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Sample records for cricetidae k1 cells

  1. S6K1 controls pancreatic β cell size independently of intrauterine growth restriction.

    PubMed

    Um, Sung Hee; Sticker-Jantscheff, Melanie; Chau, Gia Cac; Vintersten, Kristina; Mueller, Matthias; Gangloff, Yann-Gael; Adams, Ralf H; Spetz, Jean-Francois; Elghazi, Lynda; Pfluger, Paul T; Pende, Mario; Bernal-Mizrachi, Ernesto; Tauler, Albert; Tschöp, Matthias H; Thomas, George; Kozma, Sara C

    2015-07-01

    Type 2 diabetes mellitus (T2DM) is a worldwide heath problem that is characterized by insulin resistance and the eventual loss of β cell function. As recent studies have shown that loss of ribosomal protein (RP) S6 kinase 1 (S6K1) increases systemic insulin sensitivity, S6K1 inhibitors are being pursued as potential agents for improving insulin resistance. Here we found that S6K1 deficiency in mice also leads to decreased β cell growth, intrauterine growth restriction (IUGR), and impaired placental development. IUGR is a common complication of human pregnancy that limits the supply of oxygen and nutrients to the developing fetus, leading to diminished embryonic β cell growth and the onset of T2DM later in life. However, restoration of placental development and the rescue of IUGR by tetraploid embryo complementation did not restore β cell size or insulin levels in S6K1-/- embryos, suggesting that loss of S6K1 leads to an intrinsic β cell lesion. Consistent with this hypothesis, reexpression of S6K1 in β cells of S6K1-/- mice restored embryonic β cell size, insulin levels, glucose tolerance, and RPS6 phosphorylation, without rescuing IUGR. Together, these data suggest that a nutrient-mediated reduction in intrinsic β cell S6K1 signaling, rather than IUGR, during fetal development may underlie reduced β cell growth and eventual development of T2DM later in life.

  2. S6K1 controls pancreatic β cell size independently of intrauterine growth restriction

    PubMed Central

    Um, Sung Hee; Sticker-Jantscheff, Melanie; Chau, Gia Cac; Vintersten, Kristina; Mueller, Matthias; Gangloff, Yann-Gael; Adams, Ralf H.; Spetz, Jean-Francois; Elghazi, Lynda; Pfluger, Paul T.; Pende, Mario; Bernal-Mizrachi, Ernesto; Tauler, Albert; Tschöp, Matthias H.; Thomas, George; Kozma, Sara C.

    2015-01-01

    Type 2 diabetes mellitus (T2DM) is a worldwide heath problem that is characterized by insulin resistance and the eventual loss of β cell function. As recent studies have shown that loss of ribosomal protein (RP) S6 kinase 1 (S6K1) increases systemic insulin sensitivity, S6K1 inhibitors are being pursued as potential agents for improving insulin resistance. Here we found that S6K1 deficiency in mice also leads to decreased β cell growth, intrauterine growth restriction (IUGR), and impaired placental development. IUGR is a common complication of human pregnancy that limits the supply of oxygen and nutrients to the developing fetus, leading to diminished embryonic β cell growth and the onset of T2DM later in life. However, restoration of placental development and the rescue of IUGR by tetraploid embryo complementation did not restore β cell size or insulin levels in S6K1–/– embryos, suggesting that loss of S6K1 leads to an intrinsic β cell lesion. Consistent with this hypothesis, reexpression of S6K1 in β cells of S6K1–/– mice restored embryonic β cell size, insulin levels, glucose tolerance, and RPS6 phosphorylation, without rescuing IUGR. Together, these data suggest that a nutrient-mediated reduction in intrinsic β cell S6K1 signaling, rather than IUGR, during fetal development may underlie reduced β cell growth and eventual development of T2DM later in life. PMID:26075820

  3. Effects of Lipopolysaccharide Biosynthesis Mutations on K1 Polysaccharide Association with the Escherichia coli Cell Surface

