to 296 in the hypervariable domain were constructed. Both mutants were flagellated, adherent to IPEC-J2 cells, and colonized BALB/c mice. These hypervariable regions may have future utility in the display of heterologous epitopes.

  13. Sphingosine kinase-1 (SphK-1 regulates Mycobacterium smegmatis infection in macrophages.

    Directory of Open Access Journals (Sweden)

    Hridayesh Prakash

    Full Text Available Sphingosine kinase-1 is known to mediate Mycobacterium smegmatis induced inflammatory responses in macrophages, but its role in controlling infection has not been reported to date. We aimed to unravel the significance of SphK-1 in controlling M. smegmatis infection in RAW 264.7 macrophages. Our results demonstrated for the first time that selective inhibition of SphK-1 by either D, L threo dihydrosphingosine (DHS; a competitive inhibitor of Sphk-1 or Sphk-1 siRNA rendered RAW macrophages sensitive to M. smegmatis infection. This was due to the reduction in the expression of iNOs, p38, pp-38, late phagosomal marker, LAMP-2 and stabilization of the RelA (pp-65 subunit of NF-kappaB. This led to a reduction in the generation of NO and secretion of TNF-alpha in infected macrophages. Congruently, overexpression of SphK-1 conferred resistance in macrophages to infection which was due to enhancement in the generation of NO and expression of iNOs, pp38 and LAMP-2. In addition, our results also unraveled a novel regulation of p38MAPK by SphK-1 during M. smegmatis infection and generation of NO in macrophages. Enhanced NO generation and expression of iNOs in SphK-1++ infected macrophages demonstrated their M-1(bright phenotype of these macrophages. These findings thus suggested a novel antimycobacterial role of SphK-1 in macrophages.

  14. 猜想M(2k,k+1)=3k-1+[(k-1)/2]的反例%Counter-Examples to the Conjecture M(2k, k + 1) = 3k - 1 + 「(k-1)/2」

    Institute of Scientific and Technical Information of China (English)

    游林; 王天明

    2002-01-01

    Brualdi 与Jung在[1]中研究了一类具有固定线和k的n×n矩阵上的最大跳跃数M(n,k),M(2k,k+1)=3k-1+[k-1/2]并提出猜想本文给出了这一猜想的两个反例.%The maximum jump number M(n, k) over a class of n × n matrices of zeros and ones with constant row and colunn sum k has been investigated by Brualdi and Jung in [1] where they proposed the conjecture M(2k, k + 1) = 3k - 1 + [k-1/2].In this note, we give two counter-exalnples to this conjecture.

  15. Vaccinia virus K1 ankyrin repeat protein inhibits NF-κB activation by preventing RelA acetylation.

    Science.gov (United States)

    Bravo Cruz, Ariana G; Shisler, Joanna L

    2016-10-01

    The vaccinia virus (VACV) K1 protein inhibits dsRNA-dependent protein kinase (PKR) activation. A consequence of this function is that K1 inhibits PKR-induced NF-κB activation during VACV infection. However, transient expression of K1 also inhibits Toll-like receptor (TLR)-induced NF-κB activation. This suggests that K1 has a second NF-κB inhibitory mechanism that is PKR-independent. This possibility was explored by expressing K1 independently of infection and stimulating NF-κB under conditions that minimized or excluded PKR activation. K1 inhibited both TNF- and phorbol 12-myristate 13-acetate (PMA)-induced NF-κB activation, as detected by transcription of synthetic (e.g. luciferase) and natural (e.g. CXCL8) genes controlled by NF-κB. K1 also inhibited NF-κB activity in PKRkd cells, cells that have greatly decreased amounts of PKR. K1 no longer prevented IκBα degradation or NF-κB nuclear translocation in the absence of PKR, suggesting that K1 acted on a nuclear event. Indeed, K1 was present in the nucleus and cytoplasm of stimulated and unstimulated cells. K1 inhibited acetylation of the RelA (p65) subunit of NF-κB, a nuclear event known to be required for NF-κB activation. Moreover, p65-CBP (CREB-binding protein) interactions were blocked in the presence of K1. However, K1 did not preclude NF-κB binding to oligonucleotides containing κB-binding sites. The current interpretation of these data is that NF-κB-promoter interactions still occur in the presence of K1, but NF-κB cannot properly trigger transcriptional activation because K1 antagonizes acetylation of RelA. Thus, in comparison to all known VACV NF-κB inhibitory proteins, K1 acts at one of the most downstream events of NF-κB activation.

  16. An Aeromonas caviae Genomic Island Is Required for both O-Antigen Lipopolysaccharide Biosynthesis and Flagellin Glycosylation ▿

    OpenAIRE

    Tabei, S. Mohammed B.; Hitchen, Paul G.; Day-Williams, Michaela J.; Merino, Susana; Vart, Richard; Pang, Poh-Choo; Horsburgh, Gavin J.; Viches, Silvia; Wilhelms, Markus; Tomás, Juan M.; Dell, Anne; Shaw, Jonathan G

    2009-01-01

    Aeromonas caviae Sch3N possesses a small genomic island that is involved in both flagellin glycosylation and lipopolysaccharide (LPS) O-antigen biosynthesis. This island appears to have been laterally acquired as it is flanked by insertion element-like sequences and has a much lower G+C content than the average aeromonad G+C content. Most of the gene products encoded by the island are orthologues of proteins that have been shown to be involved in pseudaminic acid biosynthesis and flagellin gl...

  17. Identification of an O-linked repetitive glycan chain of the polar flagellum flagellin of Azospirillum brasilense Sp7.

    Science.gov (United States)

    Belyakov, Alexei Ye; Burygin, Gennady L; Arbatsky, Nikolai P; Shashkov, Alexander S; Selivanov, Nikolai Yu; Matora, Larisa Yu; Knirel, Yuriy A; Shchyogolev, Sergei Yu

    2012-11-01

    This is the first report to have identified an O-linked repetitive glycan in bacterial flagellin, a structural protein of the flagellum. Studies by sugar analysis, Smith degradation, (1)H and (13)C NMR spectroscopy, and mass spectrometry showed that the glycan chains of the polar flagellum flagellin of the plant-growth-promoting rhizobacterium Azospirillum brasilense Sp7 are represented by a polysaccharide with a molecular mass of 7.7 kDa, which has a branched tetrasaccharide repeating unit of the following structure:

  18. P4k-1-factorization of complete bipartite graphs

    Institute of Scientific and Technical Information of China (English)

    DU; Beiliang; WANG; Jian

    2005-01-01

    Let Km,n be a complete bipartite graph with two partite sets having m and n vertices, respectively. A Pv-factorization of Km,n is a set of edge-disjoint Pv-factors of Km,n which partition the set of edges of Km,n. When v is an even number,Wang and Ushio gave a necessary and sufficient condition for the existence of Pv-factorization of Km,n. When v is an odd number, Ushio in 1993 proposed a conjecture. However, up to now we only know that Ushio Conjecture is true for v=3. In this paper wewill show that Ushio Conjecture is true when v=4k-1. That is, we shall prove that a necessary and sufficient condition for the existence of a P4k-1-factorization of Km,n is (1) (2k-1)m≤2kn, (2) (2k-1)n ≤2km, (3) m+n ≡0 (mod 4k-1), (4) (4k-1)mn/[2(2k-1)(m+n)] is an integer.integer.

  19. The K-1 Active Dispenser for Orbit Transfer

    Science.gov (United States)

    Lai, G.; Cochran, D.; Curtis, R.

    2002-01-01

    Kistler Aerospace Corporation is building the K-1, the world's first fully reusable launch vehicle. The two-stage K- 1 is designed primarily to service the market for low-earth orbit (LEO) missions, due to Kistler's need to recover both stages. For customers requiring payload delivery to high-energy orbits, Kistler can outfit the payload with a K- 1 Active Dispenser (an expendable third stage). The K-1 second stage will deploy the Active Dispenser mated with its payload into a 200 km circular LEO parking orbit. From this orbit, the Active Dispenser would use its own propulsion to place its payload into the final desired drop-off orbit or earth-escape trajectory. This approach allows Kistler to combine the low-cost launch services offered by the reusable two-stage K-1 with the versatility of a restartable, expendable upper stage. Enhanced with an Active Dispenser, the K-1 will be capable of delivering 1,500 kg to a geosynchronous transfer orbit or up to approximately 1,000 kg into a Mars rendezvous trajectory. The list price of a K-1 Active Dispenser launch is 25 million (plus the price of mission unique integration services) significantly less than the price of any launch vehicle service in the world with comparable capability.

  20. S-Layer Glycoproteins and Flagellins: Reporters of Archaeal Posttranslational Modifications

    Directory of Open Access Journals (Sweden)

    Ken F. Jarrell

    2010-01-01

    Full Text Available Many archaeal proteins undergo posttranslational modifications. S-layer proteins and flagellins have been used successfully to study a variety of these modifications, including N-linked glycosylation, signal peptide removal and lipid modification. Use of these well-characterized reporter proteins in the genetically tractable model organisms, Haloferax volcanii, Methanococcus voltae and Methanococcus maripaludis, has allowed dissection of the pathways and characterization of many of the enzymes responsible for these modifications. Such studies have identified archaeal-specific variations in signal peptidase activity not found in the other domains of life, as well as the enzymes responsible for assembly and biosynthesis of novel N-linked glycans. In vitro assays for some of these enzymes have already been developed. N-linked glycosylation is not essential for either Hfx. volcanii or the Methanococcus species, an observation that allowed researchers to analyze the role played by glycosylation in the function of both S-layers and flagellins, by generating mutants possessing these reporters with only partial attached glycans or lacking glycan altogether. In future studies, it will be possible to consider questions related to the heterogeneity associated with given modifications, such as differential or modulated glycosylation.

  1. Study in ovo immunisation with flagellin and whole cell protein antigens of Campylobacter jejuni in chickens

    Directory of Open Access Journals (Sweden)

    Susan Maphilindawati Noor

    2000-06-01

    Full Text Available In ovo immunisation of chickens with flagellin and whole cell protein antigens of Campylobacter jejuni was examined to determine Campylobacter infection. Four groups of embryonated chicken eggs (10 eggs per group were immunised in ovo at day 17 of incubation and booster was given at 7 days post-hatch. Group I was immunised in ovo and oral booster with whole cell protein of C. jejuni, group II was immunised in ovo and oral booster with C. jejuni flagellin protein, group III was immunised in ovo and intraperitoneal booster with whole cell, and group IV was treated as control. The humoral immune responses were determined by enzyme-linked immunosorbent assay (ELISA and the mucosal immune responses were examined by a direct fluorescent histology antibody technique. Immunised chickens of Group I, II, and III shown to have higher antibody titers than those of control chickens (group IV. The titres of anti-campylobacter antibodies of all isotypes in serum, bile, and intestinal scrapping after challenge were not significantly different in all groups. In addition, when immunised chickens were orally challenged with a homologous strain of viable C. jejuni organism, the chickens remained infected throughout the experiment based on cloacal swabs and caecal contents. These findings indicated that although in ovo immunisation resulted in increasing of the mucosal and humoral immune responses in chickens, it is not strong enough to protect the Campylobacter colonisation in the intestinal tract.

  2. K1 Group of Finite Dimensional Path Algebra

    Institute of Scientific and Technical Information of China (English)

    Xue Jun GUO; Li Bin LI

    2001-01-01

    In this paper, by calculating the commutator subgroup of the unit group of finite pathalgebra κ/△ and the unit group abelianized, we explicitly characterize the K1 group of finite dimensionalpath algebra over an arbitrary field.

  3. Serum endotoxins and flagellin and risk of colorectal cancer in the European prospective investigation into cancer and nutrition (EPIC) cohort

    NARCIS (Netherlands)

    Kong, So Yeon; Tran, Hao Quang; Gewirtz, Andrew T.; McKeown-Eyssen, Gail; Fedirko, Veronika; Romieu, Isabelle; Tjønneland, Anne; Olsen, Anja; Overvad, Kim; Boutron-Ruault, Marie Christine; Bastide, Nadia; Affret, Aurélie; Kühn, Tilman; Kaaks, Rudolf; Boeing, Heiner; Aleksandrova, Krasimira; Trichopoulou, Antonia; Kritikou, Maria; Vasilopoulou, Effie; Palli, Domenico; Krogh, Vittorio; Mattiello, Amalia; Tumino, Rosario; Naccarati, Alessio; Bueno-De-mesquita, H. B.; Peeters, Petra H.; Weiderpass, Elisabete; Quirós, J. Ramón; Sala, Núria; Sánchez, María José; Castanõ, José María Huerta; Barricarte, Aurelio; Dorronsoro, Miren; Werner, Marten; Wareham, Nicholas J.; Khaw, Kay Tee; Bradbury, Kathryn E.; Freisling, Heinz; Stavropoulou, Faidra; Ferrari, Pietro; Gunter, Marc J.; Cross, Amanda J.; Riboli, Elio; Bruce, W. Robert; Jenab, Mazda

    2016-01-01

    Background: Chronic inflammation and oxidative stress are thought to be involved in colorectal cancer development. These processes may contribute to leakage of bacterial products, such as lipopolysaccharide (LPS) and flagellin, across the gut barrier. The objective of this study, nested within a pro

  4. Cell surface expression system for the display of heterologous gene products using chimeric flagellin fusions of bacillus halodurans isolate

    CSIR Research Space (South Africa)

    Du Plessis, A

    2006-10-01

    Full Text Available N-terminal sequencing gave rise to homology to flagellin protein, product of the hag gene. protein, product of the hag gene. Gene was cloned by using degenerate primers and inverse PCR. The gene sequence as well as the up- and down- stream regions...

  5. Identification of gene candidates associated with huanglongbing tolerance using Candidatus Liberibacter asiaticus flagellin 22 as a proxy to challenge citrus

    Science.gov (United States)

    Plant defense elicited by pathogen-associated molecular patterns (PAMPs) is an important component of disease resistance. Previous research indicated the canker resistance in citrus correlates with responsiveness to Xcc-flg22, the 22 amino acid PAMP from the flagellin of Xanthomonas citri ssp. citri...

  6. Characterization of structural and immunological properties of a fusion protein between flagellin from Salmonella and lumazine synthase from Brucella.

    Science.gov (United States)

    Hiriart, Y; Rossi, A H; Biedma, M E; Errea, A J; Moreno, G; Cayet, D; Rinaldi, J; Blancá, B; Sirard, J C; Goldbaum, F; Berguer, P; Rumbo, M

    2017-05-01

    Aiming to combine the flexibility of Brucella lumazine synthase (BLS) to adapt different protein domains in a decameric structure and the capacity of BLS and flagellin to enhance the immunogenicity of peptides that are linked to their structure, we generated a chimeric protein (BLS-FliC131) by fusing flagellin from Salmonella in the N-termini of BLS. The obtained protein was recognized by anti-flagellin and anti-BLS antibodies, keeping the oligomerization capacity of BLS, without affecting the folding of the monomeric protein components determined by circular dichroism. Furthermore, the thermal stability of each fusion partner is conserved, indicating that the interactions that participate in its folding are not affected by the genetic fusion. Besides, either in vitro or in vivo using TLR5-deficient animals we could determine that BLS-FliC131 retains the capacity of triggering TLR5. The humoral response against BLS elicited by BLS-FliC131 was stronger than the one elicited by equimolar amounts of BLS + FliC. Since BLS scaffold allows the generation of hetero-decameric structures, we expect that flagellin oligomerization on this protein scaffold will generate a new vaccine platform with enhanced capacity to activate immune responses. © 2017 The Protein Society.

  7. S6K1 Is Required for Increasing Skeletal Muscle Force during Hypertrophy

    Directory of Open Access Journals (Sweden)

    Manuela Marabita

    2016-10-01

    Full Text Available Loss of skeletal muscle mass and force aggravates age-related sarcopenia and numerous pathologies, such as cancer and diabetes. The AKT-mTORC1 pathway plays a major role in stimulating adult muscle growth; however, the functional role of its downstream mediators in vivo is unknown. Here, we show that simultaneous inhibition of mTOR signaling to both S6K1 and 4E-BP1 is sufficient to reduce AKT-induced muscle growth and render it insensitive to the mTORC1-inhibitor rapamycin. Surprisingly, lack of mTOR signaling to 4E-BP1 only, or deletion of S6K1 alone, is not sufficient to reduce muscle hypertrophy or alter its sensitivity to rapamycin. However, we report that, while not required for muscle growth, S6K1 is essential for maintaining muscle structure and force production. Hypertrophy in the absence of S6K1 is characterized by compromised ribosome biogenesis and the formation of p62-positive protein aggregates. These findings identify S6K1 as a crucial player for maintaining muscle function during hypertrophy.

  8. Pro-inflammatory flagellin proteins of prevalent motile commensal bacteria are variably abundant in the intestinal microbiome of elderly humans.

    Directory of Open Access Journals (Sweden)

    B Anne Neville

    Full Text Available Some Eubacterium and Roseburia species are among the most prevalent motile bacteria present in the intestinal microbiota of healthy adults. These flagellate species contribute "cell motility" category genes to the intestinal microbiome and flagellin proteins to the intestinal proteome. We reviewed and revised the annotation of motility genes in the genomes of six Eubacterium and Roseburia species that occur in the human intestinal microbiota and examined their respective locus organization by comparative genomics. Motility gene order was generally conserved across these loci. Five of these species harbored multiple genes for predicted flagellins. Flagellin proteins were isolated from R. inulinivorans strain A2-194 and from E. rectale strains A1-86 and M104/1. The amino-termini sequences of the R. inulinivorans and E. rectale A1-86 proteins were almost identical. These protein preparations stimulated secretion of interleukin-8 (IL-8 from human intestinal epithelial cell lines, suggesting that these flagellins were pro-inflammatory. Flagellins from the other four species were predicted to be pro-inflammatory on the basis of alignment to the consensus sequence of pro-inflammatory flagellins from the β- and γ- proteobacteria. Many fliC genes were deduced to be under the control of σ(28. The relative abundance of the target Eubacterium and Roseburia species varied across shotgun metagenomes from 27 elderly individuals. Genes involved in the flagellum biogenesis pathways of these species were variably abundant in these metagenomes, suggesting that the current depth of coverage used for metagenomic sequencing (3.13-4.79 Gb total sequence in our study insufficiently captures the functional diversity of genomes present at low (≤1% relative abundance. E. rectale and R. inulinivorans thus appear to synthesize complex flagella composed of flagellin proteins that stimulate IL-8 production. A greater depth of sequencing, improved evenness of sequencing

  9. Ramsey numbers r(K1, 4, G) for all three-partite graphs G of order six%K1,4和六阶三部图的Ramsey数r(K1,4,G)

    Institute of Scientific and Technical Information of China (English)

    顾华; 宋洪雪; 刘向阳

    2004-01-01

    本文主要应用了组合分析的方法. 在对所有边任意地进行了红、蓝2种颜色着色的完全图KN中, 考察了完全图KN的顶点集中某点的邻域在红图或蓝图中所生成的子图的性质. 在Jayawardene和Rousseau (Ars Combinatoria, 2000, 163-173)的主要结果的启发下, 研究并确定了另一种常见的五阶图K1, 4对于所有无孤立点的六阶三部图G的Ramsey数r(K1, 4, G).%In this paper, we use a combinatorial analysis method. In the complete graph KN with edges colored arbitrarily by red or blue, we consider the proposition of the subgraph of the red graph or blue graph induced by the neighborhood of some vertex in V(KN). Inspired by the main results of Jayawardene and Rousseau (Ars Combinatoria, 2000, 163-173), we determine the Ramsey numbers of r(K1, 4, G), where G is the three-partite graph of order six without isolate vertex.

  10. Multi-satellite ocean tide modelling - the K-1 constituent

    DEFF Research Database (Denmark)

    Andersen, Ole Baltazar; Knudsen, Per

    1997-01-01

    All major ocean tide constituents are aliased into signals with periods less than 90 days from TOPEX/POSEIDON altimetry, except the K-1 constituent. The aliased K-1 has a period of 173 days. Consequently, it might be confounded with height variations caused by the semiannual cycle having a period......, where the presence of crossing tracks cannot separate K-1 from the semiannual signal from TOPEX/POSEIDON, the importance of including ERS-1 and GEOSAT observations was demonstrated. A comparison with 29 pelagic and coastal tide gauges in the Southern Ocean south of 50 degrees S gave 5.59 (M-2), 2.27 (S......-2) and 5.04 (K-1) cm RMS agreement for FES95.1 ocean tide model. The same comparison for the best empirical estimated constituents based on TOPEX/POSEIDON + ERS-1 + GEOSAT gave 4.32, 2.21, and 4.29 cm for M-2, S-2 and K-1, respectively....

  11. Development of organometallic S6K1 inhibitors.

    Science.gov (United States)

    Qin, Jie; Rajaratnam, Rajathees; Feng, Li; Salami, Jemilat; Barber-Rotenberg, Julie S; Domsic, John; Reyes-Uribe, Patricia; Liu, Haiying; Dang, Weiwei; Berger, Shelley L; Villanueva, Jessie; Meggers, Eric; Marmorstein, Ronen

    2015-01-08

    Aberrant activation of S6 kinase 1 (S6K1) is found in many diseases, including diabetes, aging, and cancer. We developed ATP competitive organometallic kinase inhibitors, EM5 and FL772, which are inspired by the structure of the pan-kinase inhibitor staurosporine, to specifically inhibit S6K1 using a strategy previously used to target other kinases. Biochemical data demonstrate that EM5 and FL772 inhibit the kinase with IC50 value in the low nanomolar range at 100 μM ATP and that the more potent FL772 compound has a greater than 100-fold specificity over S6K2. The crystal structures of S6K1 bound to staurosporine, EM5, and FL772 reveal that the EM5 and FL772 inhibitors bind in the ATP binding pocket and make S6K1-specific contacts, resulting in changes to the p-loop, αC helix, and αD helix when compared to the staurosporine-bound structure. Cellular data reveal that FL772 is able to inhibit S6K phosphorylation in yeast cells. Together, these studies demonstrate that potent, selective, and cell permeable S6K1 inhibitors can be prepared and provide a scaffold for future development of S6K inhibitors with possible therapeutic applications.

  12. Intestinal scavenger receptors are involved in vitamin K1 absorption.

    Science.gov (United States)

    Goncalves, Aurélie; Margier, Marielle; Roi, Stéphanie; Collet, Xavier; Niot, Isabelle; Goupy, Pascale; Caris-Veyrat, Catherine; Reboul, Emmanuelle

    2014-10-31

    Vitamin K1 (phylloquinone) intestinal absorption is thought to be mediated by a carrier protein that still remains to be identified. Apical transport of vitamin K1 was examined using Caco-2 TC-7 cell monolayers as a model of human intestinal epithelium and in transfected HEK cells. Phylloquinone uptake was then measured ex vivo using mouse intestinal explants. Finally, vitamin K1 absorption was compared between wild-type mice and mice overexpressing scavenger receptor class B type I (SR-BI) in the intestine and mice deficient in cluster determinant 36 (CD36). Phylloquinone uptake by Caco-2 cells was saturable and was significantly impaired by co-incubation with α-tocopherol (and vice versa). Anti-human SR-BI antibodies and BLT1 (a chemical inhibitor of lipid transport via SR-BI) blocked up to 85% of vitamin K1 uptake. BLT1 also decreased phylloquinone apical efflux by ∼80%. Transfection of HEK cells with SR-BI and CD36 significantly enhanced vitamin K1 uptake, which was subsequently decreased by the addition of BLT1 or sulfo-N-succinimidyl oleate (CD36 inhibitor), respectively. Similar results were obtained in mouse intestinal explants. In vivo, the phylloquinone postprandial response was significantly higher, and the proximal intestine mucosa phylloquinone content 4 h after gavage was increased in mice overexpressing SR-BI compared with controls. Phylloquinone postprandial response was also significantly increased in CD36-deficient mice compared with wild-type mice, but their vitamin K1 intestinal content remained unchanged. Overall, the present data demonstrate for the first time that intestinal scavenger receptors participate in the absorption of dietary phylloquinone.

  13. Intestinal Scavenger Receptors Are Involved in Vitamin K1 Absorption*

    Science.gov (United States)

    Goncalves, Aurélie; Margier, Marielle; Roi, Stéphanie; Collet, Xavier; Niot, Isabelle; Goupy, Pascale; Caris-Veyrat, Catherine; Reboul, Emmanuelle

    2014-01-01

    Vitamin K1 (phylloquinone) intestinal absorption is thought to be mediated by a carrier protein that still remains to be identified. Apical transport of vitamin K1 was examined using Caco-2 TC-7 cell monolayers as a model of human intestinal epithelium and in transfected HEK cells. Phylloquinone uptake was then measured ex vivo using mouse intestinal explants. Finally, vitamin K1 absorption was compared between wild-type mice and mice overexpressing scavenger receptor class B type I (SR-BI) in the intestine and mice deficient in cluster determinant 36 (CD36). Phylloquinone uptake by Caco-2 cells was saturable and was significantly impaired by co-incubation with α-tocopherol (and vice versa). Anti-human SR-BI antibodies and BLT1 (a chemical inhibitor of lipid transport via SR-BI) blocked up to 85% of vitamin K1 uptake. BLT1 also decreased phylloquinone apical efflux by ∼80%. Transfection of HEK cells with SR-BI and CD36 significantly enhanced vitamin K1 uptake, which was subsequently decreased by the addition of BLT1 or sulfo-N-succinimidyl oleate (CD36 inhibitor), respectively. Similar results were obtained in mouse intestinal explants. In vivo, the phylloquinone postprandial response was significantly higher, and the proximal intestine mucosa phylloquinone content 4 h after gavage was increased in mice overexpressing SR-BI compared with controls. Phylloquinone postprandial response was also significantly increased in CD36-deficient mice compared with wild-type mice, but their vitamin K1 intestinal content remained unchanged. Overall, the present data demonstrate for the first time that intestinal scavenger receptors participate in the absorption of dietary phylloquinone. PMID:25228690

  14. Incorporation of membrane-anchored flagellin or Escherichia coli heat-labile enterotoxin B subunit enhances the immunogenicity of rabies virus-like particles in mice and dogs

    Directory of Open Access Journals (Sweden)

    Yinglin eQi

    2015-03-01

    Full Text Available Rabies remains an important worldwide public health threat, so safe, effective and affordable vaccines are still being sought. Virus-like particle (VLP-based vaccines targeting various viral pathogens have been successfully produced, licensed and commercialized. Here, we designed and constructed two chimeric rabies virus-like particles (cRVLPs containing rabies virus (RABV glycoprotein (G, matrix (M protein, and membrane-anchored flagellin (EVLP-F or Escherichia coli heat-labile enterotoxin B subunit (EVLP-L as molecular adjuvants to enhance the immune response against rabies. The immunogenicity and potential of cRVLPs as novel rabies vaccine were evaluated by intramuscular vaccination in mouse and dog models. Mouse studies demonstrated that both EVLP-F and EVLP-L induced faster and larger virus-neutralizing antibodies (VNA responses and elicited greater numbers of CD4+ and CD8+ T cells secreting IFN-γ or IL-4 compared with a standard rabies VLP (sRVLP containing only G and M. Moreover, cRVLPs recruited and/or activated more B cells and dendritic cells in inguinal lymph nodes. EVLP-F induced a strong, specific IgG2a response but not an IgG1 response, suggesting the activation of Th1 class immunity; in contrast, Th2 class immunity was observed with EVLP-L. The significantly enhanced humoral and cellular immune responses induced by cRVLPs provided complete protection against lethal challenge with RABV. Most importantly, dogs vaccinated with EVLP-F or EVLP-L exhibited increased VNA titers in sera and enhanced IFN-γ and IL-4 secretion from peripheral blood mononuclear cells. Taken together, these results illustrate that when incorporated into sRVLP, membrane-anchored flagellin and LTB possess strong adjuvant activity. EVLP-F and EVLP-L induce significantly enhanced RABV-specific humoral and cellular immune responses in both mouse and dog. Therefore, these cRVLPs may be developed as safe and more efficacious rabies vaccine candidate for animals.

  15. An Algebraic Proof of Quillen's Resolution Theorem for K_1

    CERN Document Server

    Whale, Ben

    2009-01-01

    In his 1973 paper Quillen proved a resolution theorem for the K-Theory of an exact category; his proof was homotopic in nature. By using the main result of a paper by Nenashev, we are able to give an algebraic proof of Quillen's Resolution Theorem for K_1 of an exact category.

  16. Polymorphism of 41 kD Flagellin Gene and Its Human B-Cell Epitope in Borrelia burgdorferi Strains of China

    Directory of Open Access Journals (Sweden)

    Huixin Liu

    2016-01-01

    Full Text Available The 41 kD flagellin of Borrelia burgdorferi (B. burgdorferi is a major component of periplasmic flagellar filament core and a good candidate for serodiagnosis in early stage of Lyme disease. Here, we chose 89 B. burgdorferi strains in China, amplified the gene encoding the 41 kD flagellin, and compared the sequences. The results showed that genetic diversity presented in the 41 kD flagellin genes of all 89 strains among the four genotypes of B. burgdorferi, especially in the genotype of B. garinii. Some specific mutation sites for each genotype of the 41 kD flagellin genes were found, which could be used for genotyping B. burgdorferi strains in China. Human B-cell epitope analysis showed that thirteen of 15 nonsynonymous mutations occurred in the epitope region of 41 kD flagellin and thirty of 42 B-cell epitopes were altered due to all 13 nonsynonymous mutations in the epitope region, which may affect the function of the antigen. Nonsynonymous mutations and changed human B-cell epitopes exist in 41 kD flagellin of B. burgdorferi sensu lato strains; these changes should be considered in serodiagnosis of Lyme disease.

  17. Indirect Toll-like receptor 5-mediated activation of conventional dendritic cells promotes the mucosal adjuvant activity of flagellin in the respiratory tract.

    Science.gov (United States)

    Fougeron, Delphine; Van Maele, Laurye; Songhet, Pascal; Cayet, Delphine; Hot, David; Van Rooijen, Nico; Mollenkopf, Hans-Joachim; Hardt, Wolf-Dietrich; Benecke, Arndt G; Sirard, Jean-Claude

    2015-06-26

    The Toll-like receptor 5 (TLR5) agonist flagellin is an effective adjuvant for vaccination. Recently, we demonstrated that the adaptive responses stimulated by intranasal administration of flagellin and antigen were linked to TLR5 signaling in the lung epithelium. The present study sought to identify the antigen presenting cells involved in this adjuvant activity. We first found that the lung dendritic cells captured antigen very efficiently in a process independent of TLR5. However, TLR5-mediated signaling specifically enhanced the maturation of lung dendritic cells. Afterward, the number of antigen-bound and activated conventional dendritic cells (both CD11b(+) and CD103(+)) increased in the mediastinal lymph nodes in contrast to monocyte-derived dendritic cells. These data suggested that flagellin-activated lung conventional dendritic cells migrate to the draining lymph nodes. The lymph node dendritic cells, in particular CD11b(+) cells, were essential for induction of CD4 T-cell response. Lastly, neutrophils and monocytes were recruited into the lungs by flagellin administration but did not contribute to the adjuvant activity. The functional activation of conventional dendritic cells was independent of direct TLR5 signaling, thereby supporting the contribution of maturation signals produced by flagellin-stimulated airway epithelium. In conclusion, our results demonstrated that indirect TLR5-dependent stimulation of airway conventional dendritic cells is essential to flagellin's mucosal adjuvant activity.

  18. Comparative ultrastructural and functional studies of Helicobacter pylori and Helicobacter mustelae flagellin mutants: both flagellin subunits, FlaA and FlaB, are necessary for full motility in Helicobacter species.

    OpenAIRE

    Josenhans, C; Labigne, A; Suerbaum, S

    1995-01-01

    Helicobacter mustelae causes chronic gastritis and ulcer disease in ferrets. It is therefore considered an important animal model of human Helicobacter pylori infection. High motility even in a viscous environment is one of the common virulence determinants of Helicobacter species. Their sheathed flagella contain a complex filament that is composed of two distinctly different flagellin subunits, FlaA and FlaB, that are coexpressed in different amounts. Here, we report the cloning and sequence...

  19. Neighborhood Union and Connected [k, k + 1]-Factors in Graphs%图的邻域并和连通的[k,k+1]-因子

    Institute of Scientific and Technical Information of China (English)

    刘红霞; 乔贵平

    2013-01-01

    Let G be a graph of order n. A spanning subgraph F of G is called a[ k, k + 1 ] -factor if k≤dF(x) ≤ k + 1 holds for each x∈V(G). A [k, k + 1]-factor is called a connected [k, k + 1 ] -factor if it is connected. A [k, k + 1 ]-factor F is called a Hamilton [k, k + 1 ] -factor if F contains a Hamilton cycle. In this paper, sev-eralsufficient conditions related to neighborhood union for graphs to have connected [ k, k + 1 ] -factors or Hamilton [k, k + 1]-factors are given.%设G是阶为n的图.F是G的支撑子图且对所有的x∈ V(G)都有k≤dF(x)≤k+1,则称F为G的[k,k+1]-因子.一个[k,k+1]-因子如果连通,则称为连通的[k,k+1]-因子.一个[k,k+1]-因子若包含一个哈密顿圈,则称为哈密顿[k,k+1]-因子.给出了图有哈密顿[k,k+1]-因子或连通的[k,k+1]-因子关于邻域并的若干新的充分条件.

  20. New methodologies for lower-K1 EUV OPC and RET optimization

    Science.gov (United States)

    Hooker, Kevin; Kazarian, Aram; Zhou, Xibin; Tuttle, Josh; Xiao, Guangming; Zhang, Yunqiang; Lucas, Kevin

    2017-03-01

    EUV lithography is viewed as a highly desirable technology for 5nm and 7nm node patterning cost reduction and process simplicity. However, for the 5nm and 7nm nodes EUV not only needs to function in a low-K1 resolution environment but has several new and complex patterning issues which will need accurate compensation by mask synthesis tools and flows. The main new issues are: long-range flare variation across the chip, feature dependent focus offsets due to high mask topography, asymmetry inducing shadowing effects which vary across the lens slit, significantly higher lens aberrations, illumination source changes (across the lens and with time) and new resist exposure mechanisms. These solutions must be successfully deployed at low K1 values and must be integrated together to create OPC/RET flows which have high resolution, high accuracy, and are fast to deploy. Therefore, the combined requirements of low-K1 resolution, full reticle correction accuracy and process window can be even more challenging than in current optical lithography mask synthesis flows. Advanced computational methods such as ILT and model-based SRAF optimization are well known to have considerable benefits in process window and resolution for low-K1 193 lithography. However, these methods have not been well studied to understand their benefits for lower-K1 EUV lithography where fabs must push EUV resolution, 2D accuracy and process window to their limits. In this paper, we investigate where inverse lithography methods can improve EUV patterning weaknesses vs. traditional OPC/RET. We first show how ILT can be used to guide a better understanding of optimal solutions for EUV mask synthesis. We then provide detailed comparisons of ILT and traditional methods on a wide range of mask synthesis applications.

  1. Possible dual regulatory circuits involving AtS6K1 in the regulation of plant cell cycle and growth.

    Science.gov (United States)

    Shin, Yun-jeong; Kim, Sunghan; Du, Hui; Choi, Soonyoung; Verma, Desh Pal S; Cheon, Choong-Ill

    2012-05-01

    The role of Arabidopsis S6 Kinase 1 (AtS6K1), a downstream target of TOR kinase, in controlling plant growth and ribosome biogenesis was characterized after generating transgenic plants expressing AtS6K1 under auxin-inducible promoter. Down regulation of selected cell cycle regulatory genes upon auxin treatment was observed in the transgenic plants, confirming the negative regulatory role of AtS6K1 in the plant cell cycle progression reported earlier. Callus tissues established from these transgenic plants grew to larger cell masses with more number of enlarged cells than untransformed control, demonstrating functional implication of AtS6K1 in the control of plant cell size. The observed negative correlation between the expression of AtS6K1 and the cell cycle regulatory genes, however, was completely reversed in protoplasts generated from the transgenic plants expressing AtS6K1, suggesting a possible existence of dual regulatory mechanism of the plant cell cycle regulation mediated by AtS6K1. An alternative method of kinase assay, termed "substrate-mediated kinase pull down", was employed to examine the additional phosphorylation on other domains of AtS6K1 and verified the phosphorylation of both amino- and carboxy-terminal domains, which is a novel finding regarding the phosphorylation target sites on plant S6Ks by upstream regulatory kinases. In addition, this kinase assay under the stress conditions revealed the salt- and sugar-dependencies of AtS6K1 phosphorylations.

  2. On Potentially K1,4+P2-graphic Sequences%蕴含K1,4+P2的可图序列

    Institute of Scientific and Technical Information of China (English)

    王艳

    2008-01-01

    本文刻划了蕴含K1,4+P2的可图序列,其中K1,4+P2是向完全二部图K1,4添加一条被剖分的边后构成的简单图.%In this paper, we characterize the potentially K1,4+P2-graphic sequences, where K1,4+P2 be a graph obtained by adding an edge e which is subdivided to complete bipartite graph K1,4.

  3. Early diagnosis of typhoid fever by nested PCR for flagellin gene of Salmonella enterica serotype Typhi.

    Science.gov (United States)

    Khan, S; Harish, B N; Menezes, G A; Acharya, N S; Parija, S C

    2012-11-01

    Typhoid fever caused by Salmonella Typhi continues to be a major health problem in spite of the use of antibiotics and the development of newer antibacterial drugs. Inability to make an early laboratory diagnosis and resort to empirical therapy, often lead to increased morbidity and mortality in cases of typhoid fever. This study was aimed to optimize a nested PCR for early diagnosis of typhoid fever and using it as a diagnostic tool in culture negative cases of suspected typhoid fever. Eighty patients with clinical diagnosis of typhoid fever and 40 controls were included in the study. The blood samples collected were subjected to culture, Widal and nested PCR targeting the flagellin gene of S. Typhi. The sensitivity of PCR on blood was found to be 100 per cent whereas the specificity was 76.9 per cent. The positive predictive value (PPV) of PCR was calculated to be 76.9 per cent with an accuracy of 86 per cent. None of the 40 control samples gave a positive PCR. Due to its high sensitivity and specificity nested PCR can be used as a useful tool to diagnose clinically suspected, culture negative cases of typhoid fever.

  4. Exhaustive exercise increases the TNF-α production in response to flagellin via the upregulation of toll-like receptor 5 in the large intestine in mice.

    Science.gov (United States)

    Uchida, Masataka; Oyanagi, Eri; Kawanishi, Noriaki; Iemitsu, Motoyuki; Miyachi, Motohiko; Kremenik, Michael J; Onodera, Sho; Yano, Hiromi

    2014-01-01

    Although intense exercise may induce temporary immune depression, it is unclear whether exercise stimulates tumor necrosis factor-alpha (TNF-α) production in response to flagella protein flagellin (FG), which binds to toll-like receptor 5 (TLR5) and induces the production of pro-inflammatory cytokines. Male C3H/HeN mice were administered FG (1mg/kg, i.v.) after exhaustive exercise (EX), and the plasma TNF-α concentrations were examined. The production of TNF-α and the TLR5 expression in both RAW264 and Caco2 cells were measured under FG conditions in vitro. Although the plasma TNF-α concentrations were observed to significantly increase in both the EX and non-EX (N-EX) mice (pTNF-α levels in the EX mice were significantly higher than those observed in the N-EX mice (pTNF-α production and TLR5 expression on the Caco2, but not RAW264 cells. Interestingly, a high Ep-induced TLR5 expression was observed on the Caco2 cell surface, which was inhibited by an inhibitor of phosphoinositide3-kinase (PI3K), Ly294002, as well as a β-adrenergic blocker, propranolol. In addition, the EX-induced TNF-α production observed in response to FG was also attenuated by pretreatment with propranolol. Our findings suggest that exhaustive exercise upregulates the production of TNF-α in response to FG via a high expression of TLR5 on the intestinal cell surface following the stimulation of β-adrenergic receptors with exercise.

  5. Cloning and sequencing of the central region of the flagellin gene from the Gram-positive bacterium Clostridium tyrobutyricum ATCC 25755.

    Science.gov (United States)

    Arnold, F; Bédouet, L; Batina, P; Robreau, G

    1999-01-01

    The purpose of this study was to sequence the central part of the coding region of the Clostridium tyrobutyricum fiagellin gene to improve the immunoenzymatic counting of cells after milk filtration. The coding region was amplified by PCR, and the amplified products were cloned. A DNA sequence analysis of positive clones gave us 1,131 nucleotides with a partial calculated flagellin molecular mass of 40,143 Da. The flagellar filament protein sequence exhibited high levels of homology to sequences of flagellin protein from other bacteria in both N- and C-terminal parts, but little homology in the central domain. A PCR-restriction fragment length polymorphism analysis of amplified C. tyrobutyricum flagellin gene products confirmed the variability of the central domain. The flagellin mRNA was determined to be 1.1 kb in size, which suggests a monocistronic mRNA. Furthermore, the deduced protein flagellin contains eleven potential N-glycosylation sites and one sequence rich in serine, which could be modified by O-glycosylation.

  6. Finding the optimal dose of vitamin K1 to treat vitamin K deficiency and to avoid anaphylactoid reactions.

    Science.gov (United States)

    Mi, Yan-Ni; Ping, Na-Na; Li, Bo; Xiao, Xue; Zhu, Yan-Bing; Cao, Lei; Ren, Jian-Kang; Cao, Yong-Xiao

    2017-10-01

    Vitamin K1 injection induces severe dose-related anaphylactoid reactions and overdose for the treatment of vitamin K deficiency. We aimed to find an optimal and small dose of vitamin K1 injection to treat vitamin K deficiency and avoid anaphylactoid reactions in animal. Rats were administered a vitamin K-deficient diet and gentamicin to establish vitamin K deficiency model. Behaviour tests were performed in beagle dogs to observe anaphylactoid reactions. The results showed an increased protein induced by vitamin K absence or antagonist II (PIVKA-II) levels, a prolonging of prothrombin time (PT) and activated partial thromboplastin time (APTT) and a decrease in vitamin K-dependent coagulation factor (F) II, VII, IX and X activities in the model group. In vitamin K1 0.01 mg/kg group, the liver vitamin K1 levels increased fivefold and the liver vitamin K2 levels increased to the normal amount. Coagulation markers PT, APTT, FVII and FIX activities returned to normal. Both in the 0.1 and 1.0 mg/kg vitamin K1 groups, coagulation functions completely returned to normal. Moreover, the amount of liver vitamin K1 was 40 (0.1 mg/kg) or 100 (1.0 mg/kg) times as in normal. Vitamin K2 was about 4 (0.1 mg/kg) or 5 (1.0 mg/kg) times as the normal amount. There was no obvious anaphylactoid symptom in dogs with the dose of 0.03 mg/kg, which is equivalent to the dose of 0.01 mg/kg in rats. These results demonstrated that a small dose of vitamin K1 is effective to improve vitamin K deficiency and to prevent anaphylactoid reactions, simultaneously. © 2017 Société Française de Pharmacologie et de Thérapeutique.

  7. Comparative ultrastructural and functional studies of Helicobacter pylori and Helicobacter mustelae flagellin mutants: both flagellin subunits, FlaA and FlaB, are necessary for full motility in Helicobacter species.

    Science.gov (United States)

    Josenhans, C; Labigne, A; Suerbaum, S

    1995-01-01

    Helicobacter mustelae causes chronic gastritis and ulcer disease in ferrets. It is therefore considered an important animal model of human Helicobacter pylori infection. High motility even in a viscous environment is one of the common virulence determinants of Helicobacter species. Their sheathed flagella contain a complex filament that is composed of two distinctly different flagellin subunits, FlaA and FlaB, that are coexpressed in different amounts. Here, we report the cloning and sequence determination of the flaA gene of H. mustelae NCTC12032 from a PCR amplification product. The FlaA protein has a calculated molecular mass of 53 kDa and is 73% homologous to the H. pylori FlaA subunit. Isogenic flaA and flaB mutants of H. mustelae F1 were constructed by means of reverse genetics. A method was established to generate double mutants (flaA flaB) of H. mustelae F1 as well as H. pylori N6. Genotypes, motility properties, and morphologies of the H. mustelae flagellin mutants were determined and compared with those of the H. pylori flaA and flaB mutants described previously. The flagellar organizations of the two Helicobacter species proved to be highly similar. When the flaB genes were disrupted, motility decreased by 30 to 40%. flaA mutants retained weak motility by comparison with strains that were devoid of both flagellin subunits. Weakly positive motility tests of the flaA mutants correlated with the existence of short truncated flagella. In H. mustelae, lateral as well as polar flagella were present in the truncated form. flaA flaB double mutants were completely nonmotile and lacked any form of flagella. These results show that the presence of both flagellin subunits is necessary for complete motility of Helicobacter species. The importance of this flagellar organization for the ability of the bacteria to colonize the gastric mucosa and to persist in the gastric mucus remains to be proven. PMID:7768796

  8. Genetic diversity of the flagellin genes of Clostridium botulinum groups I and II.

    Science.gov (United States)

    Woudstra, Cedric; Lambert, Dominic; Anniballi, Fabrizio; De Medici, Dario; Austin, John; Fach, Patrick

    2013-07-01

    Botulinum neurotoxins (BoNTs) are produced by phenotypically and genetically different Clostridium species, including Clostridium botulinum and some strains of Clostridium baratii (serotype F) and Clostridium butyricum (serotype E). BoNT-producing clostridia responsible for human botulism encompass strains of group I (secreting proteases, producing toxin serotype A, B, or F, and growing optimally at 37°C) and group II (nonproteolytic, producing toxin serotype E, B, or F, and growing optimally at 30°C). Here we report the development of real-time PCR assays for genotyping C. botulinum strains of groups I and II based on flaVR (variable region sequence of flaA) sequences and the flaB gene. Real-time PCR typing of regions flaVR1 to flaVR10 and flaB was optimized and validated with 62 historical and Canadian C. botulinum strains that had been previously typed. Analysis of 210 isolates of European origin allowed the identification of four new C. botulinum flaVR types (flaVR11 to flaVR14) and one new flaVR type specific to C. butyricum type E (flaVR15). The genetic diversity of the flaVR among C. botulinum strains investigated in the present study reveals the clustering of flaVR types into 5 major subgroups. Subgroups 1, 3, and 4 contain proteolytic Clostridium botulinum, subgroup 2 is made up of nonproteolytic C. botulinum only, and subgroup 5 is specific to C. butyricum type E. The genetic variability of the flagellin genes carried by C. botulinum and the possible association of flaVR types with certain geographical areas make gene profiling of flaVR and flaB promising in molecular surveillance and epidemiology of C. botulinum.

  9. 莱姆病螺旋体鞭毛平截性蛋白的表达及其诊断潜力研究%Expression on specific fragement of flagellin from Borrelia burgdorferi and its diagnostic

    Institute of Scientific and Technical Information of China (English)

    仝彩玲; 吴银娟; 周勇志; 曹杰; 李培英; 周金林

    2011-01-01

    目的 原核表达伯氏疏螺旋体鞭毛蛋白Flagellin A基因特异性区段,获得重组鞭毛蛋白平截性蛋白作为诊断抗原,建立间接ELISA方法用于动物莱姆病的诊断.方法 PCR扩增获取伯氏疏螺旋体鞭毛蛋白基因的同源性较低的第394-798 bp区段,构建重组质粒pGEX-4T-(1)/tFlaA,构建好的表达质粒转化到大肠杆菌BL21( DE3)中进行表达,并纯化重组蛋白,用纯化的表达蛋白作为莱姆病诊断的抗原,用于ELISA检测实验感染小鼠莱姆病.结果 成功构建莱姆病螺旋体鞭毛平截性蛋白的表达载体,重组蛋白在宿主菌内高效、稳定表达,重组平截性蛋白显示了可作为ELISA诊断的抗原用于莱姆病的诊断价值.结论 纯化的伯氏疏螺旋体鞭毛蛋白可作为莱姆病ELISA诊断抗原用于莱姆病的诊断,为莱姆病快速诊断试剂盒的开发打下基础.%The purpose was to extract Borrelia burgdorferi DNA and use PCR method to obtain the gene coding of the Borrelia burgdorferi flagellin from 394 to 798 nucleotide that exhibited low homology with related gene from other bactdrial specises we insert Borrelia burgdorferi flagellin from 394 to 798 nucleotide to PGEX-4T-1 vector, verified by DNA sequence detection and tren thansformed it into in E. coli BL 21 (DE3) to induce target protein. Then we purified the recombination pro-tein as ELISA antigen to diagnosis Lyme disease. On other hand, we infected mouse with Borrelia burgdorferi. Two weeks, we got positive serum from mouse. We used recombinant protein as diagnostic antigen andpositive mouse serum as antibody to con-duct ELISA test and analysts. The ELISA results showed the feasibility of recombinant protein as diagnostic antigen to diag-nose Lyme disease, which lay foundations for the diagnosis of Lyme disease.

  10. 二部图的K1,4-因子分解%K1,4-factorization of bipartite graphs

    Institute of Scientific and Technical Information of China (English)

    王建

    2001-01-01

    Km,n的K1,k-因子分解问题已被多位研究者所研究,当k=2时Km,n具有K1,2-因子分解的存在性问题已被Ushio完全解决.当k=3时Wang研究了Km,n的K1,3-因子分解问题,并给出了Km,n具有K1,3-因子分解的一个充分条件.本文研究Km,n的K1,4-因子分解问题,并给出Km,n具有K1,4-因子分解的一个充分条件.%The K1,k-factorization of complete bipartite graph Km,n has been studied by severalresearchers. When k = 2 , the spectum problem for K1,2-factorization of Km,n has been com-pletely solved by Ushio. When k=3, Wang investigated K1,3-factorization of Km, n and gavea sufficient condition for such a factorization to exist. In this paper, we investigate K1,4-fac-torization of Km, n, and give a sufficient condition for such a factorization to exist.

  11. SphK1 inhibitor II (SKI-II) inhibits acute myelogenous leukemia cell growth in vitro and in vivo.

    Science.gov (United States)

    Yang, Li; Weng, Wei; Sun, Zhi-Xin; Fu, Xian-Jie; Ma, Jun; Zhuang, Wen-Fang

    2015-05-15

    Previous studies have identified sphingosine kinase 1 (SphK1) as a potential drug target for treatment of acute myeloid leukemia (AML). In the current study, we investigated the potential anti-leukemic activity of a novel and specific SphK1 inhibitor, SKI-II. We demonstrated that SKI-II inhibited growth and survival of human AML cell lines (HL-60 and U937 cells). SKI-II was more efficient than two known SphK1 inhibitors SK1-I and FTY720 in inhibiting AML cells. Meanwhile, it induced dramatic apoptosis in above AML cells, and the cytotoxicity by SKI-II was almost reversed by the general caspase inhibitor z-VAD-fmk. SKI-II treatment inhibited SphK1 activation, and concomitantly increased level of sphingosine-1-phosphate (S1P) precursor ceramide in AML cells. Conversely, exogenously-added S1P protected against SKI-II-induced cytotoxicity, while cell permeable short-chain ceramide (C6) aggravated SKI-II's lethality against AML cells. Notably, SKI-II induced potent apoptotic death in primary human AML cells, but was generally safe to the human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. In vivo, SKI-II administration suppressed growth of U937 leukemic xenograft tumors in severe combined immunodeficient (SCID) mice. These results suggest that SKI-II might be further investigated as a promising anti-AML agent.

  12. Gene-Environment Interactions Target Mitogen-activated Protein 3 Kinase 1 (MAP3K1) Signaling in Eyelid Morphogenesis*

    Science.gov (United States)

    Mongan, Maureen; Meng, Qinghang; Wang, Jingjing; Kao, Winston W.-Y.; Puga, Alvaro; Xia, Ying

    2015-01-01

    Gene-environment interactions determine the biological outcomes through mechanisms that are poorly understood. Mouse embryonic eyelid closure is a well defined model to study the genetic control of developmental programs. Using this model, we investigated how exposure to dioxin-like environmental pollutants modifies the genetic risk of developmental abnormalities. Our studies reveal that mitogen-activated protein 3 kinase 1 (MAP3K1) signaling is a focal point of gene-environment cross-talk. Dioxin exposure, acting through the aryl hydrocarbon receptor (AHR), blocked eyelid closure in genetic mutants in which MAP3K1 signaling was attenuated but did not disturb this developmental program in either wild type or mutant mice with attenuated epidermal growth factor receptor or WNT signaling. Exposure also markedly inhibited c-Jun phosphorylation in Map3k1+/− embryonic eyelid epithelium, suggesting that dioxin-induced AHR pathways can synergize with gene mutations to inhibit MAP3K1 signaling. Our studies uncover a novel mechanism through which the dioxin-AHR axis interacts with the MAP3K1 signaling pathways during fetal development and provide strong empirical evidence that specific gene alterations can increase the risk of developmental abnormalities driven by environmental pollutant exposure. PMID:26109068

  13. Comprehensive mapping of O-glycosylation in flagellin from Campylobacter jejuni 11168: A multienzyme differential ion mobility mass spectrometry approach.

    Science.gov (United States)

    Ulasi, Gloria N; Creese, Andrew J; Hui, Sam Xin; Penn, Charles W; Cooper, Helen J

    2015-08-01

    Glycosylation of flagellin is essential for the virulence of Campylobacter jejuni, a leading cause of bacterial gastroenteritis. Here, we demonstrate comprehensive mapping of the O-glycosylation of flagellin from Campylobacter jejuni 11168 by use of a bottom-up proteomics approach that incorporates differential ion mobility spectrometry (also known as high field asymmetric waveform ion mobility spectrometry or FAIMS) together with proteolysis with proteinase K. Proteinase K provides complementary sequence coverage to that achieved following trypsin proteolysis. The use of FAIMS increased the number of glycopeptides identified. Novel glycans for this strain were identified (pseudaminic acid and either acetamidino pseudaminic acid or legionaminic acid), as were novel glycosylation sites: Thr208, Ser343, Ser348, Ser349, Ser395, Ser398, Ser423, Ser433, Ser436, Ser445, Ser448, Ser451, Ser452, Ser454, Ser457 and Thr465. Multiply glycosylated peptides were observed, as well as variation at individual residues in the nature of the glycan and its presence or absence. Such extreme heterogeneity in the pattern of glycosylation has not been reported previously, and suggests a novel dimension in molecular variation within a bacterial population that may be significant in persistence of the organism in its natural environment. These results demonstrate the usefulness of differential ion mobility in proteomics investigations of PTMs.

  14. Cultivos de células CHO-K1

    Directory of Open Access Journals (Sweden)

    M.C. Nóvoa-Valiñas

    2005-01-01

    Full Text Available El uso de determinados metales pesados y pesticidas es la estrategia más empleada para el control de plagas. Estas sustancias, una vez aplicadas a los cultivos, pueden pasan al medio ambiente, permaneciendo en él como xenobióticos que van a afectar, en mayor o menor medida, a los seres vivos. En el presente estudio se ha evaluado la toxicidad basal de un metal, cobre, y un pesticida organoclorado, lindano, así como mezclas de ambos a distintas concentraciones. Para llevar a cabo este trabajo se ha utilizado la línea celular CHO-K1 (células epiteliales de ovario de hamster, usándose como criterio de citotoxicidad la muerte celular, determinada mediante la técnica del rojo neutro. Las concentraciones iniciales fueron: 0,03; 0,06 y 0,9 mM de cobre y 0,01; 0,03 y 0,1 mM de lindano. Y en las mezclas, las concentraciones estuvieron comprendidas entre 0,01-0,9 de cobre y 0,001-0,1de lindano. Como resultados, la citotoxicidad del cobre y lindano fue dosis-dependiente. En las exposiciones a mezclas se observa que a concentraciones fijas de lindano, la viabilidad desciende al aumentar la concentración de cobre, mientras que, dentro de un cierto rango, a concentraciones fijas de cobre, la viabilidad celular se incrementa al aumentar la concentración de lindano

  15. 计算n∑k=1 f(k)的一个初等方法

    Institute of Scientific and Technical Information of China (English)

    杨欣芳

    1994-01-01

    本文利用二项式定理推出前n个自然数的各次方和公式,进而将n∑k=1 f(k)=n∑k=1 (ackm+a1km-1+...+am-1k+am)写成n∑k=1 km+a1n∑k=1 km-1+...+am-1n∑k=1 K+amn∑k=1 1从而进行有关的计算。

  16. Measurement of Branching Fractions of B0 Decays to K1(1270)+ pi- and K1(1400)+ pi-

    Energy Technology Data Exchange (ETDEWEB)

    Aubert, Bernard; Bona, M.; Karyotakis, Y.; Lees, J.P.; Poireau, V.; Prencipe, E.; Prudent, X.; Tisserand, V.; /Annecy, LAPP; Garra Tico, J.; Grauges, E.; /Barcelona U., ECM; Lopez, L.; Palano, Antimo; Pappagallo, M.; /Bari U. /INFN, Bari; Eigen, G.; Stugu, Bjarne; Sun, L.; /Bergen U.; Abrams, G.S.; Battaglia, M.; Brown, D.N.; Cahn, Robert N.; Jacobsen, R.G.; /LBL, Berkeley /Birmingham U. /Ruhr U., Bochum /Bristol U. /British Columbia U. /Brunel U. /Novosibirsk, IYF /UC, Irvine /UCLA /UC, Riverside /UC, San Diego /UC, Santa Barbara /UC, Santa Cruz /Caltech /Cincinnati U. /Colorado U. /Colorado State U. /Dortmund U. /Dresden, Tech. U. /Ecole Polytechnique /Edinburgh U. /Ferrara U. /INFN, Ferrara /Frascati /Genoa U. /INFN, Genoa /Harvard U. /Heidelberg U. /Humboldt U., Berlin /Imperial Coll., London /Iowa U. /Iowa State U. /Johns Hopkins U. /Orsay, LAL /LLNL, Livermore /Liverpool U. /Queen Mary, U. of London /Royal Holloway, U. of London /Louisville U. /Mainz U., Inst. Kernphys. /Manchester U. /Maryland U. /Massachusetts U., Amherst /MIT /McGill U. /Consorzio Milano Ricerche /INFN, Milan /Mississippi U. /Montreal U. /Mt. Holyoke Coll. /Napoli Seconda U. /INFN, Naples /NIKHEF, Amsterdam /Notre Dame U. /Ohio State U. /Oregon U. /Padua U. /INFN, Padua /Paris U., VI-VII /Pennsylvania U. /Perugia U. /INFN, Perugia /INFN, Pisa /Princeton U. /Banca di Roma /Frascati /Rostock U. /Rutherford /DAPNIA, Saclay /South Carolina U. /SLAC /Stanford U., Phys. Dept. /SUNY, Albany /Tennessee U. /Texas U. /Texas U., Dallas /Turin U. /INFN, Turin /Trieste U. /INFN, Trieste /Valencia U., IFIC /Victoria U. /Warwick U. /Wisconsin U., Madison

    2008-08-04

    We present a measurement of the branching fraction of neutral B meson decaying to final states containing a K1 meson, i.e. K{sub 1}(1270) and K{sub 1}(1400), and a charged pion. The data, collected with the BABAR detector at the Stanford Linear Accelerator Center, represent 454 million B{bar B} pairs produced in e{sup +}e{sup -} annihilation. We measure the branching fraction {Beta}(B{sup 0} {yields} K{sub 1}{sup +}{pi}{sup -}) = (31.0 {+-} 2.7 {+-} 6.9) x 10{sup -6}, where the first error quoted is statistical and the second is systematic. In the framework of the K-matrix formalism used to describe these decays, we also set limits on the ratio of the production constants for the K{sub 1}(1270){sup +} and K{sub 1}(1400){sup +} mesons in B{sup 0} decays.

  17. Inter-laboratory evaluation of three flagellin PCR/RFLP methods for typing Campylobacter jejuni and C-coli: the CAMPYNET experience

    DEFF Research Database (Denmark)

    Harrington, Clare S.; Moran, L.; Ridley, A.M.

    2003-01-01

    Aims: To compare typeability, discriminatory ability, and inter-laboratory reproducibility of three flagellin PCR/RFLP(fla typing) methods previously described for Campylobacter. Methods and Results: The sample set(n = 100) was diverse, including both C. jejuni (n = 85) and C. coli (n = 15). Two...

  18. p53 modulates the effect of ribosomal protein S6 kinase1 (S6K1) on cisplatin toxicity in chronic myeloid leukemia cells.

    Science.gov (United States)

    Xiao, Ling-Yi; Kan, Wai-Ming

    2017-03-14

    Chronic myeloid leukemia (CML) is characterized by the expression of the oncoprotein, BCR-ABL. BCR-ABL inhibitors revolutionized CML chemotherapy while blast crisis (BC) CML patients are less responsive. Since suppression of ribosomal protein S6 kinase1 (S6K1) phosphorylation reverses the resistance to BCR-ABL inhibitor in CML cells and S6K1 inhibitors augment cisplatin toxicity in lung cancer cells, we speculated that combination of S6K1 inhibitor and cisplatin may be beneficial for eliminating BC CML cells. To our surprise, S6K1 inhibition decreased cisplatin-induced DNA damage and cell death only in p53(-/-) BC CML cells but not in p53(+/+) BC CML cells. During the progression of CML, p53 expression either decreases or mutates. Moreover, the expression of p53 affects drug response of CML cells. Our results confirmed that S6K1 inhibition reversed cisplatin toxicity is dependent on p53 expression in CML cells. Moreover, p53 attenuated the phosphorylation and localization of S6K1 via attenuating 3-phosphoinositide dependent protein kinase-1 (PDK1) phosphorylation. Furthermore, S6K1 acts via DNA-PKcs to regulate H2AX phosphorylation and PARP cleavage, respectively. Taken together, our results suggest that p53/PDK1/S6K1 is a novel pathway regulating cisplatin toxicity in BC CML cells.

  19. Productions of $K^*_0(1430)$ and $K_{1}$ in B decays

    CERN Document Server

    Chen, C H; Hsiao, Y K; Wei, Z T; Chen, Chuan-Hung; Geng, Chao-Qiang; Hsiao, Yu-Kuo; Wei, Zheng-Tao

    2005-01-01

    We study the productions of p-wave mesons $K^*_{0}(1430)$, $K_{1}(1270)$ and $K_{1}(1400)$ in B decays. By the generalized factorization approach, we find that the branching ratios of $B\\to K^*_{0}(1430) \\phi$ are similar to those of $B\\to K \\phi$ while the branching ratios of $B\\to K_{1}(1270) \\phi$ and $B\\to K_{1}(1400) \\phi$ are %could be $O(10^{-5})$ and $O(10^{-6})$, respectively. In terms of the observation of $B\\to K_{1}(1270) \\gamma$ by BELLE, we can remove the sign ambiguity in the mixing angle for physical states $K_1(1270)$ and $K_{1}(1400)$. In addition, we analyze annihilation contributions in the decays $B\\to K_1 \\phi$ and we conclude that they could be neglected.

  20. Effects of aldicarb and propoxur on cytotoxicity and lipid peroxidation in CHO-K1 cells.

    Science.gov (United States)

    Maran, E; Fernández-Franzón, M; Font, G; Ruiz, M J

    2010-06-01

    Cytotoxic effects of aldicarb, its sulfone and sulfoxide, and propoxur, lipid peroxidation and antioxidant parameters in Chinese Hamster Ovary (CHO-K1) cells were determined. D,L-buthionine-(S,R)-sulfoximine (BSO) was assayed to determine the role of GSH in the protection against carbamate cytotoxicity. Pre-treatment with 60 microM BSO, induced a significant decrease in the glutathione reductase (GR; 64-141%), the glutathione peroxidase (GPx; 10-30%) and the glutathione S-transferase (GST; 59-93%) activities, and its GSH levels (79-85%), while the oxidized glutathione (GSSG) levels significantly increased (64-78%) respect to experiment non-BSO-pretreated. Carbamates BSO pre-treated vs. non-BSO pre-treated showed a significant increase in malondialdehyde (MDA) production (from 13% to 52% vs. 25% to 93%). These data suggest that carbamates could injure CHO-K1 cells via oxidative stress by the increase of MDA production; moreover, BSO enhance the oxidative damage caused by carbamates. However, the glutathione system protects cells from carbamates damage.

  1. Bending elasticity modulus of giant vesicles composed of aeropyrum pernix k1 archaeal lipid.

    Science.gov (United States)

    Genova, Julia; Ulrih, Nataša Poklar; Kralj-Iglič, Veronika; Iglič, Aleš; Bivas, Isak

    2015-03-26

    Thermally induced shape fluctuations were used to study elastic properties of giant vesicles composed of archaeal lipids C25,25-archetidyl (glucosyl) inositol and C25,25-archetidylinositol isolated from lyophilised Aeropyrum pernix K1 cells. Giant vesicles were created by electroformation in pure water environment. Stroboscopic illumination using a xenon flash lamp was implemented to remove the blur effect due to the finite integration time of the camera and to obtain an instant picture of the fluctuating vesicle shape. The mean weighted value of the bending elasticity modulus kc of the archaeal membrane determined from the measurements meeting the entire set of qualification criteria was (1.89 ± 0.18) × 10-19 J, which is similar to the values obtained for a membrane composed of the eukaryotic phospholipids SOPC (1.88 ± 0.17) × 10-19 J and POPC (2.00 ± 0.21) ´ 10-19 J. We conclude that membranes composed of archaeal lipids isolated from Aeropyrum pernix K1 cells have similar elastic properties as membranes composed of eukaryotic lipids. This fact, together with the importance of the elastic properties for the normal circulation through blood system, provides further evidence in favor of expectations that archaeal lipids could be appropriate for the design of drug delivery systems.

  2. More on the degree condition for the existence of regular factors in K1,n-free graphs%关于在K1,n-free图中存在正则因子度条件的推广

    Institute of Scientific and Technical Information of China (English)

    李建湘

    2004-01-01

    图被称为 K1,n-free图,如果它不含有导出子图K1,n. 设G是一个具有顶点集V(G)的图, 并设g和f是两个定义在V(G)的函数,使得g(x)≤f(x)对所有V(G)中的点x都成立.设a=max{g(x)|x∈V(G)}, b=min{f(x)|x∈V(G)}, 并有b, a≥2, n≥b/(a-1)+1(如果存在点v∈V(G)使得f(v)≡1(mod 2), 假定b≥n-1). 证明了:每个连通的使得∑x∈V(G)f(x)为偶数的K1,n-free图G有(g,f)-因子,如果它的最小度至少是((n-1)(a+1))/(b)+1「(b+a(n-1))/(2(n-1))-(n-1)/(b)「(b+a(n-1))/(2(n-1))2+n-3.这个结果是K.Ota 和T.Tokuda(J. Graph Theory. 1996, 22:59-64.)关于在K1,n-free 图中存在正则因子度条件的推广.%A graph is called K1,n-free if it contains no K1,n as an induced subgraph. Let G be a graph with vertex set V(G), and let g and f be two integer-valued functions defined on V(G) such that g(x)≤f(x) for all x∈V(G). Let a =max {g(x)|x∈V(G)}, b=min {f(x)|x∈V(G)}, and b, a≥2, n≥b/(a-1)+1(if there exists a vertex v∈V(G) such that f(v)≡1 (mod 2), b≥n-1). We prove that every K1,n-free connected graph G with ∑x∈V(G) f(x) even has a (g, f)-factor if its minimum degree is at leastThis result is the generalization for the existence theorem of regular factors in K1,n-free graphs, which is due to K. Ota and T. Tokuda (J. Graph Theory. 1996, 22:59-64).

  3. An Eimeria vaccine candidate based on Eimeria tenella immune mapped protein 1 and the TLR-5 agonist Salmonella typhimurium FliC flagellin

    Energy Technology Data Exchange (ETDEWEB)

    Yin, Guangwen; Qin, Mei [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Liu, Xianyong [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Key Laboratory of Zoonosis, China Ministry of Agriculture and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Suo, Jingxia; Tang, Xinming; Tao, Geru [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Han, Qian [Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061 (United States); Suo, Xun [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Key Laboratory of Zoonosis, China Ministry of Agriculture and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Wu, Wenxue, E-mail: labboard@126.com [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Key Laboratory of Zoonosis, China Ministry of Agriculture and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China)

    2013-10-25

    Highlights: •We found a new protective protein – (IMPI) in Eimeria tenella. •EtIMP1-flagellin fusion protein is an effective immunogen against Eimeria infection. •Flagellin can be as an apicomplexan parasite vaccine adjuvant in chickens. -- Abstract: Immune mapped protein-1 (IMP1) is a new protective protein in apicomplexan parasites, and exits in Eimeria tenella. But its structure and immunogenicity in E. tenella are still unknown. In this study, IMPI in E. tenella was predicted to be a membrane protein. To evaluate immunogenicity of IMPI in E. tenella, a chimeric subunit vaccine consisting of E. tenella IMP1 (EtIMP1) and a molecular adjuvant (a truncated flagellin, FliC) was constructed and over-expressed in Escherichia coli and its efficacy against E. tenella infection was evaluated. Three-week-old AA broiler chickens were vaccinated with the recombinant EtIMP1-truncated FliC without adjuvant or EtIMP1 with Freund’s Complete Adjuvant. Immunization of chickens with the recombinant EtIMP1-truncated FliC fusion protein resulted in stronger cellular immune responses than immunization with only recombinant EtIMP1 with adjuvant. The clinical effect of the EtIMP1-truncated FliC without adjuvant was also greater than that of the EtIMP1 with adjuvant, which was evidenced by the differences between the two groups in body weight gain, oocyst output and caecal lesions of E. tenella-challenged chickens. The results suggested that the EtIMP1-flagellin fusion protein can be used as an effective immunogen in the development of subunit vaccines against Eimeria infection. This is the first demonstration of antigen-specific protective immunity against avian coccidiosis using a recombinant flagellin as an apicomplexan parasite vaccine adjuvant in chickens.

  4. 荧光假单胞菌GcM51 A重组鞭毛蛋白的表达及其对黑松细胞的毒性研究%Expression of Recombinant Flagellin from Pseudomonas Fluorescens GcM5 1 A and It’s Toxicity to Pinus Thunbergii Cells

    Institute of Scientific and Technical Information of China (English)

    肖莉; 张蕾; 牟卉; 李荣贵

    2013-01-01

    针对松萎蔫病的早期诊断及生物防治,本文利用PCR 技术从荧光假单胞菌 GcM51 A的基因组中克隆了鞭毛蛋白编码基因 fliC,再将该基因克隆到表达载体 pET 15 b的NcoI 和XhoI位点,构建重组质粒pET 15 b fliC,再将重组质粒导入大肠杆菌BL21(DE3),构建工程菌,IPTG诱导工程菌高效表达 C 末端具有多聚组氨酸标签的重组鞭毛蛋白,重组蛋白主要以包涵体的形式存在,包涵体经8 mol/L尿素溶解,复性并经 Ni2+螯合柱亲和层析得到了电泳纯的重组鞭毛蛋白。生测结果表明,重组鞭毛蛋白可引起黑松细胞的大量死亡,与天然鞭毛蛋白对黑松愈伤组织细胞有相似的毒性。该研究为松萎蔫病致病机理的研究奠定了基础。%In this paper,gene fliC encoding flagellin was amplified from genomic DNA of Pseudomonasflu-orescens GcM5 1A by polymerase chain reaction (PCR),and cloned into pET 15b to construct recombi-nant plasmid pET 15b fliC.This plasmid was introduced into Escherichiacoli BL21(DE3)to construct engineering bacteria,and overexpression of recombinant flagellin with a his-tag at its C-terminal was a-chieved when engineering bacteria were induced by isopropylβ D 1 thiogalactopyranoside (IPTG). The recombinant protein in engineering bacteria appeared mainly in the form of inclusion bodies.Recombi-nant flagellin in inclusion bodies,which was dissolved by 8 mol/L urea and refolded,was purified through affinity chromatography on a nickel column.Bioassay results indicated that the recombinant protein was toxic to callus cells of Pinus thunbergii,which had a similar result to native flagellin.This study laid a good foundation for further research on the mechanism of pine wilt disease.

  5. Sustained protection in mice immunized with fractional doses of Salmonella Enteritidis core and O polysaccharide-flagellin glycoconjugates.

    Directory of Open Access Journals (Sweden)

    Raphael Simon

    Full Text Available Non-typhoidal Salmonella (NTS serovars S. Enteritidis and S. Typhimurium are a major cause of invasive bacterial disease (e.g., bacteremia, meningitis in infants and young children in sub-Saharan Africa and also occasionally cause invasive disease in highly susceptible hosts (young infants, the elderly, and immunocompromised subjects in industrialized countries. No licensed vaccines exist against human NTS infections. NTS core and O polysaccharide (COPS and FliC (Phase 1 flagellin subunits each constitute protective antigens in murine models. S. Enteritidis COPS conjugated to FliC represents a promising vaccine approach that elicits binding and opsonophagocytic antibodies and protects mice against lethal challenge with virulent S. Enteritidis. We examined the protective efficacy of fractional dosages of S. Enteritidis COPS:FliC conjugate vaccines in mice, and also established that protection can be passively transferred to naïve mice by administering sera from mice immunized with conjugate. Mice were immunized with three doses of either 10 µg, 2.5 µg (full dose, 0.25 µg, or 0.025 µg S. Enteritidis COPS:FliC conjugate at 28 day intervals. Antibody titers to COPS and FliC measured by ELISA fell consonant with progressively smaller vaccine dosage levels; anti-FliC IgG responses remained robust at fractional dosages for which anti-COPS serum IgG titers were decreased. Nevertheless, >90% protection against intraperitoneal challenge was observed in mice immunized with fractional dosages of conjugate that elicited diminished titers to both FliC and COPS. Passive transfer of immune sera from mice immunized with the highest dose of COPS:FliC to naïve mice was also protective, demonstrating the role of antibodies in mediating protection. These results provide important insights regarding the potency of Salmonella glycoconjugate vaccines that use flagellin as a carrier protein.

  6. Flagellin FliC Phosphorylation Affects Type 2 Protease Secretion and Biofilm Dispersal in Pseudomonas aeruginosa PAO1

    Science.gov (United States)

    Suriyanarayanan, Tanujaa; Periasamy, Saravanan; Lin, Miao-Hsia; Ishihama, Yasushi; Swarup, Sanjay

    2016-01-01

    Protein phosphorylation has a major role in controlling the life-cycle and infection stages of bacteria. Proteome-wide occurrence of S/T/Y phosphorylation has been reported for many prokaryotic systems. Previously, we reported the phosphoproteome of Pseudomonas aeruginosa and Pseudomonas putida. In this study, we show the role of S/T phosphorylation of one motility protein, FliC, in regulating multiple surface-associated phenomena of P. aeruginosa PAO1. This is the first report of occurrence of phosphorylation in the flagellar protein, flagellin FliC in its highly conserved N-terminal NDO domain across several Gram negative bacteria. This phosphorylation is likely a well-regulated phenomenon as it is growth phase dependent in planktonic cells. The absence of phosphorylation in the conserved T27 and S28 residues of FliC, interestingly, did not affect swimming motility, but affected the secretome of type 2 secretion system (T2SS) and biofilm formation of PAO1. FliC phosphomutants had increased levels and activities of type 2 secretome proteins. The secretion efficiency of T2SS machinery is associated with flagellin phosphorylation. FliC phosphomutants also formed reduced biofilms at 24 h under static conditions and had delayed biofilm dispersal under dynamic flow conditions, respectively. The levels of type 2 secretome and biofilm formation under static conditions had an inverse correlation. Hence, increase in type 2 secretome levels was accompanied by reduced biofilm formation in the FliC phosphomutants. As T2SS is involved in nutrient acquisition and biofilm dispersal during survival and spread of P. aeruginosa, we propose that FliC phosphorylation has a role in ecological adaptation of this opportunistic environmental pathogen. Altogether, we found a system of phosphorylation that affects key surface related processes such as proteases secretion by T2SS, biofilm formation and dispersal. PMID:27701473

  7. Development of a Certified Reference Material for Lyophilized Vitamin K 1%维生素 K1冻干标准物质的研制

    Institute of Scientific and Technical Information of China (English)

    张晓光; 黄挺; 张伟; 黄亮; 全灿; 李红梅; 杨屹

    2015-01-01

    采用二甲基亚砜(DMSO)作为冻干标准物质的溶剂,以重量法准确配制维生素 K1/ DMSO 溶液,再进行分装、冷冻干燥,经定性分析、定值分析、均匀性检验、稳定性考察和不确定度评定,研制了维生素 K1冻干标准物质。以维生素 K1纯度标准物质溶液为校准品,对得到的冻干物质进行了高效液相色谱法定值分析,冻干标准物质的准确定值结果为0.96 mg/ mL,相对扩展不确定度为7%。建立的维生素 K1冻干标准物质研制方法,对于临床检验中维生素 K1的准确测定和相关疾病的正确诊断治疗以及维生素 K1长期保存具有重要意义。%Dimethyl sulfone( DMSO)is firstly applied as the solvent for a lyophilized certified reference material (CRM). Vitamin K1 / DMSO solution is prepared by gravimetry,and is sub-divided and lyophilized,the lyophilized vitamin K1 CRM is developed through qualitative analysis,quantitative analysis,homogeneity study,stability study and uncertainty evaluation. Certified value of lyophilization vitamin K1 is analyzed by high performance liquid chromatography method, using the vitamin K1 purity CRM as a calibrant. The certified value is 0. 96 mg/ mL,with the relative expanded uncertainty of 7% . The method of CRM lyophilized vitamin K1 makes great significance for the accurate determination the disease in clinical examination and the long-term preservation of vitamin K1 .

  8. Ramsey Number of K2,s+1 vs. K1,n%关于K2,s+1 VS,K1,n的Ramsey数

    Institute of Scientific and Technical Information of China (English)

    秦大伟; 沈大鹏

    2007-01-01

    It is shown that the Ramsey number r(K2,s+1, K1,n) ≤ n + √sn+ (s + 3)/2 + o(1) for large n, and r(K2,s+1, K1,n)∈{(q-1)2/s+1,-(q-1)2/s+2},wheren: (q-1)2/s-q+2 and q is a prime power such that s|(q - 1).

  9. SIRT1 overexpression antagonizes cellular senescence with activated ERK/S6k1 signaling in human diploid fibroblasts.

    Directory of Open Access Journals (Sweden)

    Jing Huang

    Full Text Available Sir2, a NAD-dependent deacetylase, modulates lifespan in yeasts, worms and flies. The SIRT1, mammalian homologue of Sir2, regulates signaling for favoring survival in stress. But whether SIRT1 has the function to influence cell viability and senescence under non-stressed conditions in human diploid fibroblasts is far from unknown. Our data showed that enforced SIRT1 expression promoted cell proliferation and antagonized cellular senescence with the characteristic features of delayed Senescence-Associated beta-galactosidase (SA-beta-gal staining, reduced Senescence-Associated Heterochromatic Foci (SAHF formation and G1 phase arrest, increased cell growth rate and extended cellular lifespan in human fibroblasts, while dominant-negative SIRT1 allele (H363Y did not significantly affect cell growth and senescence but displayed a bit decreased lifespan. Western blot results showed that SIRT1 reduced the expression of p16(INK4A and promoted phosphorylation of Rb. Our data also exposed that overexpression of SIRT1 was accompanied by enhanced activation of ERK and S6K1 signaling. These effects were mimicked in both WI38 cells and 2BS cells by concentration-dependent resveratrol, a SIRT1 activator. It was noted that treatment of SIRT1-.transfected cells with Rapamycin, a mTOR inhibitor, reduced the phosphorylation of S6K1 and the expression of Id1, implying that SIRT1-induced phosphorylation of S6K1 may be partly for the decreased expression of p16(INK4A and promoted phosphorylation of Rb in 2BS. It was also observed that the expression of SIRT1 and phosphorylation of ERK and S6K1 was declined in senescent 2BS. These findings suggested that SIRT1-promoted cell proliferation and antagonized cellular senescence in human diploid fibroblasts may be, in part, via the activation of ERK/ S6K1 signaling.

  10. Peptides-Derived from Thai Rice Bran Improves Endothelial Function in 2K-1C Renovascular Hypertensive Rats

    Directory of Open Access Journals (Sweden)

    Orachorn Boonla

    2015-07-01

    Full Text Available In recent years, a number of studies have investigated complementary medical approaches to the treatment of hypertension using dietary supplements. Rice bran protein hydrolysates extracted from rice is a rich source of bioactive peptides. The present study aimed to investigate the vasorelaxation and antihypertensive effects of peptides-derived from rice bran protein hydrolysates (RBP in a rat model of two kidney-one clip (2K-1C renovascular hypertension. 2K-1C hypertension was induced in male Sprague-Dawley rats by placing a silver clip around the left renal artery, whereas sham-operated rats were served as controls. 2K-1C and sham-operated rats were intragastrically administered with RBP (50 mg kg−1 or 100 mg kg−1 or distilled water continuously for six weeks. We observed that RBP augmented endothelium-dependent vasorelaxation in all animals. Administration of RBP to 2K-1C rats significantly reduced blood pressure and decreased peripheral vascular resistance compared to the sham operated controls (p < 0.05. Restoration of normal endothelial function and blood pressure was associated with reduced plasma angiotensin converting enzyme (ACE, decreased superoxide formation, reduced plasma malondialdehyde and increased plasma nitrate/nitrite (p < 0.05. Up-regulation of eNOS protein and down-regulation of p47phox protein were found in 2K-1C hypertensive rats-treated with RBP. Our results suggest that RBP possesses antihypertensive properties which are mainly due to the inhibition of ACE, and its vasodilatory and antioxidant activity.

  11. The K-1 reusable aerospace vehicle: managing to achieve low cost.

    Science.gov (United States)

    Mueller (HM), George E.; Lepore, Debra Facktor

    2000-03-01

    Kistler Aerospace Corporation is developing the world's first privately funded, fully reusable aerospace vehicle, the K-1. This vehicle represents a new implementation of proven technologies, designed by an elite, experienced team of engineers and managers and implemented by the best manufacturing capability in the United States. Kistler Aerospace expects to begin commercial operations of the K-1 in 2000. Market researchers predict that during the next decade telecommunications satellite ventures will require launch services for over 1,400 payloads to LEO. This prediction greatly exceeds the current available industry capacity. The K-1 was designed primarily to meet this anticipated growth in demand. Significant progress has been made in constructing the K-1 vehicle fleet. The fully reusable K-1 vehicle is designed to lower the cost of access to space, increase launch reliability, and reduce lead-time-to-launch requirements. The K-1 will offer significant cost benefits and aircraft type reliability based on a proven flight record.

  12. Cytotoxic effects of zearalenone and its metabolites and antioxidant cell defense in CHO-K1 cells.

    Science.gov (United States)

    Tatay, Elena; Font, Guillermina; Ruiz, Maria-Jose

    2016-10-01

    Zearalenone (ZEA) and its metabolites (α-zearalenol; α-ZOL, β-zearalenol; β-ZOL) are secondary metabolites of Fusarium fungi that produce cell injury. The present study explores mycotoxin-induced cell damage and cellular protection mechanisms in CHO-K1 cells. Cytotoxicity has been determined by reactive oxygen species (ROS) production and DNA damage. ROS production was determined using the fluorescein assay and DNA strand breakage by comet assay. Intracellular protection systems were glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD). The results demonstrated that all mycotoxins increased the ROS levels up to 5.3-fold the control levels in CHO-K1 cells. Zearalenone metabolites, but not ZEA, increased DNA damage 43% (α-ZOL) and 28% (β-ZOL) compared to control cells. The GSH levels decreased from 18% to 36%. The GPx and SOD activities respectively increased from 26% to 62% and from 23% to 69% in CHO-K1 cells, whereas CAT activity decreased from 14% to 52%. In addition, intracellular ROS production was induced by ZEA and its metabolites. The endogenous antioxidant system components GSH, GPx and SOD were activated against ZEA and its metabolites. These antioxidant system components thus could contribute to decrease cell injury by ZEA and its metabolites. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. [Research on progress and prospect of kinase S6K1].

    Science.gov (United States)

    Zhang, Hui; Liang, Junyu; Zhang, Ji

    2014-08-01

    Obesity is a prevalent metabolic disorder, which seriously affects human health and has become the world's public health problem. Kinase S6K1, an important downstream effector of mammalian target of rapamycin (mTOR), influences specific pathological responses, including obesity, type 2 diabetes and cancer. Presently, S6K1 has become an attractive therapeutic target in the treatment of these disorders. Here, the functions of kinase S6K1, its molecular regulation mechanisms, related pathogenesis of disease and relevant small molecular inhibitors are reviewed. Finally, the prospect of research toward S6K1 is expected as well.

  14. A non-synonymous coding variant (L616F in the TLR5 gene is potentially associated with Crohn's disease and influences responses to bacterial flagellin.

    Directory of Open Access Journals (Sweden)

    Jared Sheridan

    Full Text Available BACKGROUND AND OBJECTIVES: Although numerous studies have implicated TLR5, or its ligands, bacterial flagellins, in the pathogenesis of Crohn's disease (CD, genome-wide association studies (GWAS have not reported associations with the TLR5 gene. We aimed to examine potential CD-associated TLR5 variants and assess whether they modified inflammatory responses to bacterial flagellins. METHODS AND PRINCIPAL RESULTS: A two-stage study was carried out. In stage 1, we genotyped tagging single-nucleotide polymorphisms (tag-SNPs in the TLR5 gene in a sample of CD cases (<20 years of age, N = 566 and controls (N = 536. Single SNP and haplotype analysis was carried out. In Stage 2, we assessed the functional significance of potential CD-associated variant(s vis-à-vis effects on the inflammatory response to bacterial flagellin using HEK293T cells. We observed marginal association between a non-synonymous coding SNP rs5744174 (p = 0.05 and CD. Associations between SNP rs851139 that is in high linkage disequilibrium (LD with SNP rs5744174 were also suggested (p = 0.07. Haplotype analysis revealed that a 3 marker haplotype was significantly associated with CD (p = 0.01. Functional studies showed that the risk allele (616F (corresponding to the C allele of SNP rs5744174 conferred significantly greater production of CCL20 in response to a range of flagellin doses than the comparator allele (616L. CONCLUSIONS: Our findings suggest that a non-synonymous coding variation in the TLR5 gene may confer modest susceptibility for CD.

  15. Role of Glycosyltransferases Modifying Type B Flagellin of Emerging Hypervirulent Clostridium difficile Lineages and Their Impact on Motility and Biofilm Formation*

    Science.gov (United States)

    Valiente, Esmeralda; Bouché, Laura; Hitchen, Paul; Faulds-Pain, Alexandra; Songane, Mario; Dawson, Lisa F.; Donahue, Elizabeth; Stabler, Richard A.; Panico, Maria; Morris, Howard R.; Bajaj-Elliott, Mona; Logan, Susan M.; Dell, Anne; Wren, Brendan W.

    2016-01-01

    Clostridium difficile is the principal cause of nosocomial infectious diarrhea worldwide. The pathogen modifies its flagellin with either a type A or type B O-linked glycosylation system, which has a contributory role in pathogenesis. We study the functional role of glycosyltransferases modifying type B flagellin in the 023 and 027 hypervirulent C. difficile lineages by mutagenesis of five putative glycosyltransferases and biosynthetic genes. We reveal their roles in the biosynthesis of the flagellin glycan chain and demonstrate that flagellar post-translational modification affects motility and adhesion-related bacterial properties of these strains. We show that the glycosyltransferases 1 and 2 (GT1 and GT2) are responsible for the sequential addition of a GlcNAc and two rhamnoses, respectively, and that GT3 is associated with the incorporation of a novel sulfonated peptidyl-amido sugar moiety whose structure is reported in our accompanying paper (Bouché, L., Panico, M., Hitchen, P., Binet, D., Sastre, F., Faulds-Pain, A., Valiente, E., Vinogradov, E., Aubry, A., Fulton, K., Twine, S., Logan, S. M., Wren, B. W., Dell, A., and Morris, H. R. (2016) J. Biol. Chem. 291, 25439–25449). GT2 is also responsible for methylation of the rhamnoses. Whereas type B modification is not required for flagellar assembly, some mutations that result in truncation or abolition of the glycan reduce bacterial motility and promote autoaggregation and biofilm formation. The complete lack of flagellin modification also significantly reduces adhesion of C. difficile to Caco-2 intestinal epithelial cells but does not affect activation of human TLR5. Our study advances our understanding of the genes involved in flagellar glycosylation and their biological roles in emerging hypervirulent C. difficile strains. PMID:27703012

  16. Deletion of inositol hexakisphosphate kinase 1 (IP6K1) reduces cell migration and invasion, conferring protection from aerodigestive tract carcinoma in mice.

    Science.gov (United States)

    Jadav, Rathan S; Kumar, Dharmika; Buwa, Natasha; Ganguli, Shubhra; Thampatty, Sitalakshmi R; Balasubramanian, Nagaraj; Bhandari, Rashna

    2016-08-01

    Inositol hexakisphosphate kinases (IP6Ks), a family of enzymes found in all eukaryotes, are responsible for the synthesis of 5-diphosphoinositol pentakisphosphate (5-IP7) from inositol hexakisphosphate (IP6). Three isoforms of IP6Ks are found in mammals, and gene deletions of each isoform lead to diverse, non-overlapping phenotypes in mice. Previous studies show a facilitatory role for IP6K2 in cell migration and invasion, properties that are essential for the early stages of tumorigenesis. However, IP6K2 also has an essential role in cancer cell apoptosis, and mice lacking this protein are more susceptible to the development of aerodigestive tract carcinoma upon treatment with the oral carcinogen 4-nitroquinoline-1-oxide (4NQO). Not much is known about the functions of the equally abundant and ubiquitously expressed IP6K1 isoform in cell migration, invasion and cancer progression. We conducted a gene expression analysis on mouse embryonic fibroblasts (MEFs) lacking IP6K1, revealing a role for this protein in cell receptor-extracellular matrix interactions that regulate actin cytoskeleton dynamics. Consequently, cells lacking IP6K1 manifest defects in adhesion-dependent signaling, evident by lower FAK and Paxillin activation, leading to reduced cell spreading and migration. Expression of active, but not inactive IP6K1 reverses migration defects in IP6K1 knockout MEFs, suggesting that 5-IP7 synthesis by IP6K1 promotes cell locomotion. Actin cytoskeleton remodeling and cell migration support the ability of cancer cells to achieve their complete oncogenic potential. Cancer cells with lower IP6K1 levels display reduced migration, invasion, and anchorage-independent growth. When fed an oral carcinogen, mice lacking IP6K1 show reduced progression from epithelial dysplasia to invasive carcinoma. Thus, our data reveal that like IP6K2, IP6K1 is also involved in early cytoskeleton remodeling events during cancer progression. However, unlike IP6K2, IP6K1 is essential for 4NQO-induced

  17. A (k+1)-Slope Theorem for the k-Dimensional Infinite Group Relaxation

    CERN Document Server

    Basu, Amitabh; Köppe, Matthias; Molinaro, Marco

    2011-01-01

    We prove that any minimal valid function for the k-dimensional infinite group relaxation that is piecewise linear with at most k+1 slopes and does not factor through a linear map with non-trivial kernel is extreme. This generalizes a theorem of Gomory and Johnson for k=1, and Cornuejols and Molinaro for k=2.

  18. 完全二部图的K1,pk-因子分解%K1,pk-FACTORIZATION OF COMPLETE BIPARTITE GRAPHS

    Institute of Scientific and Technical Information of China (English)

    杜北樑

    2001-01-01

    In this paper, it is shown that a sufficient condition for the existence of a K1,pk-factorization of Km,n, whenever p is a prime number and k is a positive integer, is (1) m≤pkn,(2) n≤pkm,(3)pkn-m≡pkm- n≡0(mod(p2k - 1)) and (4)(pkn-m)(pkm- n)≡0(mod(pk - 1)pk×(p2k-1)(m+n)).

  19. Inositol Hexakisphosphate Kinase 1 (IP6K1) Regulates Inositol Synthesis in Mammalian Cells.

    Science.gov (United States)

    Yu, Wenxi; Ye, Cunqi; Greenberg, Miriam L

    2016-05-13

    myo-Inositol, the precursor of all inositol compounds, has pivotal roles in cell metabolism and signaling pathways. Although physiological studies indicate a strong correlation between abnormal intracellular inositol levels and neurological disorders, very little is known about the regulation of inositol synthesis in mammalian cells. In this study, we report that IP6K1, an inositol hexakisphosphate kinase that catalyzes the synthesis of inositol pyrophosphate, regulates inositol synthesis in mammalian cells. Ip6k1 ablation led to profound changes in DNA methylation and expression of Isyna1 (designated mIno1), which encodes the rate-limiting enzyme inositol-3-phosphate synthase. Interestingly, IP6K1 preferentially bound to the phospholipid phosphatidic acid, and this binding was required for IP6K1 nuclear localization and the regulation of mIno1 transcription. This is the first demonstration of IP6K1 as a novel negative regulator of inositol synthesis in mammalian cells.

  20. Involvement of S6K1 in mitochondria function and structure in HeLa cells.

    Science.gov (United States)

    Park, Jisoo; Tran, Quangdon; Mun, Kisun; Masuda, Kouhei; Kwon, So Hee; Kim, Seon-Hwan; Kim, Dong-Hoon; Thomas, George; Park, Jongsun

    2016-12-01

    The major biological function of mitochondria is to generate cellular energy through oxidative phosphorylation. Apart from cellular respiration, mitochondria also play a key role in signaling processes, including aging and cancer metabolism. It has been shown that S6K1-knockout mice are resistant to obesity due to enhanced beta-oxidation, with an increased number of large mitochondria. Therefore, in this report, the possible involvement of S6K1 in regulating mitochondria dynamics and function has been investigated in stable lenti-shS6K1-HeLa cells. Interestingly, S6K1-stably depleted HeLa cells showed phenotypical changes in mitochondria morphology. This observation was further confirmed by detailed image analysis of mitochondria shape. Corresponding molecular changes were also observed in these cells, such as the induction of mitochondrial fission proteins (Drp1 and Fis1). Oxygen consumption is elevated in S6K1-depeleted HeLa cells and FL5.12 cells. In addition, S6K1 depletion leads to enhancement of ATP production in cytoplasm and mitochondria. However, the relative ratio of mitochondrial ATP to cytoplasmic ATP is actually decreased in lenti-shS6K1-HeLa cells compared to control cells. Lastly, induction of mitophagy was found in lenti-shS6K1-HeLa cells with corresponding changes of mitochondria shape on electron microscope analysis. Taken together, our results indicate that S6K1 is involved in the regulation of mitochondria morphology and function in HeLa cells. This study will provide novel insights into S6K1 function in mitochondria-mediated cellular signaling. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Modulation of pathogen-induced CCL20 secretion from HT-29 human intestinal epithelial cells by commensal bacteria.

    LENUS (Irish Health Repository)

    Sibartie, Shomik

    2009-01-01

    BACKGROUND: Human intestinal epithelial cells (IECs) secrete the chemokine CCL20 in response to infection by various enteropathogenic bacteria or exposure to bacterial flagellin. CCL20 recruits immature dendritic cells and lymphocytes to target sites. Here we investigated IEC responses to various pathogenic and commensal bacteria as well as the modulatory effects of commensal bacteria on pathogen-induced CCL20 secretion. HT-29 human IECs were incubated with commensal bacteria (Bifidobacterium infantis or Lactobacillus salivarius), or with Salmonella typhimurium, its flagellin, Clostridium difficile, Mycobacterium paratuberculosis, or Mycobacterium smegmatis for varying times. In some studies, HT-29 cells were pre-treated with a commensal strain for 2 hr prior to infection or flagellin stimulation. CCL20 and interleukin (IL)-8 secretion and nuclear factor (NF)-kappaB activation were measured using enzyme-linked immunosorbent assays. RESULTS: Compared to untreated cells, S. typhimurium, C. difficile, M. paratuberculosis, and flagellin activated NF-kappaB and stimulated significant secretion of CCL20 and IL-8 by HT-29 cells. Conversely, B. infantis, L. salivarius or M. smegmatis did not activate NF-kappaB or augment CCL20 or IL-8 production. Treatment with B. infantis, but not L. salivarius, dose-dependently inhibited the baseline secretion of CCL20. In cells pre-treated with B. infantis, C. difficile-, S. typhimurium-, and flagellin-induced CCL20 were significantly attenuated. B. infantis did not limit M. Paratuberculosis-induced CCL20 secretion. CONCLUSION: This study is the first to demonstrate that a commensal strain can attenuate CCL20 secretion in HT-29 IECs. Collectively, the data indicate that M. paratuberculosis may mediate mucosal damage and that B. infantis can exert immunomodulatory effects on IECs that mediate host responses to flagellin and flagellated enteric pathogens.

  2. SphK1 inhibitor II (SKI-II) inhibits acute myelogenous leukemia cell growth in vitro and in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Li; Weng, Wei; Sun, Zhi-Xin; Fu, Xian-Jie; Ma, Jun, E-mail: majuntongrensh1@126.com; Zhuang, Wen-Fang, E-mail: wenfangzhuangmd@163.com

    2015-05-15

    Previous studies have identified sphingosine kinase 1 (SphK1) as a potential drug target for treatment of acute myeloid leukemia (AML). In the current study, we investigated the potential anti-leukemic activity of a novel and specific SphK1 inhibitor, SKI-II. We demonstrated that SKI-II inhibited growth and survival of human AML cell lines (HL-60 and U937 cells). SKI-II was more efficient than two known SphK1 inhibitors SK1-I and FTY720 in inhibiting AML cells. Meanwhile, it induced dramatic apoptosis in above AML cells, and the cytotoxicity by SKI-II was almost reversed by the general caspase inhibitor z-VAD-fmk. SKI-II treatment inhibited SphK1 activation, and concomitantly increased level of sphingosine-1-phosphate (S1P) precursor ceramide in AML cells. Conversely, exogenously-added S1P protected against SKI-II-induced cytotoxicity, while cell permeable short-chain ceramide (C6) aggravated SKI-II's lethality against AML cells. Notably, SKI-II induced potent apoptotic death in primary human AML cells, but was generally safe to the human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. In vivo, SKI-II administration suppressed growth of U937 leukemic xenograft tumors in severe combined immunodeficient (SCID) mice. These results suggest that SKI-II might be further investigated as a promising anti-AML agent. - Highlights: • SKI-II inhibits proliferation and survival of primary and transformed AML cells. • SKI-II induces apoptotic death of AML cells, but is safe to normal PBMCs. • SKI-II is more efficient than two known SphK1 inhibitors in inhibiting AML cells. • SKI-II inhibits SphK1 activity, while increasing ceramide production in AML cells. • SKI-II dose-dependently inhibits U937 xenograft growth in SCID mice.

  3. Effect of hypothermia (20-25 degrees C) on mitosis in PtK1 cells.

    Science.gov (United States)

    Rieder, C L

    1981-06-01

    PtK1 cells enter prophase and complete mitosis at 24-25 degrees C but are inhibited from entering prophase at 20-21 degrees C. Cells which have progressed up to midprophase at 24-37 degrees C return to interphase when cooled to 20-21 degrees C, but those in late prophase complete a normal, although prolonged mitosis. If prophase cells which have reverted to interphase at 20-21 degrees C are incubated at 24-37 degrees C they reenter prophase and complete mitosis. This temperature-induced prophase-interphase-prophase transition can be repeated several times on the same cell. At 24-25 degrees C the process of spindle formation (i.e. prometaphase to the initiation of anaphase) encompasses approximately 75% of the total mitotic interval, with a duration of 8-12 h, compared to about 50% of the mitotic interval and a duration of 0.5 to 1.0 h at 37 degrees C.

  4. Neonatal Escherichia coli K1 meningitis causes learning and memory impairments in adulthood.

    Science.gov (United States)

    Barichello, Tatiana; Dagostim, Valdemira S; Generoso, Jaqueline S; Simões, Lutiana R; Dominguini, Diogo; Silvestre, Cintia; Michels, Monique; Vilela, Márcia Carvalho; Jornada, Luciano K; Comim, Clarissa M; Dal-Pizzol, Felipe; Teixeira, Antonio Lucio; Quevedo, João

    2014-07-15

    Neonatal Escherichia coli meningitis continues to be an important cause of mortality and morbidity in newborns worldwide. The aim of this study was to investigate the cytokines/chemokines, brain-derived neurotrophic factor (BDNF) levels, blood-brain barrier integrity in neonatal rats following E. coli K1 experimental meningitis infection and subsequent behavioural parameters in adulthood. In the hippocampus, interleukin increased at 96 h, IL-6 at 12, 48 and 96 h, IL-10 at 96 h, cytokine-induced neutrophil chemoattractant-1 at 6, 12, 24, 48 and 96 h, and BDNF at 48 and 96 h. In the cerebrospinal fluid, tumour necrosis factor alpha levels increased at 6, 12, 24, 48 and 96 h. The BBB breakdown occurred at 12 h in the hippocampus, and at 6h in the cortex. We evaluated behavioural parameters in adulthood: habituation to the open-field, step-down inhibitory avoidance, object recognition, continuous multiple-trials step-down inhibitory avoidance and forced swimming tasks. In adulthood, the animals showed habituation and aversive memory impairment. The animals needed a significant increase in the number of training periods to learn and not had depressive-like symptoms.

  5. A Rapid and Sensitive Diagnosis of Typhoid Fever Based On Nested PCR-Voltammetric DNA Biosensor Using Flagellin Gene Fragment

    Directory of Open Access Journals (Sweden)

    Yeni Wahyuni Hartati

    2016-03-01

    Full Text Available Typhoid fever caused by Salmonella typhi is an important issue for public health in the world. Laboratory methods for rapid and sensitive diagnosis are very important for disease management. The purpose of this study was to determine the performance of nested PCR–voltammetric DNA biosensor using flagellin gene (fla of S. typhi as a marker. The differential pulse voltammetry using pencil graphite electrode was applied to measure the guanine oxidation signal of probes vs synthetic target stDNA and probes vs fla PCR product hybridizations. The probe DNA selectivity was examined by hybridized probes vs non-complementary sequence. The result showed that the first round nested PCR product can not be visualized by agarose electrophoresis, whereas using the voltammetric biosensor methods can be detected both for the first or second round nested PCR product. The average peak current of hybridized probe vs first and second round of PCR product was 2.32 and 1.47 μA respectively, at 0.9 V. Detection of the DNA sequences of the infectious diseases from PCR amplified real sample was also carried out using this voltammetric DNA biosensor methods.

  6. Linear antigenic mapping of flagellin (FliC) from Salmonella enterica serovar Enteritidis with yeast surface expression system.

    Science.gov (United States)

    Wang, Gaoling; Shi, Bingtian; Li, Tao; Zuo, Teng; Wang, Bin; Si, Wei; Xin, Jiuqing; Yang, Kongbin; Shi, Xuanlin; Liu, Siguo; Liu, Henggui

    2016-02-29

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major cause of food-borne illness around the world and can have significant health implications in humans, poultry and other animals. Flagellin (FliC) is the primary component of bacterial flagella. It has been shown that the FliC of S. Enteritidis is a significant antigenic structure and can elicit strong humoral responses against S. Enteritidis infection in chickens. Here, we constructed a FliC antigen library using a yeast surface expression system. Yeast cells expressing FliC peptide antigens were labeled with chicken sera against S. Enteritidis and sorted using FACS. The analyses of FliC peptides revealed that the FliC linear antigenicity in chickens resided on three domains which were able to elicit strong humoral responses in vivo. Animal experiments further revealed that the antibodies elicited by these antigenic domains were able to significantly inhibit the invasion of S. Enteritidis into the liver and spleen of chickens. These findings will facilitate our better understanding of the humoral responses elicited by FliC in chickens upon infection by S. Enteritidis.

  7. Integrable deformations of the $G_{k_1} \\times G_{k_2}/G_{k_1+k_2}$ coset CFTs arXiv

    CERN Document Server

    Sfetsos, Konstantinos

    We study the effective action for the integrable $\\lambda$-deformation of the $G_{k_1} \\times G_{k_2}/G_{k_1+k_2}$ coset CFTs. For unequal levels theses models do not fall into the general discussion of $\\lambda$-deformations of CFTs corresponding to symmetric spaces and have many attractive features. We show that the perturbation is driven by parafermion bilinears and we revisit the derivation of their algebra. We uncover a non-trivial symmetry of these models parametric space, which has not encountered before in the literature. Using field theoretical methods and the effective action we compute the exact in the deformation parameter $\\beta$-function and explicitly demonstrate the existence of a fixed point in the IR corresponding to the $G_{k_1-k_2} \\times G_{k_2}/G_{k_1}$ coset CFTs. The same result is verified using gravitational methods for $G=SU(2)$. We examine various limiting cases previously considered in the literature and found agreement.

  8. Host cell type-dependent translocation and PhoP-mediated positive regulation of the effector SseK1 of Salmonella enterica

    Directory of Open Access Journals (Sweden)

    Fernando eBaisón-Olmo

    2015-04-01

    Full Text Available Salmonella enterica expresses two virulence-related type III secretion systems (T3SSs encoded in Salmonella pathogenicity island 1 (SPI1 and SPI2, respectively. SseK1 is a poorly characterized substrate of the SPI2-encoded T3SS. Here, we show that this effector is essential to get full virulence both in oral and intraperitoneal mice infections, in spite of not having a role in invasion or intracellular proliferation in cultured mammalian cells. In vitro, expression of sseK1 was higher in media mimicking intracellular conditions, when SPI2 was induced, but it was also significant under SPI1 inducing conditions. A detailed analysis of translocation of SseK1 into host cells unveiled that it was a substrate of both, T3SS1 and T3SS2, although with different patterns and kinetics depending on the specific host cell type (epithelial, macrophages or fibroblasts. The regulation of the expression of sseK1 was examined using lacZ and bioluminescent lux fusions. The two-component system PhoQ/PhoP is a positive regulator of this gene. A combination of sequence analysis, directed mutagenesis and electrophoretic mobility shift assays showed that phosphorylated PhoP binds directly to the promoter region of sseK1 and revealed a PhoP binding site located upstream of the predicted -35 hexamer of this promoter.

  9. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-12-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.

  10. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-01-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent. Images PMID:6094355

  11. Data of evolutionary structure change: 1QX5K-1CLMA [Confc[Archive

    Lifescience Database Archive (English)

    Full Text Available 1QX5K-1CLMA 1QX5 1CLM K A LTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQN...seq1> LTEEQIAEFKEAFALFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLSLMARKMKEQDSEEELIEAFKVFDRDGNGL...> A 1CLMA GDGHI-NYEEF

  12. Prediction of 3-hydroxypyridin-4-one (HPO) log K1 values for Fe(III).

    Science.gov (United States)

    Chen, Yu-Lin; Barlow, Dave J; Kong, Xiao-Le; Ma, Yong-Min; Hider, Robert C

    2012-09-21

    As a means to aid in the design of 3-hydroxypyridin-4-ones (HPOs) intended for use as therapeutic Fe(3+) chelating agents, a novel methodology has been developed using quantum mechanical (QM) calculations for predicting the iron binding affinities of the compounds (more specifically, their log K(1) values). The reported/measured HPO log K(1) values were verified through their correlation with the corresponding sum of the compounds' ligating group pK(a) values. Using a training set of eleven HPOs with known log K(1) values, reliable predictions are shown to be obtained with QM calculations using the B3LYP/6-31+G(d)/CPCM model chemistry (with Bondi radii, and water as solvent). With this methodology, the observed log K(1) values for the training set compounds are closely matched by the predicted values, with the correlation between the observed and predicted values giving r(2) = 0.9. Predictions subsequently made by this method for a test set of 42 HPOs of known log K(1) values gave predicted values accurate to within ±0.32 log units. In order to further investigate the predictive power of the method, four novel HPOs were synthesised and their log K(1) values were determined experimentally. Comparison of these predicted log K(1) values against the measured values gave absolute deviations of 0.22 (13.87 vs. 14.09), 0.02 (14.31 vs. 14.29), 0.12 (14.62 vs. 14.50), and 0.13 (15.04 vs. 15.17). The prediction methodology reported here is the first to be provided for predicting the absolute log K(1) values of iron-chelating agents in the absence of pK(a) values.

  13. Lectin complement protein Collectin 11 (CL-K1 and susceptibility to urinary schistosomiasis.

    Directory of Open Access Journals (Sweden)

    Justin S Antony

    2015-03-01

    Full Text Available Urinary Schistosomiasis is a neglected tropical disease endemic in many sub Saharan -African countries. Collectin Kidney 1 (CL-K1, encoded by COLEC11 on chromosome 2p25.3, a member of the vertebrate C-type lectin super family, has recently been identified as pattern-recognition molecule (PRR of the lectin complement pathway. CL-K1 is preferentially expressed in the kidneys, but also in other organs and it is considered to play a role in host defense to some infectious agents. Schistosome teguments are fucosylated and CL-K1 has, through its collagen-like domain, a high binding affinity to fucose.We utilized a Nigerian study group consisting of 167 Schistosoma haematobium infected individuals and 186 matched healthy subjects, and investigated the contribution of CL-K1 deficiency and of COLEC11 polymorphisms to infection phenotype. Higher CL-K1 serum levels were associated with decreased risk of schistosome infection (P corr = 0.0004. CL-K1 serum levels were differentially distributed between the COLEC11 genotypes and haplotypes observed. The non-synonymous variant p.R216H was associated with the occurrence of schistosomiasis (OR = 0.44, 95%CI = 0.22-0.72, P corr = 0.0004. The reconstructed COLEC11*TCCA haplotypes were associated with higher CL-K1 serum levels (P = 0.002 and with decreased schistosomiasis (OR = 0.38, 95%CI = 0.23-0.63, P corr = 0.0001.In agreement with findings from our earlier published study, our findings support the observation that CL-K1 and their functional variants may be host factors associated with protection in schistosomiasis and may be a useful marker for further investigations.

  14. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii trophozoites and cysts.

    Science.gov (United States)

    Matin, Abdul; Jung, Suk-Yul

    2011-12-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. coli K12 was also applied. The association ratio of E. coli K1 with A. castellanii was 4.3 cfu per amoeba for 1 hr but E. coli K5 with A. castellanii was 1 cfu per amoeba for 1 hr. By invasion and survival assays, E. coli K5 was recovered less than E. coli K1 but still alive inside A. castellanii. E. coli K1 and K5 survived and multiplied intracellularly in A. castellanii. The survival assay was performed under a favourable condition for 22 hr and 43 hr with the encystment of A. castellanii. Under the favourable condition for the transformation of trophozoites into cysts, E. coli K5 multiplied significantly. Moreover, the pathogenic potential of E. coli K1 from A. castellanii cysts exhibited no changes as compared with E. coli K1 from A. castellanii trophozoites. E. coli K5 was multiplied in A. castellanii trophozoites and survived in A. castellanii cysts. Therefore, this study suggests that E. coli K5 can use A. castellanii as a reservoir host or a vector for the bacterial transmission.

  15. Characterization of $m$-Sequences of Lengths $2^{2k}-1$ and $2^k-1$ with Three-Valued Crosscorrelation

    OpenAIRE

    Helleseth, Tor; Kholosha, Alexander; Ness, Geir Jarle

    2007-01-01

    Considered is the distribution of the crosscorrelation between $m$-sequences of length $2^m-1$, where $m=2k$, and $m$-sequences of shorter length $2^k-1$. New pairs of $m$-sequences with three-valued crosscorrelation are found and the complete correlation distribution is determined. Finally, we conjecture that there are no more cases with a three-valued crosscorrelation apart from the ones proven here.

  16. KEY COMPARISON: Force key comparison CCM.F-K1.a and CCM.F-K1.b: 5 kN and 10 kN

    Science.gov (United States)

    Pusa, Aimo

    2009-01-01

    This report describes the key comparisons named CCM.F-K1.a and CCM.F-K1.b, for force with loads of 5 kN and 10 kN. The Draft A Report, reporting the measurement results of the key comparisons, has been accepted at the force expert group meeting in Pretoria on 23 March 2004. This became Part 1 of the preliminary Draft B Report. Then, there have been several discussions to find the best way for the determination of the reference value for force values 5 kN and 10 kN in the key comparisons CCM.F-K1.a and CCM.F-K1.b. Following the meeting held in Queretaro, Mexico, by CENAM, from 3 to 5 December 2007, the reference values have been calculated for each single transducer (see chapter 2), and as one reference value for 5 kN and a second reference value for 10 kN (see chapter 3). To get a better consistency a linear model for the drift of transducers has been applied. The results have been evaluated according to the paper from M G Cox, 'The evaluation of key comparison data' (2002 Metrologia 39 589-595). Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  17. Carvacrol induces heat shock protein 60 and inhibits synthesis of flagellin in Escherichia coli O157:H7

    NARCIS (Netherlands)

    Burt, S.A.; van der Zee, R.; Koets, A.P.; de Graaff, A.M.; van Knapen, F.; Gaastra, W.; Haagsman, H.P.; Veldhuizen, E.J.A.

    2007-01-01

    The essential oils of oregano and thyme are active against a number of food-borne pathogens, such as Escherichia coli O157:H7. Carvacrol is one of the major antibacterial components of these oils, and p-cymene is thought to be its precursor in the plant. The effects of carvacrol and p-cymene on prot

  18. Co-delivery of polyinosinic:polycytidylic acid and flagellin by poly(lactic-co-glycolic acid) MPs synergistically enhances immune response elicited by intranasally delivered hepatitis B surface antigen

    Science.gov (United States)

    Dai, Xiaojing; He, Jintian; Zhang, Ruxia; Wu, Guanghao; Xiong, Fangfang; Zhao, Baohua

    2017-01-01

    The aim of the present work was to investigate the synergistic effect between toll-like receptor (TLR) 3 ligand polyinosinic:polycytidylic acid (pI:C) and TLR5 ligand flagellin (FLN) on immune responses induced by nasally delivered hepatitis B virus surface antigen (HBsAg). Mannan and chitosan oligosaccharide-modified, pH-responsive poly(lactic-co-glycolic acid) (MC-PLGA) microparticles (MPs) containing HBsAg, FLN, pI:C or both ligands were prepared with a double-emulsion method. In vitro uptake experiments show that cellular uptake of MC-PLGA MPs by macrophages was through energy-dependent, receptor-mediated endocytosis mechanism. After uptake of MPs by macrophages, MC-PLGA MPs existed both in the endo-some and in the cytoplasm. FLN and pI:C in solution or MP formulation could synergize to activate macrophages and induce higher pro-inflammatory cytokines interleukin (IL)-6, IL-12, interferon-γ and anti-inflammatory cytokines IL-10 compared to single TLR ligand (P<0.05). In vivo immunogenicity studies indicated that co-delivery of FLN and pI:C within MC-PLGA MPs synergistically induced higher serum anti-HBsAg IgG levels and Th1 cytokine levels compared with MC-PLGA MPs encapsulated single TLR ligand plus MPs encapsulated HBsAg (P<0.05). These results suggest that synergic TLR3 and TLR5 stimulation might be a promising novel tool for nasally delivered HBsAg. PMID:28924346

  19. ppk1 gene relates with the pathogenesis of meningitis infected by E.coli K1%脑膜炎大肠杆菌K1株ppk1基因致病机制初探

    Institute of Scientific and Technical Information of China (English)

    彭亮; 赵铁; 罗文英; 付美芹; 曹虹; 黄胜和

    2012-01-01

    [Objective] To construct the polyphosphate kinase 1 gene deletion mutant of Es-cherichia coli (E. Coli) Kl strain E44 and explore the role of ppkl in the pathopoiesis of meningitis E. Coli Kl. [Methods] The ppkl gene was knocked out from the genome of E. Coli Kl strain E44 by using suicide vetor pCVD442 and homologous recombination, and the mutant was named Appkl. Then the survival ablities of E44 and Appkl in poor nutrition condition and oxidative stress were examined. The ability of the mutant adhering to human brain microvas-cular endothelial cells (HBMEC) was also compared with that of the wild type. The cytoge-netic toxic effects induced by Appkl and E44 were tested by using the lactic dehydrogenase testing kit. [Results] The ppkl gene of the mutant had been knocked out and was confirmed by PCR and DNA sequencing. The ppkl deletion mutant showed to be defective in adhesion ability to HBMEC and survival abilities under poor nutrition condition and oxidative stress. The damaging effects of HBMEC induced by Appkl were significantly less than that induced by E44. [Conclusion] The ppkl gene plays an important role in E. Coli Kl surviving in low nutrition and oxidative stress condition, adhering to HBMEC and inducing cell injury.%[目的]构建脑膜炎大肠杆菌K1 (Escherichia coli,E.coli K1)株E44的聚磷酸盐激酶1 (Polyphosphate kinase 1,PPK1)基因敲除株,并对其生物学功能进行初步研究,为明确ppk1基因在E.coli K1株致脑膜炎机制中的作用奠定基础.[方法]利用自杀质粒pCVD442及基因同源重组技术敲除E.coli K1株E44中的ppk1基因,构建ppk1缺失突变株Δppk1;体外比较野生株和突变株在低营养及氧化压力情况下的生存能力;考察二者对人脑微血管内皮细胞(Human brain microvascular endothelial cells,HBMEC)的黏附能力;通过测定乳酸脱氢酶(Lactic dehydrogenase,LDH)释放活性,比较野生株和突变株对HBMEC的损伤效应.[结果]PCR及序列分析

  20. Effects of oral and intramuscular vitamin K prophylaxis on vitamin K1, PIVKA-II, and clotting factors in breast fed infants.

    Science.gov (United States)

    Cornelissen, E A; Kollée, L A; De Abreu, R A; van Baal, J M; Motohara, K; Verbruggen, B; Monnens, L A

    1992-10-01

    A randomised clinical trial was conducted to establish the effects of oral and intramuscular administration of vitamin K at birth on plasma concentrations of vitamin K1, proteins induced by vitamin K absence (PIVKA-II), and clotting factors. Two groups of about 165 healthy breast fed infants who received at random 1 mg vitamin K1 orally or intramuscularly after birth were studied at 2 weeks and 1 and 3 months of age. Although vitamin K1 concentrations were statistically significantly higher in the intramuscular group, blood coagulability, activities of factors VII and X and PIVKA-II concentrations did not reveal any difference between the two groups. At 2 weeks of age vitamin K1 concentrations were raised compared with reported unsupplemented concentrations and no PIVKA-II was detectable. At 3 months vitamin K1 concentrations were back at unsupplemented values and PIVKA-II was detectable in 11.5% of infants. Therefore, a repeated oral prophylaxis will be necessary to completely prevent (biochemical) vitamin K deficiency beyond the age of 1 month.

  1. Synthesis of the tetrasaccharide repeating unit of the O-glycan from the polar flagellum flagellin of Azospirillum brasilense Sp7.

    Science.gov (United States)

    Pal, Kumar Bhaskar; Mukhopadhyay, Balaram

    2014-12-01

    Chemical synthesis of the tetrasaccharide repeating unit of the O-glycan from the polar flagellum flagellin of Azospirillum brasilense Sp7 in the form of its p-methoxyphenyl glycoside is reported. The required glycosidic linkages have been accomplished by activation of thioglycosides with N-iodosuccinimide in the presence of H2SO4-silica. H2SO4-silica was found to be an effective alternative to the classical acid promoters like TfOH or TMSOTf and it can lead to the formation of both 1,2-cis and 1,2-trans glycosidic linkages depending on the protecting group manipulation and control of the reaction condition.

  2. Deforming the Lie Superalgebra $\\mathcal{K}(1)$-Modules Of Symbols

    CERN Document Server

    Faouzi, Ammar

    2008-01-01

    We study non-trivial deformations of the natural action of the Lie superalgebra $mathcalK(1)$ of contact vector fields on the (1,1)-dimensional superspace $mathbbR^{1|1}$ of th espace of symbols. We calculate obstructions for integrability of infinitesimal multi-parameter deformation and determine the complete commutative algebra corresponding to the miniversal deformation in the sense of A. Fialowski. Besides, we compute the first even differential cohomology space $mathrmH^1_{mathrm{diff}}(cK(1);widetilde{cD}_{lambda,mu})$ of the Lie superalgebra $mathcalK(1)$ of contact vector fields on the (1,1)-dimensional superspace $mathbbR^{1|1}$ with coefficients in the superspace $widetilde{mathcalD}_{lambda,mu}$ of linear differential operators from the superspace of weighted densities $\\fF_{\\lamda}$ to $\\fF_{\\mu}$. (To appear in Journal of Generalized Lie Theory and Applications)

  3. Determination of ultimate carbonaceous BOD and the specific rate constant (K1)

    Science.gov (United States)

    Stamer, J.K.; Bennett, J.P.; McKenzie, Stuart W.

    1982-01-01

    Ultimate carbonaceous biochemical oxygen demand (BODu) and the specific rate constant (K1) at which the demand is exerted are important parameters in designing biological wastewater treatment plants and in assessing the impact of wastewater on receiving streams. An analytical method is presented which uses time-series concentrations of BOD, defined as the calculated sum of dissolved oxygen (DO) losses at each time of measurement, for determining BODu and K1. Time-series DO measurements are obtained from a water sample that is incubated in darkness at 20 degrees Celsius in the presence of nitrapyrin, a chemical nitrification inhibitor. Time-series concentrations of BOD that approximate first order kinetics can be analyzed graphically or mathematically to compute BODu and K1.

  4. Autonomous safety and reliability features of the K-1 avionics system

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, G.E.; Kohrs, D.; Bailey, R.; Lai, G. [Kistler Aerospace Corp., Kirkland, WA (United States)

    2004-03-01

    Kistler Aerospace Corporation is developing the K-1, a fully reusable, two-stage-to-orbit launch vehicle. Both stages return to the launch site using parachutes and airbags. Initial flight operations will occur from Woomera, Australia. K-1 guidance is performed autonomously. Each stage of the K- 1 employs a triplex, fault tolerant avionics architecture, including three fault tolerant computers and three radiation hardened Embedded GPS/INS units with a hardware voter. The K-1 has an Integrated Vehicle Health Management (IVHM) system on each stage residing in the three vehicle computers based on similar systems in commercial aircraft. During first-stage ascent, the IVHM system performs an Instantaneous Impact Prediction (IIP) calculation 25 times per second, initiating an abort in the event the vehicle is outside a predetermined safety corridor for at least three consecutive calculations. In this event, commands are issued to terminate thrust, separate the stages, dump all propellant in the first-stage, and initiate a normal landing sequence. The second-stage flight computer calculates its ability to reach orbit along its state vector, initiating an abort sequence similar to the first stage if it cannot. On a nominal mission, following separation, the second-stage also performs calculations to assure its impact point is within a safety corridor. The K-1's guidance and control design is being tested through simulation with hardware-in-the-loop at Draper Laboratory. Kistler's verification strategy assures reliable and safe operation of the K-1. (author)

  5. Des-gamma-carboxyprothrombin (PIVKA II) and plasma vitamin K1 in newborns and their mothers.

    Science.gov (United States)

    von Kries, R; Shearer, M J; Widdershoven, J; Motohara, K; Umbach, G; Göbel, U

    1992-10-01

    Assessments of the vitamin K status in newborns and their mothers by means of des-gamma-carboxy-prothrombin (PIVKA II) measurement have given equivocal results. Part of the variability could be attributed to differences in sensitivity (i.e. the ability to detect small concentrations) and validity (i.e. ability to detect vitamin K deficiency) of the methods applied. None of these methods have yet been validated with respect to plasma vitamin K1. In 22 healthy mother/infant pairs PIVKA II was determined using three different assays including ratio Xa/ecarin (Xa/ec), crossed immunoelectrophoresis (CIE), and an ELISA with a monoclonal antibody (MAB). The results were compared with conventional clotting tests and plasma vitamin K1. The following results were obtained: Cord blood: Clotting tests within age-related normal ranges; PIVKA II detection rates: 0/22 (Xa/ec), 1/22 (CIE), 4/22 (MAB); plasma vitamin K1: undetectable in 20/22. Mothers: Clotting tests all within normal range; PIVKA II detection rates: 1/22 (Xa/ec), 0/22 (CIE), 5/22 (MAB); plasma vitamin K1 (pg/ml) for all mothers (median; range): 186; 55-833; for PIVKA II positive mothers: 213; 59-699. PIVKA II detectability in newborns and mothers was not correlated. The results show an increase in sensitivity for PIVKA II detection in the order of MAB > CIE > Xa/ec. Due to the very low plasma vitamin K1 at birth, no correlation was possible between cord PIVKA II detectability and plasma vitamin K1. However, in mothers at term PIVKA II MAB appears to be unrelated to the vitamin K status.

  6. Real-time visualization of conformational changes within single MloK1 cyclic nucleotide-modulated channels.

    Science.gov (United States)

    Rangl, Martina; Miyagi, Atsushi; Kowal, Julia; Stahlberg, Henning; Nimigean, Crina M; Scheuring, Simon

    2016-09-20

    Eukaryotic cyclic nucleotide-modulated (CNM) ion channels perform various physiological roles by opening in response to cyclic nucleotides binding to a specialized cyclic nucleotide-binding domain. Despite progress in structure-function analysis, the conformational rearrangements underlying the gating of these channels are still unknown. Here, we image ligand-induced conformational changes in single CNM channels from Mesorhizobium loti (MloK1) in real-time, using high-speed atomic force microscopy. In the presence of cAMP, most channels are in a stable conformation, but a few molecules dynamically switch back and forth (blink) between at least two conformations with different heights. Upon cAMP depletion, more channels start blinking, with blinking heights increasing over time, suggestive of slow, progressive loss of ligands from the tetramer. We propose that during gating, MloK1 transitions from a set of mobile conformations in the absence to a stable conformation in the presence of ligand and that these conformations are central for gating the pore.

  7. Emergence of Carbapenem-Resistant Serotype K1 Hypervirulent Klebsiella pneumoniae Strains in China.

    Science.gov (United States)

    Zhang, Rong; Lin, Dachuan; Chan, Edward Wai-Chi; Gu, Danxia; Chen, Gong-Xiang; Chen, Sheng

    2015-11-16

    We report the emergence of five carbapenem-resistant K1 hypervirulent Klebsiella pneumoniae (hvKP) strains which caused fatal infections in hospital patients in Zhejiang Province, China, upon entry through surgical wounds. Genotyping results revealed the existence of three genetically related strains which exhibited a new sequence type, ST1797, and revealed that all strains harbored the magA and wcaG virulence genes and a plasmid-borne bla(KPC-2) gene. These findings indicate that K1 hvKP is simultaneously hypervirulent, multidrug resistant, and transmissible.

  8. Form factors in the massless coset models su(2)_k+1 \\otimes su(2)_k /su(2)_2k+1 - Part II

    CERN Document Server

    Grinza, P

    2004-01-01

    Massless flows from the coset model su(2)_k+1 \\otimes su(2)_k /su(2)_2k+1 to the minimal model M_k+2 are studied from the viewpoint of form factors. These flows include in particular the flow from the Tricritical Ising model to the Ising model. By analogy with the magnetization operator in the flow TIM -> IM, we construct all form factors of an operator that flows to \\Phi_1,2 in the IR. We make a numerical estimation of the difference of conformal weights between the UV and the IR thanks to the \\Delta-sum rule; the results are consistent with the conformal weight of the operator \\Phi_2,2 in the UV. By analogy with the energy operator in the flow TIM -> IM, we construct all form factors of an operator that flows to \\Phi_2,1. We propose to identify the operator in the UV with \\sigma_1\\Phi_1,2.

  9. 一类整群环的K1群的计算%Calculation of K1 group of some integral group rings

    Institute of Scientific and Technical Information of China (English)

    刘佳奇; 高玉彬

    2011-01-01

    The integral group ring (Z)G is an important ring structure.This paper focuses on the calculation of the K1 groups of integral group rings.The key point of the problem lies in the calculation of their corresponding SK1 group rings.The SK1 groups of the integral group rings of p groups are relatively easy to calculate, but the SK1 groups of the integral group rings of non-p groups are relatively diffcult to calculate.This article discusses the basic theory of the problem.Furthermore, this article also treats the groups whose orders are p1q1 and discusses the calculation of the SK1 group rings of their integral group rings.%着重讨论整群环的K1群的计算,问题的关键点在于其对应的SK1群的计算,其中,P群的整群环的SK1群的结果是已知的.讨论了计算SK1群的基本理论,并证明了一类特殊的非P群(P1q1阶群)的整群环的SK1群是平凡的.