    PubMed Central

    Jiménez, Natalia; Senchenkova, Sofya N.; Knirel, Yuriy A.; Pieretti, Giuseppina; Corsaro, Maria M.; Aquilini, Eleonora; Regué, Miguel; Merino, Susana

    2012-01-01

    The presence of cell-bound K1 capsule and K1 polysaccharide in culture supernatants was determined in a series of in-frame nonpolar core biosynthetic mutants from Escherichia coli KT1094 (K1, R1 core lipopolysaccharide [LPS] type) for which the major core oligosaccharide structures were determined. Cell-bound K1 capsule was absent from mutants devoid of phosphoryl modifications on l-glycero-d-manno-heptose residues (HepI and HepII) of the inner-core LPS and reduced in mutants devoid of phosphoryl modification on HepII or devoid of HepIII. In contrast, in all of the mutants, K1 polysaccharide was found in culture supernatants. These results were confirmed by using a mutant with a deletion spanning from the hldD to waaQ genes of the waa gene cluster to which individual genes were reintroduced. A nuclear magnetic resonance (NMR) analysis of core LPS from HepIII-deficient mutants showed an alteration in the pattern of phosphoryl modifications. A cell extract containing both K1 capsule polysaccharide and LPS obtained from an O-antigen-deficient mutant could be resolved into K1 polysaccharide and core LPS by column chromatography only when EDTA and deoxycholate (DOC) buffer were used. These results suggest that the K1 polysaccharide remains cell associated by ionically interacting with the phosphate-negative charges of the core LPS. PMID:22522903

  4. eIF3 controls cell size independently of S6K1-activity

    PubMed Central

    Schipany, Katharina; Rosner, Margit; Ionce, Loredana; Hengstschläger, Markus; Kovacic, Boris

    2015-01-01

    All multicellular organisms require a life-long regulation of the number and the size of cells, which build up their organs. mTOR acts as a signaling nodule for the regulation of protein synthesis and growth. To activate the translational cascade, mTOR phosphorylates S6 kinase (S6K1), which is liberated from the eIF3-complex and mobilized for activation of its downstream targets. How S6K1 regulates cell size remains unclear. Here, we challenged cell size control through S6K1 by specifically depleting its binding partner eIF3 in normal and transformed cell lines. We show that loss of eIF3 leads to a massive reduction of cell size and cell number accompanied with an unexpected increase in S6K1-activity. The hyperactive S6K1-signaling was rapamycin-sensitive, suggesting an upstream mTOR-regulation. A selective S6K1 inhibitor (PF-4708671) was unable to interfere with the reduced size, despite efficiently inhibiting S6K1-activity. Restoration of eIF3 expression recovered size defects, without affecting the p-S6 levels. We further show that two, yet uncharacterized, cancer-associated mutations in the eIF3-complex, have the capacity to recover from reduced size phenotype, suggesting a possible role for eIF3 in regulating cancer cell size. Collectively, our results uncover a role for eIF3-complex in maintenance of normal and neoplastic cell size - independent of S6K1-signaling. PMID:26172298

  5. eIF3 controls cell size independently of S6K1-activity.

    PubMed

    Schipany, Katharina; Rosner, Margit; Ionce, Loredana; Hengstschläger, Markus; Kovacic, Boris

    2015-09-15

    All multicellular organisms require a life-long regulation of the number and the size of cells, which build up their organs. mTOR acts as a signaling nodule for the regulation of protein synthesis and growth. To activate the translational cascade, mTOR phosphorylates S6 kinase (S6K1), which is liberated from the eIF3-complex and mobilized for activation of its downstream targets. How S6K1 regulates cell size remains unclear. Here, we challenged cell size control through S6K1 by specifically depleting its binding partner eIF3 in normal and transformed cell lines. We show that loss of eIF3 leads to a massive reduction of cell size and cell number accompanied with an unexpected increase in S6K1-activity. The hyperactive S6K1-signaling was rapamycin-sensitive, suggesting an upstream mTOR-regulation. A selective S6K1 inhibitor (PF-4708671) was unable to interfere with the reduced size, despite efficiently inhibiting S6K1-activity. Restoration of eIF3 expression recovered size defects, without affecting the p-S6 levels. We further show that two, yet uncharacterized, cancer-associated mutations in the eIF3-complex, have the capacity to recover from reduced size phenotype, suggesting a possible role for eIF3 in regulating cancer cell size. Collectively, our results uncover a role for eIF3-complex in maintenance of normal and neoplastic cell size - independent of S6K1-signaling.