  10. Immunogenicity and efficacy of flagellin-fused vaccine candidates targeting 2009 pandemic H1N1 influenza in mice.

    Directory of Open Access Journals (Sweden)

    Ge Liu

    Full Text Available We have previously demonstrated that the globular head of the hemagglutinin (HA antigen fused to flagellin of Salmonella typhimurium fljB (STF2, a TLR5 ligand elicits protective immunity to H1N1 and H5N1 lethal influenza infections in mice (Song et al., 2008, PLoS ONE 3, e2257; Song et al., 2009, Vaccine 27, 5875-5888. These fusion proteins can be efficiently and economically manufactured in E. coli fermentation systems as next generation pandemic and seasonal influenza vaccines. Here we report immunogenicity and efficacy results of three vaccine candidates in which the HA globular head of A/California/07/2009 (H1N1 was fused to STF2 at the C-terminus (STF2.HA1, in replace of domain 3 (STF2R3.HA1, or in both positions (STF2R3.2xHA1. For all three vaccines, two subcutaneous immunizations of BALB/c mice with doses of either 0.3 or 3 µg elicit robust neutralizing (HAI antibodies, that lead to > = 2 Log(10 unit reduction in day 4 lung virus titer and full protection against a lethal A/California/04/2009 challenge. Vaccination with doses as low as 0.03 µg results in partial to full protection. Each candidate, particularly the STF2R3.HA1 and STF2R3.2xHA1 candidates, elicits robust neutralizing antibody responses that last for at least 8 months. The STF2R3.HA1 candidate, which was intermediately protective in the challenge models, is more immunogenic than the H1N1 components of two commercially available trivalent inactivated influenza vaccines (TIVs in mice. Taken together, the results demonstrate that all three vaccine candidates are highly immunogenic and efficacious in mice, and that the STF2R3.2xHA1 format is the most effective candidate vaccine format.

  11. 76 FR 7847 - Glenn A. Baxter, Application To Renew License for Amateur Radio Service Station K1MAN

    Science.gov (United States)

    2011-02-11

    ... COMMISSION Glenn A. Baxter, Application To Renew License for Amateur Radio Service Station K1MAN AGENCY... renew the license for Amateur Radio Service Station K1MAN filed by Glenn A. Baxter should be granted.... Baxter for renewal of his license for Amateur Radio Station K1MAN should be granted. As discussed below...

  12. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Science.gov (United States)

    2010-04-01

    ... deductibility of certain dividend distributions. (Temporary) 1.404(k)-1T Section 1.404(k)-1T Internal Revenue... Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.404(k)-1T Questions and answers relating to the deductibility of certain dividend distributions. (Temporary) Q-1: What does section 404(k) provide? A-1: Section...

  13. K-1 Teachers' Visual Arts Beliefs and Their Role in the Early Childhood Classroom

    Science.gov (United States)

    Goodman-Schanz, Blythe Annette

    2012-01-01

    The purpose of this qualitative study was to explore and describe the visual arts beliefs and practices of eight K-1 teachers in four schools and in two different school districts in a southern state. Using a phenomenological framework (Creswell, 2007; Leedy & Ormrod, 2005), the research revealed the teachers' understandings of beliefs and how…

  14. 26 CFR 1.168(k)-1 - Additional first year depreciation deduction.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 2 2010-04-01 2010-04-01 false Additional first year depreciation deduction. 1... Corporations § 1.168(k)-1 Additional first year depreciation deduction. (a) Scope and definitions—(1) Scope. This section provides the rules for determining the 30-percent additional first year...

  15. Inhibition of p70 S6 kinase (S6K1) activity by A77 1726 and its effect on cell proliferation and cell cycle progress.

    Science.gov (United States)

    Doscas, Michelle E; Williamson, Ashley J; Usha, Lydia; Bogachkov, Yedida; Rao, Geetha S; Xiao, Fei; Wang, Yimin; Ruby, Carl; Kaufman, Howard; Zhou, Jingsong; Williams, James W; Li, Yi; Xu, Xiulong

    2014-10-01

    Leflunomide is a novel immunomodulatory drug prescribed for treating rheumatoid arthritis. It inhibits the activity of protein tyrosine kinases and dihydroorotate dehydrogenase, a rate-limiting enzyme in the pyrimidine nucleotide synthesis pathway. Here, we report that A77 1726, the active metabolite of leflunomide, inhibited the phosphorylation of ribosomal protein S6 and two other substrates of S6K1, insulin receptor substrate-1 and carbamoyl phosphate synthetase 2, in an A375 melanoma cell line. A77 1726 increased the phosphorylation of AKT, p70 S6 (S6K1), ERK1/2, and MEK through the feedback activation of the IGF-1 receptor-mediated signaling pathway. In vitro kinase assay revealed that leflunomide and A77 1726 inhibited S6K1 activity with IC50 values of approximately 55 and 80 μM, respectively. Exogenous uridine partially blocked A77 1726-induced inhibition of A375 cell proliferation. S6K1 knockdown led to the inhibition of A375 cell proliferation but did not potentiate the antiproliferative effect of A77 1726. A77 1726 stimulated bromodeoxyuridine incorporation in A375 cells but arrested the cell cycle in the S phase, which was reversed by addition of exogenous uridine or by MAP kinase pathway inhibitors but not by rapamycin and LY294002 (a phosphoinositide 3-kinase inhibitor). These observations suggest that A77 1726 accelerates cell cycle entry into the S phase through MAP kinase activation and that pyrimidine nucleotide depletion halts the completion of the cell cycle. Our study identified a novel molecular target of A77 1726 and showed that the inhibition of S6K1 activity was in part responsible for its antiproliferative activity. Our study also provides a novel mechanistic insight into A77 1726-induced cell cycle arrest in the S phase.

  16. Inhibition of p70 S6 Kinase (S6K1 Activity by A77 1726 and Its Effect on Cell Proliferation and Cell Cycle Progress

    Directory of Open Access Journals (Sweden)

    Michelle E. Doscas

    2014-10-01

    Full Text Available Leflunomide is a novel immunomodulatory drug prescribed for treating rheumatoid arthritis. It inhibits the activity of protein tyrosine kinases and dihydroorotate dehydrogenase, a rate-limiting enzyme in the pyrimidine nucleotide synthesis pathway. Here, we report that A77 1726, the active metabolite of leflunomide, inhibited the phosphorylation of ribosomal protein S6 and two other substrates of S6K1, insulin receptor substrate-1 and carbamoyl phosphate synthetase 2, in an A375 melanoma cell line. A77 1726 increased the phosphorylation of AKT, p70 S6 (S6K1, ERK1/2, and MEK through the feedback activation of the IGF-1 receptor–mediated signaling pathway. In vitro kinase assay revealed that leflunomide and A77 1726 inhibited S6K1 activity with IC50 values of approximately 55 and 80 μM, respectively. Exogenous uridine partially blocked A77 1726–induced inhibition of A375 cell proliferation. S6K1 knockdown led to the inhibition of A375 cell proliferation but did not potentiate the antiproliferative effect of A77 1726. A77 1726 stimulated bromodeoxyuridine incorporation in A375 cells but arrested the cell cycle in the S phase, which was reversed by addition of exogenous uridine or by MAP kinase pathway inhibitors but not by rapamycin and LY294002 (a phosphoinositide 3-kinase inhibitor. These observations suggest that A77 1726 accelerates cell cycle entry into the S phase through MAP kinase activation and that pyrimidine nucleotide depletion halts the completion of the cell cycle. Our study identified a novel molecular target of A77 1726 and showed that the inhibition of S6K1 activity was in part responsible for its antiproliferative activity. Our study also provides a novel mechanistic insight into A77 1726–induced cell cycle arrest in the S phase.

  17. Inhibition of p70 S6 Kinase (S6K1) Activity by A77 1726 and Its Effect on Cell Proliferation and Cell Cycle Progress12

    Science.gov (United States)

    Doscas, Michelle E.; Williamson, Ashley J.; Usha, Lydia; Bogachkov, Yedida; Rao, Geetha S.; Xiao, Fei; Wang, Yimin; Ruby, Carl; Kaufman, Howard; Zhou, Jingsong; Williams, James W.; Li, Yi; Xu, Xiulong

    2014-01-01

    Leflunomide is a novel immunomodulatory drug prescribed for treating rheumatoid arthritis. It inhibits the activity of protein tyrosine kinases and dihydroorotate dehydrogenase, a rate-limiting enzyme in the pyrimidine nucleotide synthesis pathway. Here, we report that A77 1726, the active metabolite of leflunomide, inhibited the phosphorylation of ribosomal protein S6 and two other substrates of S6K1, insulin receptor substrate-1 and carbamoyl phosphate synthetase 2, in an A375 melanoma cell line. A77 1726 increased the phosphorylation of AKT, p70 S6 (S6K1), ERK1/2, and MEK through the feedback activation of the IGF-1 receptor–mediated signaling pathway. Invitro kinase assay revealed that leflunomide and A77 1726 inhibited S6K1 activity with IC50 values of approximately 55 and 80 μM, respectively. Exogenous uridine partially blocked A77 1726–induced inhibition of A375 cell proliferation. S6K1 knockdown led to the inhibition of A375 cell proliferation but did not potentiate the antiproliferative effect of A77 1726. A77 1726 stimulated bromodeoxyuridine incorporation in A375 cells but arrested the cell cycle in the S phase, which was reversed by addition of exogenous uridine or by MAP kinase pathway inhibitors but not by rapamycin and LY294002 (a phosphoinositide 3-kinase inhibitor). These observations suggest that A77 1726 accelerates cell cycle entry into the S phase through MAP kinase activation and that pyrimidine nucleotide depletion halts the completion of the cell cycle. Our study identified a novel molecular target of A77 1726 and showed that the inhibition of S6K1 activity was in part responsible for its antiproliferative activity. Our study also provides a novel mechanistic insight into A77 1726–induced cell cycle arrest in the S phase. PMID:25379019

  18. Cytoskeleton, endoplasmic reticulum and nucleus alterations in CHO-K1 cell line after Crotalus durissus terrificus (South American rattlesnake venom treatment

    Directory of Open Access Journals (Sweden)

    B. P. Tamieti

    2007-01-01

    Full Text Available Snake venoms are toxic to a variety of cell types. However, the intracellular damages and the cell death fate induced by venom are unclear. In the present work, the action of the South American rattlesnake Crotalus durissus terrificus venom on CHO-K1 cell line was analyzed. The cells CHO-K1 were incubated with C. d. terrificus venom (10, 50 and 100g/ml for 1 and 24 hours, and structural alterations of actin filaments, endoplasmic reticulum and nucleus were assessed using specific fluorescent probes and agarose gel electrophoresis for DNA fragmentation. Significant structural changes were observed in all analyzed structures. DNA fragmentation was detected suggesting that, at the concentrations used, the venom induced apoptosis.

  19. Presynaptic facilitation of glutamate release in the basolateral amygdala: a mechanism for the anxiogenic and seizurogenic function of GluK1 receptors.

    Science.gov (United States)

    Aroniadou-Anderjaska, V; Pidoplichko, V I; Figueiredo, T H; Almeida-Suhett, C P; Prager, E M; Braga, M F M

    2012-09-27

    Kainate receptors containing the GluK1 subunit (GluK1Rs; previously known as GluR5 kainate receptors) are concentrated in certain brain regions, where they play a prominent role in the regulation of neuronal excitability, by modulating GABAergic and/or glutamatergic synaptic transmission. In the basolateral nucleus of the amygdala (BLA), which plays a central role in anxiety as well as in seizure generation, GluK1Rs modulate GABAergic inhibition via postsynaptic and presynaptic mechanisms. However, the role of these receptors in the regulation of glutamate release, and the net effect of their activation on the excitability of the BLA network are not well understood. Here, we show that in amygdala slices from 35- to 50-day-old rats, the GluK1 agonist (RS)-2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA) (300 nM) increased the frequency of spontaneous excitatory postsynaptic currents (sEPSCs) and miniature EPSCs (mEPSCs) recorded from BLA principal neurons, and decreased the rate of failures of evoked EPSCs. The GluK1 antagonist (S)-1-(2-amino-2-carboxyethyl)-3-(2-carboxybenzyl) pyrimidine-2,4-dione (UBP302) (25 or 30 μM) decreased the frequency of mEPSCs, reduced evoked field potentials, and increased the "paired-pulse ratio" of the field potential amplitudes. Taken together, these results suggest that GluK1Rs in the rat BLA are present on presynaptic terminals of principal neurons, where they mediate facilitation of glutamate release. In vivo bilateral microinjections of ATPA (250 pmol) into the rat BLA increased anxiety-like behavior in the open field test, while 2 nmol ATPA induced seizures. Similar intra-BLA injections of UBP302 (20 nmol) had anxiolytic effects in the open field and the acoustic startle response tests, without affecting pre-pulse inhibition. These results suggest that although GluK1Rs in the rat BLA facilitate both GABA and glutamate release, the facilitation of glutamate release prevails, and these receptors can have an

  20. The mTORC1 effectors S6K1 and 4E-BP play different roles in CNS axon regeneration.

    Science.gov (United States)

    Yang, Liu; Miao, Linqing; Liang, Feisi; Huang, Haoliang; Teng, Xiuyin; Li, Shaohua; Nuriddinov, Jaloliddin; Selzer, Michael E; Hu, Yang

    2014-11-10

    Using mouse optic nerve (ON) crush as a CNS injury model, we and others have found that activation of the mammalian target of rapamycin complex 1 (mTORC1) in mature retinal ganglion cells by deletion of the negative regulators, phosphatase and tensin homologue (PTEN), and tuberous sclerosis 1 promotes ON regeneration. mTORC1 activation inhibits eukaryotic translation initiation factor 4E-binding protein (4E-BP) and activates ribosomal protein S6 kinase 1 (S6K1), both of which stimulate translation. We reasoned that mTORC1's regeneration-promoting effects might be separable from its deleterious effects by differential manipulation of its downstream effectors. Here we show that S6K1 activation, but not 4E-BP inhibition, is sufficient to promote axon regeneration. However, inhibition of 4E-BP is required for PTEN deletion-induced axon regeneration. Both activation and inhibition of S6K1 decrease the effect of PTEN deletion on axon regeneration, implicating a dual role of S6K1 in regulating axon growth.

  1. Functional subtypes of renal α1-adrenoceptor in diabetic and non-dia-betic 2K1C Goldblatt renovascular hypertension

    Institute of Scientific and Technical Information of China (English)

    A ARMENIA; Munavvar Abdul SATTAR; Nor Azizan ABDULLAH; Md Abdul Hye KHAN; Edward James JOHNS

    2008-01-01

    Aim: This study investigates the subtypes of the α-adrenoceptor mediating the adrenergically-induced renal vasoconstrictor responses in streptozotocin-induced diabetic and non-diabetic 2-kidney one clip (2K1C) Goldblatt hypertensive rats. Methods: The renal blood flow responses to renal nerve stimulation, noradrenaline, phenylephrine, and methoxamine were measured in the absence and presence of nitrendipine, 5-methylurapidil, chloroethylclonidine and BMY 7378. Results: The renal vasoconstrictor responses were markedly attenuated by nitrendipine and 5-methylurapidil in the diabetic rats (all P<0.05). In the non-diabetic rats, these responses were markedly attenuated by nitrendipine, 5-methylurapidil, and BMY 7378 (all P<0.05). In both experimental groups, chloroethylclonidine markedly accentuated the renal vasoconstrictions caused by all the adrenergic stimuli (all P<0.05). Conclusion: These observations indicate that α1A-adrenoceptor subtypes play a major role in mediating adrenergically-induced renal vasoconstriction in the dia-betic 2K1C Goldblatt hypertensive rats. In the non-diabetic 2KIC Goldblatt hy-pertensive rats, contributions of α1A and α1D-adrenoceptor subtypes were proposed. Apart from post-synaptic α1-adrenoceptors, both in the diabetic and non-diabetic 2K1C Goldblatt hypertensive rats, the potential involvement of presynaptic α1-adrenoceptors is also suggested.

  2. Going for baroque at the Escherichia coli K1 cell surface.

    Science.gov (United States)

    King, Michael R; Steenbergen, Susan M; Vimr, Eric R

    2007-05-01

    Phase variation is usually thought of as the stochastic switching between alternatively expressed ('on') and unexpressed ('off') phenotypic states. However, coupling synthesis of a monotonous homopolysaccharide to a mechanism of random but incomplete chemical modification produces almost infinite structural variation. Potentially limitless variability implies that evolution can produce highly ornate or extravagant flourishes reminiscent of the baroque style. Here, we describe an analysis of capsular polysialic acid form variation in Escherichia coli K1, demonstrating that the large number of variant structures is controlled by a single contingency locus. The mechanism for generating maximum structural diversity from maximal genetic parsimony is conferred by a simple translational switch carried on a K1-specific prophage.

  3. The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line

    DEFF Research Database (Denmark)

    Xu, Xun; Pan, Shengkai; Liu, Xin;

    2011-01-01

    Chinese hamster ovary (CHO)-derived cell lines are the preferred host cells for the production of therapeutic proteins. Here we present a draft genomic sequence of the CHO-K1 ancestral cell line. The assembly comprises 2.45 Gb of genomic sequence, with 24,383 predicted genes. We associate most...... of the assembled scaffolds with 21 chromosomes isolated by microfluidics to identify chromosomal locations of genes. Furthermore, we investigate genes involved in glycosylation, which affect therapeutic protein quality, and viral susceptibility genes, which are relevant to cell engineering and regulatory concerns....... Homologs of most human glycosylation-associated genes are present in the CHO-K1 genome, although 141 of these homologs are not expressed under exponential growth conditions. Many important viral entry genes are also present in the genome but not expressed, which may explain the unusual viral resistance...

  4. The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line

    DEFF Research Database (Denmark)

    Xu, Xun; Pan, Shengkai; Liu, Xin

    2011-01-01

    Chinese hamster ovary (CHO)-derived cell lines are the preferred host cells for the production of therapeutic proteins. Here we present a draft genomic sequence of the CHO-K1 ancestral cell line. The assembly comprises 2.45 Gb of genomic sequence, with 24,383 predicted genes. We associate most...... of the assembled scaffolds with 21 chromosomes isolated by microfluidics to identify chromosomal locations of genes. Furthermore, we investigate genes involved in glycosylation, which affect therapeutic protein quality, and viral susceptibility genes, which are relevant to cell engineering and regulatory concerns....... Homologs of most human glycosylation-associated genes are present in the CHO-K1 genome, although 141 of these homologs are not expressed under exponential growth conditions. Many important viral entry genes are also present in the genome but not expressed, which may explain the unusual viral resistance...

  5. Friedreich's Ataxia, Frataxin, PIP5K1B: Echo of a Distant Fracas

    Directory of Open Access Journals (Sweden)

    Aurélien Bayot

    2013-01-01

    Full Text Available “Frataxin fracas” were the words used when referring to the frataxin-encoding gene (FXN burst in as a motive to disqualify an alternative candidate gene, PIP5K1B, as an actor in Friedreich's ataxia (FRDA (Campuzano et al., 1996; Cossee et al., 1997; Carvajal et al., 1996. The instrumental role in the disease of large triplet expansions in the first intron of FXN has been thereafter fully confirmed, and this no longer suffers any dispute (Koeppen, 2011. On the other hand, a recent study suggests that the consequences of these large expansions in FXN are wider than previously thought and that the expression of surrounding genes, including PIP5K1B, could be concurrently modulated by these large expansions (Bayot et al., 2013. This recent observation raises a number of important and yet unanswered questions for scientists and clinicians working on FRDA; these questions are the substratum of this paper.

  6. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii Trophozoites and Cysts

    OpenAIRE

    Matin, Abdul; Jung, Suk-Yul

    2011-01-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. ...

  7. Non-Traditional Aspects of Renal Diets: Focus on Fiber, Alkali and Vitamin K1 Intake

    Science.gov (United States)

    Cupisti, Adamasco; D’Alessandro, Claudia; Gesualdo, Loreto; Cosola, Carmela; Gallieni, Maurizio; Egidi, Maria Francesca; Fusaro, Maria

    2017-01-01

    Renal diets for advanced chronic kidney disease (CKD) are structured to achieve a lower protein, phosphate and sodium intake, while supplying adequate energy. The aim of this nutritional intervention is to prevent or correct signs, symptoms and complications of renal insufficiency, delaying the start of dialysis and preserving nutritional status. This paper focuses on three additional aspects of renal diets that can play an important role in the management of CKD patients: the vitamin K1 and fiber content, and the alkalizing potential. We examined the energy and nutrients composition of four types of renal diets according to their protein content: normal diet (ND, 0.8 g protein/kg body weight (bw)), low protein diet (LPD, 0.6 g protein/kg bw), vegan diet (VD, 0.7 g protein/kg bw), very low protein diet (VLPD, 0.3 g protein/kg bw). Fiber content is much higher in the VD and in the VLPD than in the ND or LPD. Vitamin K1 content seems to follow the same trend, but vitamin K2 content, which could not be investigated, might have a different pattern. The net endogenous acid production (NEAP) value decreases from the ND and LPD to the vegetarian diets, namely VD and VLPD; the same finding occurred for the potential renal acid load (PRAL). In conclusion, renal diets may provide additional benefits, and this is the case of vegetarian diets. Namely, VD and VLPD also provide high amounts of fibers and Vitamin K1, with a very low acid load. These features may have favorable effects on Vitamin K1 status, intestinal microbiota and acid-base balance. Hence, we can speculate as to the potential beneficial effects on vascular calcification and bone disease, on protein metabolism, on colonic environment and circulating levels of microbial-derived uremic toxins. In the case of vegetarian diets, attention must be paid to serum potassium levels. PMID:28468236

  8. K1.33Mn8O16 as an electrocatalyst and a cathode

    Science.gov (United States)

    Jalili, Seifollah; Moharramzadeh Goliaei, Elham; Schofield, Jeremy

    2017-02-01

    Density functional theory (DFT) calculations are carried out to investigate the electronic, magnetic and thermoelectric properties of bulk and nanosheet K1.33Mn8O16 materials. The catalytic activity and cathodic performance of bulk and nanosheet structures are examined using the Tran-Blaha modified Becke-Johnson (TB-mBJ) exchange potential. Electronic structure calculations reveal an anti-ferromagnetic ground state, with a TB-mMBJ band gap in bulk K1.33Mn8O16 that is in agreement with experimental results. Density of state plots indicate a partial reduction of Mn4+ ions to Mn3+, without any obvious sign of Jahn-Teller distortion. Moreover, use of the O p-band center as a descriptor of catalytic activity suggests that the nanosheet has enhanced catalytic activity compared to the bulk structure. Thermoelectric parameters such as the Seebeck coefficient, electrical conductivity, and thermal conductivity are also calculated, and it is found that the Seebeck coefficients decrease with increasing temperature. High Seebeck coefficients for both spin-up and spin-down states are found in the nanosheet relative to their value in the bulk K1.33Mn8O16 structure, whereas the electrical and thermal conductivity are reduced relative to the bulk. In addition, figures of merit values are calculated as a function of the chemical potential and it is found that the nanosheet has a figure of merit of 1 at room temperature, compared to 0.5 for the bulk material. All results suggest that K1.33Mn8O16 nanosheets can be used both as a material in waste heat recovery and as an electrocatalyst in fuel cells and batteries.

  9. Doping dependence of correlation effects in K1 - x Fe2 - y Se2 superconductors: LDA' + DMFT investigation

    Science.gov (United States)

    Nekrasov, I. A.; Pavlov, N. S.; Sadovskii, M. V.

    2013-11-01

    We present a detailed LDA' + DMFT investigation of the doping dependence of correlation effects in the novel K1 - x Fe2 - y Se2 superconductor. Calculations are performed at four different hole doping levels, starting from a hypothetical stoichiometric composition with the total number of electrons equal to 29 per unit cell through 28 and 27.2 electrons toward the case of 26.52, which corresponds to the chemical composition K0.76Fe1.72Se2 studied in recent ARPES experiments. In the general case, the increase in hole doping leads to quasiparticle bands in a wide energy window ±2 eV around the Fermi level becoming more broadened by lifetime effects, while correlation-induced compression of Fe-3 d LDA' bandwidths stays almost the same, of the order of 1.3 for all hole concentrations. However, close to the Fermi level, the situation is more complicated. In the energy interval from -1.0 eV to 0.4 eV, the bare Fe-3 d LDA' bands are compressed by significantly larger renormalization factors up to 5 with increased hole doping, while the value of Coulomb interaction remains the same. This fact manifests the increase in correlation effects with hole doping in the K1 - x Fe2 - y Se2 system. Moreover, in contrast to typical pnictides, K1 - x Fe2 - y Se2 does not have well-defined quasiparticle bands on the Fermi levels, but has a "pseudogap"-like dark region instead. We also find that with the growth of hole doping, Fe-3 d orbitals of various symmetries are affected by correlations differently in different parts of the Brillouin zone. To illustrate this, we determine the quasiparticle mass renormalization factors and energy shifts that transform the bare Fe-3 d LDA' bands of various symmetries into LDA' + DMFT quasiparticle bands. These renormalization factors effectively mimic more complicated energy-dependent self-energy effects and can be used to analyze the available ARPES data.

  10. Identification of new flagellin-encoding fliC genes in Escherichia coli isolated from domestic animals using RFLP-PCR and sequencing methods

    Directory of Open Access Journals (Sweden)

    Cláudia de Moura

    2013-04-01

    Full Text Available Identification of Escherichia coli requires knowledge regarding the prevalent serotypes and virulence factors profiles allows the classification in pathogenic/non-pathogenic. However, some of these bacteria do not express flagellar antigen invitro. In this case the PCR-restriction fragment length polymorphism (RFLP-PCR and sequencing of the fliC may be suitable for the identification of antigens by replacing the traditional serology. We studied 17 samples of E. coli isolated from animals and presenting antigen H nontypeable (HNT. The H antigens were characterized by PCR-RFLP and sequencing of fliC gene. Three new flagellin genes were identified, for which specific antisera were obtained. The PCR-RFLP was shown to be faster than the serotyping H antigen in E. coli, provided information on some characteristics of these antigens and indicated the presence of new genes fliC.

  11. PPS nanoparticles as versatile delivery system to induce systemic and broad mucosal immunity after intranasal administration.

    Science.gov (United States)

    Stano, Armando; van der Vlies, André J; Martino, Mikael M; Swartz, Melody A; Hubbell, Jeffrey A; Simeoni, Eleonora

    2011-01-17

    Degradable polymer nanoparticles (NPs, 50 nm) based on polypropylene sulfide (PPS) were conjugated to thiolated antigen and adjuvant proteins by reversible disulfide bonds and evaluated in mucosal vaccination. Ovalbumin was used as a model antigen, and antigen-conjugated NPs were administered intranasally in the mouse. We show penetration of nasal mucosae, transit via M cells, and uptake by antigen-presenting cells in the nasal-associated lymphoid tissue. Ovalbumin-conjugated NPs induced cytotoxic T lymphocytic responses in lung and spleen tissues, as well as humoral response in mucosal airways. Co-conjugation of the TLR5 ligand flagellin further enhanced humoral responses in the airways as well as in the distant vaginal and rectal mucosal compartments and induced cellular immune responses with a Th1 bias, in contrast with free flagellin. The PPS NP platform thus appears interesting as a platform for intranasally-administered mucosal vaccination for inducing broad mucosal immunity.

  12. Expression and characterization of recombinant flagellin FlgE of Pseudomonas aeruginosa%绿脓杆菌鞭毛蛋白FlgE的重组表达及初步活性鉴定

    Institute of Scientific and Technical Information of China (English)

    林丹丹; 赵格; 王斌; 王宜强

    2013-01-01

    ligated to obtain the recombinant plasmid pET24a-FlgE.DNA sequencing of positive clone confirmed that the target gene and the junctions with vectors were all correct.The plasmid pET24a-FlgE was transformed into BL21 bacteria.The culture conditions like temperature,rotation speed,inducer concentrations,time length were optimized to achieve maximal expression of the target recombinant FlgE with 6×His tag at C terminal.FlgE-His proteins were purified using His-Trap affinity chromatography columns and identified by SDS-PAGE.The purified proteins were further subjected to endotoxin elimination with proper kits.The purified recombinant FlgE was added to cultured corneal epithelial cells for 4 h and the expression of several inflammation-related molecules was examined by using real-time quantitative PCR.Results The recombinant plasmid pET24a-FlgE was successfully constructed and high level FlgE expression was achieved in BL21 with rotation at 16℃ and 1 mmol/L isopropyl-β-D-glucopyranosyl-galactosidase induction for 20 h.Purified recombinant FlgE-His was obtained and used for primary bioactivity assay.After treatment of corneal epithelial cells with 20 μg/ml FlgE for 4 h,the expression of inflammatory cytokines IL-6,IL-8 were significantly increased.Inactivation of the FlgE with ethanol abolished its stimulatory activity.Conclusion The prokaryotic expression and purification system for recombinant Pseudomonas aeruginosa flagellin FlgE was set up,and the recombinant FlgE stimulated the expression of inflammatory factors in corneal epithelial cells.

  13. Unexpected roles of plastoglobules (plastid lipid droplets) in vitamin K1 and E metabolism.

    Science.gov (United States)

    Spicher, Livia; Kessler, Felix

    2015-06-01

    Tocopherol (vitamin E) and phylloquinone (vitamin K1) are lipid-soluble antioxidants that can only be synthesized by photosynthetic organisms. These compounds function primarily at the thylakoid membrane but are also present in chloroplast lipid droplets, also known as plastoglobules (PG). Depending on environmental conditions and stage of plant development, changes in the content, number and size of PG occur. PG are directly connected to the thylakoid membrane via the outer lipid leaflet. Apart from storage, PG are active in metabolism and likely trafficking of diverse lipid species. This review presents recent advances on how plastoglobules are implicated in the biosynthesis and metabolism of vitamin E and K.

  14. The K1.8BR spectrometer system at J-PARC

    CERN Document Server

    Agari, Keizo; Beer, George; Bhang, Hyoungchan; Bragadireanu, Mario; Buehler, Paul; Busso, Luigi; Cargnelli, Michael; Choi, Seonho; Curceanu, Catalina; Enomoto, Shun; Faso, Diego; Fujioka, Hiroyuki; Fujiwara, Yuya; Fukuda, Tomokazu; Guaraldo, Carlo; Hashimoto, Tadashi; Hayano, Ryugo S; Hiraiwa, Toshihiko; Hirose, Erina; Ieiri, Masaharu; Iio, Masami; Iliescu, Mihai; Inoue, Kentaro; Ishiguro, Yosuke; Ishikawa, Takashi; Ishimoto, Shigeru; Ishiwatari, Tomoichi; Itahashi, Kenta; Iwai, Masaaki; Iwasaki, Masahiko; Kakiguchi, Yutaka; Katoh, Yohji; Kawasaki, Shingo; Kienle, Paul; Kou, Hiroshi; Marton, Johann; Matsuda, Yasuyuki; Minakawa, Michifumi; Mizoi, Yutaka; Morra, Ombretta; Muto, Ryotaro; Nagae, Tomofumi; Naruki, Megumi; Noumi, Hiroyuki; Ohnishi, Hiroaki; Okada, Shinji; Outa, Haruhiko; Piscicchia, Kristian; Lener, Marco Poli; Vidal, Antonio Romero; Sada, Yuta; Sakaguchi, Atsushi; Sakuma, Fuminori; Sato, Masaharu; Sato, Yoshinori; Sawada, Shin'ya; Scordo, Alessandro; Sekimoto, Michiko; Shi, Hexi; Shirakabe, Yoshihisa; Sirghi, Diana; Sirghi, Florin; Suzuki, Ken; Suzuki, Shoji; Suzuki, Takatoshi; Suzuki, Yoshihiro; Takahashi, Hitoshi; Tanaka, Kazuhiro; Tanaka, Nobuaki; Tatsuno, Hideyuki; Tokuda, Makoto; Toyoda, Akihisa; Tomono, Dai; Toyoda, Akihisa; Tsukada, Kyo; Doce, Oton Vazquez; Watanabe, Hiroaki; Widmann, Eberhard; Wunschek, Barbara K; Yamanoi, Yutaka; Yamazaki, Toshimitsu; Yim, Heejoong; Zmeskal, Johann

    2012-01-01

    A new spectrometer system was designed and constructed at the secondary beam-line K1.8BR in the hadron hall of J-PARC, in order to investigate $\\bar K N$ interactions and $\\bar K$-nuclear bound systems. The spectrometer consists of a high precision beam line spectrometer, a liquid helium target system, a Cylindrical Detector System that surrounds the target to detect the decay particles from the target region, and a neutron time-of-flight counter array located $\\sim$15 m away from the target position. Details of the design, construction, and performance of the detector components are described.

  15. 关于k=1∑nkα(α≠1)的表示公式

    Institute of Scientific and Technical Information of China (English)

    杨必成

    1993-01-01

    本文利用改进的Euler—Maclaurin公式,导出如下k=1∑nKα(α≠1)的表示公式:i)当a∈R,a≠-1时,对于自然数q〉a+1,有k-1∑nkα=1/a+1na+1+k-1∑1(-1)k/K(a k-1)Bkna-k+1+γa+O(1/n1-a-1)ii)当a=m)O为整数时,有k-1∑nKm=k-0∑m(-1)km1/(m-k+1|k|Bknm-k+1这里,BK(k=1,2,……)Bernoulli数,γa为与a有关的常数,规定组合数(a k-1)=a(a-1)…(a-k+2)/(k-1)1(k为自然数)

  16. Observation and polarization measurements of B+/- -->phiK1 +/- and B +/- -->phiK2 *+/-.

    Science.gov (United States)

    Aubert, B; Bona, M; Karyotakis, Y; Lees, J P; Poireau, V; Prencipe, E; Prudent, X; Tisserand, V; Garra Tico, J; Grauges, E; Lopez, L; Palano, A; Pappagallo, M; Eigen, G; Stugu, B; Sun, L; Abrams, G S; Battaglia, M; Brown, D N; Cahn, R N; Jacobsen, R G; Kerth, L T; Kolomensky, Yu G; Kukartsev, G; Lynch, G; Osipenkov, I L; Ronan, M T; Tackmann, K; Tanabe, T; Hawkes, C M; Soni, N; Watson, A T; Koch, H; Schroeder, T; Walker, D; Asgeirsson, D J; Fulsom, B G; Hearty, C; Mattison, T S; McKenna, J A; Barrett, M; Khan, A; Teodorescu, L; Blinov, V E; Bukin, A D; Buzykaev, A R; Druzhinin, V P; Golubev, V B; Onuchin, A P; Serednyakov, S I; Skovpen, Yu I; Solodov, E P; Todyshev, K Yu; Bondioli, M; Curry, S; Eschrich, I; Kirkby, D; Lankford, A J; Lund, P; Mandelkern, M; Martin, E C; Stoker, D P; Abachi, S; Buchanan, C; Gary, J W; Liu, F; Long, O; Shen, B C; Vitug, G M; Yasin, Z; Zhang, L; Sharma, V; Campagnari, C; Hong, T M; Kovalskyi, D; Mazur, M A; Richman, J D; Beck, T W; Eisner, A M; Flacco, C J; Heusch, C A; Kroseberg, J; Lockman, W S; Schalk, T; Schumm, B A; Seiden, A; Wang, L; Wilson, M G; Winstrom, L O; Cheng, C H; Doll, D A; Echenard, B; Fang, F; Hitlin, D G; Narsky, I; Piatenko, T; Porter, F C; Andreassen, R; Mancinelli, G; Meadows, B T; Mishra, K; Sokoloff, M D; Bloom, P C; Ford, W T; Gaz, A; Hirschauer, J F; Kreisel, A; Nagel, M; Nauenberg, U; Smith, J G; Ulmer, K A; Wagner, S R; Ayad, R; Soffer, A; Toki, W H; Wilson, R J; Altenburg, D D; Feltresi, E; Hauke, A; Jasper, H; Karbach, M; Merkel, J; Petzold, A; Spaan, B; Wacker, K; Kobel, M J; Mader, W F; Nogowski, R; Schubert, K R; Schwierz, R; Sundermann, J E; Volk, A; Bernard, D; Bonneaud, G R; Latour, E; Thiebaux, Ch; Verderi, M; Clark, P J; Gradl, W; Playfer, S; Watson, J E; Andreotti, M; Bettoni, D; Bozzi, C; Calabrese, R; Cecchi, A; Cibinetto, G; Franchini, P; Luppi, E; Negrini, M; Petrella, A; Piemontese, L; Santoro, V; Baldini-Ferroli, R; Calcaterra, A; de Sangro, R; Finocchiaro, G; Pacetti, S; Patteri, P; Peruzzi, I M; Piccolo, M; Rama, M; Zallo, A; Buzzo, A; Contri, R; Lo Vetere, M; Macri, M M; Monge, M R; Passaggio, S; Patrignani, C; Robutti, E; Santroni, A; Tosi, S; Chaisanguanthum, K S; Morii, M; Marks, J; Schenk, S; Uwer, U; Klose, V; Lacker, H M; Bard, D J; Dauncey, P D; Nash, J A; Panduro Vazquez, W; Tibbetts, M; Behera, P K; Chai, X; Charles, M J; Mallik, U; Cochran, J; Crawley, H B; Dong, L; Meyer, W T; Prell, S; Rosenberg, E I; Rubin, A E; Gao, Y Y; Gritsan, A V; Guo, Z J; Lae, C K; Denig, A G; Fritsch, M; Schott, G; Arnaud, N; Béquilleux, J; D'Orazio, A; Davier, M; Firmino da Costa, J; Grosdidier, G; Höcker, A; Lepeltier, V; Le Diberder, F; Lutz, A M; Pruvot, S; Roudeau, P; Schune, M H; Serrano, J; Sordini, V; Stocchi, A; Wormser, G; Lange, D J; Wright, D M; Bingham, I; Burke, J P; Chavez, C A; Fry, J R; Gabathuler, E; Gamet, R; Hutchcroft, D E; Payne, D J; Touramanis, C; Bevan, A J; Clarke, C K; George, K A; Di Lodovico, F; Sacco, R; Sigamani, M; Cowan, G; Flaecher, H U; Hopkins, D A; Paramesvaran, S; Salvatore, F; Wren, A C; Brown, D N; Davis, C L; Alwyn, K E; Bailey, D S; Barlow, R J; Barlow, R J; Chia, Y M; Edgar, C L; Lafferty, G D; West, T J; Yi, J I; Anderson, J; Chen, C; Jawahery, A; Roberts, D A; Simi, G; Tuggle, J M; Dallapiccola, C; Li, X; Salvati, E; Saremi, S; Cowan, R; Dujmic, D; Fisher, P H; Koeneke, K; Sciolla, G; Spitznagel, M; Taylor, F; Yamamoto, R K; Zhao, M; Patel, P M; Robertson, S H; Lazzaro, A; Lombardo, V; Palombo, F; Bauer, J M; Cremaldi, L; Eschenburg, V; Godang, R; Kroeger, R; Sanders, D A; Summers, D J; Zhao, H W; Simard, M; Taras, P; Viaud, F B; Nicholson, H; De Nardo, G; Lista, L; Monorchio, D; Onorato, G; Sciacca, C; Raven, G; Snoek, H L; Jessop, C P; Knoepfel, K J; LoSecco, J M; Wang, W F; Benelli, G; Corwin, L A; Honscheid, K; Kagan, H; Kass, R; Morris, J P; Rahimi, A M; Regensburger, J J; Sekula, S J; Wong, Q K; Blount, N L; Brau, J; Frey, R; Igonkina, O; Kolb, J A; Lu, M; Rahmat, R; Sinev, N B; Strom, D; Strube, J; Torrence, E; Castelli, G; Gagliardi, N; Margoni, M; Morandin, M; Posocco, M; Rotondo, M; Simonetto, F; Stroili, R; Voci, C; del Amo Sanchez, P; Ben-Haim, E; Briand, H; Calderini, G; Chauveau, J; David, P; Del Buono, L; Hamon, O; Leruste, Ph; Ocariz, J; Perez, A; Prendki, J; Gladney, L; Biasini, M; Covarelli, R; Manoni, E; Angelini, C; Batignani, G; Bettarini, S; Carpinelli, M; Cervelli, A; Forti, F; Giorgi, M A; Lusiani, A; Marchiori, G; Morganti, M; Neri, N; Paoloni, E; Rizzo, G; Walsh, J J; Biesiada, J; Lopes Pegna, D; Lu, C; Olsen, J; Smith, A J S; Telnov, A V; Anulli, F; Baracchini, E; Cavoto, G; del Re, D; Di Marco, E; Faccini, R; Ferrarotto, F; Ferroni, F; Gaspero, M; Jackson, P D; Li Gioi, L; Mazzoni, M A; Morganti, S; Piredda, G; Polci, F; Renga, F; Voena, C; Ebert, M; Hartmann, T; Schröder, H; Waldi, R; Adye, T; Franek, B; Olaiya, E O; Roethel, W; Wilson, F F; Emery, S; Escalier, M; Esteve, L; Gaidot, A; Ganzhur, S F; Hamel de Monchenault, G; Kozanecki, W; Vasseur, G; Yèche, Ch; Zito, M; Chen, X R; Liu, H; Park, W; Purohit, M V; White, R M; Wilson, J R; Allen, M T; Aston, D; Bartoldus, R; Bechtle, P; Benitez, J F; Cenci, R; Coleman, J P; Convery, M R; Dingfelder, J C; Dorfan, J; Dubois-Felsmann, G P; Dunwoodie, W; Field, R C; Gabareen, A M; Gowdy, S J; Graham, M T; Grenier, P; Hast, C; Innes, W R; Kaminski, J; Kelsey, M H; Kim, H; Kim, P; Kocian, M L; Leith, D W G S; Li, S; Lindquist, B; Luitz, S; Luth, V; Lynch, H L; MacFarlane, D B; Marsiske, H; Messner, R; Muller, D R; Neal, H; Nelson, S; O'Grady, C P; Ofte, I; Perazzo, A; Perl, M; Ratcliff, B N; Roodman, A; Salnikov, A A; Schindler, R H; Schwiening, J; Snyder, A; Su, D; Sullivan, M K; Suzuki, K; Swain, S K; Thompson, J M; Va'vra, J; Wagner, A P; Weaver, M; West, C A; Wisniewski, W J; Wittgen, M; Wright, D H; Wulsin, H W; Yarritu, A K; Yi, K; Young, C C; Ziegler, V; Burchat, P R; Edwards, A J; Majewski, S A; Miyashita, T S; Petersen, B A; Wilden, L; Ahmed, S; Alam, M S; Ernst, J A; Pan, B; Saeed, M A; Zain, S B; Spanier, S M; Wogsland, B J; Eckmann, R; Ritchie, J L; Ruland, A M; Schilling, C J; Schwitters, R F; Drummond, B W; Izen, J M; Lou, X C; Bianchi, F; Gamba, D; Pelliccioni, M; Bomben, M; Bosisio, L; Cartaro, C; Della Ricca, G; Lanceri, L; Vitale, L; Azzolini, V; Lopez-March, N; Martinez-Vidal, F; Milanes, D A; Oyanguren, A; Albert, J; Banerjee, Sw; Bhuyan, B; Choi, H H F; Hamano, K; Kowalewski, R; Lewczuk, M J; Nugent, I M; Roney, J M; Sobie, R J; Gershon, T J; Harrison, P F; Ilic, J; Latham, T E; Mohanty, G B; Band, H R; Chen, X; Dasu, S; Flood, K T; Pan, Y; Pierini, M; Prepost, R; Vuosalo, C O; Wu, S L

    2008-10-17

    With the full BABAR data sample of 465 x 10(6) B(over)B pairs, we observe the decays B+/- -->phiK_(1)(1270) +/- and B +/- -->phiK*_(2)(1430)+/-. We measure the branching fractions (6.1+/-1.6+/-1.1) x 10(-6) and (8.4+/-1.8+/-1.0) x 10(-6) and the fractions of longitudinal polarization 0.46 (+0.12+0.06) _(-0.13-0.07) and 0.80(+0.09)_(-0.10)+/-0.03, respectively. We also report on the B+/- -->phiK*_(0)(1430)+/- decay branching fraction of (7.0+/-1.3+/-0.9) x 10(-6) and several parameters sensitive to CP violation and interference in the above three decays. Upper limits are placed on the B+/- decay rates to final states with phi and K_1(1400)+/-, K*(1410)+/-, K2(1770)+/-, or K_2(1820)+/-. Understanding the observed polarization pattern requires amplitude contributions from an uncertain source.

  17. Genetically similar isolates of Klebsiella pneumoniae serotype K1 causing liver abscesses in three continents.

    Science.gov (United States)

    Turton, Jane F; Englender, Hilary; Gabriel, Samantha N; Turton, Sarah E; Kaufmann, Mary E; Pitt, Tyrone L

    2007-05-01

    The magA gene was sought in hypermucoviscous isolates of Klebsiella spp., the Klebsiella K serotype reference strains and in isolates of the K1 serotype of Klebsiella pneumoniae from the UK, Hong Kong, Israel, Taiwan and Australia. Only K1 isolates were PCR positive for magA; this gene was found in all such isolates tested. Hypermucoviscosity was not confined to magA positive isolates, nor was it found in all magA positive isolates. Comparison of XbaI PFGE profiles revealed that most (19/23) of the magA positive isolates clustered within 72 % similarity, with a further subcluster of isolates, from three different continents, clustering within >80 %. All of the 16 isolates tested within the main cluster had the same sequence type (ST 23) by multilocus sequence typing, with the exception of one isolate, which had a single nucleotide difference at one of the seven loci. This study indicates that a genotype strongly associated with highly invasive disease in Taiwan, where large numbers of cases have been reported, is geographically very widespread.

  18. Vitamins K1 and K2: The Emerging Group of Vitamins Required for Human Health

    Directory of Open Access Journals (Sweden)

    Gerry Kurt Schwalfenberg

    2017-01-01

    Full Text Available Objective. To review the evidence for the use of vitamin K supplementation in clinical conditions such as osteoporosis, vascular calcification, arthritis, cancer, renal calculi, diabetes, and warfarin therapy. Quality of Evidence. PubMed was searched for articles on vitamin K (K1 and K2 along with books and conference proceedings and health conditions listed above. Level I and II evidence supports the use of vitamins K1 and K2 in osteoporosis and Level II evidence supports vitamin K2 in prevention of coronary calcification and cardiovascular disease. Evidence is insufficient for use in diabetes, arthritis, renal calculi, and cancer. Main Message. Vitamin K2 may be a useful adjunct for the treatment of osteoporosis, along with vitamin D and calcium, rivaling bisphosphonate therapy without toxicity. It may also significantly reduce morbidity and mortality in cardiovascular health by reducing vascular calcification. Vitamin K2 appears promising in the areas of diabetes, cancer, and osteoarthritis. Vitamin K use in warfarin therapy is safe and may improve INR control, although a dosage adjustment is required. Conclusion. Vitamin K supplementation may be useful for a number of chronic conditions that are afflicting North Americans as the population ages. Supplementation may be required for bone and cardiovascular health.

  19. K 1-6: an asymmetric planetary nebula with a binary central star

    CERN Document Server

    Frew, David J; Fitzgerald, Michael; Parker, Quentin; Danaia, Lena; McKinnon, David; Guerrero, Martín A; Hedberg, John; Hollow, Robert; An, Yvonne; Bor, Shu Han; Colman, Isabel; Graham-White, Claire; Li, Qing Wen; Mai, Juliette; Papadakis, Katerina; Picone-Murray, Julia; Hoang, Melanie Vo; Yean, Vivian

    2010-01-01

    We present new imaging data and archival multiwavelength observations of the little studied emission nebula K 1-6 and its central star. Narrow-band images in H-alpha (+ [NII]) and [OIII] taken with the Faulkes Telescope North reveal a stratified, asymmetric, elliptical nebula surrounding a central star which has the colours of a late G- or early K-type subgiant or giant. GALEX ultraviolet images reveal a very hot subdwarf or white dwarf coincident in position with this star. The cooler, optically dominant star is strongly variable with a period of 21.312 +/- 0.008 days, and is possibly a high amplitude member of the RS CVn class, although an FK Com classification is also possible. Archival ROSAT data provide good evidence that the cool star has an active corona. We conclude that K 1-6 is most likely an old bona fide planetary nebula at a distance of ~1.0 kpc, interacting with the interstellar medium, and containing a binary or ternary central star. The observations and data analyses reported in this paper wer...

  20. Path representation of su(2)_k states I: Operators and particles for k=1,2

    CERN Document Server

    Lamy-Poirier, Joel

    2010-01-01

    This is the first of two articles devoted to the analysis of the path description of the states in integrable modules of su(2)_k. In this first article, the cases k=1,2 are treated in detail, emphasizing a different description in each case (operators vs particles). Our main result for k=1 is a proof of equivalence of two known path representations for the finitized su(2)_1 states, obtained by displaying an explicit bijection. An immediate offshoot is the gain of a new and simple weighting for the (Kyoto) path representation that generalizes to level k. The bijection also suggests two operator constructions for the su(2)_1 paths, a local and a nonlocal one, both interrelated. The nonlocal operator description is the starting point for a direct derivation of the su(2)_1 spinon character. The second part presents an extensive study of the su(2)_2 paths from their particle point of view, where the particles are defined as the path building blocks. The resulting generating functions appear to provide new (at leas...

  1. Neuroprotection of GluK1 kainate receptor agonist ATPA against ischemic neuronal injury through inhibiting GluK2 kainate receptor-JNK3 pathway via GABA(A) receptors.

    Science.gov (United States)

    Lv, Qian; Liu, Yong; Han, Dong; Xu, Jing; Zong, Yan-Yan; Wang, Yao; Zhang, Guang-Yi

    2012-05-25

    It is well known that GluK2-containing kainate receptors play essential roles in seizure and cerebral ischemia-induced neuronal death, while GluK1-containing kainate receptors could increase tonic inhibition of post-synaptic pyramidal neurons. This research investigated whether GluK1 could inhibit activation of c-Jun N-terminal kinase 3 (JNK3) signaling pathway mediated by the GluK2 in cerebral ischemia-reperfusion. The results show that GluK1 activation by (RS)-2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA) at 1nmol per rat could inhibit the assembly of GluK2·Postsynaptic density 95·mixed lineage kinase 3 signaling module, activation of JNK3 and its downstream signal molecules. However, the inhibition of ATPA could be prevented by GluK1 antagonist NS3763, GluK1 antisense, and GABA(A) receptor antagonist bicuculline. In addition, ATPA played a neuroprotective role against cerebral ischemia. In sum, the findings indicate that activation of GluK1 by ATPA at specific dosages may promote GABA release, which then suppresses post-synaptic GluK2-JNK3 signaling-mediated cerebral ischemic injury via GABA(A)R.

  2. Regulation of Toll-like receptor 2 interaction with Ecgp96 controls Escherichia coli K1 invasion of brain endothelial cells

    Science.gov (United States)

    Krishnan, Subramanian; Chen, Shuang; Turcatel, Gianluca; Arditi, Moshe; Prasadarao, Nemani V.

    2012-01-01

    SUMMARY The interaction of outer membrane protein A (OmpA) with its receptor, Ecgp96 (a homologue of Hsp90β) is critical for the pathogenesis of E. coli K1 meningitis. Since Hsp90 chaperones Toll-like receptors (TLRs), we examined the role of TLRs in E. coli K1 infection. Herein, we show that newborn TLR2−/− mice are resistant to E. coli K1 meningitis, while TLR4−/− mice succumb to infection sooner. In vitro, OmpA+ E. coli infection selectively upregulates Ecgp96 and TLR2 in human brain microvascular endothelial cells (HBMEC), whereas OmpA− E. coli upregulates TLR4 in these cells. Furthermore, infection with OmpA+ E. coli causes Ecgp96 and TLR2 translocate to the plasma membrane of HBMEC as a complex. Immunoprecipitation studies of the plasma membrane fractions from infected HBMEC reveal that the C-termini of Ecgp96 and TLR2 are critical for OmpA+ E. coli invasion. Knockdown of TLR2 using siRNA results in inefficient membrane translocation of Ecgp96 and significantly reduces invasion. In addition, the interaction of Ecgp96 and TLR2 induces a bipartite signal, one from Ecgp96 through PKC-α while the other from TLR2 through MyD88, ERK1/2 and NF-κB. This bipartite signal ultimately culminates in the efficient production of NO, which in turn promotes E. coli K1 invasion of HBMEC. PMID:22963587

  3. Cell growth stimulating effect of Ganoderma lucidum spores and their potential application for Chinese hamster ovary K1 cell cultivation.

    Science.gov (United States)

    Li, Ding; Zhong, Qi; Liu, Tingting; Wang, Jufang

    2016-06-01

    In this work, water-soluble extracts of Ganoderma lucidum spores (Gls), a Chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for Chinese hamster ovary (CHO) cell cultivation. The Gls extract was prepared and supplemented to CHO K1 cell culture media with various serum levels. Our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount Gls extract effectively promoted cell growth and suppressed cell apoptosis induced by serum deprivation with normal cell cycle maintained in a low-serum medium. The low-serum medium containing 1 % (v/v) fetal bovine serum (FBS) and 0.01 % (w/v) Gls extract showed a comparable performance on both cell growth and fusion protein productivity with the conventional CHO culture medium containing 10 % (v/v) FBS and a commercial serum-free medium. This is the first study of the potential of Gls extracts for use as an alternative cell growth factor and nutrient for CHO cells. The findings have presented a new approach to economic cultivation of CHO cells for therapeutic protein production.

  4. Blocking Effect and Moments of Inertia of K = 1/2 Rotational Band in 171Yb

    Institute of Scientific and Technical Information of China (English)

    YU Shao-Ying; LIU Yan-Xin; LIU Shu-Xin; HE Xiao-Tao

    2004-01-01

    The K = 1/2 rotational band in 171yb is investigated using the particle number conserving (PNC) method for treating the cranked shell model with monopole and quadrupole pairing interactions. The experimental moments of inertia of 171 Yb [521]1/2 (signature α = +1/2) are reproduced well by the PNC calculation, in which no free parameter is involved. The difference in the contribution to the moment of inertia between protons and neutrons is mainly due to the blocking effect of neutron normal orbitals. The ω variation of the occupation probability of each cranked orbital and the contribution to the moment of inertia from each major shell and from each cranked orbital are investigated.

  5. Blocking Effect and Moments of Inertia of K = 1/2 Rotational Band in 171Yb

    Institute of Scientific and Technical Information of China (English)

    YUShao-Ying; LIUYan-Xin; LIUShu-Xin; HEXiao-Tao

    2004-01-01

    The K = 1/2 rotational band in 171Yb is investigated using the particle number conserving (PNC) method for treating the cranked shell model with monopole and quadrupole pairing interactions. The experimental moments of inertia of 171Yb [52111/2 (signature α = ±1/2) are reproduced well by the P N C calculation, in which no free parameter is involved. The difference in the contribution to the moment of inertia between protons and neutrons is mainly due to the blocking effect of neutron normal orbitals. The ω variation of the occupation probability of each cranked orbital and the contribution to the moment of inertia from each major shell and from each cranked orbital are investigated.

  6. Cell survival and chromosomal aberrations in CHO-K1 cells irradiated by carbon ions.

    Science.gov (United States)

    Czub, J; Banaś, D; Błaszczyk, A; Braziewicz, J; Buraczewska, I; Choiński, J; Górak, U; Jaskóła, M; Korman, A; Lankoff, A; Lisowska, H; Łukaszek, A; Szefliński, Z; Wójcik, A

    2009-03-01

    Chinese hamster ovary CHO-K1 cells were exposed to high LET (12)C-beam (LET: 830 keV/microm) in the dose range of 0-6 Gy and to (60)Co irradiation and the RBE value was obtained. Effects of (12)C-beam exposure on cell survival and chromosomal aberrations were calculated. The chromosomal aberration data were fitted with linear equation. The distribution of aberration in cells was examined with a standard u-test and used to evaluate the data according to Poisson probabilities. The variance to the mean ratio sigma(2)/Y and the dispersion index (u) were determined. Overdispersion was significant (p<0.05) when the value of u exceeded 1.96.

  7. Performance evaluation of CHO-K1 cell in culture medium supplemented with hemolymph

    Directory of Open Access Journals (Sweden)

    Tássia Raffoul

    2005-06-01

    Full Text Available The aim of this work was to evaluate the potential of hemolymph utilization as a culture medium supplement to cultivate the animal cell CHO-K1. For this purpose 1% v/v of hemolymph was added to DMEM medium containing 10% v/v of FBS and 1 or 4.5 g/L of glucose. The culture was grown in spinner flasks incubated in a 10% v/v CO2 environment, at 37ºC, with the Cytodex 1 microcarrier. Comparing the results obtained from the culture with hemolymph against those without hemolymph, a positive influence of the hemolymph was observed, as the experiment with hemolymph presented a 52% higher cell concentration and a higher productivity of up to 40%.Desenvolvimento de meios de cultura isentos de soro fetal bovino (SFB é uma das grandes prioridades de pesquisa em desenvolvimento de processos com célula animal. O objetivo do presente trabalho foi realizar uma análise do potencial de uso da hemolinfa como suplemento do meio utilizado no cultivo da célula animal ancorante CHO-K1. Para isso, foi adicionado 1% v/v de extrato de hemolinfa ao meio DMEM contendo 10% v/v de SFB e 1,0 ou 4,5 g/L de glicose. O cultivo foi realizado em frascos tipo spinner em um ambiente de 10% v/v de CO2, a 37ºC, utilizando o microcarregador Cytodex 1. Comparando os resultados obtidos no ensaio com hemolinfa com um sem hemolinfa pode-se notar uma influência positiva da hemolinfa no cultivo, já que o ensaio com hemolinfa apresentou uma concentração máxima de células 52% maior e uma produtividade máxima de até 40% maior.

  8. Sulfhydryl compounds inhibit the cyto- and geno-toxicity of o-phenylphenol metabolites in CHO-K1 cells.

    Science.gov (United States)

    Tayama, S; Nakagawa, Y

    1991-01-01

    The effects of cysteine and reduced glutathione (GSH) on the genotoxicity of o-phenylphenol (OPP) and its metabolites, phenylhydroquinone (PHQ) and phenylbenzoquinone (PBQ), were examined using the frequency of sister-chromatid exchanges (SCEs) and chromosome aberrations in CHO-K1 cells as parameters. Cytotoxic (cell-progression delay) and cytogenetic effects induced by a 3-h treatment with OPP, PHQ (100 micrograms/ml) or PBQ (50 micrograms/ml) with S9 mix after a 27-h expression time were inhibited by cysteine or GSH (3-10 mM). Materials corresponding to the cysteine or GSH adducts were found by HPLC in each incubation mixture. In the culture without S9 mix, PHQ and PBQ showed severe cytotoxicity since no metaphases could be obtained at doses over 25 and 5 micrograms/ml, respectively, and the sulfhydryl compounds inhibited the toxicity by the formation of adducts with PBQ and by inhibiting the formation of PBQ in the case of PHQ. With PHQ, the sulfhydryl compounds appeared to inhibit autooxidation. However, the sulfhydryl compounds did not inhibit the cytotoxic and cytogenetic effects caused by OPP in the cell mixture without S9 mix, but on the contrary intensified them. No adduct formation was detected in the incubation solution. On the basis of these results, it is considered that electrophilic quinone (PBQ) and/or semiquinone (phenylsemiquinone, PSQ) radicals, capable of binding to nucleophilic small molecules (such as cysteine and GSH) or (biological) macromolecules, are produced from metabolite PHQ in metabolic oxidation of OPP, and induce cyto- and geno-toxic effects in the cells. The cyto- and geno-toxic effects of OPP itself to the cells are clearly independent of any electrophilic radical reaction.

  9. SseK1 and SseK3 Type III Secretion System Effectors Inhibit NF-κB Signaling and Necroptotic Cell Death in Salmonella-Infected Macrophages

    Science.gov (United States)

    Günster, Regina A.; Matthews, Sophie A.; Holden, David W.

    2017-01-01

    ABSTRACT Within host cells such as macrophages, Salmonella enterica translocates virulence (effector) proteins across its vacuolar membrane via the SPI-2 type III secretion system. Previously, it was shown that when expressed ectopically, the effectors SseK1 and SseK3 inhibit tumor necrosis factor alpha (TNF-α)-induced NF-κB activation. In this study, we show that ectopically expressed SseK1, SseK2, and SseK3 suppress TNF-α-induced, but not Toll-like receptor 4- or interleukin-induced, NF-κB activation. Inhibition required a DXD motif in SseK1 and SseK3, which is essential for the transfer of N-acetylglucosamine to arginine residues (arginine-GlcNAcylation). During macrophage infection, SseK1 and SseK3 inhibited NF-κB activity in an additive manner. SseK3-mediated inhibition of NF-κB activation did not require the only known host-binding partner of this effector, the E3-ubiquitin ligase TRIM32. SseK proteins also inhibited TNF-α-induced cell death during macrophage infection. Despite SseK1 and SseK3 inhibiting TNF-α-induced apoptosis upon ectopic expression in HeLa cells, the percentage of infected macrophages undergoing apoptosis was SseK independent. Instead, SseK proteins inhibited necroptotic cell death during macrophage infection. SseK1 and SseK3 caused GlcNAcylation of different proteins in infected macrophages, suggesting that these effectors have distinct substrate specificities. Indeed, SseK1 caused the GlcNAcylation of the death domain-containing proteins FADD and TRADD, whereas SseK3 expression resulted in weak GlcNAcylation of TRADD but not FADD. Additional, as-yet-unidentified substrates are likely to explain the additive phenotype of a Salmonella strain lacking both SseK1 and SseK3. PMID:28069818

  10. Investigation on the suitable dose of vitamin k1 in treating hemorrhagic disease of newborn%维生素k1治疗新生儿出血症适宜剂量探讨

    Institute of Scientific and Technical Information of China (English)

    张敏

    2008-01-01

    目的 观察不同剂量维生素k1治疗新生儿出血症的临床疗效.方法 将100例新生儿出血症患儿随机分为两组,大剂量维生素k1治疗(治疗组)50例;小剂量维生素k1治疗(对照组)50例.结果 治疗组与对照组患儿凝血酶原时间恢复正常时间和症状消失时间无显著差异(P>0.05).结论 维生毒素k1治疗新生儿出血症选用小剂量即可,无须大剂量使用.

  11. Single Photon K-2 and K-1K-1 Double Core Ionization in C2H2n (n=1-3), CO, and N2 as a Potential New Tool for Chemical Analysis

    Science.gov (United States)

    Nakano, M.; Penent, F.; Tashiro, M.; Grozdanov, T. P.; Žitnik, M.; Carniato, S.; Selles, P.; Andric, L.; Lablanquie, P.; Palaudoux, J.; Shigemasa, E.; Iwayama, H.; Hikosaka, Y.; Soejima, K.; Suzuki, I. H.; Kouchi, N.; Ito, K.

    2013-04-01

    We have observed single photon double K-shell photoionization in the C2H2n (n=1-3) hydrocarbon sequence and in N2 and CO, using synchrotron radiation and electron coincidence spectroscopy. Our previous observations of the K-2 process in these molecules are extended by the observations of a single photon double photoionization with one core hole created at each of the two neighboring atoms in the molecule (K-1K-1 process). In the C2H2n sequence, the spectroscopy of K-1K-1 states is much more sensitive to the bond length than conventional electron spectroscopy for chemical analysis spectroscopy based on single K-shell ionization. The cross section variation for single photon K-1K-1 double core ionization in the C2H2n sequence and in the isoelectronic C2H2, N2 and CO molecules validates a knock-out mechanism in which a primary ionized 1s photoelectron ejects another 1s electron of the neighbor atom. The specific Auger decay from such states is clearly observed in the CO case.

  12. KLF5 regulates infection- and inflammation-induced pro-labour mediators in human myometrium.

    Science.gov (United States)

    Lappas, Martha

    2015-05-01

    The transcription factor Kruppel-like factor 5 (KLF5) has been shown to associate with nuclear factor kappa B (NFκB) to regulate genes involved in inflammation. However, there are no studies on the expression and regulation of KLF5 in the processes of human labour and delivery. Thus, the aims of this study were to determine the effect of i) human labour on KLF5 expression in both foetal membranes and myometrium; ii) the pro-inflammatory cytokine interleukin 1 beta (IL1β), bacterial product flagellin and the viral dsRNA analogue poly(I:C) on KLF5 expression and iii) KLF5 knockdown by siRNA in human myometrial primary cells on pro-inflammatory and pro-labour mediators. In foetal membranes, there was no effect of term or preterm labour on KLF5 expression. In myometrium, the term labour was associated with an increase in nuclear KLF5 protein expression. Moreover, KLF5 expression was also increased in myometrial cells treated with IL1β, flagellin or poly(IC), likely factors contributing to preterm birth. KLF5 silencing in myometrial cells significantly decreased IL1β-induced cytokine expression (IL6 and IL8 mRNA expression and release), COX2 mRNA expression, and subsequent release of prostaglandins PGE2 and PGF2 α. KLF5 silencing also significantly reduced flagellin- and poly(I:C)-induced IL6 and IL8 mRNA expression. Lastly, IL1β-, flagellin- and poly(I:C)-stimulated NFκB transcriptional activity was significantly suppressed in KLF5-knockout myometrial cells. In conclusion, this study describes novel data in which KLF5 is increased in labouring myometrium, and KLF5 silencing decreased inflammation- and infection-induced pro-labour mediators. © 2015 Society for Reproduction and Fertility.

  13. No effect of vitamin K1 intake on bone mineral density and fracture risk in perimenopausal women

    DEFF Research Database (Denmark)

    Rejnmark, L; Vestergaard, P; Charles, P

    2006-01-01

    INTRODUCTION: Vitamin K functions as a co-factor in the post-translational carboxylation of several bone proteins, including osteocalcin. AIM: The aim of this study was to investigate the relationship between vitamin K(1) intake and bone mineral density (BMD) and fracture risk in a perimenopausal...... Danish population. DESIGN: The study was performed within the Danish Osteoporosis Prevention Study (DOPS), including a population-based cohort of 2,016 perimenopausal women. During the study approximately 50% of the women received hormone replacement therapy (HRT). Associations between vitamin K(1......) intake and BMD were assessed at baseline and after 5-years of follow-up (cross-sectional design). Moreover, associations between vitamin K(1) intake and 5-year and 10-year changes in BMD were studied (follow-up design). Finally, fracture risk was assessed in relation to vitamin K(1) intake (nested case...

  14. The {P,Q,k+1}-Reflexive Solution to System of Matrix Equations AX=C, XB=D

    Directory of Open Access Journals (Sweden)

    Chang-Zhou Dong

    2015-01-01

    Full Text Available Let P∈Cm×m and Q∈Cn×n be Hermitian and {k+1}-potent matrices; that is, Pk+1=P=P⁎ and Qk+1=Q=Q⁎, where ·⁎ stands for the conjugate transpose of a matrix. A matrix X∈Cm×n is called {P,Q,k+1}-reflexive (antireflexive if PXQ=X (PXQ=-X. In this paper, the system of matrix equations AX=C and XB=D subject to {P,Q,k+1}-reflexive and antireflexive constraints is studied by converting into two simpler cases: k=1 and k=2. We give the solvability conditions and the general solution to this system; in addition, the least squares solution is derived; finally, the associated optimal approximation problem for a given matrix is considered.

  15. Inhibition of S6K1 accounts partially for the anti-inflammatory effects of the arginase inhibitor L-norvaline

    Directory of Open Access Journals (Sweden)

    Ruffieux Jean

    2009-03-01

    Full Text Available Abstract Background Pharmacological inhibition of endothelial arginase-II has been shown to improve endothelial nitric oxide synthase (eNOS function and reduce atherogenesis in animal models. We investigated whether the endothelial arginase II is involved in inflammatory responses in endothelial cells. Methods Human endothelial cells were isolated from umbilical veins and stimulated with TNFα (10 ng/ml for 4 hours. Endothelial expression of the inflammatory molecules i.e. vascular cell adhesion molecule-1 (VCAM-1, intercellular adhesion molecule-1 (ICAM-1, and E-selectin were assessed by immunoblotting. Results The induction of the expression of endothelial VCAM-1, ICAM-1 and E-selectin by TNFα was concentration-dependently reduced by incubation of the endothelial cells with the arginase inhibitor L-norvaline. However, inhibition of arginase by another arginase inhibitor S-(2-boronoethyl-L-cysteine (BEC had no effects. To confirm the role of arginase-II (the prominent isoform expressed in HUVECs in the inflammatory responses, adenoviral mediated siRNA silencing of arginase-II knocked down the arginase II protein level, but did not inhibit the up-regulation of the adhesion molecules. Moreover, the inhibitory effect of L-norvaline was not reversed by the NOS inhibitor L-NAME and L-norvaline did not interfere with TNFα-induced activation of NF-κB, JNK, p38mapk, while it inhibited p70s6k (S6K1 activity. Silencing S6K1 prevented up-regulation of E-selectin, but not that of VCAM-1 or ICAM-1 induced by TNFα. Conclusion The arginase inhibitor L-norvaline exhibits anti-inflammatory effects independently of inhibition of arginase in human endothelial cells. The anti-inflammatory properties of L-norvaline are partially attributable to its ability to inhibit S6K1.

  16. Approximating common fixed points of asymptotically quasi-nonexpansive mappings by a k+1-step iterative scheme with error terms

    Science.gov (United States)

    Xiao, Jian-Zhong; Sun, Jing; Huang, Xuan

    2010-02-01

    In this paper a k+1-step iterative scheme with error terms involving k+1 asymptotically quasi-nonexpansive mappings is studied. In usual Banach spaces, some sufficient and necessary conditions are given for the iterative scheme to approximate a common fixed point. In uniformly convex Banach spaces, power equicontinuity for a mapping is introduced and a series of new convergence theorems are established. Several known results in the current literature are extended and refined.

  17. An Approach to a Method of Construction of (F, K, 1) Optical Orthogonal Codes from Block Design

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    (F, K, 1) Optical orthogonal codes (OOC) are the best address codes applied to optical code division multiple access (OCDMA) communication systems, but the construction of the codes is very complex. In this paper, a method of construction of the OOC from block design is discussed and a method of computer aid design is presented, by which we can construct desired (F, K, 1) OOC easily.

  18. Minor abnormalities of testis development in mice lacking the gene encoding the MAPK signalling component, MAP3K1.

    Directory of Open Access Journals (Sweden)

    Nick Warr

    Full Text Available In mammals, the Y chromosome is a dominant male determinant, causing the bipotential gonad to develop as a testis. Recently, cases of familial and spontaneous 46,XY disorders of sex development (DSD have been attributed to mutations in the human gene encoding mitogen-activated protein kinase kinase kinase 1, MAP3K1, a component of the mitogen-activated protein kinase (MAPK signal transduction pathway. In individuals harbouring heterozygous mutations in MAP3K1, dysregulation of MAPK signalling was observed in lymphoblastoid cell lines, suggesting a causal role for these mutations in disrupting XY sexual development. Mice lacking the cognate gene, Map3k1, are viable and exhibit the eyes open at birth (EOB phenotype on a mixed genetic background, but on the C57BL/6J genetic background most mice die at around 14.5 dpc due to a failure of erythropoiesis in the fetal liver. However, no systematic examination of sexual development in Map3k1-deficient mice has been described, an omission that is especially relevant in the case of C57BL/6J, a genetic background that is sensitized to disruptions to testis determination. Here, we report that on a mixed genetic background mice lacking Map3k1 are fertile and exhibit no overt abnormalities of testis development. On C57BL/6J, significant non-viability is observed with very few animals surviving to adulthood. However, an examination of development in Map3k1-deficient XY embryos on this genetic background revealed no significant defects in testis determination, although minor abnormalities were observed, including an increase in gonadal length. Based on these observations, we conclude that MAP3K1 is not required for mouse testis determination. We discuss the significance of these data for the functional interpretation of sex-reversing MAP3K1 mutations in humans.

  19. Investigation of the utility of complementary electrochemical detection techniques to examine the in vitro affinity of bacterial flagellins for a toll-like receptor 5 biosensor.

    Science.gov (United States)

    She, Zhe; Topping, Kristin; Shamsi, Mohtashim H; Wang, Nan; Chan, Nora W C; Kraatz, Heinz-Bernhard

    2015-04-21

    An initial investigation of the fabrication of a novel biosensor utilizing toll-like receptor 5 (TLR5) has been conducted. The detection assay using this sensor platform has been carried out using two complementary electrochemical techniques. The electrochemical properties of the modified bare gold surface following TLR5 immobilization were characterized. The electrochemical response to changes in the sensor film resistance and electron charge-transfer permittivity triggered by independent exposures to flagellins from Salmonella typhimurium (S. typhimurium) and Bacillus subtilis (B. subtilis) were examined and observed. The quantified film resistance data gathered using electrochemical impedance spectroscopy (EIS) over a macroscopic scale are in significant agreement with the corresponding electron charge-transfer permittivity measured locally by scanning electrochemical microscopy (SECM). Unlike other sensors that exploit pathogen recognition elements, TLR5 biosensors have the potential to carry out broad-spectrum detection of flagellated bacterial pathogens in near real time. This broad-spectrum detection platform is a significant step toward the development of fast, inexpensive clinical tools for early warning diagnoses and immediate on-site treatment.

  20. TLR5-dependent immunogenicity of a recombinant fusion protein containing an immunodominant epitope of malarial circumsporozoite protein and the FliC flagellin of Salmonella Typhimurium

    Directory of Open Access Journals (Sweden)

    Ariane Guglielmi Ariza Camacho

    2011-08-01

    Full Text Available Recently, we described the improved immunogenicity of new malaria vaccine candidates based on the expression of fusion proteins containing immunodominant epitopes of merozoites and Salmonella enterica serovar Typhimurium flagellin (FliC protein as an innate immune agonist. Here, we tested whether a similar strategy, based on an immunodominant B-cell epitope from malaria sporozoites, could also generate immunogenic fusion polypeptides. A recombinant His6-tagged FliC protein containing the C-terminal repeat regions of the VK210 variant of Plasmodium vivax circumsporozoite (CS protein was constructed. This recombinant protein was successfully expressed in Escherichia coli as soluble protein and was purified by affinity to Ni-agarose beads followed by ion exchange chromatography. A monoclonal antibody specific for the CS protein of P. vivax sporozoites (VK210 was able to recognise the purified protein. C57BL/6 mice subcutaneously immunised with the recombinant fusion protein in the absence of any conventional adjuvant developed protein-specific systemic antibody responses. However, in mice genetically deficient in expression of TLR5, this immune response was extremely low. These results extend our previous observations concerning the immunogenicity of these recombinant fusion proteins and provide evidence that the main mechanism responsible for this immune activation involves interactions with TLR5, which has not previously been demonstrated for any recombinant FliC fusion protein.

  1. Introduction of a carbon paste electrode based on nickel carbide for investigation of interaction between warfarin and vitamin K1.

    Science.gov (United States)

    Torkashvand, Maryam; Gholivand, Mohammad Bagher; Taherpour, Avat Arman; Boochani, Arash; Akhtar, Arsalan

    2017-05-30

    In this paper a novel electrochemical sensor based on nickel carbide (Ni3C) nanoparticles as a new modifier was constructed. Ni3C nanoparticle was synthesized and characterized by scanning electron microscopy, X-ray diffraction and first-principles study. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies confirmed the electrode modification. Afterwards, the new electrode for the first time was used for interaction study between vitamin K1 and warfarin as an anticoagulant drug by differential pulse voltammetry. The adduct formation between the drug and vitamin K1 was improved by decreasing in anodic peak current of warfarin in the presence of different amounts of vitamin K1. The binding constant between warfarin and vitamin K1 was obtained by voltammetric and UV-vis and fluorescence spectroscopic methods. The molecular modeling method was also performed to explore the structural features and binding mechanism of warfarin to vitamin K1. The different aspects of modeling of vitamin K1 and warfarin and their adduct structures confirmed the adduct formation by hydrogen bonding. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense

    Science.gov (United States)

    He, Xiaolong; Zeng, Qing; Puthiyakunnon, Santhosh; Zeng, Zhijie; Yang, Weijun; Qiu, Jiawen; Du, Lei; Boddu, Swapna; Wu, Tongwei; Cai, Danxian; Huang, Sheng-He; Cao, Hong

    2017-01-01

    The objective of this study was to determine whether Lactobacillus rhamnosus GG culture supernatant (LCS) has a preventive effect against gut-derived systemic neonatal Escherichia coli (E. coli) K1 infection. The preventive effects were evaluated in human colonic carcinoma cell line Caco-2 and neonatal rat models. Our in vitro results showed that LCS could block adhesion, invasion and translocation of E. coli K1 to Caco-2 monolayer via up-regulating mucin production and maintaining intestinal integrity. In vivo experiments revealed that pre-treatment with LCS significantly decrease susceptibility of neonatal rats to oral E. coli K1 infection as reflected by reduced bacterial intestinal colonization, translocation, dissemination and systemic infections. Further, we found that LCS treated neonatal rats have higher intestinal expressions of Ki67, MUC2, ZO-1, IgA, mucin and lower barrier permeability than those in untreated rats. These results indicated that LCS could enhance neonatal resistance to systemic E. coli K1 infection via promoting maturation of neonatal intestinal defense. In conclusions, our findings suggested that LCS has a prophylactic effect against systemic E. coli K1 infection in neonates. Future studies aimed at identifying the specific active ingredients in LCS will be helpful in developing effective pharmacological strategies for preventing neonatal E. coli K1 infection. PMID:28262688

  3. Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria.

    Science.gov (United States)

    Miao, Edward A; Leaf, Irina A; Treuting, Piper M; Mao, Dat P; Dors, Monica; Sarkar, Anasuya; Warren, Sarah E; Wewers, Mark D; Aderem, Alan

    2010-12-01

    Macrophages mediate crucial innate immune responses via caspase-1-dependent processing and secretion of interleukin 1β (IL-1β) and IL-18. Although infection with wild-type Salmonella typhimurium is lethal to mice, we show here that a strain that persistently expresses flagellin was cleared by the cytosolic flagellin-detection pathway through the activation of caspase-1 by the NLRC4 inflammasome; however, this clearance was independent of IL-1β and IL-18. Instead, caspase-1-induced pyroptotic cell death released bacteria from macrophages and exposed the bacteria to uptake and killing by reactive oxygen species in neutrophils. Similarly, activation of caspase-1 cleared unmanipulated Legionella pneumophila and Burkholderia thailandensis by cytokine-independent mechanisms. This demonstrates that activation of caspase-1 clears intracellular bacteria in vivo independently of IL-1β and IL-18 and establishes pyroptosis as an efficient mechanism of bacterial clearance by the innate immune system.

  4. Expression, Purification and Crystal Structure of a Truncated Acylpeptide Hydrolase from Aeropyrum pernix K1

    Institute of Scientific and Technical Information of China (English)

    Hai-Feng ZHANG; Bai-Song ZHENG; Ying PENG; Zhi-Yong LOU; Yan FENG; Zi-He RAO

    2005-01-01

    Acylpeptide hydrolase (APH) catalyzes the N-terminal hydrolysis of Nα-acylpeptides to release Nα-acylated amino acids. The crystal structure of recombinant APH from the thermophilic archaeon Aeropyrum pernix K1 (apAPH) was reported recently to be at a resolution of 2.1 A using X-ray diffraction. A truncated mutant of apAPH that lacks the first short α-helix at the N-terminal, apAPH-△(1-21), was cloned, expressed,characterized and crystallized. Data from biochemical experiments indicate that the optimum temperature of apAPH is decreased by 15 ℃ with the deletion of the N-terminal α-helix. However, the enzyme activity at the optimal temperature does not change. It suggests that this N-terminal α-helix is essential for thermostability. Here, the crystal structure of apAPH-△(1-21) has been determined by molecular replacement to 2.5A. A comparison between the two structures suggests a difference in thermostability, and it can be concluded that by adding or deleting a linking structure (located over different domains), the stability or even the activity of an enzyme can be modified.

  5. Crystal structure of a lactosucrose-producing enzyme, Arthrobacter sp. K-1 β-fructofuranosidase.

    Science.gov (United States)

    Tonozuka, Takashi; Tamaki, Akiko; Yokoi, Gaku; Miyazaki, Takatsugu; Ichikawa, Megumi; Nishikawa, Atsushi; Ohta, Yukari; Hidaka, Yuko; Katayama, Kinya; Hatada, Yuji; Ito, Tetsuya; Fujita, Koki

    2012-12-10

    Arthrobacter sp. K-1 β-fructofuranosidase (ArFFase), a glycoside hydrolase family 68 enzyme, catalyzes the hydrolysis and transfructosylation of sucrose. ArFFase is useful for producing a sweetener, lactosucrose (4(G)-β-D-galactosylsucrose). The primary structure of ArFFase is homologous to those of levansucrases, although ArFFase catalyzes mostly hydrolysis when incubated with sucrose alone, even at high concentration. Here, we determined the crystal structure of ArFFase in unliganded form and complexed with fructose. ArFFase consisted of a five-bladed β-propeller fold as observed in levansucrases. The structure of ArFFase was most similar to that of Gluconacetobacter diazotrophicus levansucrase (GdLev). The structure of the catalytic cleft of ArFFase was also highly homologous to that of GdLev. However, two amino acid residues, Tyr232 and Pro442 in ArFFase, were not conserved between them. A tunnel observed at the bottom of the catalytic cleft of ArFFase may serve as a water drain or its reservoir.

  6. Phase diagram and quantum order by disorder in the Kitaev K1-K2 honeycomb magnet

    Science.gov (United States)

    Rousochatzakis, Ioannis; Reuther, Johannes; Thomale, Ronny; Rachel, Stephan; Perkins, Natalia

    We show that the topological Kitaev spin liquid on the honeycomb lattice is extremely fragile against the second neighbor Kitaev coupling K2, which has been recently identified as the dominant perturbation away from the nearest neighbor model in iridate Na2IrO3, and may also play a role in α-RuCl3. This coupling explains naturally the zig-zag ordering and the special entanglement between real and spin space observed recently in Na2IrO3. The minimal K1-K2 model that we present here holds in addition the unique property that the classical and quantum phase diagrams and their respective order-by-disorder mechanisms are qualitatively different due to their fundamentally different symmetry structure. Nsf DMR-1511768; Freie Univ. Berlin Excellence Initiative of German Research Foundation; European Research Council, ERC-StG-336012; DFG-SFB 1170; DFG-SFB 1143, DFG-SPP 1666, and Helmholtz association VI-521.

  7. Antisolar differential rotation of the K1-giant sigma Geminorum revisited

    CERN Document Server

    Kovari, Zs; Künstler, A; Carroll, T A; Strassmeier, K G; Vida, K; Olah, K; Bartus, J; Weber, M

    2014-01-01

    Context. Surface differential rotation and other global surface flows on magnetically active stars are among the observable manifestations of the stellar dynamo working underneath. Therefore, such observations are important for stellar dynamo theory and useful constraints for solar dynamo studies as well. Aims. The active K1-giant component of the long-period RS CVn-type binary system sigma Gem and its global surface flow pattern is revisited. Methods. We refine the differential rotation law from recovering the spot migration pattern. We apply a detailed cross-correlation technique to a unique set of 34 time-series Doppler images recovered using data from 1996/97. By increasing the number of the available cross-correlation function maps from the formerly used 4 to 17 we expect a more robust determination of the differential surface rotation law. In addition, we present a new time-series Doppler imaging study of sigma Gem using our advanced surface reconstruction code iMap for a dataset collected in 2006/07. R...

  8. Denaturing Effects of Urea and Guanidine Hydrochloride on Hyperthermophilic Esterase from Aeropyrum pernix K1

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The changes in the activity and the conformation of the hyperthermophilic esterase derived from aerobic thermophilic Aeropyrumpernix K1 (APE1547) were studied during denaturation by guanidine hydrochloride (GdnHCl)and urea. The denaturation course of APE1547 was followed by the steady-state and time resolved fluorescence methods. An increase in the denaturant concentration in the denatured system can significantly enhance the inactivation and unfolding of APE1547. The enzyme can be completely inactivated with a urea concentration of 2. 7 mol/L or a GdnHCl concentration of 7.5 mol/L. The fluorescence emission maximum of the enzyme protein red shifts in magnitude to a maximum value(355 nm) when the concentration of GdnHCl is 5.1 mol/L. The experimental results indicate that APE1547 has a high resistance to urea. Unfolding of APE1547 in GdnHCl(4.2-6.0 mol/L) was shown to be an irreversible process. The present results indicate that the ion pairs in this protein may be a key factor for the stability of this esterase.

  9. Citotoxicidad del fungicida mancozeb en cultivos de CHO-K1

    Directory of Open Access Journals (Sweden)

    A.E. Bayoumi

    2002-01-01

    Full Text Available Se ha determinado la citotoxicidad del fungicida ditiocarbámico mancozeb, en cultivos celulares de ovario de hámster (CHO-K1, usando los bioensayos estandarizados de incorporación de rojo neutro (RN y del contenido total de proteínas (PT. Las dos técnicas mostraron ser comparables en la determinación del efecto citotóxico, mostrando valores de RN50 menores de 15 mg/ml después de 24 h de exposición al plaguicida. La citotoxicidad fue mayor cuanto mayor fue el tiempo de exposición al mancozeb, en ausencia de suero fetal bovino en el medio de cultivo. La preincubación del mancozeb con diferentes concentraciones de fracción submitocondrial de hígado de rata, originó metabolitos menos tóxicos que el compuesto de origen, lo que indica una cierta protección metabólica proporcionada por la fracción S9. Igualmente, el metabolito final de su degradación, la etilentiourea (ETU mostró menor citotoxicidad que el compuesto original a los tiempos de exposición cortos.

  10. SOFIA Infrared Spectrophotometry of Comet C/2012 K1 (Pan-STARRS)

    CERN Document Server

    Woodward, Charles E; Harker, David E; Ryan, Erin L; Wooden, Diane H; Sitko, Michael L; Russell, Ray W; Reach, William T; de Pater, Imke; Kolokolova, Ludmilla; Gehrz, Robert D

    2015-01-01

    We present pre-perihelion infrared 8 to 31 micron spectrophotometric and imaging observations of comet C/2012 K1 (Pan-STARRS), a dynamically new Oort Cloud comet, conducted with NASA's Stratospheric Observatory for Infrared Astronomy (SOFIA) facility (+FORCAST) in 2014 June. As a "new" comet (first inner solar system passage), the coma grain population may be extremely pristine, unencumbered by a rime and insufficiently irradiated by the Sun to carbonize its surface organics. The comet exhibited a weak 10 micron silicate feature ~1.18 +/- 0.03 above the underlying best-fit 215.32 +/- 0.95 K continuum blackbody. Thermal modeling of the observed spectral energy distribution indicates that the coma grains are fractally solid with a porosity factor D = 3 and the peak in the grain size distribution, a_peak = 0.6 micron, large. The sub-micron coma grains are dominated by amorphous carbon, with a silicate-to-carbon ratio of 0.80 (+0.25) (- 0.20). The silicate crystalline mass fraction is 0.20 (+0.30) (-0.10), simila...

  11. Enhancement in xylose utilization using Kluyveromyces marxianus NIRE-K1 through evolutionary adaptation approach.

    Science.gov (United States)

    Sharma, Nilesh Kumar; Behera, Shuvashish; Arora, Richa; Kumar, Sachin

    2016-05-01

    The evolutionary adaptation was carried out on the thermotolerant yeast Kluyveromyces marxianus NIRE-K1 at 45 °C up to 60 batches to enhance its xylose utilization capability. The adapted strain showed higher specific growth rate and 3-fold xylose uptake rate and short lag phase as compared to the native strain. During aerobic growth adapted yeast showed 2.81-fold higher xylose utilization than that of native. In anaerobic batch fermentation, adapted yeast utilized about 91% of xylose in 72 h and produced 2.88 and 18.75 g l⁻¹ of ethanol and xylitol, respectively, which were 5.11 and 5.71-fold higher than that of native. Ethanol yield, xylitol yield and specific sugar consumption rate obtained by the adapted cells were found to be 1.57, 1.65 and 4.84-fold higher than that of native yeast, respectively. Aforesaid results suggested that the evolutionary adaptation will be a very effective strategy in the near future for economic lignocellulosic ethanol production.

  12. The Leaderless Bacteriocin Enterocin K1 Is Highly Potent against Enterococcus faecium: A Study on Structure, Target Spectrum and Receptor

    Directory of Open Access Journals (Sweden)

    Kirill V. Ovchinnikov

    2017-05-01

    Full Text Available Enterocin K1 (EntK1, enterocin EJ97 (EntEJ97, and LsbB are three sequence related leaderless bacteriocins. Yet LsbB kills only lactococci while EntK1 and EntEJ97 target wider spectra with EntK1 being particularly active against Enterococcus faecium, including nosocomial multidrug resistant isolates. NMR study of EntK1 showed that it had a structure very similar to LsbB – both having an amphiphilic N-terminal α-helix and an unstructured C-terminus. The α-helix in EntK1 is, however, about 3–4 residues longer than that of LsbB. Enterococcal mutants highly resistant to EntEJ97 and EntK1 were found to have mutations within rseP, a gene encoding a stress response membrane-bound Zn-dependent protease. Heterologous expression of the enterococcal rseP rendered resistant cells of Streptococcus pneumoniae sensitive to EntK1 and EntEJ97, suggesting that RseP likely serves as the receptor for EntK1 and EntEJ97. It was also shown that the conserved proteolytic active site in E. faecalis RseP is partly required for EntK1 and EntEJ97 activity, since alanine substitutions of its conserved residues (HExxH reduced the sensitivity of the clones to the bacteriocins. RseP is known to be involved in bacterial stress response. As expected, the growth of resistant mutants with mutations within rseP was severely affected when they were exposed to higher (stressing growth temperatures, e.g., at 45°C, at which wild type cells still grew well. These findings allow us to design a hurdle strategy with a combination of the bacteriocin(s and higher temperature that effectively kills bacteriocin sensitive bacteria and prevents the development of resistant cells.

  13. The microbiota regulates neutrophil homeostasis and host resistance to Escherichia coli K1 sepsis in neonatal mice.

    Science.gov (United States)

    Deshmukh, Hitesh S; Liu, Yuhong; Menkiti, Ogechukwu R; Mei, Junjie; Dai, Ning; O'Leary, Claire E; Oliver, Paula M; Kolls, Jay K; Weiser, Jeffrey N; Worthen, G Scott

    2014-05-01

    Neonatal colonization by microbes, which begins immediately after birth, is influenced by gestational age and the mother's microbiota and is modified by exposure to antibiotics. In neonates, prolonged duration of antibiotic therapy is associated with increased risk of late-onset sepsis (LOS), a disorder controlled by neutrophils. A role for the microbiota in regulating neutrophil development and susceptibility to sepsis in the neonate remains unclear. We exposed pregnant mouse dams to antibiotics in drinking water to limit transfer of maternal microbes to the neonates. Antibiotic exposure of dams decreased the total number and composition of microbes in the intestine of the neonates. This was associated with decreased numbers of circulating and bone marrow neutrophils and granulocyte/macrophage-restricted progenitor cells in the bone marrow of antibiotic-treated and germ-free neonates. Antibiotic exposure of dams reduced the number of interleukin-17 (IL-17)-producing cells in the intestine and production of granulocyte colony-stimulating factor (G-CSF). Granulocytopenia was associated with impaired host defense and increased susceptibility to Escherichia coli K1 and Klebsiella pneumoniae sepsis in antibiotic-treated neonates, which could be partially reversed by administration of G-CSF. Transfer of a normal microbiota into antibiotic-treated neonates induced IL-17 production by group 3 innate lymphoid cells (ILCs) in the intestine, increasing plasma G-CSF levels and neutrophil numbers in a Toll-like receptor 4 (TLR4)- and myeloid differentiation factor 88 (MyD88)-dependent manner and restored IL-17-dependent resistance to sepsis. Specific depletion of ILCs prevented IL-17- and G-CSF-dependent granulocytosis and resistance to sepsis. These data support a role for the intestinal microbiota in regulation of granulocytosis, neutrophil homeostasis and host resistance to sepsis in neonates.

  14. PMA Induces Vaccine Adjuvant Activity by the Modulation of TLR Signaling Pathway

    Directory of Open Access Journals (Sweden)

    Dool-Ri Oh

    2014-01-01

    Full Text Available Toll-like receptor (TLR ligands are being developed for use as vaccine adjuvants and as immunomodulators because of their ability to stimulate innate and adaptive immune responses. Flagellin, a TLR5 ligand, was reported to show potent mucosal vaccine adjuvant activity. To identify ligands that potentiate the adjuvant activity of flagellin, we screened a plant library using HEK293T cells transiently cotransfected with phTLR5 and pNF-κB-SEAP plasmids. The 90% EtOH extract from Croton tiglium showed significant NF-κB transactivation in a TLR5-independent manner along with the increase of a flagellin activity. We have studied to characterize an active component from Croton tiglium and to elucidate the action mechanisms. Phorbol 12-myristate 13-acetate (PMA was isolated as an active component of Croton tiglium by activity-guided fractionation, column chromatography, HPLC, NMR, and MS. PMA at a range of nM induced PKC-dependent NF-κB activation and IL-8 production in both TLR5− and TLR5+ assay systems. In in vivo mouse vaccination model, PMA induced antigen-specific IgG and IgA antibody responses and increased IL-12 production corresponding to T cell responses in spleen lymphocytes. These results suggest that PMA would serve as an efficacious mucosal vaccine adjuvant.

  15. Dietary vitamin K1 requirement and comparison of biopotency of different vitamin K sources for young turkeys.

    Science.gov (United States)

    Jin, S; Sell, J L

    2001-05-01

    In a preliminary experiment, the inclusion of vitamin K1 (K1) at a dietary level of 0.1 mg/kg was as effective as 1 or 2 mg/kg in reducing plasma prothrombin time (PT). To obtain an estimate of the dietary K1 requirement and to compare the biopotency of different vitamin K sources for poults, three additional experiments were conducted. In Experiment 1, an incomplete factorial arrangement of treatments was used in which five dietary concentrations of K1 (0, 0.1, 0.25, 0.5, or 2.0 mg/kg) were tested and two concentrations of neomycin (0 or 75 mg/L) in drinking water were used in conjunction with 0, 0.1, and 0.5 mg of K1/kg of diet. Thus, we used a total of eight treatments. Each treatment was given to two pens of poults, with eight poults per pen. Prothrombin time and prothrombin concentration (PC) in plasma were not influenced by inclusion of neomycin in drinking water. The K1 requirement was estimated, on the basis of PT and PC, to be 0.099 and 0.13 mg/kg, respectively, in Experiment 1. Dietary K1 concentrations tested in Experiment 2 were 0, 0.08, 0.31, or 0.44 mg/kg. A similar protocol to that of Experiment 1 was used in this experiment. The results of Experiment 2 indicated that the dietary K1 requirement was 0.079 mg, based on the influence of dietary K1 on PT. In Experiment 3, dietary treatments consisted of the equivalent of 0.22, 0.55, or 1.11 microM of menadione equivalent/kg from vitamin K1, menadione dimethypyrimidinol bisulfite (MPB) or menadione nicotinamide bisulfite (MNB), respectively, and a control without supplementation of any vitamin K source. The results of Experiment 3 showed that the biopotency of K1 was greater than that of MPB or MNB. The biopotencies of MPB and MNB were similar, although MNB was more potent in reducing plasma PT when supplemented at the level of 0.1 mg of menadione/kg. A nadir of PT and a plateau of PC were evident with a dietary supplementation of MPB or MNB at a level of 0.25 mg of menadione/kg. Results of this research

  16. Spatially Resolved Spectroscopy Across HD189733 (K1V) Using Exoplanet Transits

    Science.gov (United States)

    Gustavsson, Martin; Dravins, Dainis; Ludwig, Hans-Günter

    2016-06-01

    For testing 3-dimensional models of stellar atmospheres, spectroscopy across spatially resolved stellar surfaces would be desired with a spectral resolution of(R = 100,000) or more. Hydrodynamic models predict variations in line profile shapes, strengths, wavelength positions and asymmetries. These variations vary systematically between disk center and limb and as a function of line strength, excitation potential and wavelength region. However, except for a few supergiants and the Sun, current telescopes are not yet capable of resolving any stellar surfaces. One alternative method to resolve distant stellar surfaces, feasible already now, is differential spectroscopy of transiting exoplanet systems. By subtracting in-transit spectra from the spectrum outside of transit, the spectra from stellar surface portions temporarily hidden behind the planet can be disentangled. Since transiting planets cover only a small portion of the stellar surface, the method requires a very high signal-to-noise ratio, obtainable by averaging numerous similar spectral lines. We apply such differential spectroscopy on the 7.7 mag K1V star HD 189733 ('Alopex'*); its transiting planet covers ˜ 3% of its host star's surface, which is the deepest known transit among the brighter systems. Archival data from the ESO HARPS spectrometerare used to construct averaged profiles of photospheric Fe I lines, with the aim of comparing spatially resolved profiles to analogous synthetic line profiles computed from the 3-dimensional hydrodynamic CO5BOLD model. * We refer to HD 189733 as 'Alopex' (from the Greek 'αλɛπού'), denoting a fox related to the one that gave name to its constellation of Vulpecula.

  17. Coordination-resolved local bond strain and 3p energy entrapment of K atomic clusters and K(1 1 0) skin

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Ting; Bo, Maolin; Guo, Yongling [Key Laboratory of Low-Dimensional Materials and Application Technologies (Ministry of Education), Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Hunan Provincial Key Laboratory of Thin Film Materials and Devices, Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Chen, Hefeng [United Superconductive Institution, Shanghai Jiaotong University, Shanghai 200240 (China); Wang, Yan [School of Information and Electronic Engineering, Hunan University of Science and Technology, Hunan 411201 (China); Huang, Yongli, E-mail: huangyongli@xtu.edu.cn [Key Laboratory of Low-Dimensional Materials and Application Technologies (Ministry of Education), Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Hunan Provincial Key Laboratory of Thin Film Materials and Devices, Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Sun, Chang Q., E-mail: ecqsun@ntu.edu.sg [Key Laboratory of Low-Dimensional Materials and Application Technologies (Ministry of Education), Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Hunan Provincial Key Laboratory of Thin Film Materials and Devices, Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); School of Information and Electronic Engineering, Hunan University of Science and Technology, Hunan 411201 (China); NOVITAS, School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore)

    2015-09-15

    Graphical abstract: - Highlights: • Coordination environment resolves electron binding-energy shift of K{sub 44}, K{sub 46}, K{sub 55} clusters. • Predict the effective coordination number of K nanoclusters when we get the atomic number N. • Atomic under coordination shortens the local bonds and entrapment. • XPS derives core level of an isolated atom and its bulk shift. - Abstract: We have examined the atomic coordination effect on the local bond strain and the 3p core-level shift of K(1 1 0) skin and nanoclusters using a combination of the bond order–length–strength correlation notion, tight-binding approach, density functional theory calculations, and photoelectron spectroscopy measurements. It turns out that: (i) the 3p core-level shifts from 15.595 ± 0.003 eV for an isolated K atom by 2.758 eV to the bulk value of 18.353 eV; (ii) the effective atomic coordination number reduces from the bulk value of 12 to 3.93 for the first layer and to 5.81 for the second layer of K(1 1 0) skin associated with the local lattice strain of 12.76%, a binding energy density 72.67%, and atomic cohesive energy −62.46% for the skin; and (iii) K cluster size reduction lowers the effective atomic coordination number and enhances further the skin electronic attribution. Results have revealed that the 3p core-level shifts of K(1 1 0) and nanoclusters originate from perturbation of the Hamiltonian by under-coordination induced charge densification and quantum entrapment.

  18. Comparison of different inner diameter silver clip impact on 2K1C hypertension model%不同内径银夹诱导2K1C高血压模型的比较

    Institute of Scientific and Technical Information of China (English)

    黄玲玲; 俞婷婷; 郭焜; 兰潮棕; 刘彪; 宋佳治; 刘有静; 汪星辉; 高杉

    2013-01-01

    AIM:To compare the effect of three different inner diameter silver clips on 2K1C hypertensive rats.METHODS:A 2K1C hypertensive rats were established by narrow the right renal artery with different inner diameter of U-shaped silver clips.Animals were divided into 4 groups:the sham-operate group,2K1C-0.2 mm group,2K1C-0.25 mm group and 2K1C-0.3 mm group.The tail-cuff apparatus was employed to measure the rat tail arterial blood pressure during the 0-8 weeks.At the end of 8th wk,the hemodynamics index recorded.The cardiac and kidney hypertrophy index were expressed as heart weight/body weight (HW/BW),left kidney weight/body weight (LKW/BW) and right kidney weight/body weight (RKW/BW).The aorta remodeling was investigated by HE stain.The content of hydroxyproline in myocardial tissue was measured by colorimetric determination.Radioimmunoassay method was used to detected the AngⅡ content in serum.RESULTS:The 2K1C-0.25 group and 2K1C-0.3 group showed the higher model successful rate,less pyknotic kidneys,significant impairment of cardiac function,markedly increasing of HW/BW,vascular remodeling index,the content of hydroxyproline and the AngⅡ content compared with sham-operate group.However,the 2K1C-0.2 group performed not well in model successful rate,pyknotic kidneys and the AngⅡ content in serum.CONCLUSION:The 2K1C renovascular hypertension model that narrow the renal artery with 0.25 or 0.3 mm inner diameter of U-shaped silver clips proves to be the optimal one.%目的:比较不同内径银夹缩窄大鼠肾动脉诱导的2K1C高血压模型效果.方法:用U型银夹缩窄大鼠右侧肾动脉建立2K1C高血压模型,分为4组:假手术组、2K1C-0.2 mm组、2K1C-0.25 mm组和2K1C-0.3 mm组.0~8周内,用无创尾动脉压测量仪测量大鼠尾动脉压.第8周末,BL-420S生物机能实验系统监测各组动物的心功能,计算大鼠心重量指数(HW/BW)、左和右肾脏重量指数(LKW/BW和RKW/BW),HE染色观察胸主动脉重构情况,比色法

  19. Multi-mode q-oscillator algebras with q2(k+1)= 1 and related thermo field dynamics

    Institute of Scientific and Technical Information of China (English)

    高亚军

    2002-01-01

    A type of multi-mode q-oscillator algebra with q2(k+1) = 1 is set up and the associated qk-thermo field dynamics is constructed for all k = 1, 2, …, ∞ in a unified form. It is demonstrated that these qk-thermo field dynamics can all be nicely fitted into the algebraic formulation of statistical mechanics (axiomatized form for statistical physics). This means that we obtain infinitely many realizations of the algebraic scheme, which extend the consideration of Ojima[1981 Ann. Phys. 137 1] and contain the usual thermo field dynamics for the fermionic (k = 1) and bosonic (k = ∞)systems as special cases. As simple applications, the qk-statistical average of some operators are given.

  20. Immune response of broiler chickens immunized orally with the recombinant proteins flagellin and the subunit B of cholera toxin associated with Lactobacillus spp.

    Science.gov (United States)

    Baptista, A A S; Donato, T C; Garcia, K C O D; Gonçalves, G A M; Coppola, M P; Okamoto, A S; Sequeira, J L; Andreatti Filho, R L

    2014-01-01

    This study investigated the immune response of broiler chickens with oral treatment of a Lactobacillus spp. pool (PL) associated with microencapsulated recombinant proteins flagellin (FliC) and the subunit B of cholera toxin (CTB). Immune responses were evaluated by measuring IgA from intestinal fluid, serum IgY, and immunostaining of CD8(+) T lymphocytes present in the cecum. The evaluations were performed on d 0, 7, 14, 21, and 28 posttreatment. A significant increase (P < 0.05) was observed in IgA levels in all immunized groups, especially 3 wk after immunization. Treatments 2 (recombinant CTB) and 3 (recombinant FliC+CTB) showed the highest concentrations. Similarly, serum concentrations IgY (μg/mL) increased along the experiment, and the means for treatments 2 and 3 showed significant differences (P < 0.05) compared with controls, reaching concentrations of 533 and 540 μg/mL, respectively. The number of CD8(+) T lymphocytes in all treatments greatly differed (P < 0.05) compared with the negative control at 21 d posttreatment. However, only treatment 2 (recombinant CTB), 4 (PL), and 5 (recombinant FliC+ recombinant CTB + PL) remained significantly (P < 0.05) different from the control at 28 d posttreatment. Thus, it is concluded that the microencapsulated recombinant proteins administered orally to broiler chickens are capable of stimulating humoral and cellular immune response, and the combinations of these antigens with Lactobacillus spp. can influence the population of CD8(+) T cells residing in the cecum.

  1. Direct and individual analysis of stress-related phytohormone dispersion in the vascular system of Cucurbita maxima after flagellin 22 treatment.

    Science.gov (United States)

    Furch, Alexandra C U; Zimmermann, Matthias R; Kogel, Karl-Heinz; Reichelt, Michael; Mithöfer, Axel

    2014-03-01

    • The stress-related phytohormones, salicylic acid (SA) and abscisic acid (ABA), and the three jasmonates, jasmonic acid (JA), cis-12-oxo-phytodienoic acid (cis-OPDA), and (+)-7-iso-jasmonoyl-L-isoleucine (JA-Ile), were investigated in phloem and xylem exudates of Cucurbita maxima. • Phloem and xylem exudates were separately collected and analysed via liquid chromatography-mass spectrometry. • We show direct evidence for all three jasmonates, ABA, and SA in both phloem and xylem exudates of C. maxima. JA and JA-Ile concentrations are higher in xylem (JA: c(xylem) ≈ 199.5 nM, c(phloem) ≈ 43.9 nM; JA-Ile: c(xylem) ≈ 7.9 nM, c(phloem) ≈ 1.6 nM), whereas ABA and SA concentrations are higher in phloem exudates (ABA: c(xylem) ≈ 37.1 nM, c(phloem) ≈ 142.6 nM; SA: c(xylem) ≈ 61.6 nM, c(phloem) ≈ 1319 nM). During bacteria-derived flagellin 22 (flg22)-triggered remote root-to-shoot signalling, phytohormone concentration changed rapidly both in phloem and xylem. • The unequal distribution of phytohormones suggests that phloem and xylem have distinct roles in defence responses. Our data shed light on systemic phytohormone signalling and help explain how plants cope with environmental challenges by lateral exchange between phloem and xylem. Our analysis is a starting point for further investigations of how phytohormones contribute to phloem- and xylem-based defence signalling. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  2. Induction of 8-hydroxy-2'-deoxyguanosine in CHO-K1 cells exposed to phenyl-hydroquinone, a metabolite of ortho-phenylphenol.

    Science.gov (United States)

    Nakagawa, Y; Tayama, S

    1996-03-29

    The induction of 8-hydroxy-2'-deoxyguanosine (8-OHdG), an index of oxidative DNA modification, was investigated in CHO-K1 cells exposed to phenyl-hydroquinone (PHQ), a major metabolite of ortho-phenylphenol (OPP), an antimicrobial. Addition of PHQ at a concentration of 50 microM to CHO cell suspensions (10(6) cells/ml) induced slight elevation of intracellular 8-OHdG levels. Pretreatment of CHO cells with 3-amino-1,2,4-triazole (AT, 20 mM) enhanced PHQ-induced 8-OHdG formation which was accompanied by cell death. Pretreatment of CHO-K1 cells with AT (20 mM) and deferoxamine (DeFe, 20 mM) inhibited the formation of 8-OHdG as well as cell death caused by PHQ. Neither AT nor DeFe affected cell viability or the formation of 8-OHdG in untreated CHO cells during the incubation period. The loss of cellular glutathione induced by the addition of PHQ alone was enhanced by the pretreatment of CHO cells with AT or AT plus DeFe. When PHQ was added to AT-pretreated cell suspensions, the concentration of PHQ decreased with time. This decrease was accompanied by the formation of phenyl-benzoquinone (PBQ). These results suggest that the reactive oxygen species derived from autoxidation of PHQ which converts to PBQ via phenyl-semiquinone elicit DNA damage in CHO cells, especially when the activity of cellular catalase is inhibited.

  3. IMPACT OF NLRP3 INFLAMMASOME ON THE DEVELOPMENT OF HYPERTENSION AND RENAL AND CARDIAC HYPERTROPHY IN 2K1C AND DOCA/SALT MICE

    Directory of Open Access Journals (Sweden)

    Q. Wang

    2012-06-01

    Full Text Available NLRP3 inflammasome is formed by NLRP3, the adaptor ASC, and caspase-1, functions as a sensor of danger signals and triggers processing and release of interleukin-1beta (IL-1β. Recent animal studies demonstrate that NLRP3 inflammasome promotes renal inflammation. However, little is known on the role of NLRP3 in hypertension. We have investigated the effect of NLRP3 inflammasome on blood pressure, plasma renin activity and concentration (PRA and PRC, renal and cardiac hypertrophy in NALP3 and ASC deficient mice on which the two-kidney, one clip (2K1C and the deoxycorticosterone-acetate (DOCA/salt models were applied. MBP, PRA, PRC, and HW/BW were significantly increased at week 12 in WT-2K1C mice compared to the sham. Neither NLAP3-KO nor ASC-KO 2K1C treated mice developed hypertension and had lower circulating levels of PRA and PRC and serum amyloid A (SAA and IL6 compared to the control. RNA levels of SAA, NLRP3, IL1β and IL1α were increased in the ischemic kidney in C57BL/6J mice. Administration of anti-IL1β antibody to the WT mice attenuated the increases of blood pressure and renin in 2K1C mice. With chronic administration of DOCA/salt, MBPs in both NLRP3-KO and WT mice were comparable and not significantly increased compared to tap water group. PRA and PRC in both NALP3-KO and WT mice were significantly suppressed by DOCA/salt. HW/BW and KW/BW in both DOCA/salt treated NALP3-KO and WT mice were significantly increased. DOCA/salt induced hypokalemia was comparable between Nlrp3 KO and WT groups. However, the heart and kidney index in DOCA/salt NLRP3-KO mice was significantly lower than that in DOCA/salt WT mice. Our data show that NLRP3 mediated IL1β is linked to development of renovascular hypertension and suggest that a novel target for the treatment of hypertension. The results also implicate that NLRP3 contributes to the development of cardiac and renal hypertrophy independently of blood pressure in the DOCA/salt model.

  4. Comparison of five different in vitro assays for assessment of sodium metavanadate cytotoxicity in Chinese hamster ovary cells (CHO-K1 line).

    Science.gov (United States)

    Zwolak, Iwona

    2015-08-01

    This investigation was undertaken to compare five different in vitro cytotoxicity assays for their power in revealing vanadium-mediated toxicity in Chinese hamster ovary (CHO)-K1 cells. The cells were exposed to sodium metavanadate (NaVO(3)) in the range of 10-1000 µM for 24 h and thereafter the cytotoxic effects of NaVO(3) were measured by colorimetric in vitro assays: the neutral red (NR) test, the 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide inner salt (XTT) assay, the resazurin assay, the sulforhodamine B (SR-B) assay, and by microscopic assessment of cell viability using the trypan blue (TB) staining method. Among the assays used, the NR test was the most sensitive, since it revealed metavanadate cytotoxicity at the lowest NaVO(3) dose (=50 µM). Also, NaVO(3) cytotoxicity expressed as inhibitory concentration (IC) showed the lowest values for the NR test. Three other tests XTT, resazurin, and SR-B assays showed intermediate sensitivity revealing the cytotoxicity of NaVO(3) at 100 µM. The corresponding IC10 and IC50 values calculated for the XTT, resazurin, and SR-B tests were similar. The TB staining method was the least sensitive, since it recorded metavanadate cytotoxicity at the highest NaVO(3) concentration tested (=600 µM). Based on the cytotoxicity end points measured with the above assays, it can be concluded that lysosomal/Golgi apparatus damage (measured by NR assay) may be the primary effect of NaVO(3) on CHO-K1 cells. The disintegration of mitochondria (assessed with the XTT and resazurin assays) probably follows lysosomal impairment. Plasma membrane permeability (staining with TB) occurs at a late stage of NaVO(3)-induced cytotoxicity on CHO-K1 cells. The results obtained in this research work show that the NR test can be recommended as a very sensitive assay for the assessment of NaVO(3) cytotoxicity in the CHO-K1 cell culture model. Considering the convenience of assay performance along with adequate sensitivity

  5. Soluble components of the flagellar export apparatus, FliI, FliJ, and FliH, do not deliver flagellin, the major filament protein, from the cytosol to the export gate.

    Science.gov (United States)

    Sajó, Ráchel; Liliom, Károly; Muskotál, Adél; Klein, Agnes; Závodszky, Péter; Vonderviszt, Ferenc; Dobó, József

    2014-11-01

    Flagella, the locomotion organelles of bacteria, extend from the cytoplasm to the cell exterior. External flagellar proteins are synthesized in the cytoplasm and exported by the flagellar type III secretion system. Soluble components of the flagellar export apparatus, FliI, FliH, and FliJ, have been implicated to carry late export substrates in complex with their cognate chaperones from the cytoplasm to the export gate. The importance of the soluble components in the delivery of the three minor late substrates FlgK, FlgL (hook-filament junction) and FliD (filament-cap) has been convincingly demonstrated, but their role in the transport of the major filament component flagellin (FliC) is still unclear. We have used continuous ATPase activity measurements and quartz crystal microbalance (QCM) studies to characterize interactions between the soluble export components and flagellin or the FliC:FliS substrate-chaperone complex. As controls, interactions between soluble export component pairs were characterized providing Kd values. FliC or FliC:FliS did not influence the ATPase activity of FliI alone or in complex with FliH and/or FliJ suggesting lack of interaction in solution. Immobilized FliI, FliH, or FliJ did not interact with FliC or FliC:FliS detected by QCM. The lack of interaction in the fluid phase between FliC or FliC:FliS and the soluble export components, in particular with the ATPase FliI, suggests that cells use different mechanisms for the export of late minor substrates, and the major substrate, FliC. It seems that the abundantly produced flagellin does not require the assistance of the soluble export components to efficiently reach the export gate. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. 鲁梅克斯K-1杂交酸模传粉生物学研究%Study on Pollination Biology in Rumex patientia ×R. tianscha nicus cv.Rumex K-1

    Institute of Scientific and Technical Information of China (English)

    鞠晓峰; 于开源; 曲善民

    2012-01-01

    Rumex k-1's pollination habit is researched by castrating and covering with sack and manual pollination.The result showed that Rumex k-1 can produce seed normally by either self pollination or cross pollination,the setting rate is more 95%.The best time of cross pollination is 1-6 hour after blooming.The quantity of pollen is larger.The pollens are spreader by wind,not by insect.%利用去雄、套袋、人工授粉方法研究不同授粉方式和授粉时间对鲁梅克斯K-1杂交酸模结实率的影响。结果表明,鲁梅克斯K-1杂交酸模自花授粉、异花授粉都能正常结实,结实率达95%以上。异花授粉最佳时间为花开放后1~6 h。花粉量大。风媒传粉,未见昆虫访问。

  7. IL-2, IL-4, IFN-γ or TNF-α enhances BAFF-stimulated cell viability and survival by activating Erk1/2 and S6K1 pathways in neoplastic B-lymphoid cells.

    Science.gov (United States)

    Gui, Lin; Zeng, Qingyu; Xu, Zhigang; Zhang, Hai; Qin, Shanshan; Liu, Chunxiao; Xu, Chong; Qian, Zhou; Zhang, Shuangquan; Huang, Shile; Chen, Long

    2016-08-01

    B-cell activating factor of the TNF family (BAFF) has been documented to act as a critical factor in the development of aggressive B lymphocytes and autoimmune diseases. However, the effect of various cytokines on BAFF-elicited neoplastic B-lymphoid cells is not known. In this study, we exhibited that administration of human soluble BAFF (hsBAFF), IL-2, IL-4, IFN-γ, or TNF-α alone increased cell viability and survival in Raji cells concentration-dependently, yet a more robust viability/survival was seen in the cells co-treatment of IL-2, IL-4, IFN-γ, or TNF-α with hsBAFF, respectively. Further research revealed that both Erk1/2 and S6K1 signaling pathways were essential for IL-2, IL-4, IFN-γ, or TNF-α enhancement of the viability/survival in the hsBAFF-stimulated cells, as inhibition of Erk1/2 with U0126 or down-regulation of Erk1/2, or blockage of S6K1 with rapamycin or silencing S6K1, or silencing S6K1/Erk1/2, respectively, reduced the cell viability/survival in the cells treated with/without hsBAFF±IL-2, IL-4, IFN-γ, or TNF-α. These findings indicate that IL-2, IL-4, IFN-γ or TNF-α enhances BAFF-stimulated cell viability/survival by activating Erk1/2 and S6K1 signaling in neoplastic B-lymphoid cells. Our data suggest that modulation of IL-2, IL-4, IFN-γ and/or TNF-α levels, or inhibitors of Erk1/2 or S6K1 may be a new approach to prevent BAFF-induced aggressive B-cell malignancies.

  8. Understanding the dynamics of Toll-like Receptor 5 response to flagellin and its regulation by estradiol

    Science.gov (United States)

    Caballero, Ignacio; Boyd, James; Almiñana, Carmen; Sánchez-López, Javier A.; Basatvat, Shaghayegh; Montazeri, Mehrnaz; Maslehat Lay, Nasim; Elliott, Sarah; Spiller, David G.; White, Michael R. H.; Fazeli, Alireza

    2017-01-01

    Toll-like receptors (TLRs) are major players of the innate immune system. Once activated, they trigger a signalling cascade that leads to NF-κB translocation from the cytoplasm to the nucleus. Single cell analysis shows that NF-κB signalling dynamics are a critical determinant of transcriptional regulation. Moreover, the outcome of innate immune response is also affected by the cross-talk between TLRs and estrogen signalling. Here, we characterized the dynamics of TLR5 signalling, responsible for the recognition of flagellated bacteria, and those changes induced by estradiol in its signalling at the single cell level. TLR5 activation in MCF7 cells induced a single and sustained NF-κB translocation into the nucleus that resulted in high NF-κB transcription activity. The overall magnitude of NF-κB transcription activity was not influenced by the duration of the stimulus. No significant changes are observed in the dynamics of NF-κB translocation to the nucleus when MCF7 cells are incubated with estradiol. However, estradiol significantly decreased NF-κB transcriptional activity while increasing TLR5-mediated AP-1 transcription. The effect of estradiol on transcriptional activity was dependent on the estrogen receptor activated. This fine tuning seems to occur mainly in the nucleus at the transcription level rather than affecting the translocation of the NF-κB transcription factor.

  9. Rapid, high performance method for the determination of vitamin K(1), menaquinone-4 and vitamin K(1) 2,3-epoxide in human serum and plasma using liquid chromatography-hybrid quadrupole linear ion trap mass spectrometry.

    Science.gov (United States)

    Gentili, Alessandra; Cafolla, Arturo; Gasperi, Tecla; Bellante, Simona; Caretti, Fulvia; Curini, Roberta; Fernández, Virginia Pérez

    2014-04-18

    Unlike the other fat-soluble vitamins, vitamin K circulates in the human bloodstream at very low levels because of a low intake in the diet. Mammals have developed an efficient recycling system, known as vitamin K-epoxide cycle, which involve quinone, hydroquinone and epoxide forms of the vitamin. Phylloquinone (K(1)) is the main homologue, while menaquinone-4 (MK-4) is both a member of the vitamin K(2) family and metabolite of K(1) in extra-hepatic tissues. Notwithstanding the recent advances, many aspects of the complex vitamin K physiology still remain to be investigated. Therefore, there is a critical need to develop more reliable analytical methods for determining the vitamin K and its metabolites in biological fluids and tissues. Nevertheless, relatively low concentrations, unavailability of some authentic standards and occurrence of interfering lipids make this a challenging task. The method proposed in the present paper can directly and accurately estimate K(1), K(1) 2,3-epoxide (K(1)O), and MK-4 in human serum and plasma at concentrations in the ng/L-μg/L range, using labelled internal standards and a quadrupole linear ion trap instrument operated in multiple reaction monitoring (MRM) mode. High sensitivity was achieved by removing signal "endogenous suppressors" and making the composition of the non-aqueous mobile phase suitable to support the positive atmospheric pressure chemical ionization of the analytes. An excellent selectivity resulted from the combination of some factors: the MRM acquisition, the adoption of an identification point system, an extraction optimized to remove most of the lipids and a tandem-C18 column-system necessary to separate isobaric interferences from analytes. The method was validated according to the Food and Drug Administration (FDA) guidelines and its accuracy was assessed by analysing 9 samples from the Vitamin K External Quality Assessment Scheme (KEQAS). Its feasibility in evaluating vitamin K status in human serum was

  10. Expression of LRP1 by human osteoblasts: a mechanism for the delivery of lipoproteins and vitamin K1 to bone

    DEFF Research Database (Denmark)

    Niemeier, Andreas; Kassem, Moustapha; Toedter, Klaus

    2005-01-01

    Accumulating clinical and experimental data show the importance of dietary lipids and lipophilic vitamins, such as vitamin K1, for bone formation. The molecular mechanism of how they enter the osteoblast is unknown. Here we describe the expression of the multifunctional LRP1 by human osteoblasts...

  11. A repetitive control scheme aiming to compensate the 6k+1 harmonics for three-phase hybrid active filter

    DEFF Research Database (Denmark)

    Luo, Zhaoxu; Su, Mei; Yang, Jian;

    2016-01-01

    in stationary reference frame, which only compensates the 6k+1 harmonics (e.g. -5, +7, -11, +13) in three-phase systems and reduces the time delay to T0/6 . So compared with the earlier reduced delay time repetitive controllers, the robustness and transient performance is further improved, the waste of control...

  12. CL-L1 and CL-K1 and other complement associated pattern recognition molecules in systemic lupus erythematosus

    DEFF Research Database (Denmark)

    Troldborg, Anne; Thiel, Steffen; Jensen, Lisbeth

    2015-01-01

    The objective of this study was to explore the involvement of collectin liver 1 (CL-L1) and collectin kidney 1 (CL-K1) and other pattern recognition molecules (PRMs) of the lectin pathway of the complement system in a cross-sectional cohort of systemic lupus erythematosus (SLE) patients...

  13. Counting Quadratic Nonresidues in Shifted Subsets of the Set of Quadratic Nonresidues for Primes p=4k+1

    Directory of Open Access Journals (Sweden)

    Rocío Meza-Moreno

    2015-01-01

    Full Text Available Let p=4k+1 be a prime number and Fp the finite field with p elements. For x∈1,n, Nx will denote the set of quadratic nonresidues less than or equal to x. In this work we calculate the number of quadratic nonresidues in the shifted set N(p-1/2+a.

  14. Topiramate reduces excitability in the basolateral amygdala by selectively inhibiting GluK1 (GluR5) kainate receptors on interneurons and positively modulating GABAA receptors on principal neurons.

    Science.gov (United States)

    Braga, Maria F M; Aroniadou-Anderjaska, Vassiliki; Li, He; Rogawski, Michael A

    2009-08-01

    Topiramate [2,3:4,5-bis-O-(1-methylethylidene)-beta-D-fructopyranose sulfamate] is a structurally novel antiepileptic drug that has broad efficacy in epilepsy, but the mechanisms underlying its therapeutic activity are not fully understood. We have found that topiramate selectively inhibits GluK1 (GluR5) kainate receptor-mediated excitatory postsynaptic responses in rat basolateral amygdala (BLA) principal neurons and protects against seizures induced by the GluK1 kainate receptor agonist (R,S)-2-amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl)propanoic acid (ATPA). Here, we demonstrate that topiramate also modulates inhibitory function in the BLA. Using whole-cell recordings in rat amygdala slices, we found that 0.3 to 10 microM topiramate 1) inhibited ATPA-evoked postsynaptic currents recorded from BLA interneurons; 2) suppressed ATPA-induced enhancement of spontaneous inhibitory postsynaptic currents (IPSCs) recorded from BLA pyramidal cells; and 3) blocked ATPA-induced suppression of evoked IPSCs, which is mediated by presynaptic GluK1 kainate receptors present on BLA interneurons. Topiramate (10 microM) had no effect on the AMPA [(R,S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid]-induced enhancement of spontaneous activity of BLA neurons. Thus, although topiramate inhibits GluK1 kainate receptor-mediated enhancement of interneuron firing, it promotes evoked GABA release, leading to a net inhibition of circuit excitability. In addition, we found that topiramate (0.3-10 microM) increased the amplitude of evoked, spontaneous, and miniature IPSCs in BLA pyramidal neurons, indicating an enhancement of postsynaptic GABA(A) receptor responses. Taken together with our previous findings, we conclude that topiramate protects against hyperexcitability in the BLA by suppressing the GluK1 kainate receptor-mediated excitation of principal neurons by glutamatergic afferents, blocking the suppression of GABA release from interneurons mediated by presynaptic GluK1

  15. PI3K-Akt-mTORC1-S6K1/2 Axis Controls Th17 Differentiation by Regulating Gfi1 Expression and Nuclear Translocation of RORγ

    Directory of Open Access Journals (Sweden)

    Yutaka Kurebayashi

    2012-04-01

    Full Text Available The PI3K-Akt-mTORC1 axis contributes to the activation, survival, and proliferation of CD4+ T cells upon stimulation through TCR and CD28. Here, we demonstrate that the suppression of this axis by deletion of p85α or PI3K/mTORC1 inhibitors as well as T cell-specific deletion of raptor, an essential component of mTORC1, impairs Th17 differentiation in vitro and in vivo in a S6K1/2-dependent fashion. Inhibition of PI3K-Akt-mTORC1-S6K1 axis impairs the downregulation of Gfi1, a negative regulator of Th17 differentiation. Furthermore, we demonstrate that S6K2, a nuclear counterpart of S6K1, is induced by the PI3K-Akt-mTORC1 axis, binds RORγ, and carries RORγ to the nucleus. These results point toward a pivotal role of PI3K-Akt-mTORC1-S6K1/2 axis in Th17 differentiation.

  16. Chronic proliferative rhinitis associated with Salmonella enterica subspecies diarizonae serovar 61:k:1, 5, (7) in sheep in Spain.

    Science.gov (United States)

    Lacasta, D; Ferrer, L M; Ramos, J J; Bueso, J P; Borobia, M; Ruiz de Arcaute, M; Figueras, L; González-Sainz, J M; De Las Heras, M

    2012-11-01

    A chronic proliferative rhinitis in sheep associated with Salmonella enterica subspecies diarizonae serovar 61:k:1, 5, (7) is described. Ten adult sheep from eight traditionally managed Rasa Aragonesa flocks located in Aragon (Spain) were studied. Clinical signs began with bilateral thick mucus nasal discharge, wheezing and snoring and progressed to partial or complete obstruction of the nostrils. Necropsy examination revealed swollen ventral nasal turbinates with a roughened mucosal surface partially covered by small polyps. Histopathology revealed chronic proliferative rhinitis with a predominant population of neutrophils that infiltrated the mucosal epithelium. Plasma cells and macrophages were present in the lamina propria. Organisms expressing Salmonella antigen immunohistochemically were detected within epithelial cells. Salmonella enterica subspecies diarizonae serovar 61:k:1, 5, (7) was isolated as a sole microorganism from nasal swabs taken from five animals. The implication of finding this bacterium in various diseases in sheep and its role as a potential zoonosis are discussed.

  17. Stars war之跑跑卡丁车K1总决赛圆满落幕

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    10月15日下午5时,随着个人竞速选手王京登上最高的领奖台,历时三个多月, 由上海邮通科技和Replays.net联合主办,游戏风云GamesTV独家现场全程直播,技嘉科技,可口可乐和优派联合赞助的Stars war之跑跑卡丁车K1锦标赛圆满落幕,来自全国23个赛区的114名选手在今天决出了跑跑卡丁车K1锦标赛全国总决赛的最终车神。

  18. An improved high-quality draft genome sequence of Carnobacterium inhibens subsp. inhibens strain K1(T).

    Science.gov (United States)

    Nicholson, Wayne L; Davis, Christina L; Shapiro, Nicole; Huntemann, Marcel; Clum, Alicia; Reddy, T B K; Pillay, Manoj; Markowitz, Victor; Varghese, Neha; Pati, Amrita; Ivanova, Natalia; Kyrpides, Nikos; Woyke, Tanja

    2016-01-01

    Despite their ubiquity and their involvement in food spoilage, the genus Carnobacterium remains rather sparsely characterized at the genome level. Carnobacterium inhibens K1(T) is a member of the Carnobacteriaceae family within the class Bacilli. This strain is a Gram-positive, rod-shaped bacterium isolated from the intestine of an Atlantic salmon. The present study determined the genome sequence and annotation of Carnobacterium inhibens K1(T). The genome comprised 2,748,608 bp with a G + C content of 34.85 %, which included 2621 protein-coding genes and 116 RNA genes. The strain contained five contigs corresponding to presumptive plasmids of sizes: 19,036; 24,250; 26,581; 65,272; and 65,904 bp.

  19. Growth and dielectric properties of KTiOPO4 and K1 - x Rb x TiOPO4 crystals

    Science.gov (United States)

    Gurtovoĭ, V. G.; Sheleg, A. U.; Guretskiĭ, S. A.; Kalanda, N. A.

    2008-07-01

    Methods of growth of KTiOPO4 and K1 - x Rb x TiOPO4 crystals of high optical quality have been optimized. The dielectric properties (permittivity and conductivity) of the crystals grown have been investigated at frequencies from 102 to 106 Hz in the temperature range from 100 to 350 K, along the [001] crystallographic direction. It is established that partial substitution of K+ ions with Rb+ ions leads to a decrease in the permittivity and conductivity.

  20. Quantum cosmology on (k = -1)-Friedmann-Robertson-Walker Universe evolving from stiff matter era to the dust dominated one

    Science.gov (United States)

    Dariescu, Marina-Aura; Dariescu, Ciprian

    2017-01-01

    This work is devoted to the spatially open Friedmann-Robertson-Walker (FRW) Universe evolving from the stiff matter era to the dust dominated one. Within the quantum analysis based on the Wheeler-DeWitt equation, we derive the wave function of the (k = -1)-FRW Universe with combined matter sources. On the classical level, one has to deal with the Friedmann equation which leads on a dependence of the scale function on time generally expressed from functional relations involving elliptic integrals.

  1. Comparative genomic analysis shows that avian pathogenic Escherichia coli isolate IMT5155 (O2:K1:H5; ST complex 95, ST140 shares close relationship with ST95 APEC O1:K1 and human ExPEC O18:K1 strains.

    Directory of Open Access Journals (Sweden)

    Xiangkai Zhu Ge

    Full Text Available Avian pathogenic E. coli and human extraintestinal pathogenic E. coli serotypes O1, O2 and O18 strains isolated from different hosts are generally located in phylogroup B2 and ST complex 95, and they share similar genetic characteristics and pathogenicity, with no or minimal host specificity. They are popular objects for the study of ExPEC genetic characteristics and pathogenesis in recent years. Here, we investigated the evolution and genetic blueprint of APEC pathotype by performing phylogenetic and comparative genome analysis of avian pathogenic E. coli strain IMT5155 (O2:K1:H5; ST complex 95, ST140 with other E. coli pathotypes. Phylogeny analyses indicated that IMT5155 has closest evolutionary relationship with APEC O1, IHE3034, and UTI89. Comparative genomic analysis showed that IMT5155 and APEC O1 shared significant genetic overlap/similarities with human ExPEC dominant O18:K1 strains (IHE3034 and UTI89. Furthermore, the unique PAI I5155 (GI-12 was identified and found to be conserved in APEC O2 serotype isolates. GI-7 and GI-16 encoding two typical T6SSs in IMT5155 might be useful markers for the identification of ExPEC dominant serotypes (O1, O2, and O18 strains. IMT5155 contained a ColV plasmid p1ColV5155, which defined the APEC pathotype. The distribution analysis of 10 sequenced ExPEC pan-genome virulence factors among 47 sequenced E. coli strains provided meaningful information for B2 APEC/ExPEC-specific virulence factors, including several adhesins, invasins, toxins, iron acquisition systems, and so on. The pathogenicity tests of IMT5155 and other APEC O1:K1 and O2:K1 serotypes strains (isolated in China through four animal models showed that they were highly virulent for avian colisepticemia and able to cause septicemia and meningitis in neonatal rats, suggesting zoonotic potential of these APEC O1:K1 and O2:K1 isolates.

  2. Near-Full Genome Characterisation of Two Natural Intergenotypic 2k/1b Recombinant Hepatitis C Virus Isolates

    Directory of Open Access Journals (Sweden)

    Victoria L. Demetriou

    2011-01-01

    Full Text Available Few natural intergenotypic hepatitis C virus (HCV recombinants have been characterised, and only RF1_2k/1b has demonstrated widespread transmission. The near-full length genome sequences for two cases of 2k/1b recombinants (CYHCV037 and CYHCV093 sampled in Cyprus were obtained using strain-specific RT-PCR amplification and sequencing protocols. Sequence analysis confirmed their similarity with the original RF1_2k/1b strain from St. Petersburg, N687. These two isolates significantly contribute to the sequence data available on this recombinant and confirm its increasing spread among individuals from Eastern Europe, and its association with transmission through intravenous drug use. Phylogenetic analyses reveal clustering of the sequence 3′ to the recombination point, not seen in the topology of the 5′ sequences, implying a more complicated evolutionary history than that held to date. The increasing cases of HCV recombinant strains underline the requirement of their contribution to the standardised rules of HCV classification and nomenclature, molecular epidemiology, diagnosis, and treatment.

  3. Inositol hexakisphosphate kinase 1 (IP6K1) activity is required for cytoplasmic dynein-driven transport

    Science.gov (United States)

    Chanduri, Manasa; Rai, Ashim; Malla, Aushaq Bashir; Wu, Mingxuan; Fiedler, Dorothea; Mallik, Roop; Bhandari, Rashna

    2016-01-01

    Inositol pyrophosphates, such as diphosphoinositol pentakisphosphate (IP7), are conserved eukaryotic signaling molecules that possess pyrophosphate and monophosphate moieties. Generated predominantly by inositol hexakisphosphate kinases (IP6Ks), inositol pyrophosphates can modulate protein function by posttranslational serine pyrophosphorylation. Here, we report inositol pyrophosphates as novel regulators of cytoplasmic dynein-driven vesicle transport. Mammalian cells lacking IP6K1 display defects in dynein-dependent trafficking pathways, including endosomal sorting, vesicle movement, and Golgi maintenance. Expression of catalytically active but not inactive IP6K1 reverses these defects, suggesting a role for inositol pyrophosphates in these processes. Endosomes derived from slime mold lacking inositol pyrophosphates also display reduced dynein-directed microtubule transport. We demonstrate that Ser51 in the dynein intermediate chain (IC) is a target for pyrophosphorylation by IP7, and this modification promotes the interaction of the IC N-terminus with the p150Glued subunit of dynactin. IC–p150Glued interaction is decreased, and IC recruitment to membranes is reduced in cells lacking IP6K1. Our study provides the first evidence for the involvement of IP6Ks in dynein function and proposes that inositol pyrophosphate-mediated pyrophosphorylation may act as a regulatory signal to enhance dynein-driven transport. PMID:27474409

  4. Distinct roles of ComK1 and ComK2 in gene regulation in Bacillus cereus.

    Directory of Open Access Journals (Sweden)

    Aleksandra M Mirończuk

    Full Text Available The B. subtilis transcriptional factor ComK regulates a set of genes coding for DNA uptake from the environment and for its integration into the genome. In previous work we showed that Bacillus cereus expressing the B. subtilis ComK protein is able to take up DNA and integrate it into its own genome. To extend our knowledge on the effect of B. subtilis ComK overexpression in B. cereus we first determined which genes are significantly altered. Transcriptome analysis showed that only part of the competence gene cluster is significantly upregulated. Two ComK homologues can be identified in B. cereus that differ in their respective homologies to other ComK proteins. ComK1 is most similar, while ComK2 lacks the C-terminal region previously shown to be important for transcription activation by B. subtilis ComK. comK1 and comK2 overexpression and deletion studies using transcriptomics techniques showed that ComK1 enhances and ComK2 decreases expression of the comG operon, when B. subtilis ComK was overexpressed simultaneously.

  5. Efficacy of avian pneumovirus vaccines against an avian pneumovirus/Escherichia coli O2:K1 dual infection in turkeys.

    Science.gov (United States)

    Van de Zande, S; Nauwynck, H; Pensaert, M

    2002-03-16

    The clinical, pathological and microbiological outcome of a challenge with avian pneumovirus (APV) and Escherichia coli O2:K1 was evaluated in turkeys vaccinated with an attenuated APV vaccine and with or without maternally derived antibodies. Two groups of two-week-old poults, one with and one without maternally derived antibodies against APV, were vaccinated oculonasally with attenuated APV subtype A or B. A third group remained unvaccinated. Eleven weeks later, the turkeys were inoculated intranasally with either virulent APV subtype A, or E. coli O2:K1, or with both agents three days apart. After the dual infection, birds vaccinated with attenuated subtype A or B, and with or without maternally derived antibodies, had lower mean clinical scores than the unvaccinated birds. In the vaccinated birds, virus replication was significantly reduced and no bacteria were isolated, except from the birds vaccinated with attenuated subtype B. In the unvaccinated turkeys, large numbers of E. coli O2:K1 were isolated from the turbinates of the dually infected birds between one-and-a-half and seven days after they were inoculated.

  6. The clinical, pathological and microbiological outcome of an Escherichia coli O2:K1 infection in avian pneumovirus infected turkeys.

    Science.gov (United States)

    Van de Zande, S; Nauwynck, H; Pensaert, M

    2001-08-20

    The purpose of this study was to evaluate the effect of an Escherichia coli infection in avian pneumovirus (APV)-infected turkeys. One group of 2-week-old specific pathogen-free (SPF) and two groups of 3-week-old conventional (CON) turkeys were inoculated oculonasally with virulent APV subtype A alone, with E. coli O2:K1 alone or with both agents at varying intervals (1, 3, 5 or 7 days) between the two inoculations. The birds were followed clinically and examined for macroscopic lesions at necropsy. Titres of APV were determined in the turbinates, trachea, lungs and air sacs. The number of E. coli O2:K1were assessed in the turbinates, trachea, lungs, air sacs, liver and heart. In both SPF and CON turkeys, dual infection resulted in an increased morbidity and a higher incidence of gross lesions compared to the groups given single infections, especially with a time interval between APV and E. coli inoculations of 3 and 5 days. APV was isolated from the respiratory tract of all APV-infected groups between 3 and 7 days post inoculation. E. coli O2:K1 was isolated only from turkeys that received a dual infection. It was recovered from the turbinates, trachea, lungs, heart and liver. These results show that APV may act as a primary agent predisposing to E. coli colonization and invasion.

  7. The acute effects of strength, endurance and concurrent exercises on the Akt/mTOR/p70S6K1 and AMPK signaling pathway responses in rat skeletal muscle

    Directory of Open Access Journals (Sweden)

    E.O.de Souza

    Full Text Available The activation of competing intracellular pathways has been proposed to explain the reduced training adaptations after concurrent strength and endurance exercises (CE. The present study investigated the acute effects of CE, strength exercises (SE, and endurance exercises (EE on phosphorylated/total ratios of selected AMPK and Akt/mTOR/p70S6K1 pathway proteins in rats. Six animals per exercise group were killed immediately (0 h and 2 h after each exercise mode. In addition, 6 animals in a non-exercised condition (NE were killed on the same day and under the same conditions. The levels of AMPK, phospho-Thr172AMPK (p-AMPK, Akt, phospho-Ser473Akt (p-Akt, p70S6K1, phospho-Thr389-p70S6K1 (p-p70S6K1, mTOR, phospho-Ser2448mTOR (p-mTOR, and phospho-Thr1462-TSC2 (p-TSC2 expression were evaluated by immunoblotting in total plantaris muscle extracts. The only significant difference detected was an increase (i.e., 87% in Akt phosphorylated/total ratio in the CE group 2 h after exercise compared to the NE group (P = 0.002. There were no changes in AMPK, TSC2, mTOR, or p70S6K1 ratios when the exercise modes were compared to the NE condition (P ≥ 0.05. In conclusion, our data suggest that low-intensity and low-volume CE might not blunt the training-induced adaptations, since it did not activate competing intracellular pathways in an acute bout of strength and endurance exercises in rat skeletal muscle.

  8. Effects of 2.45-GHz Electromagnetic Fields with a Wide Range of SARs on Micronucleus Formation in CHO-K1 Cells

    Directory of Open Access Journals (Sweden)

    S. Koyama

    2004-01-01

    Full Text Available There has been considerable discussion about the influence of high-frequency electromagnetic fields (HFEMF on the human body. In particular, HFEMF used for mobile phones may be of great concern for human health. In order to investigate the properties of HFEMF, we have examined the effects of 2.45-GHz EMF on micronucleus (MN formation in Chinese hamster ovary (CHO-K1 cells. MN formation is induced by chromosomal breakage or inhibition of spindles during cell division and leads to cell damage. We also examined the influence of heat on MN formation, since HFEMF exposure causes a rise in temperature. CHO-K1 cells were exposed to HFEMF for 2 h at average specific absorption rates (SARs of 5, 10, 20, 50, 100, and 200 W/kg, and the effects on these cells were compared with those in sham-exposed control cells. The cells were also treated with bleomycin alone as a positive control or with combined treatment of HFEMF exposure and bleomycin. Heat treatment was performed at temperatures of 37, 38, 39, 40, 41, and 42°C.The MN frequency in cells exposed to HFEMF at a SAR of lower than 50 W/kg did not differ from the sham-exposed controls, while those at SARs of 100 and 200 W/kg were significantly higher when compared with the sham-exposed controls. There was no apparent combined effect of HFEMF exposure and bleomycin treatment. On heat treatment at temperatures from 38–42°C, the MN frequency increased in a temperature-dependent manner. We also showed that an increase in SAR causes a rise in temperature and this may be connected to the increase in MN formation generated by exposure to HFEMF.

  9. Determination of IGF-1-Producing CHO-K1 Growth Phases Using GCMS-Based Global Metabolite Analysis

    Directory of Open Access Journals (Sweden)

    S. E. M. SABERI

    2011-12-01

    Full Text Available Mammalian cell lines, in particular CHO-K1 is vital for the multibillion dollar biotechnology industry. The majority of large scale bioprocessing of commercially valuable protein biopharmaceuticals is produced using this type of cell. An ideal mammalian cell system as host for biologics production should retain efficient use of energy sources in order to boost productivity at minimum cost. Various analyses such as cell counting and monitoring of specific biochemical responses are used to provide data to enable bioprocess control in order to achieve the ideal system. Our study aimed to see whether global metabolite analysis using Gas Chromatography Mass Spectrometry (GCMS would be a potential alternative approach in providing data for bioprocess control. In this study, we analyzed metabolites of CHO-K1 cells at different growth phases using GCMS. CHO-K1 cells producing insulin like growth factor-I (IGF1 were obtained from ATCC. Cells were grown in T-flask and incubated at 37°C/ 5% CO2 until 70-80% confluent in RPMI 1640 media. Samples (cells and spent/conditioned media were taken at designated intervals for routine cell counting (Trypan Blue dye exclusion method; glucose, glutamine and lactate determination (YSI 2700; IGF-1 production (ELISA kit R&D Sstems, Inc; and global metabolite analysis (GCMS. Conditioned media from each time point were spun down before subjecting into GCMS. Data from GCMS was then transferred to SIMCA P+12.0 for chemometric evaluation using Principal Component Analysis (PCA. The first component, PC1 results was able to explain 36% of the variation of the data with clear separation between exponential phase and other phases (initial and death phase. This suggests that GCMS-based global metabolite analysis has the ability to capture cell growth behaviour and offered insights of factors that may influence the biological system.ABSTRAK: Produk yang berupa sel kekal mamalia, terutamnya CHO-K1 adalah penting dan menguntungkan

  10. In vitro genotoxic and cytotoxic effects of ivermectin and its formulation ivomec on Chinese hamster ovary (CHO{sub K1}) cells

    Energy Technology Data Exchange (ETDEWEB)

    Molinari, G.; Soloneski, S.; Reigosa, M.A. [Catedra de Citologia, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, La Plata (Argentina); Larramendy, M.L., E-mail: m_larramendy@hotmail.com [Catedra de Citologia, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, La Plata (Argentina)

    2009-06-15

    The effects of ivermectin (IVM) and its commercial formulation ivomec (IVM 1.0%) were studied on Chinese hamster ovary (CHO{sub K1}) cells by several genotoxicity [sister chromatid exchange (SCE) and single cell gel electrophoresis (SCGE)] and cytotoxicity [cell-cycle progression (CCP), mitotic index (MI), proliferative replication index (PRI), 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and neutral red (NR)] bioassays within the 1.0-250 {mu}g/ml concentration-range. While IVM and ivomec did not modified SCE frequencies, they induced DNA-strand breaks revealed by SCGE. An enhancement of slightly damaged cells and a decrease in undamaged cells were observed in IVM-treated cultures with 5.0-50.0 {mu}g/ml. In ivomec-treated cells, while an increase in slightly damaged cells was induced with 5.0-50.0 {mu}g/ml, the damaged and undamaged cells increased and decreased only with 50.0 {mu}g/ml. Both compounds exerted a delay in CCP and a reduction in PRI when 25.0 {mu}g/ml was employed whereas cytotoxicity was observed at higher concentration than 50.0 {mu}g/ml. No MI alteration was observed with 1.0-10.0 and 1.0-5.0 {mu}g/ml of IVM and ivomec, respectively. A concentration-related trend to an increase in MI was achieved within 1.0-10.0 {mu}g/ml. An increase in the MI was induced in 10.0 {mu}g/ml ivomec-treated cultures. A marked reduction of about 89% and 62% in regard to controls was observed with 25.0 {mu}g/ml of IVM and ivomec, respectively. NR and MTT assays revealed a cell growth inhibition when 0.25-250.0 {mu}g/ml of both compounds was employed. The results highlighted that IVM and ivomec exert both genotoxicity and cytotoxicity in mammalian cells in vitro, at least in CHO{sub K1} cells.

  11. Cytotoxicity Evaluation of Anatase and Rutile TiO2 Thin Films on CHO-K1 Cells in Vitro

    Directory of Open Access Journals (Sweden)

    Blanca Cervantes

    2016-07-01

    Full Text Available Cytotoxicity of titanium dioxide (TiO2 thin films on Chinese hamster ovary (CHO-K1 cells was evaluated after 24, 48 and 72 h of culture. The TiO2 thin films were deposited using direct current magnetron sputtering. These films were post-deposition annealed at different temperatures (300, 500 and 800 °C toward the anatase to rutile phase transformation. The root-mean-square (RMS surface roughness of TiO2 films went from 2.8 to 8.08 nm when the annealing temperature was increased from 300 to 800 °C. Field emission scanning electron microscopy (FESEM results showed that the TiO2 films’ thickness values fell within the nanometer range (290–310 nm. Based on the results of the tetrazolium dye and trypan blue assays, we found that TiO2 thin films showed no cytotoxicity after the aforementioned culture times at which cell viability was greater than 98%. Independently of the annealing temperature of the TiO2 thin films, the number of CHO-K1 cells on the control substrate and on all TiO2 thin films was greater after 48 or 72 h than it was after 24 h; the highest cell survival rate was observed in TiO2 films annealed at 800 °C. These results indicate that TiO2 thin films do not affect mitochondrial function and proliferation of CHO-K1 cells, and back up the use of TiO2 thin films in biomedical science.

  12. Synergistic effects of atenolol and amlodipine for lowering and stabilizing blood pressure in 2K1Crenovascular hypertensive rats

    Institute of Scientific and Technical Information of China (English)

    Fu-ming SHEN; He-hui XIE; Gang LING; Li-ping XU; Ding-feng SU

    2005-01-01

    Aim: To test the synergistic effects of atenolol and amlodipine on lowering blood pressure (BP) and reducing blood pressure variability (BPV) in 2-kidney, one-clip(2K1C) renovascular hypertensive rats. Methods: Forty-eight 2K1C renovascular hypertensive rats were randomly divided into 6 groups. They were respectively given 0.8% carboxymethylcellulose sodium (control), atenolol (10.0 mg/kg),amlodipine (1.0 mg/kg), and combined atenolol and amlodipine (low dose: 5.0+0.5mg/kg; intermediate dose: 10.0+ 1.0 mg/kg; high dose: 20.0+2.0 mg/kg). The drugs were given via a catheter in a gastric fistula. BP was recorded for 25 h from 1 h before drug administration to 24 h after administration. Results: Compared with BP before medication, all 3 doses of combined atenolol and amlodipine significantly decreased the BP at 24 h after administration, except for the low dose on diastolic BP. Compared with the control group, all 3 doses of combined atenolol and amlodipine significantly reduced the average BP levels for the 24 h period after administration; furthermore, the high and intermediate doses also significantly decreased the BPV levels for the same period. The q values calculated by probability sum analysis for systolic and diastolic BP for the 24 h period after administration were 2.29 and 1.45, respectively, and for systolic and diastolic BPV for the same period they were 1.41 and 1.60, respectively. Conclusion: There is significant synergism between atenolol and amlodipine in lowering and stabilizing BP in 2K1C renovascular hypertensive rats.

  13. 婴儿口服维生素K1预防婴儿维生素K缺乏出血症的研究%A Study on Effects of Infants Oral Vitamin K1 to Prevent VKDB

    Institute of Scientific and Technical Information of China (English)

    周凤荣; 何守森; 王锡鲁

    2002-01-01

    目的探讨预防婴儿维生素K缺乏性出血症的有效措施.方法婴儿出生后即口服维生素K1,每10天1次,每次2.5mg,共10次,并采脐带血、1个月和2个月静脉血,统一检测.结果群体干预研究服药组维生素K缺乏性出血症发生率明显低于对照组(P<0.01).小样本的实验室观察结果显示了口服组1个月和2个月静脉血维生素K缺乏诱导蛋白(PIVKA-Ⅱ)已经完全转阴.结论婴儿口服维生素K1能有效地预防典型及晚发性维生素K缺乏性出血症.

  14. 对GB5413.10-2010中维生素K1检测的改进研究%Improvement research on the determination of vitamin K1 in the GB 5413.27-2010

    Institute of Scientific and Technical Information of China (English)

    姜瑞清; 周琳; 胡桂林; 陈军; 李梅; 常建军

    2011-01-01

    通过对不同种酶进行研究对比之后发现,改用胰脂肪酶能够很好的使脂肪降解,避免了对维生素K1检测的干扰,而且其加标回收率和方法重现性都能满足GB/T 27404-2008的要求.%In the GB 513.10-2010, the fat was decomposed by the lipase incompletely, so the vitamin K, peaks was not separated with the fat peak in the chromatography diagram, and then the peak area of the vitamin K1 was inaccurate. In the present study, three kinds of enzymes that include the lipase from porcine pancreas, lipase from Candida rugosa and pancreatin from porcine pancreas, were tested through detecting the different milk power sample, the pancreatin from porcine pancreas was considered to be the best one by the test results. Especially the value of testing after the improvement was more close to the amount of addition for the milk product.

  15. 伟大与胜负无关——记2007K-1MAX总决赛

    Institute of Scientific and Technical Information of China (English)

    沈诚

    2007-01-01

    2007年10月3日,经过K-1MAX全球赛区激战产生的8位70公斤级别最强战士聚集到日本东京日本武道馆。他们是东欧的"美丽死神"阿特(Artur Kyshenko),希腊小光头"铁拳"赞比迪(Mike Zambidis),日本"格斗大物"佐藤嘉洋,"钢铁难民"迪拉哥(Gago Drago),02

  16. An enzyme-linked immunosorbent assay (ELISA) for quantification of human collectin 11 (CL-11, CL-K1)

    DEFF Research Database (Denmark)

    Selman, L; Henriksen, M L; Brandt, J;

    2012-01-01

    Collectin 11 (CL-11), also referred to as collectin kidney 1 (CL-K1), is a pattern recognition molecule that belongs to the collectin group of proteins involved in innate immunity. It interacts with glycoconjugates on pathogen surfaces and has been found in complex with mannose-binding lectin-ass...... and thawing to a certain extent did not influence the ELISA. This ELISA offers a convenient and reliable method for studying CL-11 levels in relation to a variety of human diseases and syndromes....

  17. Fast intracellular dissolution and persistent cellular uptake of silver nanoparticles in CHO-K1 cells: implication for cytotoxicity

    DEFF Research Database (Denmark)

    Jiang, Xiumei; Miclaus, Teodora; Wang, Liming

    2015-01-01

    species, and cytotoxicity of Ag NPs (15.9 ± 7.6 nm) in Chinese hamster ovary cell subclone K1 cells, a cell line recommended by the OECD for genotoxicity studies. Quantification of intracellular nanoparticle uptake and ion release was performed through inductively coupled plasma mass spectrometry. X....... Subsequent cytotoxicity studies show that Ag NPs decrease cell viability and increase ROS production. Pre-incubation with N-acetyl-l-cysteine, an efficient antioxidant and Ag+ chelator, diminished the cytotoxicity caused by Ag NPs or Ag+ exposure. Our study suggests that the cytotoxicity mechanism of Ag NPs...

  18. Structure and function of collectin liver 1 (CL-L1) and collectin 11 (CL-11, CL-K1)

    DEFF Research Database (Denmark)

    Selman, Lana; Hansen, Soren

    2012-01-01

    The collectins are a group of innate immune proteins structurally characterized by their content of a carbohydrate recognition domain and a collagen-like region. Collectin liver 1 (CL-L1) and collectin 11 (CL-11, alias collectin kidney 1, CL-K1) are the more recently described members of this group...... to deficiencies have recently been identified as causative for 3MC syndrome. The 3MC syndrome is associated with a wide spectrum of developmental features including facial dysmorphism, cognitive impairment, hearing loss and vesicorenal anomalies. Similar polymorphic associations were reported for the mannan...

  19. Effect of scavengers of active oxygen species on cell damage caused in CHO-K1 cells by phenylhydroquinone, an o-phenylphenol metabolite.

    Science.gov (United States)

    Tayama, S; Nakagawa, Y

    1994-07-01

    Phenylhydroquinone (PHQ), a metabolite of o-phenylphenol (OPP), is easily autoxidized to phenylbenzoquinone (PBQ) via the semiquinone (phenylsemiquinone, PSQ) with concomitant production of superoxide anion radicals (O2-.). We have used scavengers of active oxygen species to examine whether or not O2-. produced during oxidation of PHQ is related to cell damage in CHO-K1 cells. PHQ at 10 micrograms/ml (3-h treatment) induced sister-chromatid exchange (SCE), endoreduplication (ERD) and cell-cycle delay in CHO-K1 cells. These effects were inhibited by catalase (280 U/ml), a scavenger of hydrogen peroxide (H2O2), as well as by the reductants, ascorbate (3 mM) and GSH (1 mM). Mannitol (50 mM), a scavenger of hydroxyl radical (OH.), was ineffective and superoxide dismutase (SOD, 150 U/ml), a scavenger of O2-., or SOD plus catalase rather intensified the toxicity as did aminotriazole (20 mM), an inhibitor of catalase. Analyses of incubation solutions by HPLC showed that the extent of cell damage is correlated with PHQ loss; catalase suppressed PHQ loss, whereas SOD promoted it. The correlation was more clearly seen in the time courses of cell death and PHQ loss during incubation of PHQ with each of the scavengers of active oxygen species. These results show that neither O2-. nor OH. participates in the cell damage, but rather H2O2 generated via dismutation of O2-. may participate, probably by accelerating the autoxidation of PHQ and thus causing an increase in the production of toxic intermediates. In fact, conversion of PHQ to PBQ, a reactive product, was demonstrated during incubation with PHQ in phosphate-buffered saline by following the changes in UV-visible spectra of PHQ. Inclusion of H2O2 (0.2 or 1 mM) in the incubation mixture accelerated the PHQ loss. The present results can be explained in terms of the autoxidation mechanism of hydroquinone proposed by O'Brien (1991). Different from the results in the absence of S9 mix, the cell damage induced by 50 micrograms

  20. A rat model of nerve agent exposure applicable to the pediatric population: The anticonvulsant efficacies of atropine and GluK1 antagonists

    Energy Technology Data Exchange (ETDEWEB)

    Miller, Steven L., E-mail: stevenmiller17@gmail.com [Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Program in Neuroscience, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Aroniadou-Anderjaska, Vassiliki, E-mail: vanderjaska@usuhs.edu [Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Department of Psychiatry, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Program in Neuroscience, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Figueiredo, Taiza H., E-mail: taiza.figueiredo.ctr@usuhs.edu [Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Prager, Eric M., E-mail: eric.prager683@gmail.com [Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Program in Neuroscience, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Almeida-Suhett, Camila P., E-mail: camilapalmeida@gmail.com [Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Program in Neuroscience, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814 (United States); Apland, James P., E-mail: james.p.apland.civ@mail.mil [Neurotoxicology Branch, U.S. Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD 21010 (United States); and others

    2015-04-15

    Inhibition of acetylcholinesterase (AChE) after nerve agent exposure induces status epilepticus (SE), which causes brain damage or death. The development of countermeasures appropriate for the pediatric population requires testing of anticonvulsant treatments in immature animals. In the present study, exposure of 21-day-old (P21) rats to different doses of soman, followed by probit analysis, produced an LD{sub 50} of 62 μg/kg. The onset of behaviorally-observed SE was accompanied by a dramatic decrease in brain AChE activity; rats who did not develop SE had significantly less reduction of AChE activity in the basolateral amygdala than rats who developed SE. Atropine sulfate (ATS) at 2 mg/kg, administered 20 min after soman exposure (1.2 × LD{sub 50}), terminated seizures. ATS at 0.5 mg/kg, given along with an oxime within 1 min after exposure, allowed testing of anticonvulsants at delayed time-points. The AMPA/GluK1 receptor antagonist LY293558, or the specific GluK1 antagonist UBP302, administered 1 h post-exposure, terminated SE. There were no degenerating neurons in soman-exposed P21 rats, but both the amygdala and the hippocampus were smaller than in control rats at 30 and 90 days post-exposure; this pathology was not present in rats treated with LY293558. Behavioral deficits present at 30 days post-exposure, were also prevented by LY293558 treatment. Thus, in immature animals, a single injection of atropine is sufficient to halt nerve agent-induced seizures, if administered timely. Testing anticonvulsants at delayed time-points requires early administration of ATS at a low dose, sufficient to counteract only peripheral toxicity. LY293558 administered 1 h post-exposure, prevents brain pathology and behavioral deficits. - Highlights: • The LD{sub 50} of soman was determined in postnatal-day-21 rats. • Rats with no seizures after 1.2XLD{sub 50} soman had less reduction of AChE in the amygdala. • Atropine sulfate (ATS) at 2 mg/kg, given at 20 min after

  1. Lipopeptides from the Banyan Endophyte, Bacillus subtilis K1: Mass Spectrometric Characterization of a Library of Fengycins

    Science.gov (United States)

    Pathak, Khyati V.; Keharia, Haresh; Gupta, Kallol; Thakur, Suman S.; Balaram, Padmanabhan

    2012-10-01

    Mass spectrometric analysis of a banyan endophyte, Bacillus subtilis K1, extract showing broad spectrum antifungal activity revealed a complex mixture of lipopeptides, iturins, surfactins, and fengycins. Fractionation by reversed-phase high performance liquid chromatography (HPLC) facilitated a detailed analysis of fengycin microheterogeneity. Matrix assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) mass spectrometric studies permitted the identification of several new fengycin variants. Four major sites of heterogeneity are identified: (1) N-terminus β-hydroxy fatty acid moiety, where chain length variation and the presence of unsaturation occur, (2) position 6 (Ala/Val/Ile/Leu), (3) position 10 (Val/Ile) within the macrocyclic ring, and (4) Gln to Glu replacement at position 8, resulting in fengycin variants that differ in mass by 1 Da. Diagnostic fragment ions provide a quick method for localizing the sites of variation in the macrocycle or the linear segment. Subsequent establishment of the sequences is achieved by MS/MS analysis of linear fengycin species produced by hydrolysis of the macrocyclic lactone. Unsaturation in the fatty acid chain and the presence of linear precursors in the B. subtilis K1 extract are also established by mass spectrometry. The anomalous distribution of intensities within isotopic multiplets is a diagnostic for Gln/Glu replacements. High resolution mass spectrometry facilitates the identification of fengycin species differing by 1 Da by localizing the variable position (Gln8/Glu8) in the fengycin variants.

  2. Biosynthesis of the Escherichia coli K1 group 2 polysialic acid capsule occurs within a protected cytoplasmic compartment.

    Science.gov (United States)

    Steenbergen, Susan M; Vimr, Eric R

    2008-06-01

    Capsular polysaccharides are important virulence determinants in a wide range of invasive infectious diseases. Although capsule synthesis has been extensively investigated, understanding polysaccharide export from the cytoplasm to the external environment has been more difficult. Here we present the results of a novel protection assay indicating that synthesis and export of the Escherichia coli K1 group 2 capsular polysialic acid (K1 antigen) occur within a protected subcellular compartment designated the sialisome. In addition to the polymerase encoded by neuS, localization and complementation analyses indicated that the sialisome includes the accessory membrane protein NeuE. The requirement for NeuE was suppressed by overproducing NeuS, suggesting that NeuE functions by stabilizing the polymerase or facilitating its assembly in the sialisome. Although an interaction between NeuE and NeuS could not be demonstrated with a bacterial two-hybrid system that reconstitutes an intracellular cell-signalling pathway, interactions between NeuS and KpsC as well as other sialisome components were detected. The combined results provide direct evidence for specific protein-protein interactions in the synthesis and export of group 2 capsular polysaccharides under in vivo conditions. The approaches developed here will facilitate further dissection of the sialisome, suggesting similar methodology for understanding the biosynthesis of other group 2 capsules.

  3. BIPM comparison BIPM.RI(II)-K1.Th-228 of activity measurements of the radionuclide 228Th

    Science.gov (United States)

    Michotte, C.; Ratel, G.; Courte, S.; Lucas, L.; Kossert, K.; Nähle, O.; Ott, O.

    2016-01-01

    Since 1986, two national metrology institutes (NMI) have submitted two samples of known activity of 228Th to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM), with comparison identifier BIPM.RI(II)-K1.Th-228. The values of the activity submitted were about 300 kBq and 2 MBq. A key comparison reference value (KCRV) has been evaluated for the first time for 228Th. There is only one result remaining in the BIPM.RI(II)-K1.Th-228 comparison, the 1986 NIST result being outdated. The degrees of equivalence between each equivalent activity measured in the SIR and the KCRV have been calculated and the results are given in the form of a table. A graphical presentation is also given. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  4. Synthesis of LaxK1-xCoO3 nanorod and their catalytic performances for CO oxidation

    Institute of Scientific and Technical Information of China (English)

    刘坚; 王季秋; 赵震; 徐春明; 韦岳长; 段爱军; 姜桂元

    2014-01-01

    A series of LaxK1-xCoO3 nanorod oxides with perovskite structure were synthesized by sol-gel method using polyvinyl al-cohol (PVA) as additive. These perovskite-type complex oxide catalysts were characterized by the techniques of X-ray diffraction (XRD), infrared (IR), Brumauer-Emmett-Teller (BET) and scanning electron microscopy (SEM). And the results showed that nano-rods of La1-xKxCoO3 perovskite-type complex oxides were fabricated by sol-gel method when the mass concentration of PVA was 4%and the calcined temperature kept at 700 ℃ for 4 h. The catalytic results of CO oxidation showed that the LaxK1-xCoO3 catalysts had high activity. LaCoO3 nanorods exposed more{110}plane than LaCoO3 nanoparticles, which was beneficial to the catalytic oxi-dation of CO. LaCoO3 nanorods had the best catalytic performance for the oxidation of CO. At 200 ºC, the CO conversion could reach 100%.

  5. Effects of Iron-Oxide Nanoparticle Surface Chemistry on Uptake Kinetics and Cytotoxicity in CHO-K1 Cells

    Directory of Open Access Journals (Sweden)

    Camille C. Hanot

    2015-12-01

    Full Text Available Superparamagnetic iron-oxide nanoparticles (SPIONs show great promise for multiple applications in biomedicine. While a number of studies have examined their safety profile, the toxicity of these particles on reproductive organs remains uncertain. The goal of this study was to evaluate the cytotoxicity of starch-coated, aminated, and PEGylated SPIONs on a cell line derived from Chinese Hamster ovaries (CHO-K1 cells. We evaluated the effect of particle diameter (50 and 100 nm and polyethylene glycol (PEG chain length (2k, 5k and 20k Da on the cytotoxicity of SPIONs by investigating cell viability using the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT and sulforhodamine B (SRB assays. The kinetics and extent of SPION uptake by CHO-K1 cells was also studied, as well as the resulting generation of intracellular reactive oxygen species (ROS. Cell toxicity profiles of SPIONs correlated strongly with their cellular uptake kinetics, which was strongly dependent on surface properties of the particles. PEGylation caused a decrease in both uptake and cytotoxicity compared to aminated SPIONs. Interestingly, 2k Da PEG-modifed SPIONs displayed the lowest cellular uptake and cytotoxicity among all studied particles. These results emphasize the importance of surface coatings when engineering nanoparticles for biomedical applications.

  6. K1YT METAR

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — METAR is a routine scheduled observation and is the primary observation code used in the United States to satisfy requirements for reporting surface meteorological...

  7. EPR studies of the vitamin K 1 semiquinone radical anion. Comparison to the electron acceptor A 1 in green plant photosystem I

    Science.gov (United States)

    Thurnauer, Marion C.; Brown, James W.; Gast, P.; Feezel, Laura L.

    Suggestions that the electron acceptor, A 1, in Photosystem I is a quinone have come from both optical and epr experiments. Vitamin K 1 (phylloquinone) is present in the PSI complex with a stoichiometry of two molecules per reaction center. In order to determine if A 1 can be identified with vitamin K 1, X-band and Q-band epr properties of the vitamin K 1 radical anion in frozen alcohol solutions are examined. The results are compared to the epr properties that have been observed for the reduced A 1 acceptor in vivo. The g-values obtained for the vitamin K 1 radical anion are consistent with identifying A 1 with vitamin K 1.

  8. Molecular characterization of Legionella pneumophila-induced interleukin-8 expression in T cells

    Directory of Open Access Journals (Sweden)

    Mukaida Naofumi

    2010-01-01

    Full Text Available Abstract Background Legionella pneumophila is the causative agent of human Legionnaire's disease. During infection, the bacterium invades macrophages and lung epithelial cells, and replicates intracellularly. However, little is known about its interaction with T cells. We investigated the ability of L. pneumophila to infect and stimulate the production of interleukin-8 (IL-8 in T cells. The objective of this study was to assess whether L. pneumophila interferes with the immune system by interacting and infecting T cells. Results Wild-type L. pneumophila and flagellin-deficient Legionella, but not L. pneumophila lacking a functional type IV secretion system Dot/Icm, replicated in T cells. On the other hand, wild-type L. pneumophila and Dot/Icm-deficient Legionella, but not flagellin-deficient Legionella or heat-killed Legionella induced IL-8 expression. L. pneumophila activated an IL-8 promoter through the NF-κB and AP-1 binding regions. Wild-type L. pneumophila but not flagellin-deficient Legionella activated NF-κB, p38 mitogen-activated protein kinase (MAPK, Jun N-terminal kinase (JNK, and transforming growth factor β-associated kinase 1 (TAK1. Transfection of dominant negative mutants of IκBα, IκB kinase, NF-κB-inducing kinase, TAK1, MyD88, and p38 MAPK inhibited L. pneumophila-induced IL-8 activation. Inhibitors of NF-κB, p38 MAPK, and JNK blocked L. pneumophila-induced IL-8 expression. In addition, c-Jun, JunD, cyclic AMP response element binding protein, and activating transcription factor 1, which are substrates of p38 MAPK and JNK, bound to the AP-1 site of the IL-8 promoter. Conclusions Taken together, L. pneumophila induced a flagellin-dependent activation of TAK1, p38 MAPK, and JNK, as well as NF-κB and AP-1, which resulted in IL-8 production in human T cells, presumably contributing to the immune response in Legionnaire's disease.

  9. 维生素K1辅助治疗百日咳综合征的疗效分析%Efficacy of vitamin K1 in the adjuvant treatment of pertussis syndrome

    Institute of Scientific and Technical Information of China (English)

    吴国红; 高振文; 马杰; 王迪彬; 黄秀铭

    2016-01-01

    目的:观察维生素 K1辅助治疗百日咳综合征患儿痉挛性咳嗽的效果及对住院时间的影响。方法将87例百日咳综合征患儿采用数字表法随机分成对照组41例、治疗组46例,对照组予以抗感染、呼吸道支持、解痉平喘、祛痰等治疗;治疗组在观察组治疗基础上加用维生素 K1治疗,比较两组痉挛性咳嗽持续时间及住院时间。结果治疗组痉挛性咳嗽消失时间、住院时间分别为(7.22±1.33)d、(9.52±1.84)d,对照组分别为(15.51±2.73)d、(18.71±3.30)d,两组差异均有统计学意义(t =-18.310、-16.269,均 P <0.01)。治疗组显效18例,有效25例,无效3例,总有效率为93.5%;对照组显效7例,有效23例,无效11例,总有效率为73.1%。两组总有效率差异有统计学有意义(χ2=4.285,P <0.05)。结论维生素 K1对缓解小儿痉挛性咳嗽有明显效果,可缩短百日咳综合征患儿痉挛性咳嗽持续时间及住院时间,提高临床疗效。%Objective To investigate the effects of vitamin K1 in the adjuvant treatment of spasmodic cough in pertussis syndrome children,and its impacts on length of hospital stay.Methods 87 pertussis syndrome children were randomly divided into two groups,among which 41 cases in the control group were treated with the therapies such as anti -infection,respiratory support,antispasmodic,expectorant et al,while 46 cases in the treatment group were treated additionally with vitamin K1 on the basis of the control group.Then compared the duration and hospital stay of the two groups with spasmodic cough.Results The disappeared time of spasmodic cough in the treatment group was (7.22 ±1.33)d and hospital stay was (9.52 ±1.84)d,which in the control group were (15.51 ±2.73)d,(18.71 ± 3.30)d respectively,and there were statistically significant differences(t =-18.310,-16.269,all P <0.01).In the treatment group

  10. SphK1及其抑制剂对人胃癌裸鼠移植瘤的作用%Role of SphK1 in the growth of human gastric cancer xenograft in nude mice

    Institute of Scientific and Technical Information of China (English)

    苗怡然; 黄广清; 蔡红星; 王云; 单海霞; 朱正秋

    2013-01-01

    Objective To investigate the role of SphK 1 in the growth of human gastric cancer xenograft in nude mice and to explore its mechanism. Methods Human gastric cancer cells SGC -7901 were cultured in RPMI 1640 medium to exponential phase of growth and then transplanted under the skin of BALB /c nude mice to develop the tumor model of hu -man gastric cancer. After the model was successfully developed , 38 mice were randomly divided into four groups : normal saline (NS) control group, cisplantin (DDP) group, SKI - Ⅱ group, and SKI - Ⅱ combined with DDP group. All rats were given intraperitoneal injection of drugs on seven days for 3 times. The tumor mass volume was observed every 3 days and inhibition rate of tumor growth was also calculated . All nude mice in each group were killed at the 7th day after the injection of drugs and tumor were dislodged. Immunohistochemistry staining was used to detect the protein expression of SphKl, Bax and Bel - 2. The apoptosis of tumor was measured by terminal dUTP nick - end labeling ( TUNEL). Results The tumor model of human gastric cancer was successfully developed . SphKl and Bcl - 2 protein expression in SKI - Ⅱ combined with DDP group significantly decreased (P < 0.05 ). Compared with DDP group and SKI - Ⅱ group, Bax protein expression in SKI - Ⅱ combined with DDP group significantly increased (P < 0.05 ). As a result, SKI - Ⅱ combined with DDP could raise the rate of apoptosis and inhibit the growth of tumor model of human gastric cancer . Conclusion Sphkl can promote tumor cell proliferation through down -regulating the ratio of Bax/Bcl -2. SKI - Ⅱ has synergistic anti - tumor effect of cisplatin.%目的 研究鞘氨醇激酶-1(sphingosine kinase-1,SphK1)及其抑制剂在人胃癌裸鼠移植瘤生长中的作用,并探讨其作用机制.方法 对数生长期SGC7901细胞注射于裸鼠皮下建立人胃癌裸鼠移植瘤模型,建模成功后将裸鼠随机分为4组(每组8只),分别用生理盐水、SKI-

  11. Interactive effects of zearalenone and its metabolites on cytotoxicity and metabolization in ovarian CHO-K1 cells.

    Science.gov (United States)

    Tatay, Elena; Meca, Giuseppe; Font, Guillermina; Ruiz, Maria-Jose

    2014-02-01

    Zearalenone (ZEA) is a non-steroidal estrogen mycotoxin with high binding affinity to estrogen receptors. ZEA is rapidly absorbed and metabolized in vivo to α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL). So, mixtures of them may be present in biological systems and suppose a hazard to animals and human health. The aims of this study were to determine the cytotoxic effects of ZEA and its metabolites, alone and in combination in ovarian (CHO-K1) cells during 24, 48 and 72h by the MTT assay; and to investigate the metabolism of the CHO-K1 cells on ZEA, and its conversion into α-ZOL and β-ZOL by CHO-K1 cell after 24 and 48h of exposure. The IC50 value obtained for individual mycotoxins range from 60.3 to >100.0μM, from 30.0 to 33.0μM and from 55.0 to >75.0μM for ZEA, α-ZOL and β-ZOL, respectively. Cytotoxic interactions were assayed by the isobologram method, which provides a combination index (CI) value as a quantitative measure of the degree of the three mycotoxin interaction. The CI values for binary combinations ranged from 0.56±0.15 (synergism at low concentrations) to 5.25±5.10 (addition at high concentrations) and tertiary combinations from 2.95±0.75 (antagonism at low concentrations) to 0.41±0.23 (synergism at high concentrations). The concentration of ZEA and its metabolites was determined with liquid chromatography coupled to the mass spectrometer detector-linear ion trap (LC-MS-LIT). The percentage of ZEA degradation ranged from 4% (24h) to 81% (48h). In the same conditions, α-ZOL and β-ZOL concentration decreased from 8% to 85%. No conversion of ZEA in α-ZOL and β-ZOL was detected. However, at 24h of exposure other degradation products of ZEA and its derived were detected. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Assessment of cytotoxic and cytogenetic effects of a 1,2,5-thiadiazole derivative on CHO-K1 cells. Its application as corrosion inhibitor

    Energy Technology Data Exchange (ETDEWEB)

    Grillo, C.A. [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA, CCT La Plata-CONICET), Facultad de Ciencias Exactas, Departamento de Quimica, Universidad Nacional de La Plata, Casilla de Correo 16, Sucursal 4, 1900 La Plata (Argentina); Mirifico, M.V. [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA, CCT La Plata-CONICET), Facultad de Ciencias Exactas, Departamento de Quimica, Universidad Nacional de La Plata, Casilla de Correo 16, Sucursal 4, 1900 La Plata (Argentina); Facultad de Ingenieria, Areas Departamentales Ingenieria Quimica and Mecanica, Universidad Nacional de La Plata, Calle 47 y 1, 1900 La Plata (Argentina); Morales, M.L. [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA, CCT La Plata-CONICET), Facultad de Ciencias Exactas, Departamento de Quimica, Universidad Nacional de La Plata, Casilla de Correo 16, Sucursal 4, 1900 La Plata (Argentina); Reigosa, M.A. [IMBICE (Instituto Multidisciplinario de Biologia Celular), CICPBA, CONICET, Calle 526 entre 10 y 11, 1900 La Plata (Argentina); Mele, M. Fernandez Lorenzo de, E-mail: mmele@inifta.unlp.edu.ar [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA, CCT La Plata-CONICET), Facultad de Ciencias Exactas, Departamento de Quimica, Universidad Nacional de La Plata, Casilla de Correo 16, Sucursal 4, 1900 La Plata (Argentina); Facultad de Ingenieria, Areas Departamentales Ingenieria Quimica and Mecanica, Universidad Nacional de La Plata, Calle 47 y 1, 1900 La Plata (Argentina)

    2009-10-30

    This work focuses on the possible use of phenanthro[9,10-c]-1,2,5-thiadiazole 1,1-dioxide (TDZ) as a harmless corrosion inhibitor. TDZ range-dose providing minimum adverse effects to the environment and human health, with satisfactory corrosion-inhibiting properties was evaluated. Cytotoxicity and genotoxicity of TDZ at 0.57-12.50 {mu}M concentration range were tested by neutral red, chromosomal aberrations, mitotic index, and colony formation assays. Results showed a significant increase of chromatid-type aberrations for the highest concentration of TDZ assayed (12.50 {mu}M). Additionally, a reduction in the proliferative rate for lower concentrations was detected by the MI assay. We concluded that TDZ should be used at concentrations lower than 1.16 {mu}M. Corrosion assays performed showed good inhibition effect (ca. 50%) at low (0.65 {mu}M) TDZ concentration. Consequently, our results indicated that TDZ induced a time- and dose-dependent genotoxic and cytotoxic response on CHO-K1 cells. Short assays should be complemented with long exposure tests to simulate chronic contact with TDZ since lower threshold levels may be found for shorter exposures and a wrong safety range could be determined.

  13. Fabrication and Electrical Properties of Multilayer Ceramic Actuator Using Lead-Free (Bi1/2K1/2)TiO3

    Science.gov (United States)

    Nagata, Hajime; Tabuchi, Kazuya; Takenaka, Tadashi

    2013-09-01

    A prototype of a multilayer actuator (MLA) was fabricated using (Bi1/2K1/2)TiO3 (BKT) ceramics as active layers and Ag-Pd (7:3) as inner electrodes. Then, their microstructures and electrical properties were investigated. From the scanning electron microscope (SEM) observation, the thickness of the BKT active layers between electrodes was estimated to be approximately 60 µm for the body co-fired at 1045 °C for 10 h. Also, SEM images showed homogeneous and dense microstructures on the BKT active layers. Additionally, the interface between BKT and the Ag-Pd electrode was clearly distinguished and there are no significant signs of reaction or diffusion between them at least from the SEM micrographs. The resistivity ρ of BKT sample co-fired at 1045 °C for 10 h was about 1011 Ω.cm on average, which is sufficient for applying a high DC electric field to the sample. From the measurement of the field-induce strain S under unipolar driving at 100 kV/cm, a large strain of 0.14% at 100 kV/cm was obtained and the longitudinal dynamic displacement was 1 µm at the same electric field. The normalized d33* was calculated to be 138 pm/V, which is similar to that in the bulk samples. This result indicates that the BKT-based multilayered prototype efficiently functioned under unipolar driving up to 100 kV/cm.

  14. Effect of thermal history on stability of the relaxor state in (Bi1/2K1/2)TiO3 ceramics

    Science.gov (United States)

    Hagiwara, Manabu; Fujihara, Shinobu

    2017-10-01

    Bismuth potassium titanate (Bi1/2K1/2)TiO3 (BKT) with disordered A-site Bi3+ and K+ ions is known to undergo a spontaneous phase transition between a high-temperature relaxor (R) state and a low-temperature ferroelectric (FE) state at around 300 °C. Here, we demonstrate that the thermal history of BKT ceramics has a great effect on the stability of the R and FE states. To modulate the distribution of Bi3+ and K+ ions, as-sintered BKT ceramics were postannealed at 900 °C and then cooled to 500 °C at a very low cooling rate of ‑0.1 °C/min. The slowly cooled samples showed a normal-relaxor-like temperature dependence of dielectric permittivity without any anomaly attributed to the spontaneous R-FE transition. The slow cooling treatment changed the room-temperature crystal system of the BKT ceramics from tetragonal to cubic, although it did not induce the cationic ordering of Bi3+ and K+. It was also observed that the macroscopic domain structure disappeared after the slow cooling treatment. These observations demonstrated that the slow cooling treatment stabilized the R state in BKT to inhibit the spontaneous phase transition into the FE state. The possible reason for the stabilization of the R state in the slowly cooled BKT is discussed on the basis of the changes in cationic distribution and local compositional heterogeneity.

  15. Electrical Properties of Textured (Bi1/2K1/2)TiO3-BaTiO3 Lead-Free Piezoelectric Ceramics

    Science.gov (United States)

    Nemoto, Masahiro; Hiruma, Yuji; Nagata, Hajime; Takenaka, Tadashi

    2009-07-01

    The electrical properties of (1-x)(Bi1/2K1/2)TiO3-xBaTiO3 (BKT-BT100x) ceramics prepared by an ordinary solid-state reaction and a reactive templated grain growth (RTGG) method using platelike Bi4Ti3O12 particles as a template were investigated. The degree of orientation, F, which was determined by the Lotgering method using an X-ray diffraction pattern, achieved ≈83% in the textured BKT-BT10 ceramic. A textured BKT-BT10 ceramic with a homogeneous microstructure was obtained after a long sintering time. The field-induced strains and piezoelectric properties of the BKT-BT10 ceramic were improved by improving grain orientation. In particular, the piezoelectric strain constant, d33, and the normalized strain, d33*, of the textured BKT-BT10 ceramic in the direction parallel (∥) to the tape stacking direction were 121 pC/N and 211 pm/V (at 80 kV/cm), respectively.

  16. Fabrication and Piezoelectric Properties of Grain-Oriented (Bi1/2K1/2)TiO3-BaTiO3 Ceramics

    Science.gov (United States)

    Nemoto, Masahiro; Hiruma, Yuji; Nagata, Hajime; Takenaka, Tadashi

    2008-05-01

    The piezoelectric properties of (1 - x)(Bi1/2K1/2)TiO3-xBaTiO3 (x = 0.1) [abbreviated as BKT-BT10] ceramics prepared by ordinary solid-state reaction and reactive templated grain growth (RTGG) using platelike Bi4Ti3O12 particles as a template were investigated. All samples showed a single-phase perovskite structure, and relative densities compared with theoretical densities were all higher than 95%. The degree of orientation F was determined by the Lotgering method using an X-ray diffraction (XRD) pattern. The F values of textured BKT-BT10 ceramics were relatively low at ˜35%. However, field-induced strains and piezoelectric properties of BKT-BT10 ceramics were improved by grain orientation. In particular, the piezoelectric strain constant d33 and the strain of textured BKT-BT10 ceramics in a direction parallel (∥) to the tape stacking direction were 84.5 pC/N and 168 pm/V (at 80 kV/cm), respectively.

  17. mToR/S6K1信号通路研究进展

    Institute of Scientific and Technical Information of China (English)

    刘效磊; 牛燕媚; 傅力

    2010-01-01

    @@ 哺乳动物雷帕霉素靶蛋白(mammalian target of ra-pamycin,mTOR)是哺乳动物细胞内感受细胞外营养、能量水平以及生长因子等信号变化的一种丝/苏氨酸蛋白激酶.当细胞外环境发生变化时,mTOR通过激活下游效应蛋白S6K1(ribosomal protein S6 kinase,polypeptide 1)参与调节细胞生长、分化增殖以及蛋白质合成等过程.

  18. COOMET.RI(I)-K1 comparison of national measurement standards of air kerma for {sup 60}Co {gamma} radiation

    Energy Technology Data Exchange (ETDEWEB)

    Buermann, L. [Physikalisch-Technische Bundesanstalt (PTB), Braunschweig (Germany); Oborin, A.V. [D I Mendeleyev Institute for Metrology (VNIIM), St Petersburg (Russian Federation); Dobrovosky, J. [3 Slovensky Metrologicky Ustav (SMU), Bratislava (Slovakia); Milevsky, V.S. [Belarussian State Institute for Metrology (BelGIM), Minsk (Belarus); Walwyn Salas, G. [Centro de Proteccion e Higiene de las Radiaciones (CPHR), Habana (Cuba); Lapenas, A. [Radiation Metrology and Testing Centre of the Latvian National Metrology Centre Ltd (RMTC), Salaspils (Latvia)

    2009-12-15

    Results are presented of the COOMET key comparison of the national measurement standards of air kerma for {sup 60}Co {gamma} radiation. Participants of the comparison were PTB (Germany, pilot institute), VNIIM (Russia), SMU (Slovakia), BelGIM (Belarus), CPHR (Cuba) and RMTC (Latvia). PTB, VNIIM and SMU had previously taken part in a key comparison with the Bureau International de Poids et Mesures (BIPM) and operated as link laboratories in order to evaluate the degree of equivalence of the participants' results with the key comparison reference value. These data form the basis of the results entered into the BIPM key comparison database for comparison COOMET.RI(I)-K1. (authors)

  19. 扣人心弦的“强大”赛事——K-1max 16实录

    Institute of Scientific and Technical Information of China (English)

    平常心

    2009-01-01

    2009年4月21日,一年一度的K-1 max 16"强大"赛在日本福冈体育馆举行。16位在分赛区杀出的搏击高手汇聚一堂。除了08年亚军"美丽死神"阿特(Artur Kyshenko)、2005年和07年2届王者安迪·苏瓦(?Andy Souwer)、2004年和2006年2届冠军"泰拳王子"播求、初代王者阿尔伯特·克劳斯(Albert Kraus)、钢铁难民迪拉哥(Gago

  20. Ribosomal S6K1 in POMC and AgRP Neurons Regulates Glucose Homeostasis but Not Feeding Behavior in Mice

    Directory of Open Access Journals (Sweden)

    Mark A. Smith

    2015-04-01

    Full Text Available Hypothalamic ribosomal S6K1 has been suggested as a point of convergence for hormonal and nutrient signals in the regulation of feeding behavior, bodyweight, and glucose metabolism. However, the long-term effects of manipulating hypothalamic S6K1 signaling on energy homeostasis and the cellular mechanisms underlying these roles are unclear. We therefore inactivated S6K1 in pro-opiomelanocortin (POMC and agouti-related protein (AgRP neurons, key regulators of energy homeostasis, but in contrast to the current view, we found no evidence that S6K1 regulates food intake and bodyweight. In contrast, S6K1 signaling in POMC neurons regulated hepatic glucose production and peripheral lipid metabolism and modulated neuronal excitability. S6K1 signaling in AgRP neurons regulated skeletal muscle insulin sensitivity and was required for glucose sensing by these neurons. Our findings suggest that S6K1 signaling is not a general integrator of energy homeostasis in the mediobasal hypothalamus but has distinct roles in the regulation of glucose homeostasis by POMC and AgRP neurons.

  1. Ribosomal S6K1 in POMC and AgRP Neurons Regulates Glucose Homeostasis but Not Feeding Behavior in Mice.

    Science.gov (United States)

    Smith, Mark A; Katsouri, Loukia; Irvine, Elaine E; Hankir, Mohammed K; Pedroni, Silvia M A; Voshol, Peter J; Gordon, Matthew W; Choudhury, Agharul I; Woods, Angela; Vidal-Puig, Antonio; Carling, David; Withers, Dominic J

    2015-04-21

    Hypothalamic ribosomal S6K1 has been suggested as a point of convergence for hormonal and nutrient signals in the regulation of feeding behavior, bodyweight, and glucose metabolism. However, the long-term effects of manipulating hypothalamic S6K1 signaling on energy homeostasis and the cellular mechanisms underlying these roles are unclear. We therefore inactivated S6K1 in pro-opiomelanocortin (POMC) and agouti-related protein (AgRP) neurons, key regulators of energy homeostasis, but in contrast to the current view, we found no evidence that S6K1 regulates food intake and bodyweight. In contrast, S6K1 signaling in POMC neurons regulated hepatic glucose production and peripheral lipid metabolism and modulated neuronal excitability. S6K1 signaling in AgRP neurons regulated skeletal muscle insulin sensitivity and was required for glucose sensing by these neurons. Our findings suggest that S6K1 signaling is not a general integrator of energy homeostasis in the mediobasal hypothalamus but has distinct roles in the regulation of glucose homeostasis by POMC and AgRP neurons. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  2. Active site of Zn2+-dependent sn-glycerol-1-phosphate dehydrogenase from Aeropyrum pernix K1

    Directory of Open Access Journals (Sweden)

    Jin-Suk Han

    2005-01-01

    Full Text Available The enzyme sn-glycerol-1-phosphate dehydrogenase (Gro1PDH, EC 1.1.1.261 is key to the formation of the enantiomeric configuration of the glycerophosphate backbone (sn-glycerol-1-phosphate of archaeal ether lipids. This enzyme catalyzes the reversible conversion between dihydroxyacetone phosphate and glycerol-1-phosphate. To date, no information about the active site and catalytic mechanism of this enzyme has been reported. Using the sequence and structural information for glycerol dehydrogenase, we constructed six mutants (D144N, D144A, D191N, H271A, H287A and D191N/H271A of Gro1PDH from Aeropyrum pernix K1 and examined their characteristics to clarify the active site of this enzyme. The enzyme was found to be a zinc-dependent metalloenzyme, containing one zinc ion for every monomer protein that was essential for activity. Site-directed mutagenesis of D144 increased the activity of the enzyme. Mutants D144N and D144A exhibited low affinity for the substrates and higher activity than the wild type, but their affinity for the zinc ion was the same as that of the wild type. Mutants D191N, H271A and H287A had a low affinity for the zinc ion and a low activity compared with the wild type. The double mutation, D191N/ H271A, had no enzyme activity and bound no zinc. From these results, it was clarified that residues D191, H271 and H287 participate in the catalytic activity of the enzyme by binding the zinc ion, and that D144 has an effect on substrate binding. The structure of the active site of Gro1PDH from A. pernix K1 seems to be similar to that of glycerol dehydrogenase, despite the differences in substrate specificity and biological role.

  3. MAP3K1 function is essential for cytoarchitecture of the mouse organ of Corti and survival of auditory hair cells

    Directory of Open Access Journals (Sweden)

    Rizwan Yousaf

    2015-12-01

    Full Text Available MAP3K1 is a serine/threonine kinase that is activated by a diverse set of stimuli and exerts its effect through various downstream effecter molecules, including JNK, ERK1/2 and p38. In humans, mutant alleles of MAP3K1 are associated with 46,XY sex reversal. Until recently, the only phenotype observed in Map3k1tm1Yxia mutant mice was open eyelids at birth. Here, we report that homozygous Map3k1tm1Yxia mice have early-onset profound hearing loss accompanied by the progressive degeneration of cochlear outer hair cells. In the mouse inner ear, MAP3K1 has punctate localization at the apical surface of the supporting cells in close proximity to basal bodies. Although the cytoarchitecture, neuronal wiring and synaptic junctions in the organ of Corti are grossly preserved, Map3k1tm1Yxia mutant mice have supernumerary functional outer hair cells (OHCs and Deiters' cells. Loss of MAP3K1 function resulted in the downregulation of Fgfr3, Fgf8, Fgf10 and Atf3 expression in the inner ear. Fgfr3, Fgf8 and Fgf10 have a role in induction of the otic placode or in otic epithelium development in mice, and their functional deficits cause defects in cochlear morphogenesis and hearing loss. Our studies suggest that MAP3K1 has an essential role in the regulation of these key cochlear morphogenesis genes. Collectively, our data highlight the crucial role of MAP3K1 in the development and function of the mouse inner ear and hearing.

  4. Deletion of C7L and K1L Genes Leads to Significantly Decreased Virulence of Recombinant Vaccinia Virus TianTan

    OpenAIRE

    Zheng Liu; Shuhui Wang; Qicheng Zhang; Meijuan Tian; Jue Hou; Rongmin Wang; Chang Liu; Xu Ji; Ying Liu; Yiming Shao

    2013-01-01

    The vaccinia virus TianTan (VTT) has been modified as an HIV vaccine vector in China and has shown excellent performance in immunogenicity and safety. However, its adverse effects in immunosuppressed individuals warrant the search for a safer vector in the following clinic trails. In this study, we deleted the C7L and K1L genes of VTT and constructed six recombinant vaccinia strains VTT△C7L, VTT△K1L, VTT△C7LK1L, VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag. The pathogenicity and immunogenicity o...

  5. The Effect of Rumex K-1 on Milk Performance in Guanzhong Dairy Goats%鲁梅克斯K-1对关中奶山羊产奶性能的影响

    Institute of Scientific and Technical Information of China (English)

    李瑞香

    2012-01-01

    为了研究鲁梅克斯K-1对关中奶山羊产奶性能的影响.选取50只健康、体质量、产奶量接近(P>0.5)的关中奶山羊,试验共分5个处理,每个处理10只,各处理每只羊每天均饲喂3kg粗饲料和1kg精饲料,对照组粗饲料全喂玉米秸杆青贮,A、B、C、D组用鲁梅克斯青贮分别替代0.5、1、1.5、2 kg玉米秸杆青贮,进行为期50天泌乳试验.结果表明:(1)整个试验期,处理C组共产羊奶1850 kg,处理C组分别比CK、A、B、D组多产奶490、325、260、310 kg,差异极显著(P<0.01);(2)处理C组比CK、A、B、D组奶山羊的乳干物质、乳脂率、乳蛋白及乳糖含量均有所改善,但差异不显著(P>0.5);(3)C组的纯收入为5675元,C组分别比CK、A、B、D组多收入2035、1350、1065、1215元,差异极显著(P<0.01).鲁梅克斯营养丰富,与玉米秸杆混合青贮能有效提高关中奶山羊产奶量,改善乳蛋白、乳糖及乳脂率,其在粗饲料中添加量在50%为宜.%In this study the milk performance was tested from Guanzhong dairy goat fed with Rumex K-1 Fifty healthy Guanzhong dairy goats with a similar index on body weight and milk production (P>0.5) were randomly divided into five groups, 10 goats per group. Each of experimental group was fed coarse fodder (3 kg/d) with concentrated feed (1 kg/d), respectively. The goats in normal control group was fed silage maize straw as coarse fodder, and other A, B, C, D groups were fed silage maize straw combined with Rumex K-1 with four dosage of 0.5, 1, 1.5, 2 kg, respectively for 50 days. Results showed that: (1) Goats in C group had a niore milk production which totally produced 1850 kg milk during experimental time. Compared to the C group, the milk production had a significant decrease by about 490, 325, 260, 310 kg for CK, A, B and D (P 0.5). (3) Net income of goats in C group was 5675¥, which was reduced by about 2035¥, 1350¥, 1065¥ and 1215¥ for CK, A, B and D (P<0.01). These

  6. Innate sensing of microbial products promotes wound-induced skin cancer.

    Science.gov (United States)

    Hoste, Esther; Arwert, Esther N; Lal, Rohit; South, Andrew P; Salas-Alanis, Julio C; Murrell, Dedee F; Donati, Giacomo; Watt, Fiona M

    2015-01-09

    The association between tissue damage, chronic inflammation and cancer is well known. However, the underlying mechanisms are unclear. Here we characterize a mouse model in which constitutive epidermal extracellular-signal-regulated kinase-MAP-kinase signalling results in epidermal inflammation, and skin wounding induces tumours. We show that tumour incidence correlates with wound size and inflammatory infiltrate. Ablation of tumour necrosis factor receptor (TNFR)-1/-2, Myeloid Differentiation primary response gene 88 or Toll-like receptor (TLR)-5, the bacterial flagellin receptor, but not other innate immune sensors, in radiosensitive leukocytes protects against tumour formation. Antibiotic treatment inhibits, whereas injection of flagellin induces, tumours in a TLR-5-dependent manner. TLR-5 is also involved in chemical-induced skin carcinogenesis in wild-type mice. Leukocytic TLR-5 signalling mediates upregulation of the alarmin HMGB1 (High Mobility Group Box 1) in wound-induced papillomas. HMGB1 is elevated in tumours of patients with Recessive Dystrophic Epidermolysis Bullosa, a disease characterized by chronic skin damage. We conclude that in our experimental model the combination of bacteria, chronic inflammation and wounding cooperate to trigger skin cancer.

  7. Characteristics of bile salt hydrolase by Kluyveromyces marxianus K1 from Tibetan kefir%藏灵菇源克鲁维酵母K1菌株胆盐水解酶的特性研究

    Institute of Scientific and Technical Information of China (English)

    刘慧; 何欢; 熊利霞; 张雪娇; 张红星

    2010-01-01

    探讨藏灵菇马克斯克鲁维酵母K1菌株胆盐水解酶作用底物的反应条件与不同化学试剂对酶活性的影响及其发酵动力学类型.采用单因素多水平试验方法,在pH值4~8、温度31℃~43℃、底物浓度4mmol/L~8mmol/L及不同化学试剂(SDS、EDTA、尿素、Cu2+、Mg2+、Ca2+、F3+、Al3+、Mn2+)的条件下,胆盐水解酶与底物反应30min,检测酶活力;平板活菌计数法分析细胞生长与产酶的关系;双倒数作图法求得酶促反应动力学常数Km.结果表明,胆盐水解酶最适反应条件为:pH值为6.0、底物浓度7mmol/L、温度37℃,Fe3+、Ca2+、Mn2+及尿素对酶活性有较大提高作用,Mg2+、Al3+及SDS对酶的激活作用次之,Cu2+及EDTA对酶活性影响不大.K1菌株在18h~21h进入稳定期,于21h对数生长期的末期时酶活性达到最高,表明其胆盐水解酶发酵动力学类型为生长偶联合成型.其酶促反应动力学常数Km为2.10mmol/L,说明该酶与最适底物的亲和力较大.

  8. The Escherichia coli argW-dsdCXA genetic island is highly variable, and E. coli K1 strains commonly possess two copies of dsdCXA.

    Science.gov (United States)

    Moritz, Rebecca L; Welch, Rodney A

    2006-11-01

    The genome sequences of Escherichia coli pathotypes reveal extensive genetic variability in the argW-dsdCXA island. Interestingly, the archetype E. coli K1 neonatal meningitis strain, strain RS218, has two copies of the dsdCXA genes for d-serine utilization at the argW and leuX islands. Because the human brain contains d-serine, an epidemiological study emphasizing K1 isolates surveyed the dsdCXA copy number and function. Forty of 41 (97.5%) independent E. coli K1 isolates could utilize d-serine. Southern blot hybridization revealed physical variability within the argW-dsdC region, even among 22 E. coli O18:K1:H7 isolates. In addition, 30 of 41 K1 strains, including 21 of 22 O18:K1:H7 isolates, had two dsdCXA loci. Mutational analysis indicated that each of the dsdA genes is functional in a rifampin-resistant mutant of RS218, mutant E44. The high percentage of K1 strains that can use d-serine is in striking contrast to our previous observation that only 4 of 74 (5%) isolates in the diarrheagenic E. coli (DEC) collection have this activity. The genome sequence of diarrheagenic E. coli isolates indicates that the csrRAKB genes for sucrose utilization are often substituted for dsdC and a portion of dsdX present at the argW-dsdCXA island of extraintestinal isolates. Among DEC isolates there is a reciprocal pattern of sucrose fermentation versus d-serine utilization. The ability to use d-serine is a trait strongly selected for among E. coli K1 strains, which have the ability to infect a wide range of extraintestinal sites. Conversely, diarrheagenic E. coli pathotypes appear to have substituted sucrose for d-serine as a potential nutrient.

  9. Binding mode of an α-amino acid-linked quinoxaline-2,3-dione analogue at glutamate receptor subtype GluK1.

    Science.gov (United States)

    Demmer, Charles S; Møller, Charlotte; Brown, Patricia M G E; Han, Liwei; Pickering, Darryl S; Nielsen, Birgitte; Bowie, Derek; Frydenvang, Karla; Kastrup, Jette S; Bunch, Lennart

    2015-06-17

    Two α-amino acid-functionalized quinoxalines, 1a (CNG-10301) and 1b (CNG-10300), of a quinoxaline moiety coupled to an amino acid moiety were designed, synthesized, and characterized pharmacologically. While 1a displayed low affinity at native AMPA, KA, and NMDA receptors, and at homomeric GluK1,3 receptors, the affinity for GluK2 was in the midmicromolar range (Ki = 136 μM), 1b displayed low to midmicromolar range binding affinity at all the iGluRs (Ki = 9-126 μM). In functional experiments (outside-out patches excised from transfected HEK293T cells), 100 μM 1a partially blocked GluK1 (33% peak response), while GluK2 was unaffected (96% peak response). Furthermore, 1a was shown not to be an agonist at GluK1 and GluK2 at 100 μM. On the other hand, 100 μM 1b fully antagonized GluK1 (8% peak response) but only partially blocked GluK2 (33% peak response). An X-ray structure at 2.3 Å resolution of 1b in the GluK1-LBD (ligand-binding domain) disclosed an unexpected binding mode compared to the predictions made during the design phase; the quinoxaline moiety remains to act as an amino acid bioisostere, but the amino acid moiety is oriented into a new area within the GluK1 receptor. The structure of the GluK1-LBD with 1b showed a large variation in domain openings of the three molecules from 25° to 49°, demonstrating that the GluK1-LBD is capable of undergoing major domain movements.

  10. APMP key comparison for the measurement of air kerma for 60Co (APMP.RI(I)-K1.1)

    Science.gov (United States)

    Webb, D. V.; Lee, J.-H.; Budiantari, C. T.; Laban, J.; Saito, N.; Srimanoroth, S.; Khaled, N. E.

    2016-01-01

    The results are reported for an APMP.R(I)-K1.1 comparison that extends the regional comparison of standards for air kerma APMP.R(I)-K1 to several laboratories unable to participate earlier. The comparison was conducted with the goal of supporting the relevant calibration and measurement capabilities (CMCs) planned for publication by the participant laboratories. The comparison was conducted by the pilot laboratory, the Australian Radiation Protection and Nuclear Safety (ARPANSA), Australia, supported by the Institute of Nuclear Energy Research (INER), Taiwan, in a modified ring-shaped arrangement from September 2009 to November 2010, in parallel with an APMP.R(I)-K4 comparison being piloted by the INER. The laboratories that took part in the comparison were the ARPANSA, the Centre of Technology of Radiation Safety and Metrology (PTKMR-BATAN), Indonesia, the Division of Radiation and Medical Devices (DMSC), Thailand, the INER, the National Centre for Radiation Science (NCRS), New Zealand, the National Institute for Standards (NIS), Egypt and the National Metrology Institute of Japan (NMIJ/AIST), Japan. The two primary laboratories, ARPANSA and NMIJ, were chosen as the linking laboratories. Three ionization chambers were used as transfer instruments to be calibrated in terms of air kerma in 60Co radiotherapy beams. The comparison result is based on the ratio between the air kerma calibration coefficients (NK) determined by the participants and the mean of the results of the linking laboratories. The mean comparison ratio was found to be within 0.5 % of the key comparison reference value KCRV. The largest deviation between any two comparison ratios for the three chambers in terms of air kerma was 2.0 %. An analysis of the participant uncertainty budgets enabled the calculation of degrees of equivalence (DoE) in terms of the deviations of the results and their associated uncertainties. As a result of this APMP comparison, the BIPM key comparison database (KCDB) should

  11. Targeting MUC1-C inhibits the AKT-S6K1-elF4A pathway regulating TIGAR translation in colorectal cancer.

    Science.gov (United States)

    Ahmad, Rehan; Alam, Maroof; Hasegawa, Masanori; Uchida, Yasumitsu; Al-Obaid, Omar; Kharbanda, Surender; Kufe, Donald

    2017-02-02

    Colorectal cancer is third most common malignancy and is the second most common cause of cancer-related death. The MUC1 heterodimeric protein is aberrantly overexpressed in colorectal cancer and has been linked to poor outcomes in this disease. Here, we investigate the effects of the MUC1-C subunit inhibitor (GO-203), which disrupts MUC1-C homo-oligomerization, on human colorectal cancer cells. TIGAR mRNA level was determined using qRT-PCR. Western blotting was used to measure TIGAR protein level and AKT-mTOR-S6K1 pathways. Reactive oxygen species and apoptosis were measured by flow cytometry. Effect of MUC1-C peptide, GO-203 was studied on colorectal xenograft tumors. Immunohistochemistry was utilized for TIGAR staining. Treatment of MUC1-overexpressing SKCO-1 and Colo-205 colon cancer cells with GO-203 was associated with downregulation of the TP53-inducible glycolysis and apoptosis regulator (TIGAR) protein. TIGAR promotes the shunting of glycolytic intermediates into the pentose phosphate pathway and thus is of importance for maintaining redox balance. We show that GO-203-induced suppression of TIGAR is mediated by inhibition of AKT and the downstream mTOR pathway. The results also demonstrate that targeting MUC1-C blocks eIF4A cap-dependent translation of TIGAR. In concert with these results, GO-203-induced suppression of TIGAR was associated with decreases in GSH levels. GO-203 treatment also resulted in increases in reactive oxygen species (ROS) and loss of mitochondrial transmembrane potential. Consistent with these results, GO-203 inhibited the growth of colon cancer cells in vitro and as xenografts in nude mice. Inhibition of MUC1-C also downregulated TIGAR expression in xenograft tissues. These findings indicate that MUC1-C is a potential target for the treatment of colorectal cancer. Colorectal cancer patients who overexpress MUC1-C may be candidates for treatment with the MUC1-C inhibitor alone or in combination therapy with other agents.

  12. Interference harmonics and rigorous EM spectrum analysis method for low-k1 CD Bossung tilt correction

    Science.gov (United States)

    Chou, Shuo-Yen; Ng, Hoi-Tou; Chen, Yi-Yin; Lee, Chien-Fu; Liu, Ru-Gun; Gau, Tsai-Sheng

    2013-04-01

    This paper discusses the CD Bossung tilt phenomena in low-k1 lithography using interference harmonics and rigorous EM spectrum analysis. Interference harmonics analysis is introduced to explain the interaction of diffraction orders in the focal region leading to this abnormal CD behavior. This method decomposes the vector image formula into a superposition of cosine components to describe the interference of diffraction orders. The symmetry properties of components of an optical projection system were investigated to find out three potential sources for the asymmetric Bossung behavior, namely mask 3D (M3D) effect, lens aberration, and wafer reflectivity. Under good lens aberration and substrate reflectivity controls, the M3D effect accounts for most of the CD Bossung tilt. A rigorous EM mask spectral analysis was performed to reveal the impact of mask topography on the near-field intensity of mask transmission and the far-field image formation. From the analysis, the asymmetric phase distribution in the mask spectrum is the root cause for CD Bossung tilt. Using both the interference harmonics and the rigorous EM spectrum analysis, the effect of various resolution enhancement techniques (RET) to the Bossung tilt is also studied to find the best RET combination for M3D immunity. In addition, a pupil optimization algorithm based on these two analyses is proposed to generate the phase compensation map for M3D effect counteraction.

  13. LDA'+DMFT investigation of electronic structure of K1 - x Fe2 - y Se2 superconductor

    Science.gov (United States)

    Nekrasov, I. A.; Pavlov, N. S.; Sadovskii, M. V.

    2013-03-01

    We investigate electronic structure of the new iron chalcogenide high temperature superconductor K1- x Fe2- y Se2 (hole doped case with x = 0.24, y = 0.28) in the normal phase using the novel LDA'+DMFT computational approach. We show that this iron chalcogenide is more correlated in a sense of bandwidth renormalization (energy scale compression by factor about 5 in the interval ±1.5 eV), than typical iron pnictides (compression factor about 2), though the Coulomb interaction strength is almost the same in both families. Our results for spectral densities are in general agreement with recent ARPES data on this system. It is found that all Fe-3 d( t 2 g ) bands crossing the Fermi level have equal renormalization, in contrast to some previous interpretations. Electronic states at the Fermi level are of predominantly xy symmetry. Also we show that LDA'+DMFT results are in better agreement with experimental spectral function maps, than the results of conventional LDA+DMFT. Finally we make predictions for photoemission spectra lineshape for K0.76Fe1.72Se2.

  14. 大肠杆菌44277(02:K1)中rmlB基因功能%Function of rmlB in the pathogenic Escherichia coli 44277( O2: k1)

    Institute of Scientific and Technical Information of China (English)

    丁卫平; 罗元明; 艾国民; 金城

    2011-01-01

    [ Objective ] To identify the role of rmlB in synthesizing L-rhamnose in the pathogenic Escherichia coli 44277 (O2:K1:H4). [Methods] The rmlB gene was expressed and the activity of the recombinant protein was assayed by measuring the quantity of reaction product. The rmlB gene was deleted by homologous recombination, then phenotypic changes of the △rmlB mutant was analyzed by Electron Microscope, Tricine SDS-PAGE and immunological methods. Further, various methods including MALDI-TOF-MS/MS, GC-MS and NMR was used to investigate the 0 antigen structure of the ArmlB mutant. [Results] RmlB was confirmed to be a protein harboring the activity of dTDP-D-glucose 4,6-dehydratase through enzyme assay. The ArmlB mutant was successfully constructed and no phenotypic change was observed after compared with the wild type strain. L-rhamnose still existed in the ArmlB mutant, indicating that there may be isoenzyme of RmlB presenting in the mutant or there was a novel way synthesizing L-rhamnose in the mutant. [Conclusion] RmlB has the activity of dTDP-D-glucose 4,6-dehydratase but it is not essential for the synthesis of L-rhamnose.%[目的]确定rmlB基因在大肠杆菌(02:K1)L-型鼠李糖合成中的作用.[方法]将基因rmlB进行原核表达并测定酶活;用同源重组的方法将rmlB基因敲除,分析表型变化,并运用质谱,以及核磁共振等手段分析脂多糖0侧链的结构,以确定rmlB在O抗原合成中的作用.[结果]成功对rmlB基因进行了表达并测定了重组蛋白的酶活,确定蛋白RmlB具有dTDP-D-glucose 4,6-dehydratase活性.成功构建了rmlB基因缺失突变株,对突变株进行表型分析发现突变株的表型与野生株相比无变化.对突变株分析发现突变株中的O抗原仍含有L-型鼠李糖,说明在该菌株中可能存在RmlB的同功能酶或者存在其它的L-型鼠李糖合成途径.[结论]rmlB基因编码的蛋白具有dTDP-D-glucose 4,6-dehydratase活性但此基因对于L-型鼠李糖的合成不是必需的.

  15. Comprehensive gene and microRNA expression profiling reveals a role for miRNAs in the oncogenic roles of SphK1 in papillary thyroid cancer.

    Science.gov (United States)

    Liang, Weiwei; Xie, Zhiwei; Cui, Weiling; Guo, Yan; Xu, Lijuan; Wu, Jueheng; Guan, Hongyu

    2017-04-01

    The oncogenic roles of sphingosine kinase 1 (SphK1) in various cancers, including thyroid cancer, have been well demonstrated. However, the microRNAs (miRNAs) associated with the oncogenic roles of SphK1 remain largely unknown. Global gene and miRNA expression in TPC1-Vector and TPC1-SphK1 cells was analyzed using digital gene expression (DGE) analysis and small RNA-seq, respectively. miRNA-mRNA interactions were explored by microT-CDS, and the predicted networks were visualized using CytoScape(®). Cell invasion and migration were assessed by performing Transwell invasion and wound-healing assays. Luciferase reporter and immunoblot assays were used to evaluate the targeting of fibronectin 1 (FN1) by miR-144-3p. In this study, we found that overexpression of SphK1 differentially regulates the expression of 46 miRNAs and 506 mRNAs in papillary thyroid cancer (PTC) TPC1 cells. Combining bioinformatics predictions of mRNA targets with DGE data on mRNA expression allowed us to identify the mRNA targets of deregulated miRNAs. The direct interaction between miR-144-3p and FN1, which mediates the pro-invasive role of SphK1 in PTC cells, was experimentally validated. Our results demonstrated that SphK1 overexpression drives a regulatory network governing miRNA and mRNA expression in PTC cells. We also demonstrated the roles played by miR-144-3p and FN1 in mediating the oncogenic function of SphK1, which enhanced the understanding of the etiology of PTC.

  16. Mutations in Argonaute5 Illuminate Epistatic Interactions of the K1 and I Loci Leading to Saddle Seed Color Patterns in Glycine max.

    Science.gov (United States)

    Cho, Young B; Jones, Sarah I; Vodkin, Lila O

    2017-04-01

    The soybean (Glycine max) seed coat has distinctive, genetically programmed patterns of pigmentation, and the recessive k1 mutation can epistatically overcome the dominant I and i(i) alleles, which inhibit seed color by producing small interfering RNAs (siRNAs) targeting chalcone synthase (CHS) mRNAs. Small RNA sequencing of dissected regions of immature seed coats demonstrated that CHS siRNA levels cause the patterns produced by the i(i) and i(k) alleles of the I locus, which restrict pigment to the hilum or saddle region of the seed coat, respectively. To identify the K1 locus, we compared RNA-seq data from dissected regions of two Clark isolines having similar saddle phenotypes mediated by CHS siRNAs but different genotypes (homozygous i(k) K1 versus homozygous i(i) k1). By examining differentially expressed genes, mapping information, and genome resequencing, we identified a 129-bp deletion in Glyma.11G190900 encoding Argonaute5 (AGO5), a member of the Argonaute family. Amplicon sequencing of several independent saddle pattern mutants from different genetic backgrounds revealed independent lesions affecting AGO5, thus establishing Glyma.11G190900 as the K1 locus. Nonfunctional AGO5 from k1 alleles leads to altered distributions of CHS siRNAs, thus explaining how the k1 mutation reverses the phenotype of the seed coat regions from yellow to pigmented, even in the presence of the normally dominant I or i(i) alleles. © 2017 American Society of Plant Biologists. All rights reserved.

  17. Fast intracellular dissolution and persistent cellular uptake of silver nanoparticles in CHO-K1 cells: implication for cytotoxicity.

    Science.gov (United States)

    Jiang, Xiumei; Miclăuş, Teodora; Wang, Liming; Foldbjerg, Rasmus; Sutherland, Duncan S; Autrup, Herman; Chen, Chunying; Beer, Christiane

    2015-03-01

    Toxicity of silver nanoparticles (Ag NPs) has been reported both in vitro and in vivo. However, the intracellular stability and chemical state of Ag NPs are still not very well studied. In this work, we systematically investigated the cellular uptake pathways, intracellular dissolution and chemical species, and cytotoxicity of Ag NPs (15.9 ± 7.6 nm) in Chinese hamster ovary cell subclone K1 cells, a cell line recommended by the OECD for genotoxicity studies. Quantification of intracellular nanoparticle uptake and ion release was performed through inductively coupled plasma mass spectrometry. X-ray absorption near-edge structure (XANES) was employed to assess the chemical state of intracellular silver. The toxic potential of Ag NPs and Ag(+) was evaluated by cell viability, reactive oxygen species (ROS) production and live-dead cell staining. The results suggest that cellular uptake of Ag NPs involves lipid-raft-mediated endocytosis and energy-independent diffusion. The degradation study shows that Ag NPs taken up into cells dissolved quickly and XANES results directly indicated that the internalized Ag was oxidized to Ag-O- species and then stabilized in silver-sulfur (Ag-S-) bonds within the cells. Subsequent cytotoxicity studies show that Ag NPs decrease cell viability and increase ROS production. Pre-incubation with N-acetyl-L-cysteine, an efficient antioxidant and Ag(+) chelator, diminished the cytotoxicity caused by Ag NPs or Ag(+) exposure. Our study suggests that the cytotoxicity mechanism of Ag NPs is related to the intracellular release of silver ions, followed by their binding to SH-groups, presumably coming from amino acids or proteins, and affecting protein functions and the antioxidant defense system of cells.

  18. Effects of lunar and mars dust simulants on HaCaT keratinocytes and CHO-K1 fibroblasts

    Science.gov (United States)

    Rehders, Maren; Grosshäuser, Bianka B.; Smarandache, Anita; Sadhukhan, Annapurna; Mirastschijski, Ursula; Kempf, Jürgen; Dünne, Matthias; Slenzka, Klaus; Brix, Klaudia

    2011-04-01

    Exposure to lunar dust during Apollo missions resulted in occasional reports of ocular, respiratory and dermal irritations which showed that lunar dust has a risk potential for human health. This is caused by its high reactivity as well as its small size, leading to a wide distribution also inside habitats. Hence, detailed information regarding effects of extraterrestrial lunar dusts on human health is required to best support future missions to moon, mars or other destinations. In this study, we used several methods to assess the specific effects of extraterrestrial dusts onto mammalian skin by exposing HaCaT keratinocytes and CHO-K1 fibroblasts to dusts simulating lunar or mars soils. These particular cell types were chosen because the skin protects the human body from potentially harmful substances and because a well orchestrated program ensures proper wound healing. Keratinocytes and fibroblasts were exposed to the dusts for different durations of time and their effects on morphology and viability of the cells were determined. Cytotoxicity was measured using the MTT assay and by monitoring culture impedance, while phalloidin staining of the actin cytoskeleton was performed to address structural integrity of the cells which was also investigated by propidium iodide intake. It was found that the effects of the two types of dust simulants on the different features of both cell lines varied to a considerable extent. Moreover, proliferation of HaCaT keratinocytes, as analyzed by Ki67 labeling, was suppressed in sub-confluent cultures exposed to lunar dust simulant. Furthermore, experimental evidence is provided for a delay in regeneration of keratinocyte monolayers from scratch-wounding when exposed to lunar dust simulant. The obtained results will facilitate further investigations of dust exposure during wound healing and will ease risk assessment studies e.g., for lunar lander approaches. The investigations will help to determine safety measures to be taken during

  19. Effects of lunar and mars dust on HaCaT keratinocytes and CHO-K1 fibroblasts

    Science.gov (United States)

    Brix, Klaudia; Slenzka, Klaus; Rehders, Maren; Sadhukhan, Annapurna; Mistry, Rima; Duenne, Matthias; Kempf, Juergen

    Exposure to lunar dust during Apollo missions resulted in occasional reports of ocular, respira-tory and dermal irritations which showed that lunar dust has a risk potential for human health. This is caused by its high reactivity as well as its small size, leading to a wide distribution also inside habitats. Hence, detailed information regarding effects of lunar dust on human health is required to best support future missions to moon. In this study, we used different methods to assess the specific effects of lunar dust onto mammalian skin by exposing HaCaT keratinocytes and CHO-K1 fibroblasts to dusts simulating lunar or mars soils. These particular cell types were chosen because the skin protects the human body from potentially harmful substances and since a well orchestrated program ensures proper repair in cases of wounding. Keratinocytes and fibroblasts were exposed to the dusts for different durations of time and their effects on morphology, metabolic state, survival and proliferation of the cells were determined. Cytotoxi-city and proliferation were measured using the MTT assay, metabolic activity was analyzed by vital staining of mitochondria, and phalloidin staining of the actin cytoskeleton was performed to address structural integrity of the cells. It was found that the effects of the two types of soils on the different features of both cell lines varied to considerable extent, and that lunar and mars dust were specific in their effects. The obtained results will facilitate detailed inves-tigations of dust exposure during wound healing and will ease risk assessment studies for e.g. lunar lander approaches. The investigations will help to assess the risks and to determine safety measures to be taken during extraterrestrial expeditions in order to minimize risks to human health associated with exposure of human skin to dust contaminants.

  20. KEY COMPARISON: Final report on key comparison APMP.QM-K1.d: sulphur dioxide in nitrogen

    Science.gov (United States)

    Kato, Kenji; Maruyama, Masaaki; Seog Kim, Jin; Hyub Oh, Sang; Kim, Byung Moon; Han, Qiao; Zhou, Zeyi

    2008-01-01

    The aim of this comparison was to compare capabilities for the preparation and value assignment of gas mixture standards for sulphur dioxide in air (or nitrogen), maintained at national metrology institutes (NMIs). The measurements in this key comparison took place in 2006. There were three participants and one coordinating laboratory. The key comparison reference value (KCRV) was based on the gravimetric preparation for all components. The range of the nominal amount-of-substance fractions of the comparison standard is 90 µmol/mol to 100 µmol/mol. The Chemical Evaluation Research Institute (CERI, Japan) prepared gravimetric mixture samples of sulfur dioxide in nitrogen in 10 L aluminium cylinders with a passivated inner surface. The homogeneity and long term stability of these samples were evaluated at CERI before and after the shipment of the cylinders to the participants. The uncertainty of the standard gases including the long term stability was estimated to be 0.1 %. Each participating laboratory was shipped one cylinder to be analyzed and then returned to the CERI. Three participants submitted results that were within 0.6% of the relevant reference value. In these cases, the estimated uncertainties were larger than the deviation from the reference value. The agreement of the results in this key comparison is good. CERI is the link laboratory between CCQM-K1.d and APMP.QM-K4. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  1. Investigation of superparamagnetic (Fe3O4) nanoparticles and magnetic field exposures on CHO-K1 cell line

    Science.gov (United States)

    Coker, Zachary; Estlack, Larry; Hussain, Saber; Choi, Tae-Youl; Ibey, Bennett L.

    2016-03-01

    Rapid development in nanomaterial synthesis and functionalization has led to advanced studies in actuation and manipulation of cellular functions for biomedical applications. Often these actuation techniques employ externally applied magnetic fields to manipulate magnetic nanomaterials inside cell bodies in order to drive or trigger desired effects. While cellular interactions with low-frequency magnetic fields and nanoparticles have been extensively studied, the fundamental mechanisms behind these interactions remain poorly understood. Additionally, modern investigations on these concurrent exposure conditions have been limited in scope, and difficult to reproduce. This study presents an easily reproducible method of investigating the biological impact of concurrent magnetic field and nanoparticle exposure conditions using an in-vitro CHO-K1 cell line model, with the purpose of establishing grounds for in-depth fundamental studies of the mechanisms driving cellular-level interactions. Cells were cultured under various nanoparticle and magnetic field exposure conditions from 0 to 500 μg/ml nanoparticle concentrations, and DC, 50 Hz, or 100 Hz magnetic fields with 2.0 mT flux density. Cells were then observed by confocal fluorescence microscopy, and subject to biological assays to determine the effects of concurrent extreme-low frequency magnetic field and nanoparticle exposures on cellnanoparticle interactions, such as particle uptake and cell viability by MTT assay. Current results indicate little to no variation in effect on cell cultures based on magnetic field parameters alone; however, it is clear that deleterious synergistic effects of concurrent exposure conditions exist based on a significant decrease in cell viability when exposed to high concentrations of nanoparticles and concurrent magnetic field.

  2. Selective Modulation of Protein Kinase C α over Protein Kinase C ε by Curcumin and Its Derivatives in CHO-K1 Cells.

    Science.gov (United States)

    Pany, Satyabrata; Majhi, Anjoy; Das, Joydip

    2016-04-12

    Members of the protein kinase C (PKC) family of serine/threonine kinases regulate various cellular functions, including cell growth, differentiation, metabolism, and apoptosis. Modulation of isoform-selective activity of PKC by curcumin (1), the active constituent of Curcuma L., is poorly understood, and the literature data are inconsistent and obscure. The effect of curcumin (1) and its analogues, 4-[(2Z,6E)-3-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-5-oxohepta-2,6-dien-1-yl]-2-methoxyphenyl oleate (2), (9Z,12Z)-4-[(2Z,6E)-3-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-5-oxohepta-2,6-dien-1-yl]-2-methoxyphenyl octadeca-9,12-dienoate (3), (9Z,12Z,15Z)-4-[(2Z,6E)-3-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-5-oxohepta-2,6-dien-1-yl]-2-methoxyphenyl octadeca-9,12,15-trienoate (4), and (1E,6E)-1-[4-(hexadecyloxy)-3-methoxyphenyl]-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (5), and didemethylcurcumin (6) on the membrane translocation of PKCα, a conventional PKC, and PKCε, a novel PKC, has been studied in CHO-K1 cells, in which these PKC isoforms are endogenously expressed. Translocation of PKC from the cytosol to the membrane was measured using immunoblotting and confocal microscopy. 1 and 6 inhibited the TPA-induced membrane translocation of PKCα but not of PKCε. Modification of the hydroxyl group of curcumin with a long aliphatic chain containing unsaturated double bonds in 2-4 completely abolished this inhibition property. Instead, 2-4 showed significant translocation of PKCα but not of PKCε to the membrane. No membrane translocation was observed with 1, 6, or the analogue 5 having a saturated long chain for either PKCα or PKCε. 1 and 6 inhibited TPA-induced activation of ERK1/2, and 2-4 activated it. ERK1/2 is the downstream readout of PKC. These results show that the hydroxyl group of curcumin is important for PKC activity and the curcumin template can be useful in developing isoform specific PKC modulators for regulating a particular disease state.

  3. Biophysical characterization of inwardly rectifying potassium currents (I(K1) I(K,ACh), I(K,Ca)) using sinus rhythm or atrial fibrillation action potential waveforms

    DEFF Research Database (Denmark)

    Tang, Chuyi; Skibsbye, Lasse; Yuan, Lei

    2015-01-01

    to voltage protocols adapted from atrial action potentials recorded in human tissue at 1 and 3 Hz. The current recordings were performed in the HEK-293 heterologous cell system expressing either I(K1), I(K,ACh) or I(K,Ca) to establish the individual contribution of each of these currents during the voltage...... changes of atrial action potential waveforms. I(K1) primarily contributes to the atrial electrophysiology at the latter part of repolarization and during the diastolic phase, while both I(K,Ca) under high [Ca2+]i and I(K,ACh) contribute relatively most during repolarization.......Although several physiological, pathophysiological and regulatory properties of classical inward rectifier K+ current I(K1), G-protein coupled inwardly-rectifying K+ current I(K,ACh) and the small-conductance Ca2+ activated K+ current I(K,Ca) have been identified, quantitative biophysical details...

  4. Deletion of C7L and K1L genes leads to significantly decreased virulence of recombinant vaccinia virus TianTan.

    Science.gov (United States)

    Liu, Zheng; Wang, Shuhui; Zhang, Qicheng; Tian, Meijuan; Hou, Jue; Wang, Rongmin; Liu, Chang; Ji, Xu; Liu, Ying; Shao, Yiming

    2013-01-01

    The vaccinia virus TianTan (VTT) has been modified as an HIV vaccine vector in China and has shown excellent performance in immunogenicity and safety. However, its adverse effects in immunosuppressed individuals warrant the search for a safer vector in the following clinic trails. In this study, we deleted the C7L and K1L genes of VTT and constructed six recombinant vaccinia strains VTT△C7L, VTT△K1L, VTT△C7LK1L, VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag. The pathogenicity and immunogenicity of these recombinants were evaluated in mouse and rabbit models. Comparing to parental VTT, VTT△C7L and VTT△K1L showed significantly decreased replication capability in CEF, Vero, BHK-21 and HeLa cell lines. In particular, replication of VTT△C7LK1L decreased more than 10-fold in all four cell lines. The virulence of all these mutants were decreased in BALB/c mouse and rabbit models; VTT△C7LK1L once again showed the greatest attenuation, having resulted in no evident damage in mice and erythema of only 0.4 cm diameter in rabbits, compared to 1.48 cm for VTT. VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag elicited as strong cellular and humoral responses against HIV genes as did VTKgpe, while humoral immune response against the vaccinia itself was reduced by 4-8-fold. These data show that deletion of C7L and K1L genes leads to significantly decreased virulence without compromising animal host immunogenicity, and may thus be key to creating a more safe and effective HIV vaccine vector.

  5. Deletion of C7L and K1L genes leads to significantly decreased virulence of recombinant vaccinia virus TianTan.

    Directory of Open Access Journals (Sweden)

    Zheng Liu

    Full Text Available The vaccinia virus TianTan (VTT has been modified as an HIV vaccine vector in China and has shown excellent performance in immunogenicity and safety. However, its adverse effects in immunosuppressed individuals warrant the search for a safer vector in the following clinic trails. In this study, we deleted the C7L and K1L genes of VTT and constructed six recombinant vaccinia strains VTT△C7L, VTT△K1L, VTT△C7LK1L, VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag. The pathogenicity and immunogenicity of these recombinants were evaluated in mouse and rabbit models. Comparing to parental VTT, VTT△C7L and VTT△K1L showed significantly decreased replication capability in CEF, Vero, BHK-21 and HeLa cell lines. In particular, replication of VTT△C7LK1L decreased more than 10-fold in all four cell lines. The virulence of all these mutants were decreased in BALB/c mouse and rabbit models; VTT△C7LK1L once again showed the greatest attenuation, having resulted in no evident damage in mice and erythema of only 0.4 cm diameter in rabbits, compared to 1.48 cm for VTT. VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag elicited as strong cellular and humoral responses against HIV genes as did VTKgpe, while humoral immune response against the vaccinia itself was reduced by 4-8-fold. These data show that deletion of C7L and K1L genes leads to significantly decreased virulence without compromising animal host immunogenicity, and may thus be key to creating a more safe and effective HIV vaccine vector.

  6. PPIP5K1 modulates ligand competition between diphosphoinositol polyphosphates and PtdIns(3,4,5)P3 for polyphosphoinositide-binding domains.

    Science.gov (United States)

    Gokhale, Nikhil A; Zaremba, Angelika; Janoshazi, Agnes K; Weaver, Jeremy D; Shears, Stephen B

    2013-08-01

    We describe new signalling consequences for PPIP5K1 (diphosphoinositol pentakisphosphate kinase type 1)-mediated phosphorylation of InsP6 and 5-InsP7 to 1-InsP7 and InsP8. In NIH 3T3 cells, either hyperosmotic stress or receptor activation by PDGF (platelet-derived growth factor) promoted translocation of PPIP5K1 from the cytoplasm to the plasma membrane. The PBD1 (polyphosphoinositide-binding domain) in PPIP5K1 recapitulated that translocation. Mutagenesis of PBD1 to reduce affinity for PtdIns(3,4,5)P3 prevented translocation. Using surface plasmon resonance, we found that PBD1 association with vesicular PtdIns(3,4,5)P3 was inhibited by InsP6 and diphosphoinositol polyphosphates. However, the inhibition by PPIP5K1 substrates (IC50: 5-InsP7=5 μM and InsP6=7 μM) was substantially more potent than that of the PPIP5K1 products (IC50: InsP8=32 μM and 1-InsP7=43 μM). This rank order of ligand competition with PtdIns(3,4,5)P3 was also exhibited by the PH (pleckstrin homology) domains of Akt (also known as protein kinase B), GRP1 (general receptor for phosphoinositides 1) and SIN1 (stress-activated protein kinase-interaction protein 1). We propose that, in vivo, PH domain binding of InsP6 and 5-InsP7 suppresses inappropriate signalling ('noise') from stochastic increases in PtdIns(3,4,5)P3. That restraint may be relieved by localized depletion of InsP6 and 5-InsP7 at the plasma membrane following PPIP5K1 recruitment. We tested this hypothesis in insulin-stimulated L6 myoblasts, using mTOR (mechanistic/mammalian target of rapamycin)-mediated phosphorylation of Akt on Ser473 as a readout for SIN1-mediated translocation of mTORC (mTOR complex) 2 to the plasma membrane [Zoncu, Efeyan and Sabatini (2011) Nat. Rev. Mol. Cell Biol. 12, 21-35]. Knockdown of PPIP5K1 expression was associated with a 40% reduction in Ser473 phosphorylation. A common feature of PtdIns(3,4,5)P3-based signalling cascades may be their regulation by PPIP5K1.

  7. A liver abscess deprived a healthy adult of eyesight: endogenous endophthalmitis associated with a pyogenic liver abscess caused by serotype K1 Klebsiella pneumonia.

    Science.gov (United States)

    Maruno, Takahisa; Ooiwa, Yoko; Takahashi, Ken; Kodama, Yuzo; Takakura, Shunji; Ichiyama, Satoshi; Chiba, Tsutomu

    2013-01-01

    Klebsiella pneumonia usually causes urinary tract infections, pneumonia, and other infectious diseases in hospitalized and immunocompromised patients. Among the types of Klebsiella pneumonia, serotype K1 is known to be a highly virulent pathogen. We herein report the case of a healthy 63-year-old man with a pyogenic liver abscess and bilateral endogenous endophthalmitis caused by serotype K1 Klebsiella pneumonia. Although the patient received percutaneous abscess drainage and antibiotic therapy, he lost his eyesight. To improve the poor prognoses of ocular complications, providing both an earlier diagnosis and treatment is critical.

  8. Rapidly fatal bacteremic pneumonia caused by Klebsiella pneumoniae with K1 hypermucoviscosity phenotype in a previously healthy young man receiving levofloxacin treatment.

    Science.gov (United States)

    Chuang, Tzu-Yi; Lin, Chou-Jui; Chi, Chun-Lin; Liu, An-Yu; Lee, Shih-Wei; Lin, T L; Wang, Jin-Town; Hsueh, Po-Ren

    2009-10-01

    Fatal bacteremic Klebsiella pneumoniae pneumonia is commonly encountered in alcoholic and diabetic patients. This report describes a previously healthy young man with rapidly fatal bacteremic pneumonia caused by K. pneumoniae serotype K1, complicated by septic shock and multiple organ dysfunction.

  9. RELATIONSHIP BETWEEN ANTIOXIDANT POWER OF PLASMA WITH LIPID PEROXIDE FORMATION IN PLASMA AND LIVER DAMAGES CAUSED BY OVERDOSE OF VITAMIN K1 IN ADULT AND WEANLING RATS

    Directory of Open Access Journals (Sweden)

    H. Ansari Hadipour

    2003-08-01

    Full Text Available In this study the plasma levels of lipid peroxidation (LP products, protein carbonyls and antioxidant capacity of plasma as judged by ferric reducing ability of plasma (FRAP assay were compared in adult and weanling rats treated with vitamin K1 phylloquinone.

  10. Analysis of the capsular polysaccharide biosynthesis locus of Porphyromonas gingivalis and development of a K1-specific polymerase chain reaction-based serotyping assay

    NARCIS (Netherlands)

    Brunner, J.; Crielaard, W.; Winkelhoff A.J. van

    2008-01-01

    Background and Objective: Porphyromonas gingivalis is a gram-negative obligate anaerobe that is strongly associated with severe periodontitis. Previous reports showed an association of P. gingivalis capsular polysaccharide with virulence. The K1 capsular polysaccharide was found to be more immunosti

  11. Binding Mode of an α-Amino Acid-Linked Quinoxaline-2,3-dione Analogue at Glutamate Receptor Subtype GluK1

    DEFF Research Database (Denmark)

    Demmer, Charles S; Møller, Charlotte; Brown, Patricia M G E;

    2015-01-01

    Two α-amino acid-functionalized quinoxalines, 1a (CNG-10301) and 1b (CNG-10300), of a quinoxaline moiety coupled to an amino acid moiety were designed, synthesized, and characterized pharmacologically. While 1a displayed low affinity at native AMPA, KA, and NMDA receptors, and at homomeric GluK1,...

  12. Whole-Genome Sequences of the Archetypal K1 Escherichia coli Neonatal Isolate RS218 and Contemporary Neonatal Bacteremia Clinical Isolates SCB11, SCB12, and SCB15.

    Science.gov (United States)

    Day, Michael W; Jackson, Lydgia A; Akins, Darrin R; Dyer, David W; Chavez-Bueno, Susana

    2015-02-26

    Neonatal bacteremia Escherichia coli strains commonly belong to the K1 capsular type. Their ability to cause invasive neonatal disease appears to be determined by other virulence factors that have yet to be identified. We report here the genome sequences of four E. coli neonatal bacteremia isolates, including that of the archetypal strain RS218.

  13. Mode of Action Temu Kunci (Kaempferia pandurata Essential Oil on E. coli K1.1 Cell Determined by Leakage of Material Cell and Salt Tolerance Assays

    Directory of Open Access Journals (Sweden)

    MIKSUSANTI

    2008-06-01

    Full Text Available The essential oil of Kaempferia pandurata consist of terpen and oxygenated terpen that exhibits broad-spectrum antimicrobial activity. It’s mode of action against the gram-negative bacterium E. coli K1.1 has been investigated using a range of treatments. The mode action of the essential oil were analyzed by it’s ability to leakage E. coli K1.1 cell, to change permeability of the cell, and to alter salt tolerance of the cell. Ion leakage from the cell were analyzed by atomic absorption spectrophotometer. Salt tolerance assays was conducted by investigating the ability of E. coli K1.1 treated with temu kunci essential oil to grow on NA supplemented with NaCl. Protein and acid nucleic leakage were analyzed by UV spectrophotometer. There were inorganic compound leakage (potassium, calcium ion and organic compound leakage (nucleic acid, protein from cytoplasmic membrane, after exposing this organism to essential oil of Kaempferia pandurata. The more concentration of oil added, the more leakage was observed due to the loss of absorbing material such as nucleic acid (260 nm and protein (280 nm, the loss of potassium and calcium ion, and loss of the salt tolerance of E. coli K1.1.

  14. Expression of S6K1 in human visceral adipose tissue is upregulated in obesity and related to insulin resistance and inflammation.

    Science.gov (United States)

    Catalán, Victoria; Gómez-Ambrosi, Javier; Rodríguez, Amaia; Ramírez, Beatriz; Andrada, Patricia; Rotellar, Fernando; Valentí, Víctor; Moncada, Rafael; Martí, Pablo; Silva, Camilo; Salvador, Javier; Frühbeck, Gema

    2015-04-01

    The ribosomal protein S6 kinase 1 (S6K1) is a component of the insulin signalling pathway that has been proposed as a key molecular factor in insulin resistance development under conditions of nutrient overload. The aim was to evaluate the involvement of S6K1 in obesity as well as to explore their association with visceral adipose tissue (VAT) inflammation. Samples obtained from 40 subjects were used. Gene expression levels of RPS6KB1 and key inflammatory markers were analysed in VAT. The effect of insulin on transcript levels of RPS6KB1 in human differentiated adipocytes was also explored. RPS6KB1 mRNA levels in VAT were increased (P obese patients. Insulin treatment significantly enhanced (P insulin resistance was observed. Moreover, RPS6KB1 gene expression levels were positively correlated with VAT gene expression levels of the inflammatory markers CCL2, CD68, MMP2, MMP9, VEGFA and CHI3L1 as well as with mRNA levels of MTOR and MAPK8, representative players involved in signalling pathways related to S6K1. The increased levels of S6K1 in obesity and its positive association with insulin resistance and inflammation suggest a role for this protein in the changes that take place in VAT in obesity establishing a link between inflammation and a higher risk for the development of metabolic diseases.

  15. Analysis of vitamin K-1 in fruits and vegetables using accelerated solvent extraction and liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization

    DEFF Research Database (Denmark)

    Jäpelt, Rie Bak; Jakobsen, Jette

    2016-01-01

    spectrometry with atmospheric pressure chemical ionization in selected reaction monitoring mode with deuterium-labeled vitamin K1 as an internal standard. The precision was estimated as the pooled estimate of three replicates performed on three different days for spinach, peas, apples, banana, and beetroot...

  16. Development and validation of a high-content screening in vitro micronucleus assay in CHO-k1 and HepG2 cells

    NARCIS (Netherlands)

    Westerink, W.M.; Schirris, T.J.J.; Horbach, G.J.; Schoonen, W.G.

    2011-01-01

    In the present study an automated image analysis assisted in vitro micronucleus assay was developed with the rodent cell line CHO-k1 and the human hepatoma cell line HepG2, which are both commonly used in regulatory genotoxicity assays. The HepG2 cell line was chosen because of the presence in these

  17. Emergence of KPC-producing Klebsiella pneumoniae hypervirulent clone of capsular serotype K1 that belongs to sequence type 11 in Mainland China.

    Science.gov (United States)

    Wei, Dan-Dan; Wan, La-Gen; Deng, Qiong; Liu, Yang

    2016-06-01

    KPC-2 has been rarely reported in hypervirulent Klebsiella pneumoniae strains. Here, we describe a KPC-2-producing K. pneumoniae hypervirulent clone of capsular serotype K1 belonging to sequence type 11. The presence of KPC carbapenemase in hypervirulent clone could mark an evolutionary step toward its establishment as major nosocomial pathogen.

  18. Development and validation of a high-content screening in vitro micronucleus assay in CHO-k1 and HepG2 cells

    NARCIS (Netherlands)

    Westerink, W.M.; Schirris, T.J.J.; Horbach, G.J.; Schoonen, W.G.

    2011-01-01

    In the present study an automated image analysis assisted in vitro micronucleus assay was developed with the rodent cell line CHO-k1 and the human hepatoma cell line HepG2, which are both commonly used in regulatory genotoxicity assays. The HepG2 cell line was chosen because of the presence in these

  19. Legionella pneumophila Strain 130b Evades Macrophage Cell Death Independent of the Effector SidF in the Absence of Flagellin

    Science.gov (United States)

    Speir, Mary; Vogrin, Adam; Seidi, Azadeh; Abraham, Gilu; Hunot, Stéphane; Han, Qingqing; Dorn, Gerald W.; Masters, Seth L.; Flavell, Richard A.; Vince, James E.; Naderer, Thomas

    2017-01-01

    The human pathogen Legionella pneumophila must evade host cell death signaling to enable replication in lung macrophages and to cause disease. After bacterial growth, however, L. pneumophila is thought to induce apoptosis during egress from macrophages. The bacterial effector protein, SidF, has been shown to control host cell survival and death by inhibiting pro-apoptotic BNIP3 and BCL-RAMBO signaling. Using live-cell imaging to follow the L. pneumophila-macrophage interaction, we now demonstrate that L. pneumophila evades host cell apoptosis independent of SidF. In the absence of SidF, L. pneumophila was able to replicate, cause loss of mitochondria membrane potential, kill macrophages, and establish infections in lungs of mice. Consistent with this, deletion of BNIP3 and BCL-RAMBO did not affect intracellular L. pneumophila replication, macrophage death rates, and in vivo bacterial virulence. Abrogating mitochondrial cell death by genetic deletion of the effectors of intrinsic apoptosis, BAX, and BAK, or the regulator of mitochondrial permeability transition pore formation, cyclophilin-D, did not affect bacterial growth or the initial killing of macrophages. Loss of BAX and BAK only marginally limited the ability of L. pneumophila to efficiently kill all macrophages over extended periods. L. pneumophila induced killing of macrophages was delayed in the absence of capsase-11 mediated pyroptosis. Together, our data demonstrate that L. pneumophila evades host cell death responses independently of SidF during replication and can induce pyroptosis to kill macrophages in a timely manner. PMID:28261564

  20. Flax lignan concentrate attenuate hypertension and abnormal left ventricular contractility via modulation of endogenous biomarkers in two-kidney-one-clip (2K1C hypertensive rats

    Directory of Open Access Journals (Sweden)

    Sameer Hanmantrao Sawant

    Full Text Available ABSTRACT The present investigation was designed to study the effect of flax lignan concentrate obtained from Linum usitatissimum L., Linaceae, in two-kidney, one clip (2K1C hypertension model in Wistar rats. 2K1C Goldblatt model rats were divided randomly into six groups: sham, 2K1C control, captopril (30 mg/kg, flax lignan concentrate (200, 400 and 800 mg/kg. Flax lignan concentrate and captopril were administered daily for eight consecutive weeks. Sham-operated, and 2K1C control rats received the vehicle. Treatment with flax lignan concentrate (400 and 800 mg/kg significantly and dose-dependently restored the hemodynamic parameters systolic blood pressure, diastolic blood pressure, mean arterial blood pressure and left ventricular functions. The flax lignan concentrate significantly restored the elevated hepatic, renal and cardiac marker enzymes in the serum. It also restored the organs weights (kidney and heart, serum electrolyte level and histological abnormalities. Furthermore, flax lignan concentrate significantly elevated the level of biochemical markers that is enzymatic antioxidants superoxide dismutase, glutathione and decreased malondialdehyde in the heart and kidney tissues. Meanwhile, we found that plasma nitric oxide and plasma nitric oxide synthase contents were significantly increased in the flax lignan concentrate-treated group, and plasma endothelin-1 and renal angiotensin-II levels were significantly lower than 2K1C hypertensive group. In conclusion, the antihypertensive and antioxidant effect of flax lignan concentrate were dose-dependent and at the highest dose (i.e. 800 mg/kg similar to those of captopril (30 mg/kg. It is suggested that flax lignan concentrate reduced blood pressure by reduction of renal angiotensin-II level, inhibition of plasma endothelin-1 production, induction of the nitric oxide, nitric oxide synthase and in vivo antioxidant defense system.

  1. Bioluminescent imaging reveals novel patterns of colonization and invasion in systemic Escherichia coli K1 experimental infection in the neonatal rat.

    Science.gov (United States)

    Witcomb, Luci A; Collins, James W; McCarthy, Alex J; Frankel, Gadi; Taylor, Peter W

    2015-12-01

    Key features of Escherichia coli K1-mediated neonatal sepsis and meningitis, such as a strong age dependency and development along the gut-mesentery-blood-brain course of infection, can be replicated in the newborn rat. We examined temporal and spatial aspects of E. coli K1 infection following initiation of gastrointestinal colonization in 2-day-old (P2) rats after oral administration of E. coli K1 strain A192PP and a virulent bioluminescent derivative, E. coli A192PP-lux2. A combination of bacterial enumeration in the major organs, two-dimensional bioluminescence imaging, and three-dimensional diffuse light imaging tomography with integrated micro-computed tomography indicated multiple sites of colonization within the alimentary canal; these included the tongue, esophagus, and stomach in addition to the small intestine and colon. After invasion of the blood compartment, the bacteria entered the central nervous system, with restricted colonization of the brain, and also invaded the major organs, in line with increases in the severity of symptoms of infection. Both keratinized and nonkeratinized surfaces of esophagi were colonized to a considerably greater extent in susceptible P2 neonates than in corresponding tissues from infection-resistant 9-day-old rat pups; the bacteria appeared to damage and penetrate the nonkeratinized esophageal epithelium of infection-susceptible P2 animals, suggesting the esophagus represents a portal of entry for E. coli K1 into the systemic circulation. Thus, multimodality imaging of experimental systemic infections in real time indicates complex dynamic patterns of colonization and dissemination that provide new insights into the E. coli K1 infection of the neonatal rat. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  2. Time-course effects of aerobic exercise training on cardiovascular and renal parameters in 2K1C renovascular hypertensive rats

    Directory of Open Access Journals (Sweden)

    R.C.A. Maia

    2015-01-01

    Full Text Available Exercise training (Ex has been recommended for its beneficial effects in hypertensive states. The present study evaluated the time-course effects of Ex without workload on mean arterial pressure (MAP, reflex bradycardia, cardiac and renal histology, and oxidative stress in two-kidney, one-clip (2K1C hypertensive rats. Male Fischer rats (10 weeks old; 150–180 g underwent surgery (2K1C or SHAM and were subsequently divided into a sedentary (SED group and Ex group (swimming 1 h/day, 5 days/week for 2, 4, 6, 8, or 10 weeks. Until week 4, Ex decreased MAP, increased reflex bradycardia, prevented concentric hypertrophy, reduced collagen deposition in the myocardium and kidneys, decreased the level of thiobarbituric acid-reactive substances (TBARS in the left ventricle, and increased the catalase (CAT activity in the left ventricle and both kidneys. From week 6 to week 10, however, MAP and reflex bradycardia in 2K1C Ex rats became similar to those in 2K1C SED rats. Ex effectively reduced heart rate and prevented collagen deposition in the heart and both kidneys up to week 10, and restored the level of TBARS in the left ventricle and clipped kidney and the CAT activity in both kidneys until week 8. Ex without workload for 10 weeks in 2K1C rats provided distinct beneficial effects. The early effects of Ex on cardiovascular function included reversing MAP and reflex bradycardia. The later effects of Ex included preventing structural alterations in the heart and kidney by decreasing oxidative stress and reducing injuries in these organs during hypertension.

  3. 维生素K1、新鲜冰冻血浆和冷沉淀对不同出生胎龄新生儿凝血指标的影响%Effects of vitamin k1, fresh frozen plasma and cryoprecipitate on coagulation index of newborns with different gestational ages

    Institute of Scientific and Technical Information of China (English)

    张涛; 廖嘉仪; 张费通; 谭小华; 崔其亮

    2014-01-01

    Objective To explore the effect of Vitamin K1(Vit K1), fresh frozen plasma (plasma) andcryoprecipitate on prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen original(Fbg), thrombin time (TT) of newborns with different gestational ages. Methods The serum of 1,134 newbornsfrom The Third Affiliated Hospital of Guangzhou Medical University was collected from February 2009 to September2012. All newborns had been divided into four groups (according to the gestational age of 28-31+6 weeks, 32-33+6weeks, 34-36+6 weeks and gestational age≥37 weeks).The effect of various interventions (Vit K1, Vit K1+plasmaand Vit K1+cryoprecipitate) on PT, APTT, Fbg, and TT had been recorded. Results (1)The PT and APTT ofeach group with the interventions of Vit k1 were significantly improved (P < 0.05). (2)The PT, APTT, Fbg and TTof each group with the interventions of Vit k1 combined with plasma were significantly improved (P < 0.05). (3)ThePT, APTT and Fbg of each group with the interventions of Vit k1 combined with cryoprecipitate were significantlyimproved (P < 0.05). (4)With Vit k1 combined with plasma, PT and APTT were mostly improved and Fbg wasimproved mostly with Vit k1 combined with cryoprecipitate. Conclusion Vitamin K1, fresh frozen plasma andcryoprecipitate can effectively improvedin the coagulation index of newborns with different gestational ages.%目的:回顾性分析不同出生胎龄新生儿应用维生素K1(Vitamin K1,Vit K1)、新鲜冰冻血浆(血浆)和冷沉淀对凝血酶原时间(PT)、部分活化凝血活酶时间(APTT)、纤维蛋白原(Fbg)、凝血酶时间(TT)的影响。方法:收集广州医科大学附属第三医院2009年2月至2012年9月新生儿科住院的新生儿1134例,按照胎龄28-31+6周、32-33+6周、胎龄34-36+6周、胎龄≥37周进行分组,每组均分析使用Vit K1、Vit K1联合血浆以及Vit K1联合冷沉淀等干预措施前后PT、APTT

  4. Effects of heat exposure on Akt/S6K1 signaling and expression of genes related to protein and energy metabolism in chicken (Gallus gallus) pectoralis major muscle.

    Science.gov (United States)

    Boussaid-Om Ezzine, S; Everaert, N; Métayer-Coustard, S; Rideau, N; Berri, C; Joubert, R; Temim, S; Collin, A; Tesseraud, S

    2010-11-01

    In order to improve understanding of the heat-induced changes in muscle growth, we determined the expression of genes related to protein and energy metabolism in the pectoralis major muscle of chickens. We also explored the protein kinase B (PKB also called Akt)/p70 S6 kinase (S6K1)/S6 pathway that mediates anabolic signals thereby regulating metabolism and hypertrophic/atrophic balance. Four-week-old chickens were exposed to 32 or 22 degrees C for 1 week. Chickens from both groups were then fasted for 16 h or left fed, and submitted to an oral administration of glucose-arginine to induce an anabolic response (30-min treatment) or left untreated. High ambient temperature and the associated decrease in feed intake modified the expression of certain energy-related genes (e.g. -40% for PGC-1alpha) and protein metabolism (e.g. about +80% for atrogin-1), but the expression of several muscle metabolism-related genes considered here was unchanged. The capacity for muscle protein synthesis, i.e. RNA/protein ratio, was reduced in warm conditions (approximately -20%). Slightly lower activation of S6 induced by glucose-arginine treatment was found at 32 degrees C compared to 22 degrees C, which might indicate somewhat lower efficiency of mRNA translation. Analysis of glucose/insulin balance suggested changes in glucose metabolism under heat exposure. However, this remains to be characterized.

  5. Radiation induced microbial pesticide

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Ki Yup; Lee, Young Keun; Kim, Jae Sung; Kim, Jin Kyu; Lee, Sang Jae

    2000-01-01

    To control plant pathogenic fungi, 4 strains of bacteria (K1, K3, K4, YS1) were isolated from mushroom compost and hot spring. K4, K1, K3, YS1 strain showed wide antifungal spectrum and high antifungal activities against 13 kinds of fungi. Mutants of K1 and YS1 strains were induced by gamma-ray radiation and showed promising antifungal activities. These wild type and mutants showed resistant against more than 27 kinds of commercial pesticides among 30 kinds of commercial pesticides test particularly, YS1-1006 mutant strain showed resistant against hydrogen oxide. And mutants had increased antifungal activity against Botryoshaeria dothidea. These results suggested that radiation could be an useful method for the induction of functional mutants. (author)

  6. 电子竞技——Stars war之跑跑卡丁车K1总决定落幕

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    10月15日下午5时,历时3个多月,由上海邮通科技和Replays.net联合主办,游戏风云Games TV独家现场全程直播,技嘉科技,可口可乐和优派联合赞助的Stars war之跑跑卡丁车K1锦标赛圆满落幕。来自全国23个赛区的114名选手决出了跑跑卡本车K1锦标赛全国总决赛的最终车神,

  7. Electrical and microstructural properties of CaTiO3-doped K1/2Na1/2NbO3-lead free ceramics

    Indian Academy of Sciences (India)

    L Ramajo; R Parra; M A Ramírez; M S Castro

    2011-10-01

    Microstructure, electrical properties and dielectric behaviour of K1/2Na1/2NbO3 (KNN) and CaTiO3- modified K1/2Na1/2NbO3 (CTO-KNN) systems, were investigated. Discs doped with 0 to 0.55% mol of CaTiO3 (CTO) were sintered at 1125°C for 2 h. Although minority phases were found in doped samples, CaTiO3 was not detected. It was also observed that CTO changed the microstructure and grain size of KNN drastically. Also, the Curie temperature and permittivity values decreased. Addition of CTO between 0.15 and 0.45 mol% decreases the density and dielectric values. Samples prepared with higher content of CTO than 0.45 mol% showed better electrical properties.

  8. Evaluation of a multiplex PCR for detection of serotypes K1, K2 and K5 in Klebsiella sp. and comparison of isolates within these serotypes.

    Science.gov (United States)

    Turton, Jane F; Baklan, Hatice; Siu, L K; Kaufmann, Mary E; Pitt, Tyrone L

    2008-07-01

    A multiplex PCR using targets within the serotype-specific region of the capsular polysaccharide synthesis gene cluster of serotypes K1, K2 and K5 was evaluated using the 77 reference serotype strains of Klebsiella, and a panel of clinical isolates subjected previously to conventional serotyping. The PCR was highly specific for these serotypes, which are those most associated with virulence in humans and horses. PCR confirmed that isolates of the K5 serotype had cross-reacted with antiserum for other serotypes, particularly for K7. K5 isolates received by our laboratory were almost exclusively from thoroughbred horses, and were submitted for screening prior to breeding programmes. Most, including a reference strain isolated in 1955, belonged to a cluster of genetically similar isolates of sequence type (ST) 60. K1 isolates, all from humans, belonged to a previously identified cluster of ST 23.

  9. Amplitude modulated, by M1, Earth's oscillating (T = 1 day) electric field triggered by K1 tidal waves. Its relation to the occurrence time of large EQs

    CERN Document Server

    Thanassoulas, C; Verveniotis, G

    2010-01-01

    Starting from the observation that quite often the Earth's oscillating electric field varies in amplitude, a mechanism is postulated that accounts for these observations. That mechanism is the piezoelectric one driven by the M1 and K1 tidal components. It is demonstrated how the system: piezoelectricity triggered in the lithosphere - M1 and K1 tidal components is activated and produces the amplitude modulated Earth's oscillating electric field. This procedure is linked to the strain load conditions met in the seismogenic area before the occurrence of a large EQ. Peaks of the oscillating Earth's electric field are tightly connected to the M1 peak tidal component and to the timing of the occurrence of large EQs. Typical examples from real recordings of the Earth's oscillating electric field, recorded by the ATH (Greece) monitoring site, are given in order to verify the postulated detailed piezoelectric mechanism.

  10. Computational investigation of sphingosine kinase 1 (SphK1) and calcium dependent ERK1/2 activation downstream of VEGFR2 in endothelial cells

    Science.gov (United States)

    Bazzazi, Hojjat; Popel, Aleksander S.

    2017-01-01

    Vascular endothelial growth factor (VEGF) is a powerful regulator of neovascularization. VEGF binding to its cognate receptor, VEGFR2, activates a number of signaling pathways including ERK1/2. Activation of ERK1/2 is experimentally shown to involve sphingosine kinase 1 (SphK1) activation and its calcium-dependent translocation downstream of ERK1/2. Here we construct a rule-based computational model of signaling downstream of VEGFR2, by including SphK1 and calcium positive feedback mechanisms, and investigate their consequences on ERK1/2 activation. The model predicts the existence of VEGF threshold in ERK1/2 activation that can be continuously tuned by cellular concentrations of SphK1 and sphingosine 1 phosphate (S1P). The computer model also predicts powerful effects of perturbations in plasma and ER calcium pump rates and the current through the CRAC channels on ERK1/2 activation dynamics, highlighting the critical role of intracellular calcium in shaping the pERK1/2 signal. The model is then utilized to simulate anti-angiogenic therapeutic interventions targeting VEGFR2-ERK1/2 axis. Simulations indicate that monotherapies that exclusively target VEGFR2 phosphorylation, VEGF, or VEGFR2 are ineffective in shutting down signaling to ERK1/2. By simulating therapeutic strategies that target multiple nodes of the pathway such as Raf and SphK1, we conclude that combination therapy should be much more effective in blocking VEGF signaling to EKR1/2. The model has important implications for interventions that target signaling pathways in angiogenesis relevant to cancer, vascular diseases, and wound healing. PMID:28178265

  11. Murine experimental autoimmune encephalomyelitis is diminished by treatment with the angiogenesis inhibitors B20-4.1.1 and angiostatin (K1-3.

    Directory of Open Access Journals (Sweden)

    Carolyn J MacMillan

    Full Text Available Angiogenesis is the formation of new blood vessels form pre-existing vasculature whose contribution to inflammatory conditions of the Central Nervous System is being studied in order to generate novel therapeutic targets. This study is the first to investigate the impact of two particular angiogenesis inhibitors on murine Experimental Autoimmune Encephalomyelitis (EAE, an inflammatory disease that mimics aspects of the human disease Multiple Sclerosis. The inhibitors were chosen to reduce angiogenesis by complimentary means. Extrinsic factors were targeted with B20-4.1.1 through its ability to bind to murine Vascular Endothelial Growth Factor (VEGF. Vascular processes connected to angiogenesis were targeted directly with K(1-3, the first three kringle domains of angiostatin. Mice treated with B20-4.1.1 and K(1-3 from onset of signs had reduced clinical scores 18-21 days after EAE induction. Both agents suppressed spinal cord angiogenesis without effect on local VEGF expression. B20-4.1.1 reduced spinal cord vascular permeability while K(1-3 had no effect. T cell infiltration into the spinal cord at day 21 was unaffected by either treatment. B20-4.1.1 reduced peripheral T cell proliferation while K(1-3 had no effect. Lymphoid cells from treated mice produced reduced levels of the T helper-17 (Th-17 cell cytokine interleukin (IL-17 with no effect on the Th-1 cytokine interferon (IFN-γ or Th-2 cytokine IL-4. However, when both drugs were added in vitro to naive T cells or to antigen stimulated T cells from mice with untreated EAE they had no effect on proliferation or levels of IL-17 or IFN-γ. We conclude that these angiogenesis inhibitors mitigate EAE by both suppressing spinal cord angiogenesis and reducing peripheral T cell activation.

  12. Glucose repression of lactose/galactose metabolism in Kluyveromyces lactis is determined by the concentration of the transcriptional activator LAC9 (K1GAL4) [corrected

    OpenAIRE

    Zachariae, W; Kuger, P; Breunig, K D

    1993-01-01

    In the budding yeast Kluyveromyces lactis glucose repression of genes involved in lactose and galactose metabolism is primarily mediated by LAC9 (or K1GAL4) the homologue of the well-known Saccharomyces cerevisiae transcriptional activator GAL4. Phenotypic difference in glucose repression existing between natural strains are due to differences in the LAC9 gene (Breunig, 1989, Mol.Gen.Genet. 261, 422-427). Comparison between the LAC9 alleles of repressible and non-repressible strains revealed ...

  13. GSK3β Regulates Milk Synthesis in and Proliferation of Dairy Cow Mammary Epithelial Cells via the mTOR/S6K1 Signaling Pathway

    Directory of Open Access Journals (Sweden)

    Xia Zhang

    2014-07-01

    Full Text Available Glycogen synthase kinase 3 (GSK3 is a serine/threonine kinase, whose activity is inhibited by AKT phosphorylation. This inhibitory phosphorylation of GSK3β can in turn play a regulatory role through phosphorylation of several proteins (such as mTOR, elF2B to promote protein synthesis. mTOR is a key regulator in protein synthesis and cell proliferation, and recent studies have shown that both GSK3β and mTORC1 can regulate SREBP1 to promote fat synthesis. Thus far, however, the cross talk between GSK3β and the mTOR pathway in the regulation of milk synthesis and associated cell proliferation is not well understood. In this study the interrelationship between GSK3β and the mTOR/S6K1 signaling pathway leading to milk synthesis and proliferation of dairy cow mammary epithelial cells (DCMECs was analyzed using techniques including GSK3β overexpression by transfection, GSK3β inhibition, mTOR inhibition and methionine stimulation. The analyses revealed that GSK3β represses the mTOR/S6K1 pathway leading to milk synthesis and cell proliferation of DCMECs, whereas GSK3β phosphorylation enhances this pathway. Conversely, the activated mTOR/S6K1 signaling pathway downregulates GSK3β expression but enhances GSK3β phosphorylation to increase milk synthesis and cell proliferation, whereas inhibition of mTOR leads to upregulation of GSK3β and repression of GSK3β phosphorylation, which in turn decreases milk synthesis, and cell proliferation. These findings indicate that GSK3β and phosphorylated GSK3β regulate milk synthesis and proliferation of DCMECs via the mTOR/S6K1 signaling pathway. These findings provide new insight into the mechanisms of milk synthesis.

  14. Meganuclease-driven targeted integration in CHO-K1 cells for the fast generation of HTS-compatible cell-based assays.

    Science.gov (United States)

    Cabaniols, Jean-Pierre; Ouvry, Christine; Lamamy, Véronique; Fery, Isabelle; Craplet, Marie-Laure; Moulharat, Natacha; Guenin, Sophie-Pénélope; Bedut, Stéphane; Nosjean, Olivier; Ferry, Gilles; Devavry, Séverine; Jacqmarcq, Cécile; Lebuhotel, Céline; Mathis, Luc; Delenda, Christophe; Boutin, Jean A; Duchâteau, Philippe; Cogé, Francis; Pâques, Frédéric

    2010-09-01

    The development of cell-based assays for high-throughput screening (HTS) approaches often requires the generation of stable transformant cell lines. However, these cell lines are essentially created by random integration of a gene of interest (GOI) with no control over the level and stability of gene expression. The authors developed a targeted integration system in Chinese hamster ovary (CHO) cells, called the cellular genome positioning system (cGPS), based on the stimulation of homologous gene targeting by meganucleases. Five different GOIs were knocked in at the same locus in cGPS CHO-K1 cells. Further characterization revealed that the cGPS CHO-K1 system is more rapid (2-week protocol), efficient (all selected clones expressed the GOI), reproducible (GOI expression level variation of 12%), and stable over time (no change in GOI expression after 23 weeks of culture) than classical random integration. Moreover, in all cGPS CHO-K1 targeted clones, the recombinant protein was biologically active and its properties similar to the endogenous protein. This fast and robust method opens the door for creating large collections of cell lines of better quality and expressing therapeutically relevant GOIs at physiological levels, thereby enhancing the potential scope of HTS.

  15. Interactions of Neuropathogenic Escherichia coli K1 (RS218 and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Farzana Abubakar Yousuf

    2014-01-01

    Full Text Available Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin, adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (IbeA, CNF1, metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (CNF1, metabolism (D-serine catabolism abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity.

  16. 孕妇使用Vit K1预防新生儿Vit K缺乏的研究

    Institute of Scientific and Technical Information of China (English)

    黎洁芳; 何新荣

    2002-01-01

    [目的] 探索预防早发性新生儿出血的有效方法. [方法] 将134例正常孕妇分为试验组和对照组,至分娩时,分别抽取母亲血和脐带血作血清Vit K1浓度测定及脐带血PIVKA-Ⅱ检测,并在新生儿出生后24小时内检测凝血像,进行统计学分析. [结果] 试验组孕妇血清Vit K1浓度明显高于对照组,试验组PIVKA-Ⅱ阳性较对照组显著降低. [结论] 正常妊娠晚期多次口服Vit K1可降低新生儿脐血PIVKA-Ⅱ阳性率,减少新生儿Vit K缺乏,对预防Vit K缺乏中的早发性新生儿出血有效.

  17. Comparative study of the cytotoxic and genotoxic effects of titanium oxide and aluminium oxide nanoparticles in Chinese hamster ovary (CHO-K1) cells.

    Science.gov (United States)

    Di Virgilio, A L; Reigosa, M; Arnal, P M; Fernández Lorenzo de Mele, M

    2010-05-15

    The aim of this study was to analyze the cytotoxicity and genotoxicity of titanium oxide (TiO(2)) and aluminium oxide (Al(2)O(3)) nanoparticles (NPs) on Chinese hamster ovary (CHO-K1) cells using neutral red (NR), mitochondrial activity (by MTT assay), sister chromatid exchange (SCE), micronucleus (MN) formation, and cell cycle kinetics techniques. Results showed a dose-related cytotoxic effect evidenced after 24h by changes in lysosomal and mitochondrial dehydrogenase activity. Interestingly, transmission electronic microscopy (TEM) showed the formation of perinuclear vesicles in CHO-K1 cells after treatment with both NPs during 24h but no NP was detected in the nuclei. Genotoxic effects were shown by MN frequencies which significantly increased at 0.5 and 1 microg/mL TiO(2) and 0.5-10 microg/mL Al(2)O(3). SCE frequencies were higher for cells treated with 1-5 microg/mL TiO(2). The absence of metaphases evidenced cytotoxicity for higher concentrations of TiO(2). No SCE induction was achieved after treatment with 1-25 microg/mL Al(2)O(3). In conclusion, findings showed cytotoxic and genotoxic effects of TiO(2) and Al(2)O(3) NPs on CHO-K1 cells. Possible causes of controversial reports are discussed further on.

  18. Application of extracellular lipopeptide biosurfactant produced by endophytic Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) in microbially enhanced oil recovery (MEOR).

    Science.gov (United States)

    Pathak, Khyati V; Keharia, Hareshkumar

    2014-02-01

    Bacillus subtilis K1 isolated from aerial roots of banyan tree secreted mixture of surfactins, iturins and fengycins with high degree of heterogeneity. The extracellular extract consisting of mixture of these cyclic lipopeptides exhibited very good emulsification activity as well as excellent emulsion stability. The culture accumulated maximum surfactant up to 48 h of growth during batch fermentation in Luria broth. The emulsion of hexane, heptane and octane prepared using 48-h-old culture supernatant of B. subtilis K1 remained stable up to 2 days while emulsion of four stroke engine oil remained stable for more than a year. The critical micelle concentration of crude lipopeptide biosurfactant extracted by acid precipitation from 48-h-old fermentation broth of B. subtilis K1 was found to be 20.5 μg/mL. The biosurfactant activity was found to be stable at 100 °C for 2 h, over a pH range of 6-12 h and over an NaCl concentration up to 10 % (w/v). The application of biosurfactant on laboratory scale sand pack column saturated with four stroke engine oil resulted in ~43 % enhanced oil recovery, suggesting its suitability in microbially enhanced oil recovery.

  19. Estrogen regulates excitatory amino acid carrier 1 (EAAC1) expression through sphingosine kinase 1 (SphK1) transacting FGFR-mediated ERK signaling in rat C6 astroglial cells.

    Science.gov (United States)

    Huang, C; Yuan, P; Wu, J; Huang, J

    2016-04-05

    Excitatory amino acid carrier 1 (EAAC1) is one important subtype of the excitatory amino acid transporters (EAATs), and its absence can increase the vulnerability to oxidative stress in neural tissue. Enhanced expression of EAAC1 can provide neuroprotection in multiple disorders, including ischemia and multiple sclerosis. However, the mechanism regulating EAAC1 expression is not fully understood. Using rat C6 astroglial cells, which specifically express EAAC1, we found that 17β-estradiol (E2) and (±)-1-[(3aR(∗),4S(∗),9bS(∗))-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl]-ethanone (G1), an agonist of the G-protein-coupled estrogen receptor (GPR30), strongly increased EAAC1 protein levels and protected cells from hydrogen peroxide (H2O2) toxicity. We further found that E2/G1 activated sphingosine kinase 1 (SphK1) via GPR30, resulting in the transcription of fibroblast growth factor 2 (FGF2), which stimulated its receptor (FGFR) and led to the phosphorylation of FGFR substrate 2α (FRS2α). This triggered downstream ERK1/2 signaling for the expression of EAAC1. Both the knockdown of FGF2 by siRNA and the pharmacological suppression of the FGFR-ERK cascade abolished the E2/G1 effect on EAAC1 expression. Overall, our work characterizes a signaling pathway by which E2 transactivates FGFR-ERK to induce EAAC1 expression in an FGF2-dependent manner. This occurs through SphK1 activation via GPR30 and leads to a resistance to H2O2 toxicity. This signal transduction pathway may provide novel insights into our understanding of the neuroprotective effects of E2 and may reveal new therapeutic targets or drugs for regulating the oxidative toxicity effects of various neurological diseases.

  20. p57K1P2和PHLDA2蛋白表达对葡萄胎的诊断意义%The value of p57K1P2 and PHLDA2 in the differential diagnosis of placental hydropic diseases

    Institute of Scientific and Technical Information of China (English)

    方芳; 万希润; 向阳; 冯风芝; 任彤

    2012-01-01

    Objective To investigate the value of combined use of p57K1P2 and PHLDA2 immunohistochemis-try and flow cytometry in the difierential diagnosis of placental hydropic diseases. Methods Specimens of 29 cases of mole pregnancy, formerly diagnosed as complete hydatidiform moles (CHM, n=13), and partial hydatidiform moles (PHMn=16), were reviewed by a senior pathologist. p57K1P2 and PHLDA2 immunohistochemical staining and flow cytometry DNA ploidy analysis were performed in all 29 cases. Results Flow cytometry in all 29 molar cases suggested that 14 cases were triploidy or tetraploid (means these cases are PHM) and 15 cases were diploidy (means these cases are CHM) . All flow cytometrically confirmed partial moles were both p57K1P2 and PHLDA2 positive. There was strong PHLDA2 staining of the cytoplasm in virtually all cells of the villous cytotrophoblast, while p57K1P2 was localized to the nucleus in a subset of those cells. All flow cytometrically showed that complete moles cases were both p57K1P2 and PHLDA2 negative. Conclusion Immunohistochemistry for p57K1P2 and PHL-DA2 may be serves as a practical and reliable diagnostic marker for the diagnosis of complete mole from partial mole.%目的 研究母源表达的印记基因p57K1P2和PHLDA2(IPL/TSSC3)蛋白表达对葡萄胎的辅助诊断意义.方法 收集北京协和医院刮宫组织存档的石蜡包埋标本,其中病理组织学诊断完全性葡萄胎( completehydatidiform mole,CHM) 13例,部分性葡萄胎(partial hydatidiform mole,PHM) 16例.全部病例行流式细胞术DNA倍体分析,采用免疫组化二步法检测p57K1p2及PHLDA2在病理组织中表达.结果 29例病例DNA倍体分析:二倍体15例,三倍体13例,四倍体1例,诊断为CHM 15例,PHM 14例,病理诊断的符合率为86.2%.部分性葡萄胎p57K1P2和PHLDA2绒毛滋养细胞层免疫组织化学全部阳性(100%,14/14).PHLDA2染色阳性位于胞质和胞膜,表达为强阳性.p57K1P2染色阳性位于

  1. 金莲花总黄酮对人甲状腺乳头癌 K1细胞三叶因子3表达变化的影响%Effects of Trollius flavonoids on the expression of Trefoil factor 3 in human papillary thyroid cancer K1 cells in vitro

    Institute of Scientific and Technical Information of China (English)

    房涛; 张静; 吴靖芳; 张文静; 张晨磊; 张芸芸; 裴达

    2015-01-01

    Objective To investigate the inhibition effect of Trollius flavonoids on the proliferation of Trefoil factor 3 (TFF3) in human papillary thyroid cancer K1 cells in vitro, and to explore its effect on the expression of TFF3 in K1 cells.Methods The K1 cells cultured in vitro were treated by different concentrations of Trollius flavonoids for 24h,48h,72 h,respectively ,then the inhibition rate on K 1 cells growth was dectected by MTT .The changes of expression of TFF 3 protein/mRNA were detected by Western blotting and Real-time PCR, respectively.Results MTT detection showed that Trollius flavonoids had inhibition effects on proliferation and growth of K 1 cells in a dose and time dependent manner ( P <0.05). The examination results by Western blotting and Real-time PCR showed that the expression levels of TFF 3 protein and mRNA of K1 cells were decreased with the increase of flavonoids concentration ( P <0 .05 ) .Conclusion Trollius flavonoids have the inhibition effect on human throid papillary cancer cells and can inhibit the expression of TFF 3.%目的:探讨金莲花总黄酮对人甲状腺乳头癌K1细胞增殖抑制作用及三叶因子3(TFF3)表达变化。方法不同浓度金莲花总黄酮作用于体外培养K1细胞24、48、72 h,MTT检测细胞生长抑制率,Western blotting 和Real-time PCR检测不同浓度总黄酮对K1细胞TFF3蛋白及mRNA表达的影响。结果 MTT实验结果表明,同一时间总黄酮对K1细胞生长增殖抑制率随着药物浓度的增高而升高( P <0›.05),同一浓度总黄酮对K1细胞抑制率随时间延长而升高( P <0.05)。 Western blot及QPCR结果显示随着金莲花总黄酮浓度的升高K1细胞TFF3蛋白及mRNA水平下降( P <0.05)。结论金莲花总黄酮对人甲状腺乳头癌细胞具有抑制作用,并能抑制TFF3的表达。

  2. MiR-145 is downregulated in human ovarian cancer and modulates cell growth and invasion by targeting p70S6K1 and MUC1

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Huijuan [Department of Gynecological Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060 (China); Xiao, ZhengHua [Department of gynecology, Yongchuan Affiliated Hospital of Chongqing Medical University, Chongqing City 404100 (China); Wang, Ke; Liu, Wenxin [Department of Gynecological Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060 (China); Hao, Quan, E-mail: quanhao2002@163.com [Department of Gynecological Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center of Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060 (China)

    2013-11-29

    Highlights: •MiR-145 is downregulated in human ovarian cancer. •MiR-145 targets p70S6K1 and MUC1. •p70S6K1 and MUC1 are involved in miR-145 mediated tumor cell growth and cell invasion, respectively. -- Abstract: MicroRNAs (miRNAs) are a family of small non-coding RNA molecules that regulate gene expression at post-transcriptional levels. Previous studies have shown that miR-145 is downregulated in human ovarian cancer; however, the roles of miR-145 in ovarian cancer growth and invasion have not been fully demonstrated. In the present study, Northern blot and qRT-PCR analysis indicate that miR-145 is downregulated in ovarian cancer tissues and cell lines, as well as in serum samples of ovarian cancer, compared to healthy ovarian tissues, cell lines and serum samples. Functional studies suggest that miR-145 overexpression leads to the inhibition of colony formation, cell proliferation, cell growth viability and invasion, and the induction of cell apoptosis. In accordance with the effect of miR-145 on cell growth, miR-145 suppresses tumor growth in vivo. MiR-145 is found to negatively regulate P70S6K1 and MUC1 protein levels by directly targeting their 3′UTRs. Importantly, the overexpression of p70S6K1 and MUC1 can restore the cell colony formation and invasion abilities that are reduced by miR-145, respectively. MiR-145 expression is increased after 5-aza-CdR treatment, and 5-aza-CdR treatment results in the same phenotype as the effect of miR-145 overexpression. Our study suggests that miR-145 modulates ovarian cancer growth and invasion by suppressing p70S6K1 and MUC1, functioning as a tumor suppressor. Moreover, our data imply that miR-145 has potential as a miRNA-based therapeutic target for ovarian cancer.

  3. A Study on Prevention of VKDB in Infants by mothers' Oral Intake of Vitamin K1%母亲口服维生素K1预防婴儿维生素K缺乏出血症的研究

    Institute of Scientific and Technical Information of China (English)

    何守森; 周凤荣; 王锡鲁; 张秋兰; 张秀兰; 朱岩; 薛素兰; 谢鹏; 刘玉兰; 林良明; 刘玉林; 鲁杰

    2003-01-01

    [目的]探讨乳母口服维生素K1预防婴儿维生素K缺乏出血症(VKDB)的效果.[方法]干预组9 871名乳母自分娩后每10天口服维生素K1片10 mg直到满3个月,未干预组10 010人不服,均观察婴儿6个月内的出血情况;同时小样本采集乳母初乳、1月乳、2月乳汁和婴儿的脐血、1个月、2个月静脉血分别测定维生素K1和维生素K缺乏诱导蛋白(PIVKA-Ⅱ)含量.[结果]干预组发生1例VKDB,未干预组发生22例VKDB;干预组母亲1个月乳汁和2个月乳汁中维生素K1含量分别是未干预组的3.5倍和3倍,干预组婴儿1个月外周血PIVKA-Ⅱ阳性率和未干预组比较未表现出统计学差异.[结论]母亲口服维生素K1可明显提高母乳维生素K1的含量,有效减少母乳喂养儿VKDB的发生.

  4. Generation and characterization of polyclonal antibodies specific to N-terminal extension of p85 isoform of ribosomal protein S6 kinase 1 (p85 S6K1

    Directory of Open Access Journals (Sweden)

    Savinska L. O.

    2015-08-01

    Full Text Available Aim. Generation of polyclonal antibodies specific to the ribosomal protein S6 kinase isoform – p85S6K1 and directed to the N-terminal (1–23 aa extension of p85S6K1. Methods. Animal immunization with synthetic (1–23 aa peptide, ELISA, Western blot, Immunoprecipitation, immunofluorescent analysis. Results. Polyclonal antibodies have been generated, which specifically recognize only p85 but not p70 isoform of S6K1 in western blot, immunoprecipitation and immunofluorescence analysis. Conclusions. The obtained antibodies can be recommended for studies on the p85S6K1 and other S6K1 isoforms possessing the N-terminal extension – the identification of binding protein partners, analysis of subcellular localization under different physiological conditions, elucidation of the signal transduction pathways involving different S6K1 isoforms.

  5. 与lK1∪(∪Cui)的补图有相同色划分的图%Graphs with the Same Color-partition as the Complement of lK1∪(∪Cui)

    Institute of Scientific and Technical Information of China (English)

    张淑敏; 火博丰; 冶成福

    2002-01-01

    本文利用色多项式的性质讨论了lK1∪(∪Cui)的补图的色等价划分,从而在ui≠4k+2的条件下完全刻画了与此类图具有相同色等价划分的图.%In this paper, using the properties of chromatic polynomial, we discuss the color-partition of the complement of lK1∪(∪Cui),and characterize the graph with the same color-partition as the class graph under ui≠4k+2.

  6. A Study on Campylobacter jejuni and Campylobacter coli through Commercial Broiler Production Chains in Thailand: Antimicrobial Resistance, the Characterization of DNA Gyrase Subunit A Mutation, and Genetic Diversity by Flagellin A Gene Restriction Fragment Length Polymorphism.

    Science.gov (United States)

    Thomrongsuwannakij, Thotsapol; Blackall, Patrick J; Chansiripornchai, Niwat

    2017-06-01

    chain reaction-restriction fragment length polymorphism of the flagellin A gene (flaA-RFLP) to determine their genetic relationships. Ten distinct clusters were recognized by flaA-RFLP typing. The results showed that horizontal transmission was the major route of Campylobacter transmission in this study. In conclusion, the emergence of MDR and high resistance rates to several antimicrobials are major concerns identified in this study. The prudent use of these agents and active surveillance of resistance at the farm level are essential steps to reduce the public health risks identified in this work.

  7. Effect of Vitamin K1 on Perioperative Blood Coagulation Factors During Open Heart Surgery in Children%维生素K1对小儿心内直视手术围术期凝血机制的影响

    Institute of Scientific and Technical Information of China (English)

    吕小东; 龙村; 姚亚丽; 姚忠喜

    2001-01-01

    目的 小儿心内直视手术中应用维生素K1(Vitamin K1, Vit K1),观察其对凝血因子的作用。 方法 30例心脏手术患者分为3组,A组:动脉导管未闭患者;B组:使用Vit K1治疗;C组:未使用Vit K1治疗。每组各10例。分别检测A组术前、术后,B组、C组术前24小时、麻醉后开胸前、鱼精蛋白中和后10分钟、术毕、术后6小时、1天、3天和5天的血浆凝血酶原时间(PT)、激活部分凝血活酶时间(APTT)、凝血酶时间(TT)、X因子活性、纤维蛋白原含量和红细胞压积。 结果 鱼精蛋白中和后10分钟血浆凝血酶原时间、激活部分凝血活酶时间、凝血酶时间最长,X因子活性、纤维蛋白原含量最低。与C组相比,B组术后24小时内血浆凝血酶原时间明显缩短,X因子活性明显升高。 结论 术前应用Vit K1可明显提高患者术后早期凝血因子水平,有利于减少术后出血的发生。%Objective To assess the effect of vitamin K1 (Vit K1) on blood coagulation factors during open heart operation in children. Methods Thirty patients undergoing cardiac operation were assigned to three groups:group A: patent ductus arteriosus patients(n=10); group B:patients with cardiopulmonary bypass(CPB) plus Vit K1 therapy(n=10); group C: patients with CPB, with no Vit K1 administered (n=10). Blood sample were obtained for the measurement of prothrombin time, activated partial thromboplastin time, thrombin time, factor X activity, plasma fibrinogen concentration and haematocrit. Results Prothrombin time, activated partial thromboplastin time, thrombin time are the longest, factor X activity and plasma fibrinogen concentration is the lowest on 10 minutes after injected protamine. Prothrombin time of group B was significantly shortening and factor X activity of it was increased than group C in 24 hours postoperation. Conclusion Activity of coagulation factors can be increased and maintained at a higher

  8. Epigenetic regulation of nitric oxide synthase 2, inducible (Nos2) by NLRC4 inflammasomes involves PARP1 cleavage

    Science.gov (United States)

    Buzzo, Carina de Lima; Medina, Tiago; Branco, Laura M.; Lage, Silvia L.; Ferreira, Luís Carlos de Souza; Amarante-Mendes, Gustavo P.; Hottiger, Michael O.; De Carvalho, Daniel D.; Bortoluci, Karina R.

    2017-01-01

    Nitric oxide synthase 2, inducible (Nos2) expression is necessary for the microbicidal activity of macrophages. However, NOS2 over-activation causes multiple inflammatory disorders, suggesting a tight gene regulation is necessary. Using cytosolic flagellin as a model for inflammasome-dependent NOS2 activation, we discovered a surprising new role for NLRC4/caspase-1 axis in regulating chromatin accessibility of the Nos2 promoter. We found that activation of two independent mechanisms is necessary for NOS2 expression by cytosolic flagellin: caspase-1 and NF-κB activation. NF-κB activation was necessary, but not sufficient, for NOS2 expression. Conversely, caspase-1 was necessary for NOS2 expression, but dispensable for NF-κB activation, indicating that this protease acts downstream NF-κB activation. We demonstrated that epigenetic regulation of Nos2 by caspase-1 involves cleavage of the chromatin regulator PARP1 (also known as ARTD1) and chromatin accessibility of the NF-κB binding sites located at the Nos2 promoter. Remarkably, caspase-1-mediated Nos2 transcription and NO production contribute to the resistance of macrophages to Salmonella typhimurium infection. Our results uncover the molecular mechanism behind the constricted regulation of Nos2 expression and open new therapeutic opportunities based on epigenetic activities of caspase-1 against infectious and inflammatory diseases. PMID:28150715

  9. Sphingosine kinase-1 mediates androgen-induced osteoblast cell growth

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Claire [CNRS, Institut de Pharmacologie et de Biologie Structurale, Toulouse F-31000 (France); Universite de Toulouse, UPS, IPBS, Toulouse F-31000 (France); Lafosse, Jean-Michel [CHU Toulouse, Hopital Rangueil, Service d' orthopedie et Traumatologie, Toulouse F-31000 (France); Malavaud, Bernard [CNRS, Institut de Pharmacologie et de Biologie Structurale, Toulouse F-31000 (France); Universite de Toulouse, UPS, IPBS, Toulouse F-31000 (France); CHU Toulouse, Hopital Rangueil, Service d' Urologie et de Transplantation Renale, Toulouse F-31000 (France); Cuvillier, Olivier, E-mail: olivier.cuvillier@ipbs.fr [CNRS, Institut de Pharmacologie et de Biologie Structurale, Toulouse F-31000 (France); Universite de Toulouse, UPS, IPBS, Toulouse F-31000 (France)

    2010-01-01

    Herein we report that the lipid kinase sphingosine kinase-1 (SphK1) is instrumental in mediating androgen-induced cell proliferation in osteoblasts. Dihydrotestosterone (DHT) triggered cell growth in steroid-deprived MC3T3 cells, which was associated with a rapid stimulation of SphK1 and activation of both Akt and ERK signaling pathways. This mechanism relied on functional androgen receptor/PI3K/Akt nongenotropic signaling as pharmacological antagonists could block SphK1 stimulation by DHT and its consequences. Finally, SphK1 inhibition not only abrogated DHT-induced ERK activation but also blocked cell proliferation, while ERK inhibition had no impact, suggesting that SphK1 was critical for DHT signaling yet independently of the ERK.

  10. Comparative study of the cytotoxic and genotoxic effects of titanium oxide and aluminium oxide nanoparticles in Chinese hamster ovary (CHO-K1) cells

    Energy Technology Data Exchange (ETDEWEB)

    Di Virgilio, A.L. [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA), Diag. 113 y 64, Correo 16, Suc. 4, La Plata (1900) (Argentina); Reigosa, M. [Instituto Multidisciplinario de Biologia Celular (IMBICE), Calle 526 y Camino Gral. Belgrano (entre 10 y 11), La Plata 1900 (Argentina); Arnal, P.M. [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA), Diag. 113 y 64, Correo 16, Suc. 4, La Plata 1900 (Argentina); Fernandez Lorenzo de Mele, M., E-mail: mmele@inifta.unlp.edu.ar [Instituto de Investigaciones Fisicoquimicas Teoricas y Aplicadas (INIFTA), Diag. 113 y 64, Correo 16, Suc. 4, La Plata 1900 (Argentina)

    2010-05-15

    The aim of this study was to analyze the cytotoxicity and genotoxicity of titanium oxide (TiO{sub 2}) and aluminium oxide (Al{sub 2}O{sub 3}) nanoparticles (NPs) on Chinese hamster ovary (CHO-K1) cells using neutral red (NR), mitochondrial activity (by MTT assay), sister chromatid exchange (SCE), micronucleus (MN) formation, and cell cycle kinetics techniques. Results showed a dose-related cytotoxic effect evidenced after 24 h by changes in lysosomal and mitochondrial dehydrogenase activity. Interestingly, transmission electronic microscopy (TEM) showed the formation of perinuclear vesicles in CHO-K1 cells after treatment with both NPs during 24 h but no NP was detected in the nuclei. Genotoxic effects were shown by MN frequencies which significantly increased at 0.5 and 1 {mu}g/mL TiO{sub 2} and 0.5-10 {mu}g/mL Al{sub 2}O{sub 3}. SCE frequencies were higher for cells treated with 1-5 {mu}g/mL TiO{sub 2}. The absence of metaphases evidenced cytotoxicity for higher concentrations of TiO{sub 2}. No SCE induction was achieved after treatment with 1-25 {mu}g/mL Al{sub 2}O{sub 3}. In conclusion, findings showed cytotoxic and genotoxic effects of TiO{sub 2} and Al{sub 2}O{sub 3} NPs on CHO-K1 cells. Possible causes of controversial reports are discussed further on.

  11. Macrophage receptor with collagenous structure (MARCO) is a dynamic adhesive molecule that enhances uptake of carbon nanotubes by CHO-K1 Cells

    Energy Technology Data Exchange (ETDEWEB)

    Hirano, Seishiro, E-mail: seishiro@nies.go.jp [Environmental Nanotoxicology Project, RCER, National Institute for Environmental Studies (Japan); Fujitani, Yuji; Furuyama, Akiko [Environmental Nanotoxicology Project, RCER, National Institute for Environmental Studies (Japan); Kanno, Sanae [Department of Legal Medicine, St. Marianna School of Medicine (Japan)

    2012-02-15

    The toxicity of carbon nanotubes (CNTs), a highly promising nanomaterial, is similar to that of asbestos because both types of particles have a fibrous shape and are biopersistent. Here, we investigated the characteristics of macrophage receptor with collagenous structure (MARCO), a membrane receptor expressed on macrophages that recognizes environmental or unopsonized particles, and we assessed whether and how MARCO was involved in cellular uptake of multi-walled CNTs (MWCNTs). MARCO-transfected Chinese hamster ovary (CHO-K1) cells took up polystyrene beads irrespective of the particle size (20 nm–1 μm). In the culture of MARCO-transfected CHO-K1 cells dendritic structures were observed on the bottom of culture dishes, and the edges of these dendritic structures were continually renewed as the cell body migrated along the dendritic structures. MWCNTs were first tethered to the dendritic structures and then taken up by the cell body. MWCNTs appeared to be taken up via membrane ruffling like macropinocytosis, rather than phagocytosis. The cytotoxic EC{sub 50} value of MWCNTs in MARCO-transfected CHO-K1 cells was calculated to be 6.1 μg/mL and transmission electron microscopic observation indicated that the toxicity of MWCNTs may be due to the incomplete inclusion of MWCNTs by the membrane structure. -- Highlights: ►Carbon nanotubes (CNTs) were tethered to MARCO in vitro. ►CNTs were taken up rapidly into the cell body via MARCO by membrane ruffling. ►The incomplete inclusion of CNTs by membranes caused cytotoxicity.

  12. Faktor-faktor yang Memengaruhi Bidan dalam Pemberian Vitamin K1 pada Bayi Baru Lahir di Wilayah Kerja Puskesmas Simpang Limun Medan Tahun 2013

    OpenAIRE

    2015-01-01

    Vitamin K is one of the important nutrients that are soluble in fat and given to newborns routinely to prevent hemorrhagic disease. Vitamin K is a co-factor in the coagulation and is produced by bacteria in the gut. Until intestinal colonization perfect, infants are at risk of bleeding. This study aims to determine the factors that affect midwives and the most dominant factor in the provision of vitamin K1 in newborns. This type of research is a cross-sectional survey method. The populatio...

  13. Update of the BIPM.RI(II)-K1.Rn-222 comparison of activity measurements for the radionuclide {sup 222}Rn to include the LNE-LNHB, France

    Energy Technology Data Exchange (ETDEWEB)

    Michotte, C.; Ratel, G. [Bureau International de Poids et Mesures, Pavillon de Breteuil, F-92312 Sevres cedex (France); Cassette, P. [Bureau International de Poids et Mesures, Pavillon de Breteuil, F-92312 Sevres cedex (France); Laboratoire national de metrologie et d' essais -Laboratoire national Henri Becquerel, 91191 Gif-sur-Yvette cedex (France)

    2012-02-15

    In 2007, the Laboratoire national de metrologie et d'essais - Laboratoire national Henri Becquerel (LNE-LNHB), France submitted a sample of known activity of {sup 222}Rn to the International Reference System (SIR) for comparison. The value of the activity submitted was about 90 kBq. This key comparison result joins that of Switzerland and Germany in the key comparison database that now contains three results, identifier BIPM.RI(II)-K1.Rn-222. Consequently, the KCRV has been updated and the degrees of equivalence with the KCRV have been evaluated. (authors)

  14. Fine-scale mapping of the 5q11.2 breast cancer locus reveals at least three independent risk variants regulating MAP3K1.

    Science.gov (United States)

    Glubb, Dylan M; Maranian, Mel J; Michailidou, Kyriaki; Pooley, Karen A; Meyer, Kerstin B; Kar, Siddhartha; Carlebur, Saskia; O'Reilly, Martin; Betts, Joshua A; Hillman, Kristine M; Kaufmann, Susanne; Beesley, Jonathan; Canisius, Sander; Hopper, John L; Southey, Melissa C; Tsimiklis, Helen; Apicella, Carmel; Schmidt, Marjanka K; Broeks, Annegien; Hogervorst, Frans B; van der Schoot, C Ellen; Muir, Kenneth; Lophatananon, Artitaya; Stewart-Brown, Sarah; Siriwanarangsan, Pornthep; Fasching, Peter A; Ruebner, Matthias; Ekici, Arif B; Beckmann, Matthias W; Peto, Julian; dos-Santos-Silva, Isabel; Fletcher, Olivia; Johnson, Nichola; Pharoah, Paul D P; Bolla, Manjeet K; Wang, Qin; Dennis, Joe; Sawyer, Elinor J; Tomlinson, Ian; Kerin, Michael J; Miller, Nicola; Burwinkel, Barbara; Marme, Frederik; Yang, Rongxi; Surowy, Harald; Guénel, Pascal; Truong, Thérèse; Menegaux, Florence; Sanchez, Marie; Bojesen, Stig E; Nordestgaard, Børge G; Nielsen, Sune F; Flyger, Henrik; González-Neira, Anna; Benitez, Javier; Zamora, M Pilar; Arias Perez, Jose Ignacio; Anton-Culver, Hoda; Neuhausen, Susan L; Brenner, Hermann; Dieffenbach, Aida Karina; Arndt, Volker; Stegmaier, Christa; Meindl, Alfons; Schmutzler, Rita K; Brauch, Hiltrud; Ko, Yon-Dschun; Brüning, Thomas; Nevanlinna, Heli; Muranen, Taru A; Aittomäki, Kristiina; Blomqvist, Carl; Matsuo, Keitaro; Ito, Hidemi; Iwata, Hiroji; Tanaka, Hideo; Dörk, Thilo; Bogdanova, Natalia V; Helbig, Sonja; Lindblom, Annika; Margolin, Sara; Mannermaa, Arto; Kataja, Vesa; Kosma, Veli-Matti; Hartikainen, Jaana M; Wu, Anna H; Tseng, Chiu-chen; Van Den Berg, David; Stram, Daniel O; Lambrechts, Diether; Zhao, Hui; Weltens, Caroline; van Limbergen, Erik; Chang-Claude, Jenny; Flesch-Janys, Dieter; Rudolph, Anja; Seibold, Petra; Radice, Paolo; Peterlongo, Paolo; Barile, Monica; Capra, Fabio; Couch, Fergus J; Olson, Janet E; Hallberg, Emily; Vachon, Celine; Giles, Graham G; Milne, Roger L; McLean, Catriona; Haiman, Christopher A; Henderson, Brian E; Schumacher, Fredrick; Le Marchand, Loic; Simard, Jacques; Goldberg, Mark S; Labrèche, France; Dumont, Martine; Teo, Soo Hwang; Yip, Cheng Har; See, Mee-Hoong; Cornes, Belinda; Cheng, Ching-Yu; Ikram, M Kamran; Kristensen, Vessela; Zheng, Wei; Halverson, Sandra L; Shrubsole, Martha; Long, Jirong; Winqvist, Robert; Pylkäs, Katri; Jukkola-Vuorinen, Arja; Kauppila, Saila; Andrulis, Irene L; Knight, Julia A; Glendon, Gord; Tchatchou, Sandrine; Devilee, Peter; Tollenaar, Robert A E M; Seynaeve, Caroline; Van Asperen, Christi J; García-Closas, Montserrat; Figueroa, Jonine; Chanock, Stephen J; Lissowska, Jolanta; Czene, Kamila; Klevebring, Daniel; Darabi, Hatef; Eriksson, Mikael; Hooning, Maartje J; Hollestelle, Antoinette; Martens, John W M; Collée, J Margriet; Hall, Per; Li, Jingmei; Humphreys, Keith; Shu, Xiao-Ou; Lu, Wei; Gao, Yu-Tang; Cai, Hui; Cox, Angela; Cross, Simon S; Reed, Malcolm W R; Blot, William; Signorello, Lisa B; Cai, Qiuyin; Shah, Mitul; Ghoussaini, Maya; Kang, Daehee; Choi, Ji-Yeob; Park, Sue K; Noh, Dong-Young; Hartman, Mikael; Miao, Hui; Lim, Wei Yen; Tang, Anthony; Hamann, Ute; Torres, Diana; Jakubowska, Anna; Lubinski, Jan; Jaworska, Katarzyna; Durda, Katarzyna; Sangrajrang, Suleeporn; Gaborieau, Valerie; Brennan, Paul; McKay, James; Olswold, Curtis; Slager, Susan; Toland, Amanda E; Yannoukakos, Drakoulis; Shen, Chen-Yang; Wu, Pei-Ei; Yu, Jyh-Cherng; Hou, Ming-Feng; Swerdlow, Anthony; Ashworth, Alan; Orr, Nick; Jones, Michael; Pita, Guillermo; Alonso, M Rosario; Álvarez, Nuria; Herrero, Daniel; Tessier, Daniel C; Vincent, Daniel; Bacot, Francois; Luccarini, Craig; Baynes, Caroline; Ahmed, Shahana; Healey, Catherine S; Brown, Melissa A; Ponder, Bruce A J; Chenevix-Trench, Georgia; Thompson, Deborah J; Edwards, Stacey L; Easton, Douglas F; Dunning, Alison M; French, Juliet D

    2015-01-08

    Genome-wide association studies (GWASs) have revealed SNP rs889312 on 5q11.2 to be associated with breast cancer risk in women of European ancestry. In an attempt to identify the biologically relevant variants, we analyzed 909 genetic variants across 5q11.2 in 103,991 breast cancer individuals and control individuals from 52 studies in the Breast Cancer Association Consortium. Multiple logistic regression analyses identified three independent risk signals: the strongest associations were with 15 correlated variants (iCHAV1), where the minor allele of the best candidate, rs62355902, associated with significantly increased risks of both estrogen-receptor-positive (ER(+): odds ratio [OR] = 1.24, 95% confidence interval [CI] = 1.21-1.27, ptrend = 5.7 × 10(-44)) and estrogen-receptor-negative (ER(-): OR = 1.10, 95% CI = 1.05-1.15, ptrend = 3.0 × 10(-4)) tumors. After adjustment for rs62355902, we found evidence of association of a further 173 variants (iCHAV2) containing three subsets with a range of effects (the strongest was rs113317823 [pcond = 1.61 × 10(-5)]) and five variants composing iCHAV3 (lead rs11949391; ER(+): OR = 0.90, 95% CI = 0.87-0.93, pcond = 1.4 × 10(-4)). Twenty-six percent of the prioritized candidate variants coincided with four putative regulatory elements that interact with the MAP3K1 promoter through chromatin looping and affect MAP3K1 promoter activity. Functional analysis indicated that the cancer risk alleles of four candidates (rs74345699 and rs62355900 [iCHAV1], rs16886397 [iCHAV2a], and rs17432750 [iCHAV3]) increased MAP3K1 transcriptional activity. Chromatin immunoprecipitation analysis revealed diminished GATA3 binding to the minor (cancer-protective) allele of rs17432750, indicating a mechanism for its action. We propose that the cancer risk alleles act to increase MAP3K1 expression in vivo and might promote breast cancer cell survival.

  15. Characterization of pII Family (GlnK1, GlnK2, GlnB) Protein Uridylylation in Response to Nitrogen Availability for Rhodopseudomonas palustris

    Energy Technology Data Exchange (ETDEWEB)

    Connelly, Heather M [ORNL; Pelletier, Dale A [ORNL; Lu, Tse-Yuan [ORNL; Lankford, Patricia K [ORNL; Hettich, Robert {Bob} L [ORNL

    2006-01-01

    The GlnK and GlnB proteins are members of the pII signal transduction protein family, which is essential in nitrogen regulation due to this protein family's ability to sense internal cellular ammonium levels and control cellular response. The role of GlnK in nitrogen regulation has been studied in a variety of bacteria but previously has been uncharacterized in the purple nonsulfur anoxygenic phototropic bacterium Rhodopseudomonas palustris. R. palustris has tremendous metabolic versatility in its modes of energy generation and carbon metabolism, and it employs a sensitive nitrogen-ammonium regulation system that may vary from that of other commonly studied bacteria. In R. palustris, there are three annotated forms of pII proteins: GlnK1, GlnK2, and GlnB. Here we describe, for the first time, the characterization of GlnK1, GlnK2, and GlnB modifications as a response to nitrogen availability, thereby providing information about how this bacterium regulates the AmtB ammonium transporter and glutamine synthetase, which controls the rate of glutamate to glutamine conversion. Using a strategy of creating C-terminally tagged GlnK and GlnB proteins followed by tandem affinity purification in combination with top-down mass spectrometry, four isoforms of the GlnK2 and GlnB proteins and two isoforms of the GlnK1 protein were characterized at high resolution and mass accuracy. Wild-type or endogenous expression of all three proteins was also examined under normal ammonium conditions and ammonium starvation to ensure that the tagging and affinity purification methods employed did not alter the natural state of the proteins. All three proteins were found to undergo uridylylation under ammonium starvation conditions, presumably to regulate the AmtB ammonium transporter and glutamine synthetase. Under high-ammonium conditions, the GlnK1, GlnK2, and GlnB proteins are unmodified. This experimental protocol involving high-resolution mass spectrometry measurements of intact

  16. Fine-Scale Mapping of the 5q11.2 Breast Cancer Locus Reveals at Least Three Independent Risk Variants Regulating MAP3K1

    Science.gov (United States)

    Glubb, Dylan M.; Maranian, Mel J.; Michailidou, Kyriaki; Pooley, Karen A.; Meyer, Kerstin B.; Kar, Siddhartha; Carlebur, Saskia; O’Reilly, Martin; Betts, Joshua A.; Hillman, Kristine M.; Kaufmann, Susanne; Beesley, Jonathan; Canisius, Sander; Hopper, John L.; Southey, Melissa C.; Tsimiklis, Helen; Apicella, Carmel; Schmidt, Marjanka K.; Broeks, Annegien; Hogervorst, Frans B.; van der Schoot, C. Ellen; Muir, Kenneth; Lophatananon, Artitaya; Stewart-Brown, Sarah; Siriwanarangsan, Pornthep; Fasching, Peter A.; Ruebner, Matthias; Ekici, Arif B.; Beckmann, Matthias W.; Peto, Julian; dos-Santos-Silva, Isabel; Fletcher, Olivia; Johnson, Nichola; Pharoah, Paul D.P.; Bolla, Manjeet K.; Wang, Qin; Dennis, Joe; Sawyer, Elinor J.; Tomlinson, Ian; Kerin, Michael J.; Miller, Nicola; Burwinkel, Barbara; Marme, Frederik; Yang, Rongxi; Surowy, Harald; Guénel, Pascal; Truong, Thérèse; Menegaux, Florence; Sanchez, Marie; Bojesen, Stig E.; Nordestgaard, Børge G.; Nielsen, Sune F.; Flyger, Henrik; González-Neira, Anna; Benitez, Javier; Zamora, M. Pilar; Arias Perez, Jose Ignacio; Anton-Culver, Hoda; Neuhausen, Susan L.; Brenner, Hermann; Dieffenbach, Aida Karina; Arndt, Volker; Stegmaier, Christa; Meindl, Alfons; Schmutzler, Rita K.; Brauch, Hiltrud; Ko, Yon-Dschun; Brüning, Thomas; Nevanlinna, Heli; Muranen, Taru A.; Aittomäki, Kristiina; Blomqvist, Carl; Matsuo, Keitaro; Ito, Hidemi; Iwata, Hiroji; Tanaka, Hideo; Dörk, Thilo; Bogdanova, Natalia V.; Helbig, Sonja; Lindblom, Annika; Margolin, Sara; Mannermaa, Arto; Kataja, Vesa; Kosma, Veli-Matti; Hartikainen, Jaana M.; Wu, Anna H.; Tseng, Chiu-chen; Van Den Berg, David; Stram, Daniel O.; Lambrechts, Diether; Zhao, Hui; Weltens, Caroline; van Limbergen, Erik; Chang-Claude, Jenny; Flesch-Janys, Dieter; Rudolph, Anja; Seibold, Petra; Radice, Paolo; Peterlongo, Paolo; Barile, Monica; Capra, Fabio; Couch, Fergus J.; Olson, Janet E.; Hallberg, Emily; Vachon, Celine; Giles, Graham G.; Milne, Roger L.; McLean, Catriona; Haiman, Christopher A.; Henderson, Brian E.; Schumacher, Fredrick; Le Marchand, Loic; Simard, Jacques; Goldberg, Mark S.; Labrèche, France; Dumont, Martine; Teo, Soo Hwang; Yip, Cheng Har; See, Mee-Hoong; Cornes, Belinda; Cheng, Ching-Yu; Ikram, M. Kamran; Kristensen, Vessela; Zheng, Wei; Halverson, Sandra L.; Shrubsole, Martha; Long, Jirong; Winqvist, Robert; Pylkäs, Katri; Jukkola-Vuorinen, Arja; Kauppila, Saila; Andrulis, Irene L.; Knight, Julia A.; Glendon, Gord; Tchatchou, Sandrine; Devilee, Peter; Tollenaar, Robert A.E.M.; Seynaeve, Caroline; Van Asperen, Christi J.; García-Closas, Montserrat; Figueroa, Jonine; Chanock, Stephen J.; Lissowska, Jolanta; Czene, Kamila; Klevebring, Daniel; Darabi, Hatef; Eriksson, Mikael; Hooning, Maartje J.; Hollestelle, Antoinette; Martens, John W.M.; Collée, J. Margriet; Hall, Per; Li, Jingmei; Humphreys, Keith; Shu, Xiao-Ou; Lu, Wei; Gao, Yu-Tang; Cai, Hui; Cox, Angela; Cross, Simon S.; Reed, Malcolm W.R.; Blot, William; Signorello, Lisa B.; Cai, Qiuyin; Shah, Mitul; Ghoussaini, Maya; Kang, Daehee; Choi, Ji-Yeob; Park, Sue K.; Noh, Dong-Young; Hartman, Mikael; Miao, Hui; Lim, Wei Yen; Tang, Anthony; Hamann, Ute; Torres, Diana; Jakubowska, Anna; Lubinski, Jan; Jaworska, Katarzyna; Durda, Katarzyna; Sangrajrang, Suleeporn; Gaborieau, Valerie; Brennan, Paul; McKay, James; Olswold, Curtis; Slager, Susan; Toland, Amanda E.; Yannoukakos, Drakoulis; Shen, Chen-Yang; Wu, Pei-Ei; Yu, Jyh-Cherng; Hou, Ming-Feng; Swerdlow, Anthony; Ashworth, Alan; Orr, Nick; Jones, Michael; Pita, Guillermo; Alonso, M. Rosario; Álvarez, Nuria; Herrero, Daniel; Tessier, Daniel C.; Vincent, Daniel; Bacot, Francois; Luccarini, Craig; Baynes, Caroline; Ahmed, Shahana; Healey, Catherine S.; Brown, Melissa A.; Ponder, Bruce A.J.; Chenevix-Trench, Georgia; Thompson, Deborah J.; Edwards, Stacey L.; Easton, Douglas F.; Dunning, Alison M.; French, Juliet D.

    2015-01-01

    Genome-wide association studies (GWASs) have revealed SNP rs889312 on 5q11.2 to be associated with breast cancer risk in women of European ancestry. In an attempt to identify the biologically relevant variants, we analyzed 909 genetic variants across 5q11.2 in 103,991 breast cancer individuals and control individuals from 52 studies in the Breast Cancer Association Consortium. Multiple logistic regression analyses identified three independent risk signals: the strongest associations were with 15 correlated variants (iCHAV1), where the minor allele of the best candidate, rs62355902, associated with significantly increased risks of both estrogen-receptor-positive (ER+: odds ratio [OR] = 1.24, 95% confidence interval [CI] = 1.21–1.27, ptrend = 5.7 × 10−44) and estrogen-receptor-negative (ER−: OR = 1.10, 95% CI = 1.05–1.15, ptrend = 3.0 × 10−4) tumors. After adjustment for rs62355902, we found evidence of association of a further 173 variants (iCHAV2) containing three subsets with a range of effects (the strongest was rs113317823 [pcond = 1.61 × 10−5]) and five variants composing iCHAV3 (lead rs11949391; ER+: OR = 0.90, 95% CI = 0.87–0.93, pcond = 1.4 × 10−4). Twenty-six percent of the prioritized candidate variants coincided with four putative regulatory elements that interact with the MAP3K1 promoter through chromatin looping and affect MAP3K1 promoter activity. Functional analysis indicated that the cancer risk alleles of four candidates (rs74345699 and rs62355900 [iCHAV1], rs16886397 [iCHAV2a], and rs17432750 [iCHAV3]) increased MAP3K1 transcriptional activity. Chromatin immunoprecipitation analysis revealed diminished GATA3 binding to the minor (cancer-protective) allele of rs17432750, indicating a mechanism for its action. We propose that the cancer risk alleles act to increase MAP3K1 expression in vivo and might promote breast cancer cell survival. PMID:25529635

  17. The mTORC1 Effectors S6K1 and 4E-BP Play Different Roles in CNS Axon Regeneration

    OpenAIRE

    2014-01-01

    Using mouse optic nerve (ON) crush as a CNS injury model, we and others have found that activation of the mammalian target of rapamycin complex 1 (mTORC1) in mature retinal ganglion cells by deletion of the negative regulators, phosphatase and tensin homolog (PTEN) and tuberous sclerosis 1, promotes ON regeneration. mTORC1 activation inhibits eukaryotic translation initiation factor 4E-binding protein (4E-BP) and activates ribosomal protein S6 kinase 1 (S6K1), both of which stimulate translat...

  18. Rapamycin inhibition of baculovirus recombinant (BVr ribosomal protein S6 kinase (S6K1 is mediated by an event other than phosphorylation

    Directory of Open Access Journals (Sweden)

    Beigh Mushtaq A

    2012-03-01

    Full Text Available Abstract Background Ribosomal protein S6 kinase 1(S6K1 is an evolutionary conserved kinase that is activated in response to growth factors and viral stimuli to influence cellular growth and proliferation. This downstream effector of target of rapamycin (TOR signaling cascade is known to be directly activated by TOR- kinase mediated hydrophobic motif (HM phosphorylation at Threonine 412 (T412. Selective loss of this phosphorylation by inactivation of TOR kinase or activation/recruitment of a phosphatase has accordingly been implicated in mediating inhibition by rapamycin. Findings We present evidence that baculovirus driven expression of S6K1 in insect cells (Sf9 fails to activate the enzyme and instead renders it modestly active representing 4-6 folds less activity than its fully active mammalian counterpart. Contrary to the contention that viral infection activates TOR signaling pathway, we report that BVr enzyme fails to exhibit putative TOR dependent phosphorylation at the HM and the resultant phosphorylation at the activation loop (AL of the enzyme, correlating with the level of activity observed. Surprisingly, the BVr enzyme continued to exhibit sensitivity to rapamycin that remained unaffected by mutations compromised for TOR phosphorylation (T412A or deletions compromised for TOR binding (ΔNH 2-46/ΔCT104. Conclusions These data together with the ability of the BVr enzyme to resist inactivation by phosphatases indicate that inhibition by rapamycin is not mediated by any phosphorylation event in general and TOR dependent phosphorylation in particular.

  19. Structure of a putative trans-editing enzyme for prolyl-tRNA synthetase from Aeropyrum pernix K1 at 1.7 Å resolution

    Energy Technology Data Exchange (ETDEWEB)

    Murayama, Kazutaka; Kato-Murayama, Miyuki; Katsura, Kazushige; Uchikubo-Kamo, Tomomi; Yamaguchi-Hirafuji, Machiko; Kawazoe, Masahito; Akasaka, Ryogo; Hanawa-Suetsugu, Kyoko; Hori-Takemoto, Chie [RIKEN Genomic Sciences Center, Yokohama (Japan); Terada, Takaho [RIKEN Genomic Sciences Center, Yokohama (Japan); RIKEN Harima Institute at SPring-8, Hyogo (Japan); Shirouzu, Mikako [RIKEN Harima Institute at SPring-8, Hyogo (Japan); Yokoyama, Shigeyuki, E-mail: yokoyama@biochem.s.u-tokyo.ac.jp [RIKEN Genomic Sciences Center, Yokohama (Japan); RIKEN Harima Institute at SPring-8, Hyogo (Japan); Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Tokyo (Japan)

    2005-01-01

    The three-dimensional structure of the APE2540 protein from A. pernix K1 has been determined by the multiple anomalous dispersion method at 1.7 Å resolution. The structure includes two monomers in the asymmetric unit and shares structural similarity with the YbaK protein or cysteinyl-tRNA{sup Pro} deacylase from H. influenzae. The crystal structure of APE2540, the putative trans-editing enzyme ProX from Aeropyrum pernix K1, was determined in a high-throughput manner. The crystal belongs to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 47.4, b = 58.9, c = 53.6 Å, β = 106.8°. The structure was solved by the multiwavelength anomalous dispersion method at 1.7 Å and refined to an R factor of 16.8% (R{sub free} = 20.5%). The crystal structure includes two protein molecules in the asymmetric unit. Each monomer consists of eight β-strands and seven α-helices. A structure-homology search revealed similarity between the trans-editing enzyme YbaK (or cysteinyl-tRNA{sup Pro} deacylase) from Haemophilus influenzae (HI1434; 22% sequence identity) and putative ProX proteins from Caulobacter crescentus (16%) and Agrobacterium tumefaciens (21%)

  20. Lactobacillus rhamnosus GG Suppresses Meningitic E. coli K1 Penetration across Human Intestinal Epithelial Cells In Vitro and Protects Neonatal Rats against Experimental Hematogenous Meningitis

    Directory of Open Access Journals (Sweden)

    Sheng-He Huang

    2009-01-01

    Full Text Available The purpose of this study was to examine prophylactic efficacy of probiotics in neonatal sepsis and meningitis caused by E. coli K1. The potential inhibitory effect of Lactobacillus rhamnosus GG (LGG on meningitic E. coli K1 infection was examined by using (i in vitro inhibition assays with E44 (a CSF isolate from a newborn baby with E. coli meningitis, and (ii the neonatal rat model of E. coli sepsis and meningitis. The in vitro studies demonstrated that LGG blocked E44 adhesion, invasion, and transcytosis in a dose-dependent manner. A significant reduction in the levels of pathogen colonization, E. coli bacteremia, and meningitis was observed in the LGG-treated neonatal rats, as assessed by viable cultures, compared to the levels in the control group. In conclusion, probiotic LGG strongly suppresses meningitic E. coli pathogens in vitro and in vivo. The results support the use of probiotic strains such as LGG for prophylaxis of neonatal sepsis and meningitis.

  1. 婴儿口服维生素K1预防婴儿维生素K缺乏出血症的研究

    Institute of Scientific and Technical Information of China (English)

    周凤荣; 何守森; 王锡鲁

    2002-01-01

    目的 探讨预防婴儿维生素K缺乏性出血症的有效措施。方法 婴儿出生后即口服维生素K1,每10天1次,每次2.5mg,共 10次,并采脐带血、1个月和 2个月静脉血,统一检测。结果 群体干预研究服药组维生素K缺乏性出血症发生率明显低于对照组(P<0.01)。小样本的实验室观察结果显示了口服组1个月和2个月静脉血维生素K缺乏诱导蛋白(PIVKA-Ⅱ)已经完全转阴。结论 婴儿口服维生素K1能有效地预防典型及晚发性维生素K缺乏性出血症。

  2. Synthesis, biological evaluation, and structure-activity relationships of N-benzoyl-2-hydroxybenzamides as agents active against P. falciparum (K1 strain), Trypanosomes, and Leishmania.

    Science.gov (United States)

    Stec, Jozef; Huang, Qingqing; Pieroni, Marco; Kaiser, Marcel; Fomovska, Alina; Mui, Ernest; Witola, William H; Bettis, Samuel; McLeod, Rima; Brun, Reto; Kozikowski, Alan P

    2012-04-12

    In our efforts to identify novel chemical scaffolds for the development of new antiprotozoal drugs, a compound library was screened against Toxoplasma gondii tachyzoites with activity discovered for N-(4-ethylbenzoyl)-2-hydroxybenzamide 1a against T. gondii as described elsewhere. Synthesis of a compound set was guided by T. gondii SAR with 1r found to be superior for T. gondii , also active against Thai and Sierra Leone strains of Plasmodium falciparum , and with superior ADMET properties as described elsewhere. Herein, synthesis methods and details of the chemical analysis of the compounds in this series are described. Further, this series of N-benzoyl-2-hydroxybenzamides was repurposed for testing against four other protozoan parasites: Trypanosoma brucei rhodesiense , Trypanosoma cruzi , Leishmania donovani , and P. falciparum (K1 isolate). Structure-activity analyses led to the identification of compounds in this set with excellent antileishmanial activity (compound 1d). Overall, compound 1r was the best and had activity 21-fold superior to that of the standard antimalarial drug chloroquine against the K1 P. falciparum isolate. © 2012 American Chemical Society

  3. DNA display selection of peptide ligands for a full-length human G protein-coupled receptor on CHO-K1 cells.

    Directory of Open Access Journals (Sweden)

    Nobuhide Doi

    Full Text Available The G protein-coupled receptors (GPCRs, which form the largest group of transmembrane proteins involved in signal transduction, are major targets of currently available drugs. Thus, the search for cognate and surrogate peptide ligands for GPCRs is of both basic and therapeutic interest. Here we describe the application of an in vitro DNA display technology to screening libraries of peptide ligands for full-length GPCRs expressed on whole cells. We used human angiotensin II (Ang II type-1 receptor (hAT1R as a model GPCR. Under improved selection conditions using hAT1R-expressing Chinese hamster ovary (CHO-K1 cells as bait, we confirmed that Ang II gene could be enriched more than 10,000-fold after four rounds of selection. Further, we successfully selected diverse Ang II-like peptides from randomized peptide libraries. The results provide more precise information on the sequence-function relationships of hAT1R ligands than can be obtained by conventional alanine-scanning mutagenesis. Completely in vitro DNA display can overcome the limitations of current display technologies and is expected to prove widely useful for screening diverse libraries of mutant peptide and protein ligands for receptors that can be expressed functionally on the surface of CHO-K1 cells.

  4. Effect of Evolutionary Adaption on Xylosidase Activity in Thermotolerant Yeast Isolates Kluyveromyces marxianus NIRE-K1 and NIRE-K3.

    Science.gov (United States)

    Behera, Shuvashish; Sharma, Nilesh K; Arora, Richa; Kumar, Sachin

    2016-08-01

    Efficient use of xylose along with glucose is necessary for the economic production of lignocellulosic based biofuels. Xylose transporters play an important role in the microorganisms for efficient utilization of xylose. In the present study, a novel method has been developed for a rapid assay of xylose transport activity in the xylose-utilizing isolates and other known yeasts. An assay was conducted to compare the activity of β-xylosidase using p-nitrophenyl-β-D-xylopyranoside (pNPX) in the intact, intracellular, and extracellular yeasts cells showing xylose transporter. Saccharomyces cerevisiae (MTCC 170) showed no xylosidase activity, while little growth was observed in the xylose-containing medium. Although other yeasts, i.e., Kluyveromyces marxianus NIRE-K1 (MTCC 5933), K. marxianus NIRE-K3 (MTCC 5934), and Candida tropicalis (MTCC 230), showed xylosidase activity in intact, intracellular, and extracellular culture. The xylosidase activity in intact cell was higher than that of extracellular and intracellular activity in all the yeast cells. The enzyme activity was higher in case of K. marxianus NIRE-K1 and K. marxianus NIRE-K3 rather than the C. tropicalis. Further, better xylosidase activity was observed in adapted K. marxianus cells which were 2.79-28.46 % higher than that of native (non-adapted) strains, which indicates the significant improvement in xylose transportation.

  5. A novel regulatory element (E77) isolated from CHO-K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells.

    Science.gov (United States)

    Kang, Shin-Young; Kim, Yeon-Gu; Kang, Seunghee; Lee, Hong Weon; Lee, Eun Gyo

    2016-05-01

    Vectors flanked by regulatory DNA elements have been used to generate stable cell lines with high productivity and transgene stability; however, regulatory elements in Chinese hamster ovary (CHO) cells, which are the most widely used mammalian cells in biopharmaceutical production, are still poorly understood. We isolated a novel gene regulatory element from CHO-K1 cells, designated E77, which was found to enhance the stable expression of a transgene. A genomic library was constructed by combining CHO-K1 genomic DNA fragments with a CMV promoter-driven GFP expression vector, and the E77 element was isolated by screening. The incorporation of the E77 regulatory element resulted in the generation of an increased number of clones with high expression, thereby enhancing the expression level of the transgene in the stable transfectant cell pool. Interestingly, the E77 element was found to consist of two distinct fragments derived from different locations in the CHO genome shotgun sequence. High and stable transgene expression was obtained in transfected CHO cells by combining these fragments. Additionally, the function of E77 was found to be dependent on its site of insertion and specific orientation in the vector construct. Our findings demonstrate that stable gene expression mediated by the CMV promoter in CHO cells may be improved by the isolated novel gene regulatory element E77 identified in the present study.

  6. K1P1 METAR

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — METAR is a routine scheduled observation and is the primary observation code used in the United States to satisfy requirements for reporting surface meteorological...

  7. K1A6 METAR

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — METAR is a routine scheduled observation and is the primary observation code used in the United States to satisfy requirements for reporting surface meteorological...

  8. K1B7 METAR

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — METAR is a routine scheduled observation and is the primary observation code used in the United States to satisfy requirements for reporting surface meteorological...

  9. Multivariate analysis of Vitamin K1 maintenance dose in patients with anticoagulant rodenticide poisoning%影响抗凝血杀鼠剂中毒患者维生素K1维持剂量的多因素分析

    Institute of Scientific and Technical Information of China (English)

    龙剑海; 孙亚威; 邱泽武

    2016-01-01

    Objective To investigate the effect factors of Vitamin K1 maintenance dose in patients with anticoagulant rodenticide poisoning. Methods This retrospective study was analyzed 36 cases in our hospital from January 2013 to October 2014. The correlation between Vitamin K1 and types of anticoagulant rodenticide and toxic dose, prothrombin time activity (PTA), prothrombin time (PT), activated partial thromboplastin time (APTT), prehospital duration (from poisoning to hospitalization), and length of hospital stay (duration of Vitamin K1 treatment) were collected and analyzed by Spearman rank correlation analysis and multivariate regression analysis. Results The dose of Vitamin K1 was positively correlated with brodifacoum (r=0.356, P 0.05). The multivariate regression analysis also showed that prehospital duration [95% confidence interval: (-0.536, -0.103), P=0.002] and length of hospital stay [95% confidence interval: (-1.955, -0.248), P=0.013] had significant effect on the maintenance dose of vitamin K1 in the patients with coagulation function stable. Conclusion After stabilizing the coagulation function, the demanded maintenance dose of vitamin K1 can be gradually decreased with time prolonged, and have no relationship with type of anticoagulant rodenticide and toxic dose.%目的:探讨抗凝血杀鼠剂中毒患者维生素K1的维持剂量治疗的相关因素。方法收集2013年1月至2014年10月36例抗凝血杀鼠剂中毒患者的数据资料,检测并分析维生素K1使用量与抗凝血杀鼠剂种类及具体毒检剂量,凝血酶原活动度(PTA),凝血酶原时间(PT),活化部分凝血活酶时间(APTT),患者院前时间及住院时间的相关性,并对相关因素进行多因素稳健回归分析。结果经Spearman秩相关分析发现,溴鼠灵与维生素K1用量呈正相关(r=0.356, P<0.05);PTA、住院时间及院前时间与VK1用量呈负相关(r=-0.325、-0.420、-0.407,P均<0.05)。而PT、APTT及溴敌隆与维生素K

  10. 个体化干预措施在预防维生素 K1注射液过敏反应中的应用研究%Applied research of individual intervention care on prevention of anaphylaxis of Vitamin K1 Injection

    Institute of Scientific and Technical Information of China (English)

    高俊芳; 宋新宇; 朱美玉; 王静

    2016-01-01

    Objective To prevent the side effects of the Vitamin K1 Injection with the individual intervention care to the infant patients administered the injection. Methods Every infant patient was administered the dilute injection through the intravenous drip and the drug allergy history was registered. 107 infant patients administered Vitamin K1 Injection during January 1,2014 to December 31,2014 were chose as the research participants,who were randomly divided into control group (52 cases)and interventional group(55 cases). The patients in control group received the routine treatment of transfusion and careful observation,while the infants who had the first treatment and unclear history in the interventional group cannot get Vitamin K1 Injection except those who would have the negative results of allergy skin test. Results 3 anaphylactic reac-tions occurred in control group,whereas no allergy was observed in interventional group(P < 0. 05). Conclusion The indi-vidual intervention care to the infant patients administered Vitamin K1 Injection can provide the efficient prevention of ana-phylactic reactions and ensure the drug use. Furthermore,the harmonious doctor - patient relationship would come true and there would be much less medical disputes.%目的:通过对应用维生素 K1注射液的患儿实施个体化干预措施,来预防药物不良反应的发生。方法应用维生素 K1注射液患儿均采用稀释后静脉滴注方法给药,两组患儿在接受此药物前均进行药物过敏反应登记。将2014年1月1日~2014年12月31日使用维生素 K1注射液治疗的患儿107例作为研究对象,随机分为对照组52例和干预组55例。对照组仅给予常规输液治疗及严密观察,干预组对初次使用和不清楚用药史的患儿给予药物过敏实验,皮试结果为阴性方可输液治疗。结果对照组有3例发生过敏反应,干预组无过敏反应发生,两组结果的差异有统计学意义(P <0.05

  11. Update of the BIPM.RI(II)-K1.Rn-222 comparison of activity measurements for the radionuclide 222Rn to include the LNE-LNHB, France

    Science.gov (United States)

    Michotte, C.; Ratel, G.; Cassette, P.

    2012-01-01

    In 2007, the Laboratoire national de métrologie et d'essais-Laboratoire national Henri Becquerel (LNE-LNHB), France submitted a sample of known activity of 222Rn to the International Reference System (SIR) for comparison. The value of the activity submitted was about 90 kBq. This key comparison result joins that of Switzerland and Germany in the key comparison database that now contains three results, identifier BIPM.RI(II)-K1.Rn-222. Consequently, the KCRV has been updated and the degrees of equivalence with the KCRV have been evaluated. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI Section II, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  12. KEY COMPARISON: Activity measurements of the radionuclide 153Sm for the ANSTO, Australia in the ongoing comparison BIPM.RI(II)-K1.Sm-153

    Science.gov (United States)

    Ratel, G.; Michotte, C.; Reinhard, M.; Alexiev, D.; Mo, L.

    2005-01-01

    In 2004, the Australian Nuclear Science and Technology Organisation (ANSTO) submitted two samples of known activity of 153Sm to the International Reference System (SIR). The value of the activity submitted was about 920 MBq. This key comparison result has been added to the matrix of degrees of equivalence in the key comparison database that now contains five results, identifier BIPM.RI(II)-K1.Sm-153. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), according to the provisions of the Mutual Recognition Arrangement (MRA).

  13. A Study of GluK1 Kainate Receptor Polymorphisms in Down Syndrome Reveals Allelic Non-Disjunction at 1173(C/T

    Directory of Open Access Journals (Sweden)

    Debarati Ghosh

    2009-01-01

    Full Text Available Mechanisms underlying Down syndrome (DS-related mental retardation (MR remain poorly understood. In trisomic offspring, non-disjunction may result in the reduction to homozygosity of a susceptibility allele inherited from a heterozygous parent. Accordingly, we sought evidence for allelic non-disjunction in the GluK1 gene that encodes the critical kainite-binding glutamate receptor subunit-5, maps to chromosome 21q22.1 in the DS critical region and is expressed in brain regions responsible for learning and memory. Three polymorphisms of GluK1 [522(A/C rs363538; 1173(C/T rs363430 and 2705(T/C rs363504] were genotyped in 86 DS patient families by means of PCR-coupled RFLP assays and evaluated with respect to allele frequency, heterozygosity, linkage disequilibrium, stage and parental origin of allelic non-disjunction. We report that the distribution of allele frequencies is in Hardy-Weinberg equilibrium. Moderate heterozygosity (0.339 and a major allele frequency of 0.78 render the 1173(C/T marker informative. Pair-wise comparisons reveal that 522(A/C-1173(C/T [χ2 = 31.2, df = 1, p = 0.0001; D’ = 0.42] and 1173(C/T-2705(T/C [χ2 = 18.3, df = 1, p = 0.0001; D’ = 0.34] are in significant linkage disequilibrium of weak magnitude. The estimated ratio of meiosis-I to meiosis-II errors arising from allelic non-disjunction of 1173(C/T is 4:1 in maternal cases and 2:1 in paternal cases. Studies including additional markers and patient samples are warranted to further substantiate present findings.

  14. Origin of giant piezoelectric effect in lead-free K1-xNaxTa1-yNbyO3 single crystals.

    Science.gov (United States)

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-05-10

    A series of high-quality, large-sized (maximum size of 16 × 16 × 32 mm(3)) K1-xNaxTa1-yNbyO3 (x = 0.61, 0.64, and 0.70 and corresponding y = 0.58, 0.60, and 0.63) single crystals were grown using the top-seed solution growth method. The segregation of the crystals, which allowed for precise control of the individual components of the crystals during growth, was investigated. The obtained crystals exhibited excellent properties without being annealed, including a low dielectric loss (0.006), a saturated hysteresis loop, a giant piezoelectric coefficient d33 (d33 = 416 pC/N, determined by the resonance method and d33(*) = 480 pC/N, measured using a piezo-d33 meter), and a large electromechanical coupling factor, k33 (k33 = 83.6%), which was comparable to that of lead zirconate titanate. The reason the piezoelectric coefficient d33 of K0.39Na0.61Ta0.42Nb0.58O3 was larger than those of the other two crystals grown was elucidated through first-principles calculations. The obtained results indicated that K1-xNaxTa1-yNbyO3 crystals can be used as a high-quality, lead-free piezoelectric material.

  15. Origin of giant piezoelectric effect in lead-free K1-xNaxTa1-yNbyO3 single crystals

    Science.gov (United States)

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-05-01

    A series of high-quality, large-sized (maximum size of 16 × 16 × 32 mm3) K1-xNaxTa1-yNbyO3 (x = 0.61, 0.64, and 0.70 and corresponding y = 0.58, 0.60, and 0.63) single crystals were grown using the top-seed solution growth method. The segregation of the crystals, which allowed for precise control of the individual components of the crystals during growth, was investigated. The obtained crystals exhibited excellent properties without being annealed, including a low dielectric loss (0.006), a saturated hysteresis loop, a giant piezoelectric coefficient d33 (d33 = 416 pC/N, determined by the resonance method and d33* = 480 pC/N, measured using a piezo-d33 meter), and a large electromechanical coupling factor, k33 (k33 = 83.6%), which was comparable to that of lead zirconate titanate. The reason the piezoelectric coefficient d33 of K0.39Na0.61Ta0.42Nb0.58O3 was larger than those of the other two crystals grown was elucidated through first-principles calculations. The obtained results indicated that K1-xNaxTa1-yNbyO3 crystals can be used as a high-quality, lead-free piezoelectric material.

  16. Meningitic Escherichia coli K1 penetration and neutrophil transmigration across the blood-brain barrier are modulated by alpha7 nicotinic receptor.

    Directory of Open Access Journals (Sweden)

    Feng Chi

    Full Text Available Alpha7 nicotinic acetylcholine receptor (nAChR, an essential regulator of inflammation, is abundantly expressed in hippocampal neurons, which are vulnerable to bacterial meningitis. However, it is unknown whether α7 nAChR contributes to the regulation of these events. In this report, an aggravating role of α7 nAChR in host defense against meningitic E. coli infection was demonstrated by using α7-deficient (α7(-/- mouse brain microvascular endothelial cells (BMEC and animal model systems. As shown in our in vitro and in vivo studies, E. coli K1 invasion and polymorphonuclear neutrophil (PMN transmigration across the blood-brain barrier (BBB were significantly reduced in α7(-/- BMEC and α7(-/- mice. Stimulation by nicotine was abolished in the α7(-/- cells and animals. The same blocking effect was achieved by methyllycaconitine (α7 antagonist. The tight junction molecules occludin and ZO-1 were significantly reduced in the brain cortex of wildtype mice infected with E. coli and treated with nicotine, compared to α7(-/- cells and animals. Decreased neuronal injury in the hippocampal dentate gyrus was observed in α7(-/- mice with meningitis. Proinflammatory cytokines (IL-1β, IL-6, TNFα, MCP-1, MIP-1alpha, and RANTES and adhesion molecules (CD44 and ICAM-1 were significantly reduced in the cerebrospinal fluids of the α7(-/- mice with E. coli meningitis. Furthermore, α7 nAChR is the major calcium channel for nicotine- and E. coli K1-increased intracellular calcium concentrations of mouse BMEC. Taken together, our data suggest that α7 nAChR plays a detrimental role in the host defense against meningitic infection by modulation of pathogen invasion, PMN recruitment, calcium signaling and neuronal inflammation.

  17. The study of BRAF V600E and MAP2K1 gene mutations in Langerhans cell histiocytosis and its significance of clinical findings%朗格汉斯细胞组织细胞增生症中 BRAF V600E 和 MAP2K1基因突变的分析及其临床意义

    Institute of Scientific and Technical Information of China (English)

    曾凯旋; 赵丹珲; 王哲; 闫庆国; 郭双平; 郭英; 刘一雄; 张微晨; 张月华; 李明阳; 李侠; 魏洁; 吕杨

    2016-01-01

    目的:探讨我国朗格汉斯细胞组织细胞增生症(Langerhans cell histiocytosis,LCH)中 BRAF V600E 和MAP2K1基因突变发生状况及其临床意义。方法:随机选取35例 LCH 组织标本,采用桑格测序法检测其中BRAF V600E 和 MAP2K1基因突变状况,免疫组化法检测 BRAF V600E 蛋白的表达。分析 BRAF V600E、MAP2K1基因突变与 LCH 临床基本资料(年龄、性别、单/多系统)的关系。结果:在35例 LCH 患者中,男女比例为1.7∶1,82.9%侵及骨组织,97.1%是单系统 LCH(single system LCH,SS -LCH),2.9%是多系统 LCH (multi -system LCH,MS -LCH)。桑格测序法检测 BRAF V600E 基因突变率为17.1%,MAP2K1基因突变率为14.3%,MAP2K1与 BRAF V600E 基因突变有互异性;免疫组化法检测 BRAF V600E 阳性表达率为28.6%,涵盖了桑格测序法测得的突变病例。BRAF V600E 和 MAP2 K1基因突变更多出现在未成年组(35.7%和28.6%),其中 BRAF V600E 突变在未成年人组与成人组间有显著性差异(P =0.028);BRAF V600E 和MAP2K1基因突变对生存的影响无统计学差异(P >0.05)。结论:我国 LCH 患者大部分都是 SS -LCH,主要侵及的部位是骨组织,且预后良好,5年生存率为97.1%。桑格法所测的 BRAF V600E 和 MAP2K1基因突变率均低于西方报道,两者存在互异性,分别为17.1%和14.3%。所有 MAP2K1基因突变都是点突变,没有框内缺失突变,发现一个新的突变位点:c.112 G >A p.E38K;BRAF V600E 和 MAP2K1基因突变主要发生于未成年组中,提示各年龄层中 LCH 的发病机理可能不同,可能 RAS /RAF /MEK/ERK 通路在未成年人 LCH 中发挥更重要的作用;另外这两种突变对 LCH 的生存无影响。%Objective:To study BRAF V600E and MAP2K1 gene mutations in Langerhans cell histiocytosis and analyze the relationship between the

  18. TLR5激动剂鞭毛蛋白预防小鼠异基因造血干细胞移植后急性移植物抗宿主病的初步研究%Prophylactic Effect of TLR5 Agonist Flagellin on Acute Graft versus Host Disease after Allogeneic Hematopietic Stem Cell Transplantation and Its Mechanism

    Institute of Scientific and Technical Information of China (English)

    龚旭东; 马梁明; 朱镭; 郭慧敏; 任连生; 任瑞瑞; 张华屏; 卫芬; 牛燕燕

    2012-01-01

    本研究旨在探讨Toll样受体(TLR5)激动剂鞭毛蛋白(flagellin)对小鼠异基因造血干细胞移植(allo-HSCT)后急性移植物抗宿主病(aGVHD)的预防作用和可能作用机制.用主要组织相容性抗原完全不合的纯种近交系小鼠[供体鼠:雄性C57BL/6鼠;受体鼠:雌性BALB/c鼠]建立allo-HSCT的aGVHD动物模型.受鼠随机分为3组:鞭毛蛋白组,于移植前后2次尾静脉注射高纯度(纯度≥95%)的鞭毛蛋白50μl[5μg/(只·次)]预防aGVHD;单纯移植组,移植后仅给予等容量PBS;单纯照射组,仅照射不移植,亦给予等容量PBS.观察比较移植后aGVHD的表现.结果表明,移植小鼠均出现典型的aGVHD症状,单纯移植组小鼠死亡高峰在移植后第4-5天.用鞭毛蛋白作为aGVHD预防方案小鼠的aGVHD症状明显减轻,平均生存时间较单纯移植组显著延长(P<0.05).三组小鼠移植前外周血白细胞数比较均无显著性差异,但在照射后第14、21天,鞭毛蛋白组较单纯移植组外周血白细胞数显著性升高(P<0.05).鞭毛蛋白预防组移植小鼠aGVHD病理表现较单纯移植组明显减轻.流式细胞仪检测鞭毛蛋白组与单纯移植组移植前后不同时间点Treg细胞/CD4+T细胞含量结果也表明,移植后2-4周鞭毛蛋白组小鼠的Treg细胞数较单纯移植组明显增加(P<0.05).结论:鞭毛蛋白对小鼠allo-HSCT后发生aGVHD有预防作用,能减轻其症状和病理损害程度,显著延长其平均生存时间,其机制有可能通过增加移植后小鼠的Treg细胞含量,从而有效改善并减轻移植后小鼠aGVHD.%This study was aimed to investigate the prophylactic effect of Toll like receptor (TLR)5 agonist flagellin on acute graft versus host disease (aGVHD) after allogeneic hematopietic stem cell transplantation (allo-HSCT) and its possible mechemism. The animal model with allo-HSCT aGVHD was established by using purebreed mice (male mouse C57BL/6 as donor, femal mouse BALB/c as recipient

  19. The naringenin-induced exoproteome of Rhizobium etli CE3.

    Science.gov (United States)

    Meneses, Niurka; Taboada, Hermenegildo; Dunn, Michael F; Vargas, María Del Carmen; Buchs, Natasha; Heller, Manfred; Encarnación, Sergio

    2017-03-02

    Flavonoids excreted by legume roots induce the expression of symbiotically essential nodulation (nod) genes in rhizobia, as well as that of specific protein export systems. In the bean microsymbiont Rhizobium etli CE3, nod genes are induced by the flavonoid naringenin. In this study, we identified 693 proteins in the exoproteome of strain CE3 grown in minimal medium with or without naringenin, with 101 and 100 exoproteins being exclusive to these conditions, respectively. Four hundred ninety-two (71%) of the extracellular proteins were found in both cultures. Of the total exoproteins identified, nearly 35% were also present in the intracellular proteome of R. etli bacteroids, 27% had N-terminal signal sequences and a significant number had previously demonstrated or possible novel roles in symbiosis, including bacterial cell surface modification, adhesins, proteins classified as MAMPs (microbe-associated molecular patterns), such as flagellin and EF-Tu, and several normally cytoplasmic proteins as Ndk and glycolytic enzymes, which are known to have extracellular "moonlighting" roles in bacteria that interact with eukaryotic cells. It is noteworthy that the transmembrane ß (1,2) glucan biosynthesis protein NdvB, an essential symbiotic protein in rhizobia, was found in the R. etli naringenin-induced exoproteome. In addition, potential binding sites for two nod-gene transcriptional regulators (NodD) occurred somewhat more frequently in the promoters of genes encoding naringenin-induced exoproteins in comparison to those ofexoproteins found in the control condition.

  20. Integral Representation of the Pictorial Proof of Sum of [superscript n][subscript k=1]k[superscript 2] = 1/6n(n+1)(2n+1)

    Science.gov (United States)

    Kobayashi, Yukio

    2011-01-01

    The pictorial proof of the sum of [superscript n][subscript k=1] k[superscript 2] = 1/6n(n+1)(2n+1) is represented in the form of an integral. The integral representations are also applicable to the sum of [superscript n][subscript k-1] k[superscript m] (m greater than or equal to 3). These representations reveal that the sum of [superscript…

  1. Integral Representation of the Pictorial Proof of Sum of [superscript n][subscript k=1]k[superscript 2] = 1/6n(n+1)(2n+1)

    Science.gov (United States)

    Kobayashi, Yukio

    2011-01-01

    The pictorial proof of the sum of [superscript n][subscript k=1] k[superscript 2] = 1/6n(n+1)(2n+1) is represented in the form of an integral. The integral representations are also applicable to the sum of [superscript n][subscript k-1] k[superscript m] (m greater than or equal to 3). These representations reveal that the sum of [superscript…

  2. Purification and immunochemical properties of Escherichia coli B polysaccharide cross-reacting with Salmonella typhi Vi antigen: preliminary evidence for cross-reaction of the polysaccharide with Escherichia coli K1 antigen.

    OpenAIRE

    Szewczyk, B; Taylor, A

    1983-01-01

    An acidic polysaccharide of Escherichia coli B was isolated by a mild procedure and purified to homogeneity. The polysaccharide was found to react in Salmonella typhi Vi antisera and E. coli K1 antisera. Serological analysis and preliminary chemical characterization of the polysaccharide indicated that it is an aminouronic acid polymer which, although not structurally identical to either Vi or K1, appears more like the Vi antigen, both immunochemically and chemically.

  3. The 8th International Comparison of Absolute Gravimeters 2009: the first Key Comparison (CCM.G-K1) in the field of absolute gravimetry

    Science.gov (United States)

    Jiang, Z.; Pálinkáš, V.; Arias, F. E.; Liard, J.; Merlet, S.; Wilmes, H.; Vitushkin, L.; Robertsson, L.; Tisserand, L.; Pereira Dos Santos, F.; Bodart, Q.; Falk, R.; Baumann, H.; Mizushima, S.; Mäkinen, J.; Bilker-Koivula, M.; Lee, C.; Choi, I. M.; Karaboce, B.; Ji, W.; Wu, Q.; Ruess, D.; Ullrich, C.; Kostelecký, J.; Schmerge, D.; Eckl, M.; Timmen, L.; Le Moigne, N.; Bayer, R.; Olszak, T.; Ågren, J.; Del Negro, C.; Greco, F.; Diament, M.; Deroussi, S.; Bonvalot, S.; Krynski, J.; Sekowski, M.; Hu, H.; Wang, L. J.; Svitlov, S.; Germak, A.; Francis, O.; Becker, M.; Inglis, D.; Robinson, I.

    2012-12-01

    The 8th International Comparison of Absolute Gravimeters (ICAG2009) took place at the headquarters of the International Bureau of Weights and Measures (BIPM) from September to October 2009. It was the first ICAG organized as a key comparison in the framework of the CIPM Mutual Recognition Arrangement of the International Committee for Weights and Measures (CIPM MRA) (CIPM 1999). ICAG2009 was composed of a Key Comparison (KC) as defined by the CIPM MRA, organized by the Consultative Committee for Mass and Related Quantities (CCM) and designated as CCM.G-K1. Participating gravimeters and their operators came from national metrology institutes (NMIs) or their designated institutes (DIs) as defined by the CIPM MRA. A Pilot Study (PS) was run in parallel in order to include gravimeters and their operators from other institutes which, while not signatories of the CIPM MRA, nevertheless play important roles in international gravimetry measurements. The aim of the CIPM MRA is to have international acceptance of the measurement capabilities of the participating institutes in various fields of metrology. The results of CCM.G-K1 thus constitute an accurate and consistent gravity reference traceable to the SI (International System of Units), which can be used as the global basis for geodetic, geophysical and metrological observations of gravity. The measurements performed afterwards by the KC participants can be referred to the international metrological reference, i.e. they are SI-traceable. The ICAG2009 was complemented by a number of associated measurements: the Relative Gravity Campaign (RGC2009), high-precision levelling and an accurate gravity survey in support of the BIPM watt balance project. The major measurements took place at the BIPM between July and October 2009. Altogether 24 institutes with 22 absolute gravimeters (one of the 22 AGs was ultimately withdrawn) and nine relative gravimeters participated in the ICAG/RGC campaign. This paper is focused on the

  4. GM3 alpha2,8-sialyltransferase (GD3 synthase): protein characterization and sub-golgi location in CHO-K1 cells.

    Science.gov (United States)

    Daniotti, J L; Martina, J A; Giraudo, C G; Zurita, A R; Maccioni, H J

    2000-04-01

    GD3 synthase (Sial-T2) is a key enzyme of ganglioside synthesis that, in concert with GM2 synthase (GalNAc-T), regulates the ratio of a- and b-pathway gangliosides. In this work, we study the sub-Golgi location of an epitope-tagged version of chicken Sial-T2 transfected to CHO-K1 cells. The expressed protein was enzymatically active both in vitro and in vivo and showed a molecular mass of approximately 47 or approximately 95 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence or absence of, respectively, beta-mercaptoethanol. The 95-kDa form of Sial-T2 was also detected if the protein was retained in the endoplasmic reticulum (ER) due to impaired glycosylation, indicating that it was formed in the ER. Confocal immunofluorescence microscopy showed Sial-T2 localized to the Golgi complex and, within the organelle, partially co-localizing with the mannose-6-phosphate receptor, a marker of the trans-Golgi network (TGN). In cells treated with brefeldin A, a major fraction of Sial-T2 redistributed to the ER, even under controlled expression to control for mislocalization due to protein overloading. In experiments of incorporation of sugars into endogenous acceptors of Golgi membranes in vitro, GD3 molecules formed by incubation with CMP-NeuAc were converted to GD2 upon incubation with UDP-GalNAc. These results indicate that Sial-T2 localizes mainly to the proximal Golgi, although a fraction is located in the TGN functionally coupled to GalNAc-T. Consistent with this, most of the enzyme was in an endoglycosidase H (Endo-H)-sensitive, neuraminidase (NANase)-insensitive form. A minor secreted form lacking approximately 40 amino acids was Endo-H-resistant and NANase-sensitive, indicating that the cells were able to process N-glycans to Endo-H-resistant forms. Taken together, the results of these biochemical and immunocytochemical experiments indicate that in CHO-K1 cells, most Sial-T2 localizes in the proximal Golgi and that a functional fraction

  5. Role of p70S6K1-mediated phosphorylation of eIF4B and PDCD4 proteins in the regulation of protein synthesis.

    Science.gov (United States)

    Dennis, Michael D; Jefferson, Leonard S; Kimball, Scot R

    2012-12-14

    Modulation of mRNA binding to the 40 S ribosomal subunit during translation initiation controls not only global rates of protein synthesis but also regulates the pattern of protein expression by allowing for selective inclusion, or exclusion, of mRNAs encoding particular proteins from polysomes. The mRNA binding step is modulated by signaling through a protein kinase known as the mechanistic target of rapamycin complex 1 (mTORC1). mTORC1 directly phosphorylates the translational repressors eIF4E binding proteins (4E-BP) 1 and 2, releasing them from the mRNA cap binding protein eIF4E, thereby promoting assembly of the eIF4E·eIF4G complex. mTORC1 also phosphorylates the 70-kDa ribosomal protein S6 kinase 1 (p70S6K1), which subsequently phosphorylates eIF4B, and programmed cell death 4 (PDCD4), which sequesters eIF4A from the eIF4E·eIF4G complex, resulting in repressed translation of mRNAs with highly structured 5'-untranslated regions. In the present study, we compared the role of the 4E-BPs in the regulation of global rates of protein synthesis to that of eIF4B and PDCD4. We found that maintenance of eIF4E interaction with eIF4G was not by itself sufficient to sustain global rates of protein synthesis in the absence of mTORC1 signaling to p70S6K1; phosphorylation of both eIF4B and PDCD4 was additionally required. We also found that the interaction of eIF4E with eIF4G was maintained in the liver of fasted rats as well as in serum-deprived mouse embryo fibroblasts lacking both 4E-BP1 and 4E-BP2, suggesting that the interaction of eIF4G with eIF4E is controlled primarily through the 4E-BPs.

  6. Observation on the Effect of Early Puncture Drainage in the Treatment of Patients with Intracranial Hemorrhage Caused by Delayed Vitamin K1 Deficiency%早期穿刺引流在迟发维生素K1缺乏致颅内出血中的疗效观察

    Institute of Scientific and Technical Information of China (English)

    栾劲; 杨皎莹

    2014-01-01

    目的:观察早期穿刺引流在迟发维生素K1缺乏致颅内出血中的疗效。方法:选取2010年1月-2013年4月于本院进行常规治疗的27例迟发维生素K1缺乏致颅内出血患儿为对照组,27例进行早期穿刺引流治疗的患儿为观察组,然后将两组患儿的治疗总有效率、出血停止时间、住院时间、并发症发生率及不同时间的NABA评分进行比较。结果:观察组患儿的治疗总有效率高于对照组,出血停止时间及住院时间均短于对照组,并发症发生率低于对照组,不同时间的NABA评分均优于对照组,差异均有统计学意义( P<0.05)。结论:早期穿刺引流在迟发维生素K1缺乏致颅内出血中的疗效较好,可有效改善患儿的预后。%Objective:To observe the effect of early puncture drainage in the treatment of patients with intracranial hemorrhage caused by delayed vitamin K1 deficiency.Method:27 newborns with intracranial hemorrhage caused by delayed vitamin K1 deficiency who were treated with routine treatment in our hospital from January 2010 to April 2013 were the control group,27 cases who were treated with early puncture drainage were the observation group,then the total effective rate,stopping time of bleeding,hospitalization time,rate of complications and NABA score at different time of two groups were compared.Result:The total effective rate of observation group was higher than that of control group,stopping time of bleeding and hospitalization time were shorter than those of control group,rate of complications was lower than that of control group,ABA score at different time were better than those of control group,there were all significant differences(P<0.05).Conclusion:The effect of early puncture drainage in the treatment of patients with intracranial hemorrhage caused by delayed vitamin K1 deficiency is better,and it can effectively improve the prognosis of newborns.

  7. 藏灵菇酵母菌K1耐胃肠道逆环境特性及降胆固醇的实验研究%Tolerance characteristics of the gastrointestinal tract and cholesterol-lowering effects of Kluyveromyces marxianus K1 from Tibetan Kefir

    Institute of Scientific and Technical Information of China (English)

    刘慧; 李晨; 杨磊; 贾慧; 张红星

    2010-01-01

    利用从藏灵菇中筛选的马克斯克鲁维酵母K1菌株探讨胃肠道耐受性及体外降胆固醇功效.分别在pH 1.5~4.5、胆盐浓度0.3%~3.0%、NaCl浓度3.0%~6.0%的模拟胃肠道环境下,采用活菌计数法检测K1菌株的耐胃肠道逆环境特性,以邻本二甲醛法检测其降胆固醇效果.结果表明,在pH2.5~4.5作用4h活菌数超过106cfu/mL,0.3%~3.0%胆盐及3.0%~6.0%NaCl作用8h活菌数为104cfu/mL~106cfu/mL,在pH1.5~3.0(4h)及胆盐浓度0.3%~3.0%的模拟人体胃肠道环境下降胆固醇率40.0%~52.8%.显示K1菌株具有较强的耐胃肠道逆环境特性及良好的降胆固醇能力.

  8. Evaluation of the radio modifier effect of propolis on chinese hamster ovary (CHO-K1) and human prostate cancer (PC3) cells, irradiated with 60-CO; Avaliacao do efeito radiomodificador da propolis em celulas de ovario de hamster chines (CHO-K1) e em celulas tumorais de prostata (PC3), irradiadas com CO-60

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Geyza Spigoti

    2011-07-01

    In the last decades, it has been given a great interest to investigations concerning natural, effective, nontoxic compounds with radioprotective potential together with the increasing utilization of different types of ionizing radiation for various applications. Among them propolis, a resinous compound produced by honeybees (Apis mellifera), has been considered quite promising, since it presents several advantageous biological characteristics, i. e., anti-inflammatory, antimicrobial, anticarcinogenic, antioxidant and also free radical scavenging action. The purpose of the present study was to evaluate the effect of Brazilian propolis, collected in the State of Rio Grande do Sul, on Chinese hamster ovary (CHO-K1) and human prostate cancer (PC3) cells, irradiated with {sup 60}Co {gamma} radiation. For this purpose, three interlinked parameters were analyzed: micronucleus induction, cell viability and clonogenic death. The choice of these parameters was justified by their biological significance, in addition to the fact that they are readily observable and measurable in irradiated cells. The cytogenetic data obtained showed a radioprotective effect of propolis (5-100 {mu}g/ml) in the induction of DNA damage for both cell lines, irradiated with doses of 1 - 4 Gy. The cytotoxicity assay, however, showed a prominent antiproliferative effect of propolis (50 - 400{mu}/ml) in PC3 cells irradiated with 5 G{gamma}. The survival curves obtained were adequately fitted by a linear-quadratic model, where the {alpha} coefficient was higher in CHO-K1 cells. Concerning the clonogenic capacity, PC3 cells were more radiosensitive than CHO-K1 cells at the higher doses of the survival curve. Propolis at the concentrations of 30 - 100 {mu}g/ml, did not influence the clonogenic potential of PC3 cells, since the survival curves, associated or not with propolis, were found similar, although the combined treatment in CHO-K1 cells exhibited a stimulating proliferative effect. The data

  9. Radiation induced pesticidal microbes

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Ki Yup; Lee, Y. K.; Kim, J. S.; Kim, J. K.; Lee, S. J.; Lim, D. S

    2001-01-01

    To isolate pesticidal microbes against plant pathogenic fungi, 4 strains of bacteria(K1. K3, K4, YS1) were isolated from mushroom compost and hot spring. K4, K1, K3, YS1 strain showed wide antifungal spectrum and high antifungal activities against 12 kinds of fungi. Specific proteins and the specific transcribed genes were found from the YS1 and its radiation-induced mutants. And knock-out mutants of antifungal activity were derived by transposon mutagenesis. From these knock-out mutants, the antifungal activity related genes and its modification by gamma-ray radiation are going to be studied. These results suggested that radiation could be an useful tool for the induction of functional mutants.

  10. Genotypic analysis on the ORF-K1 gene of human herpesvirus 8 from patients with Kaposi's sarcoma in Xinjiang, China

    Institute of Scientific and Technical Information of China (English)

    Dezhi Zhang; Xiongming Pu; Weidong Wu; Ying Jin; Xiujuan Wu

    2008-01-01

    Human herpesvirus 8 (HH'V-8) is thought to be essential for the development of all forms of Kaposi's sarcoma (KS). HHV-8 DNA is present virtually in all KS tumor biopsy samples. Genes at both ends of the HHV-8 genome have been shown to vary considerably. Seve nmajor molecular subtypes of HHV-8 were defined based on the amino acid sequence of the open reading frame K1 (ORF-Kl), generally known as A, B, C, D, E, E and Z. Most strains collected worldwide were clustered into two subtypes (A and C). Here, the Kl/VRl region of HHV-8 was amplified by nested PCR in 22 (81.48%) of 27 cases from Xinjiang Uygur Autonomous Region, a province in northwest-ern China. Phylogenetic analysis on the basis of the KI/VRI amino acid sequence indicated that the majority of these KS patients were infected by subtype C HHV-8 (n = 18, including 15 belonging to the C2 group), and several by subtype A (n = 4, including 3 being the Al group). This is the fast report of subtype A HHV-g in China. Furthermore, the correlations between different forms and lesions of KS and different subtypes of HHV-8 were analyzed. The findings showed that subtype A HHV-8 resulted in significantly more frequent mucosal KS lesions than subtype C. However, there was no obvious correlation between different forms of KS and different subtypes of HHV-8.

  11. Investigation of diffuse phase transition in ferroelectric Pb2- x K1+ x Li x Nb5O15 (0 ≤ x ≤ 1.5) ceramics

    Science.gov (United States)

    Choukri, E.; Neqali, A.; Abkhar, Z.; Alimoussa, A.; Hajji, L.; Mezzane, D.; Belboukhari, A.; Amjoud, M.; Gagou, Y.; El Marssi, M.; Luk'yanchuk, I.

    2016-06-01

    Substitution of Pb with Li and K in the Pb2KNb5O15 phases leads to a new composition with chemical composition Pb2- x K1+ x Li x Nb5O15 which crystallizes with tetragonal tungsten bronze-type structure. Ferroelectric ceramics with different compositions were synthesized using solid-state reaction and complex dielectric permittivity measurements in these compounds were performed in a frequency and temperature range of 20 Hz-1 MHz and from 25 to 550 °C, respectively. Special attention was paid to the diffuse phase transition (DPT) that occurs close to the Curie temperature. The empirical equation proposed by Santos-Eiras for a phenomenological description of the temperature dependence of the dielectric permittivity (\\varepsilon_{{r}}^' }}) peak is used to calculate some characteristic parameters of DPT. From the results, it must be assumed that these compounds show a diffuse phase transition with non-relaxor behavior. A basic phase diagram showing the evolution of T m function of composition x is deduced from this study.

  12. The CO2 Abundance in Comets C/2012 K1 (PanSTARRS), C/2012 K5 (LINEAR), and 290P/Jager as Measured with Spitzer

    CERN Document Server

    McKay, Adam J; Cochran, Anita L; Bodewits, Dennis; DiSanti, Michael A; Russo, Neil Dello; Lisse, Carey M

    2015-01-01

    We present analysis of observations of CO2 and OI emission in three comets to measure the CO2 abundance and evaluate the possibility of employing observations of OI emission in comets as a proxy for CO2. We obtained NIR imaging sensitive to CO2 of comets C/2012 K1 (PanSTARRS), C/2012 K5 (LINEAR), and 290P/Jager with the IRAC instrument on Spitzer. We acquired observations of OI emission in these comets with the ARCES echelle spectrometer mounted on the 3.5-meter telescope at Apache Point Observatory and observations of OH with the Swift observatory (PanSTARRS) and with Keck HIRES (Jager). The CO2/H2O ratios derived from the Spitzer images are 12.6 +/- 1.3% (PanSTARRS), 28.9 +/- 3.6% (LINEAR), and 31.3 +/- 4.2% (Jager). These abundances are derived under the assumption that contamination from CO emission is negligible. The CO2 abundance for PanSTARRS is close to the average abundance measured in comets at similar heliocentric distance to date, while the abundances measured for LINEAR and Jager are significantl...

  13. Phylogenetic relationships of Cyprinidae (Teleostei: Cypriniformes) inferred from the partial S6K1 gene sequences and implication of indel sites in intron 1

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The family Cyprinidae is widely distributed in East Asia, and has the important phylogenetic signifi- cance in the fish evolution. In this study, the 5′ end partial sequences (containing exon 1, exon 2 and indel 1) of S6K1 gene were obtained from 30 representative species in Cyprinidae and outgroup using PCR amplification and sequencing. The phylogenetic relationships of Cyprinidae were reconstructed with neighbor joining (NJ), maximum parsimony (MP), maximum likelihood (ML), and Bayesian meth- ods. Myxocyprinus asiaticus (Catostomidae) was assigned to the outgroup taxon. Similar phylogenetic relationships within the family Cyprinidae were achieved with the four analyses. Leuciscini and Barbini were monophyletic lineages respectively with the high nodal supports. Leuciscini comprises Hypophthalmichthyinae, Xenocyprinae, Cultrinae, Gobioninae, Acheilognathinae and East Asian species of Leuciscinae and Danioninae. Monophyly of East Asian clade was supported with high nodal support. Barbini comprises Schizothoracinae, Barbinae, Cyprininae and Labeoninae. The monophyletic lineage consisting of Danio rerio, D. myersi, and Rasbora trilineata was basal in the tree. In addition, the large fragment indels in intron 1 were analyzed to improve the understanding of Cyprinidae relationships. The results showed that the large fragment indels were correlated with the relations among species. Some conserved regions in intron 1 were thought to be involved in the functional regulation. However, no correlation was found between sequence variations and species characteristic size.

  14. KEY COMPARISON Activity measurements of the radionuclide 111In for the LNE-LNHB, France in the ongoing comparison BIPM.RI(II)-K1.In-111

    Science.gov (United States)

    Michotte, C.; Ratel, G.; Courte, S.; Verdeau, E.; Amiot, M.-N.

    2010-01-01

    In 2006, the Laboratoire national de métrologie et d'essais-Laboratoire national Henri Becquerel (LNE-LNHB) submitted a sample of known activity of 111In to the International Reference System (SIR). The value of the activity submitted was about 14 MBq. This provides a new result for France in the matrix of degrees of equivalence in the key comparison database that enables them to maintain their degrees of equivalence for this radionuclide. The key comparison reference value has been re-evaluated and the comparison, identifier BIPM.RI(II)-K1.In-111, now contains five valid results. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI Section II, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  15. KEY COMPARISON: COOMET.RI(I)-K1 comparison of national measurement standards of air kerma for 60Co γ radiation

    Science.gov (United States)

    Büermann, L.; Oborin, A. V.; Dobrovosky, J.; Milevsky, V. S.; Walwyn Salas, G.; Lapenas, A.

    2009-01-01

    Results are presented of the COOMET key comparison of the national measurement standards of air kerma for 60Co γ radiation. Participants of the comparison were PTB (Germany, pilot institute), VNIIM (Russia), SMU (Slovakia), BelGIM (Belarus), CPHR (Cuba) and RMTC (Latvia). PTB, VNIIM and SMU had previously taken part in a key comparison with the Bureau International de Poids et Mesures (BIPM) and operated as link laboratories in order to evaluate the degree of equivalence of the participants' results with the key comparison reference value. These data form the basis of the results entered into the BIPM key comparison database for comparison COOMET.RI(I)-K1. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI Section I, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  16. Comparative study of the cytotoxicity and genotoxicity of kaurenoic acid and its semi-synthetic derivatives methoxy kaurenoic acid and kaurenol in CHO-K1 cells.

    Science.gov (United States)

    Cano, Bruno Limonti; Moreira, Monique Rodrigues; Goulart, Mirian Oliveira; Dos Santos Gonçalves, Natália; Veneziani, Rodrigo Cassio Sola; Bastos, Jairo Kenupp; Ambrósio, Sérgio Ricardo; Dos Santos, Raquel Alves

    2017-04-01

    The diterpene kaurenoic acid (KA) has vasorelaxant, antimicrobial, anti-tumoural and anti-leishmanial effects. Semi-synthetic derivatives were obtained to achieve more satisfactory responses. The assessment of genotoxicity is part of the toxicological evaluation of therapeutic compound candidates. The present study investigated the cytotoxicity and genotoxicity of KA and its semi-synthetic derivatives methoxy kaurenoic acid (MKA) and kaurenol (KRN) using the CHO-K1 cell line. The cytotoxicity evaluation demonstrated that treatments with 200 and 400 μM KA reduced cellular proliferation to 36.5 and 4.43%, respectively, and that 100 and 200 μM KA reduced the survival fraction (SF) to 48.1 and 5.5%, respectively. MKA and KRN at concentrations of 400 μM reduced proliferation to 81 and 86.8%, respectively, while 100 and 200 μM KRN reduced the SF to 50%, and 200 μM MKA reduced the SF to 74%. No genotoxicity was observed for KA or MKA. However, 100 μM KRN increased the DNA damage index, as detected by comet assay, although a micronucleus assay did not confirm these data. The results demonstrated that KA and its semi-synthetic derivative MKA were not genotoxic when tested at noncytotoxic concentrations, but KRN was genotoxic at the highest concentration that was tested, as demonstrated by the comet assay. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Isolation of a carbapenem-resistant K1 serotype Klebsiella pneumonia strain and the study of resistance mechanism%碳青霉烯类抗生素耐药K1型肺炎克雷伯菌一株的分离及耐药机制分析

    Institute of Scientific and Technical Information of China (English)

    张嵘; 王选; 吕建新

    2014-01-01

    目的 对临床分离的1株碳青霉烯类抗生素耐药K1型肺炎克雷伯菌进行毒力及耐药机制分析.方法 采用Vitek 2 Compact全自动微生物鉴定仪对细菌进行鉴定;浓度梯度法(E-test)测定细菌对临床常用抗生素的最低抑菌浓度(MIC);乙二胺四乙酸(EDTA)协同试验和改良Hodge试验分别检测金属酶和碳青霉烯酶;聚合酶链反应(PCR)扩增毒力基因K1、K2、K5、K20、K54、K57、rmpA、magA、wcaG和β内酰胺类抗生素耐药基因;接合试验检测耐药基因和毒力基因是否存在于质粒上;接合子质粒进行PBRT质粒分型;多位点序列分析(MLST)对该菌株进行分子分型.结果 药敏结果显示该菌株对临床常用的头孢类和碳青霉烯类抗生素耐药;EDTA协同试验和改良Hodge试验显示该菌株产碳青霉烯酶,不含金属酶;PCR扩增及序列分析发现该菌株携带耐药基因blaKPC-2、blaCTX-M-15、blaTEM-1、blaSHV-1,同时有毒力基因K1、magA、rmpA、wcaG;能通过接合试验将碳青霉烯耐药基因传递到大肠埃希菌EC600,接合子未发现毒力基因存在;PBRT质粒分型为Frep型质粒;MLST分型为ST23型.结论 产毒力基因的K1型肺炎克雷伯菌携带blaKPC-2碳青霉烯耐药基因,该耐药基因可以通过质粒发生转移.%Objective To study the virulence and mechanism of carbapenem resistance of a clinical isolate of carbapenem-resistant K1 serotype Klebsiella pneumonia strain.Methods Identification of isolate was carried out with VITEK-2 compact system.Antimicrobial susceptibility was determined by E-test; Metallo β-lactamases and carbapenemases screening were conducted by imipenem-EDTA double disc synergy test and modified Hodge test,respectively.Specific polymerehse chain reaction (PCR) and DNA sequencing were preformed to detect the virulence genes including K1,K2,KS,K20,K54,K57,magA,rmpA,wcaG and a series of β-lactamase resistence genes.Conjunction experiment was also performed.The plasmids of

  18. 离散时间 SM[K]/PH[K]/1/FCFS 排队系统的年龄过程改进分析%Analysis and Improvement of the Age Process in a Queuing System of Discrete Time SM[K]/PH[K]/1/FCFS

    Institute of Scientific and Technical Information of China (English)

    高卓; 徐德举

    2015-01-01

    We studied a discrete time queuing system with multiple types of customers and a first-come-first-served (FCFS) service discipline. Customers arrive according to a semi-Markov arrival process and the service times of individual customers have PH-distributions. We studied SM[K]/PH[K]/1/FCFS queue and analyzed its generalized age process particularly. We introduced some auxiliary variables to construct a Markov chain associated with ag(t) and obtained the transition probability matrix of this Markov chain.%基于一个离散时间的排队系统:顾客有着多种类型,成批到达,到达过程是一个半马尔可夫过程,按照先来先服务的准则,并且每一个顾客的服务时间服从各自的 PH 分布。对这个离散时间 SM[K]/PH[K]/1/FCFS 排队系统年龄过程进行了详细分析,引进一些附加变量构造一个关于年龄过程的马尔可夫链,从而计算出年龄过程的转移矩阵。

  19. Emergence of serotype K1 Klebsiella pneumoniae ST23 strains co-producing the plasmid-mediated AmpC beta-lactamase DHA-1 and an extended-spectrum beta-lactamase in Korea

    Directory of Open Access Journals (Sweden)

    Hae Suk Cheong

    2016-11-01

    Full Text Available Abstract Background Serotype K1 Klebsiella pneumoniae has emerged as an important community pathogen causing various infections, including liver abscesses. Although serotype K1 K. pneumoniae community isolates have been reported as susceptible to most classes of antimicrobial agents, a few cases of infection caused by extended-spectrum beta-lactamase (ESBL-producing serotype K1 K. pneumoniae have recently been reported in Asian countries. We identified three ESBL-producing strains of serotype K1 K. pneumoniae and conducted a molecular characterization of their drug resistance. Methods Three ESBL-producing serotype K1 K. pneumoniae ST23 strains were identified from strains in the Asian Bacterial Bank. Antimicrobial susceptibility testing was performed using the broth microdilution method, and ESBL production was tested by the double-disk synergy test and a confirmatory test. PCR was performed to detect the genes for plasmid-mediated ESBL and AmpC beta-lactamases. Results All three strains were resistant to cefotaxime, ceftazidime, and piperacillin/tazobactam, and all were determined to be ESBL-producers. No known ESBL genes, including bla SHV, bla TEM, bla CTX-M, bla GES, bla PER, and bla VEB, were detected among the three strains. Of all plasmid-mediated AmpC beta-lactamase (PAB genes, including bla DHA-1, bla CMY, bla FOX, and bla MOX, the bla DHA-1 gene was detected in two of the strains. The PFGE patterns revealed that the two isolates carrying bla DHA-1 were closely related (84% similarity. Conclusions No ESBL genes were detected among three ESBL-producing serotype K1 K. pneumoniae ST23 strains. Two strains contained the PAB gene bla DHA-1. The emergence of resistant strains of community-origin serotype K1 K. pneumoniae has important implications for effective treatment and infection control practices.

  20. Genetic variation of COLEC10 and COLEC11 and association with serum levels of collectin liver 1 (CL-L1 and collectin kidney 1 (CL-K1.

    Directory of Open Access Journals (Sweden)

    Rafael Bayarri-Olmos

    Full Text Available Collectin liver 1 (CL-L1, alias CL-10 and collectin kidney 1 (CL-K1, alias CL-11, encoded by the COLEC10 and COLEC11 genes, respectively, are highly homologous soluble pattern recognition molecules in the lectin pathway of complement. These proteins may be involved in anti-microbial activity and in tissue development as mutations in COLEC11 are one of the causes of the developmental defect syndrome 3MC. We studied variations in COLEC10 and COLEC11, the impact on serum concentration and to what extent CL-L1 and CL-K1 serum concentrations are correlated. We sequenced the promoter regions, exons and exon-intron boundaries of COLEC10 and COLEC11 in samples from Danish Caucasians and measured the corresponding serum levels of CL-L1 and CL-K1. The median concentration of CL-L1 and CL-K1 was 1.87 μg/ml (1.00-4.14 μg/ml and 0.32 μg/ml (0.11-0.69 μg/ml, respectively. The level of CL-L1 strongly correlated with CL-K1 (ρ = 0.7405, P T (rs3820897 was associated with decreased levels of CL-K1 (P = 0.044. In conclusion, COLEC10 and COLEC11 are highly conserved, which may reflect biological importance of CL-L1 and CL-K1. Moreover, the strong inter individual correlation between the two proteins suggests that a major proportion are found as heterooligomers or subjected to the same regulatory mechanisms.

  1. Determination of trans vitamin K1 in infant and medical nutritional products using AOAC Method 999.15 with modified preparation and extraction procedures and C30 bonded phase chromatography: single-laboratory validation.

    Science.gov (United States)

    Schimpf, Karen J; Thompson, Linda B; Schmitz, Daniel J

    2010-01-01

    Modifications were made to AOAC Official Method 999.15 to extend its applicability to specialty infant formulas containing hydrolyzed proteins and free amino acids, and to medical and adult nutritional products. Minor changes to the sample preparation procedure and chromatographic separation improved vitamin K1 recoveries and reduced chromatographic interferences in these types of matrixes. Currently AOAC Method 999.15 is applicable only to the determination of total vitamin K1 (phylloquione) in infant formula and milk (fluid, ready-to-feed, and powdered) containing > 1 microg vitamin K1/100 g solids. AOAC Method 999.15 recoveries of vitamin K1 were improved by altering sample sizes, extraction solvents and amounts, and the reagent addition order and amount of water or aqueous solutions added. The chromatographic separation of vitamin K1 in medical nutritional products containing canola and marine oils was improved, and trans vitamin K1 was separated from the biologically inactive cis isomer in all products with a C30 3 microm column and a 100% methanol mobile phase. With these modifications to the extraction procedure and chromatographic separation, AOAC Method 999.15 demonstrated acceptable precision and accuracy for the quantitation of trans vitamin K1 in specialty infant formulas containing hydrolyzed proteins and free amino acids, and medical and adult nutritional products. A single-laboratory validation of these minor modifications was completed. Fourteen different product matrixes were analyzed during validation. The intermediate precision averaged 4.15% RSD (range 2.52-5.81% RSD), and recovery data averaged 100.1% (range 92.2-109%).

  2. Flagella from five Cronobacter species induce pro-inflammatory cytokines in macrophage derivatives from human monocytes.

    Directory of Open Access Journals (Sweden)

    Ariadnna Cruz-Córdova

    Full Text Available Cronobacter spp. are opportunistic pathogens linked to lie-threatening infections in neonates and contaminated powdered infant formula that has been epidemiologically associated with these cases. Clinical symptoms of Cronobacter include necrotizing enterocolitis, bacteremia, and meningitis. Flagella from C. sakazakii are involved in biofilm formation and its adhesion to epithelial cells. We investigated the role of flagella from C. sakazakii ST1 and ST4, C. malonaticus, C. muytjensii, C. turicensis and C. dublinensis during the activation of cytokines (IL-8, TNF-α, and IL-10 in macrophage derivatives from human monocytes, which has not been extensively studied. The production and identity of flagella from the five Cronobacter species were visualized and recognized with anti-flagella antibodies by immunogold labeling through transmission electron microscopy. Purified flagella were dissociated into monomers in 12% SDS-PAGE Coomassie blue-stained gels showing a band of ∼28 kDa and, in addition, mass spectrometry revealed the presence of several peptides that correspond to flagellin. Flagella (100 ng induced the release of IL-8 (3314-6025 pg/ml, TNF-α (39-359 pg/ml, and IL-10 (2-96 pg/ml, in macrophage isolates from human monocytes and similar results were obtained when flagella were dissociated into monomers. Inhibition assays using three dilutions of anti-flagella antibodies (1∶10, 1∶100, and 1∶200 suppressed the secretion of IL-8, TNF-α, and IL-10 between 95-100% using 100 ng of protein. A transfection assay using 293-hTLR5 cells showed IL-8 release of 197 pg/ml and suppression in the secretion of IL-8 when anti-hTLR5-IgA antibodies were used at different concentrations. These observations suggest that flagella and flagellin are involved in an inflammatory response dependent on TLR5 recognition, which could contribute to the pathogenesis of the bacteria.

  3. Flagella from Five Cronobacter Species Induce Pro-Inflammatory Cytokines in Macrophage Derivatives from Human Monocytes

    Science.gov (United States)

    Cruz-Córdova, Ariadnna; Rocha-Ramírez, Luz M.; Ochoa, Sara A.; Gónzalez-Pedrajo, Bertha; Espinosa, Norma; Eslava, Carlos; Hernández-Chiñas, Ulises; Mendoza-Hernández, Guillermo; Rodríguez-Leviz, Alejandra; Valencia-Mayoral, Pedro; Sadowinski-Pine, Stanislaw; Hernández-Castro, Rigoberto; Estrada-García, Iris; Muñoz-Hernández, Onofre; Rosas, Irma; Xicohtencatl-Cortes, Juan

    2012-01-01

    Cronobacter spp. are opportunistic pathogens linked to lie-threatening infections in neonates and contaminated powdered infant formula that has been epidemiologically associated with these cases. Clinical symptoms of Cronobacter include necrotizing enterocolitis, bacteremia, and meningitis. Flagella from C. sakazakii are involved in biofilm formation and its adhesion to epithelial cells. We investigated the role of flagella from C. sakazakii ST1 and ST4, C. malonaticus, C. muytjensii, C. turicensis and C. dublinensis during the activation of cytokines (IL-8, TNF-α, and IL-10) in macrophage derivatives from human monocytes, which has not been extensively studied. The production and identity of flagella from the five Cronobacter species were visualized and recognized with anti-flagella antibodies by immunogold labeling through transmission electron microscopy. Purified flagella were dissociated into monomers in 12% SDS-PAGE Coomassie blue-stained gels showing a band of ∼28 kDa and, in addition, mass spectrometry revealed the presence of several peptides that correspond to flagellin. Flagella (100 ng) induced the release of IL-8 (3314–6025 pg/ml), TNF-α (39–359 pg/ml), and IL-10 (2–96 pg/ml), in macrophage isolates from human monocytes and similar results were obtained when flagella were dissociated into monomers. Inhibition assays using three dilutions of anti-flagella antibodies (1∶10, 1∶100, and 1∶200) suppressed the secretion of IL-8, TNF-α, and IL-10 between 95–100% using 100 ng of protein. A transfection assay using 293-hTLR5 cells showed IL-8 release of 197 pg/ml and suppression in the secretion of IL-8 when anti-hTLR5-IgA antibodies were used at different concentrations. These observations suggest that flagella and flagellin are involved in an inflammatory response dependent on TLR5 recognition, which could contribute to the pathogenesis of the bacteria. PMID:23284883

  4. Vertical Extraction Process Implemented at the 118-K-1 Burial Ground for Removal of Irradiated Reactor Debris from Silo Structures - 12431

    Energy Technology Data Exchange (ETDEWEB)

    Teachout, Douglas B. [Vista Engineering Technologies, LLC, Richland, Washington, 99352 (United States); Adamson, Clinton J.; Zacharias, Ames [Washington Closure Hanford, LLC, Richland, Washington, 99352 (United States)

    2012-07-01

    The primary objective of a remediation project is the safe extraction and disposition of diverse waste forms and materials. Remediation of a solid waste burial ground containing reactor hardware and irradiated debris involves handling waste with the potential to expose workers to significantly elevated dose rates. Therefore, a major challenge confronted by any remediation project is developing work processes that facilitate compliant waste management practices while at the same time implementing controls to protect personnel. Traditional burial ground remediation is accomplished using standard excavators to remove materials from trenches and other excavation configurations often times with minimal knowledge of waste that will be encountered at a specific location. In the case of the 118-K-1 burial ground the isotopic activity postulated in historic documents to be contained in vertical cylindrical silos was sufficient to create the potential for a significant radiation hazard to project personnel. Additionally, certain reported waste forms posed an unacceptably high potential to contaminate the surrounding environment and/or workers. Based on process knowledge, waste management requirements, historic document review, and a lack of characterization data it was determined that traditional excavation techniques applied to remediation of vertical silos would expose workers to unacceptable risk. The challenging task for the 118-K-1 burial ground remediation project team then became defining an acceptable replacement technology or modification of an existing technology to complete the silo remediation. Early characterization data provided a good tool for evaluating the location of potential high exposure rate items in the silos. Quantitative characterization was a different case and proved difficult because of the large diameter of the silos and the potential for variable density of attenuating soils and waste forms in the silo. Consequently, the most relevant

  5. Regulated expression of the rat recombinant P2X(3) receptor in stably transfected CHO-K1 tTA cells.

    Science.gov (United States)

    Lachnit, W G; Oglesby, I B; Gever, J R; Gever, M; Huang, C; Li, X C; Jin, H; McGivern, J G; Ford, A P

    2000-07-01

    In this report, the regulatable expression by tetracycline of the rat recombinant P2X(3) receptor in stably transfected Chinese hamster ovary (CHO-K1) expressing the tetracycline-controlled transactivator (tTA) is described. cDNA encoding the rat P2X(3)-receptor was subcloned into pTRE (a tetracycline-repressible expression vector) which was used to transfect stably CHO-K1 tTA cells. Using whole cell patch clamp techniques, 100 microM ATP evoked inward currents of 2.9+/-1.6 nA in transfected cells grown in the absence of tetracycline (tet-). The P2X(3) receptor protein was detectable by immunoblot as early as 24 h and protein expression levels continued to increase as much as 192 h following activation of tTA by the removal of the antibiotic. Saturation binding isotherms using [35S]ATP gamma S yielded a pK(d) of 8.2+/-0.1 and a B(max) of 31.9+/-3.5 pmol/mg protein in tet- cell membranes and a pK(d) of 8.1+/-0.1 and a B(max) of 5.8+/-0.8 pmol/mg protein in tet+ cell membranes. The agonist ligands 2MeSATP and alpha beta MeATP displaced the binding of [35S]ATP gamma S in tet- cell membranes with very high affinity, yielding pIC(50) values of 9.4+/-0.2 and 7.5+/-0. 2, respectively. In tet+ cell membrane, displacement of [35S]ATP gamma S by 2MeSATP and alpha beta MeATP was of much lower affinity (pIC(50) values of 7.8 and 6.2, respectively). ATP, ADP and UTP showed similar displacement of [35S]ATP gamma S binding in tet- and tet+ cell membranes. In other experiments, cytosolic Ca(2+) was monitored using the fluorescent indicator, fluo-3. Increases in cytosolic Ca(2+) were elicited by 100 nM alpha beta MeATP in tet- cells while no increases in cytosolic Ca(2+) were detected below 100 microM alpha beta MeATP in either tet+ cells or untransfected cells. These calcium responses to alpha beta MeATP had a pEC(50) of 6.7 and were transient, returning to baseline within 120 s. Suramin produced concentration-dependent, parallel, dextral shifts of E/[A] curves to alpha beta Me

  6. Lack of influence of etretinate on the intestinal absorption of D-xylose and of phytomenadione (vitamin K1) as representatives of hydrophilic and lipophilic compounds.

    Science.gov (United States)

    Hartmann, D; Crevoisier, C; Dubach, U C; Forgo, I; Heizmann, P

    1987-12-01

    The possible influence of the aromatic retinoid etretinate (Tigason ) on the intestinal absorption of D-xylose and phytomenadione (vitamin K1) has been studied in 7 healthy male volunteers between 22 and 25 years of age. D-xylose and phytomenadione were selected as being representative of water-soluble and fat-soluble test compounds, respectively. Following an oral dose of 25 g D-xylose the plasma levels and urinary excretion pattern of the compound were followed over 6 h. The plasma concentrations of phytomenadione after an oral dose of 20 mg of the vitamin were measured over 14 h. These absorption tests were performed before (baseline) and at the end of a 3-week treatment period with 25 mg b.i.d. etretinate. In addition some parameters on the serum lipid status pre and post etretinate treatment were monitored. The pharmacokinetic and biochemical determinants after treatment with etretinate were referenced to the pretreatment values and nonparametric confidence intervals (a less than 0.05) for these ratios were calculated. With respect to the area, AUC, under the D-xylose plasma concentration--time curve (0.79 less than or equal to RAUC = 1.00 less than or equal to 1.17) as well as to the cumulative amount excreted into urine over 6 h (0.90 less than or equal to RSUM = 1.04 less than or equal to 1.28) and to renal clearance (0.91 less than or equal to Rcl = 1.05 less than or equal to 1.25), no effect of etretinate on absorption and/or renal handling of D-xylose was discernible.(ABSTRACT TRUNCATED AT 250 WORDS)

  7. Summer (subarctic) versus winter (subtropic) production affects spinach (Spinacia oleracea L.) leaf bionutrients: vitamins (C, E, Folate, K1, provitamin A), lutein, phenolics, and antioxidants.

    Science.gov (United States)

    Lester, Gene E; Makus, Donald J; Hodges, D Mark; Jifon, John L

    2013-07-24

    Comparison of spinach (Spinacia oleracea L.) cultivars Lazio and Samish grown during the summer solstice in the subarctic versus the winter solstice in the subtropics provided insight into interactions between production environment (light intensity), cultivar, and leaf age/maturity/position affecting bionutrient concentrations of vitamins (C, E, folate, K1, provitamin A), lutein, phenolics, and antioxidants. Growing spinach during the winter solstice in the subtropics resulted in increased leaf dry matter %, oxidized (dehydro) ascorbic acid (AsA), α- and γ-tocopherol, and total phenols but lower reduced (free) AsA, α-carotene, folate, and antioxidant capacity compared to summer solstice-grown spinach in the subarctic. Both cultivars had similar bionutrients, except for higher dehydroAsA, and lower α- and γ-tocopherol in 'Samish' compared to 'Lazio'. For most bionutrients measured, there was a linear, and sometimes quadratic, increase in concentrations from bottom to top canopy leaves. However, total phenolics and antioxidant capacity increased basipetally. The current study has thus demonstrated that dehydroAsA, α-tocopherol, and γ-tocopherol were substantially lower in subarctic compared to subtropical-grown spinach, whereas the opposite relationship was found for antioxidant capacity, α-carotene, and folates (vitamin B9). The observations are consistent with previously reported isolated effects of growth environment on bionutrient status of crops. The current results clearly highlight the effect of production environment (predominantly radiation capture), interacting with genetics and plant phenology to alter the bionutrient status of crops. While reflecting the effects of changing growing conditions, these results also indicate potential alterations in the nutritive value of foods with anticipated shifts in global climatic conditions.

  8. Yellow fluorescent protein-based assay to measure GABA(A channel activation and allosteric modulation in CHO-K1 cells.

    Directory of Open Access Journals (Sweden)

    Teres Johansson

    Full Text Available The γ-aminobutyric acid A (GABA(A ion channels are important drug targets for treatment of neurological and psychiatric disorders. Finding GABA(A channel subtype selective allosteric modulators could lead to new improved treatments. However, the progress in this area has been obstructed by the challenging task of developing functional assays to support screening efforts and the generation of cells expressing functional GABA(A ion channels with the desired subtype composition. To address these challenges, we developed a yellow fluorescent protein (YFP-based assay to be able to study allosteric modulation of the GABA(A ion channel using cryopreserved, transiently transfected, assay-ready cells. We show for the first time how the MaxCyte STX electropora