  6. Intracellular boron accumulation in CHO-K1 cells using amino acid transport control.

    PubMed

    Sato, Eisuke; Yamamoto, Tetsuya; Shikano, Naoto; Ogura, Masato; Nakai, Kei; Yoshida, Fumiyo; Uemae, Yoji; Takada, Tomoya; Isobe, Tomonori; Matsumura, Akira

    2014-06-01

    BPA used in BNCT has a similar structure to some essential amino acids and is transported into tumor cells by amino acid transport systems. Previous study groups have tried various techniques of loading BPA to increase intracellular boron concentration. CHO-K1 cells demonstrate system L (LAT1) activity and are suitable for specifying the transport system of a neutral amino acid. In this study, we examined the intracellular accumulation of boron in CHO-K1 cells by amino acid transport control, which involves co-loading with L-type amino acid esters. Intracellular boron accumulation in CHO-K1 cells showed the greatest increased upon co-loading 1.0mM BPA, with 1.0mM l-Tyr-O-Et and incubating for 60min. This increase is caused by activation of a system L amino acid exchanger between BPA and l-Tyr. The amino acid esters are metabolized to amino acids by intracellular hydrolytic enzymes that increase the concentrations of intracellular amino acids and stimulate exchange transportation. We expect that this amino acid transport control will be useful for enhancing intracellular boron accumulation.

  7. Curcumin inhibits hypoxia-induced migration in K1 papillary thyroid cancer cells

    PubMed Central

    Tan, Cheng; Zhang, Li; Cheng, Xian; Lin, Xiu-Feng; Lu, Rong-Rong; Bao, Jian-Dong

    2014-01-01

    Curcumin, traditionally used as food and medicinal purposes, has recently been reported to have protective efficacy against hypoxia. Hypoxia is one of the important reactive factors in tumor metastasis, which is a key problem in clinical thyroid cancer therapy. In present study, we investigate the anti-metastatic effect of curcumin on the K1 papillary thyroid cancer cells as well as its potential mechanisms. The results show that curcumin effectively inhibits hypoxia-induced reactive oxygen species (ROS) upregulation and significantly decreases the mRNA and protein expression levels of hypoxia-inducible factor-1α (HIF-1α) in K1 cells. Curcumin also decreases the DNA binding ability of HIF-1α to hypoxia response element (HRE). Furthermore, curcumin enhances E-cadherin expression, inhibits metalloproteinase-9 (MMP-9) enzyme activity, and weakens K1 cells migration under hypoxic conditions. In summary, these results indicate that curcumin possesses a potent anti-metastatic effect and might be an effective tumoristatic agent for the treatment of aggressive papillary thyroid cancers. PMID:25349216

  8. Estrogenic regulation of S6K1 expression creates a positive regulatory loop in control of breast cancer cell proliferation.

    PubMed

    Maruani, D M; Spiegel, T N; Harris, E N; Shachter, A S; Unger, H A; Herrero-González, S; Holz, M K

    2012-12-01

    The 40S ribosomal S6 kinase 1 (S6K1) is an important regulator of cell growth. Expression of S6K1 is often elevated in breast cancer cells. However, the transcriptional mechanism of S6K1 overexpression is not understood. In this report, we demonstrate that estrogen activates expression of S6K1 via estrogen receptor (ER)α in ER-positive breast cancer cells. We also show that estrogen acts on the proximal promoter of the S6K1 gene in a mechanism involving the transcriptional factor GATA-3. Finally, we provide data that support the importance of estrogenic regulation of S6K1 expression in breast cancer cell proliferation. S6K1 directly phosphorylates and regulates ligand-independent activity of ERα, while ERα upregulates S6K1 expression. This S6K1-ERα relationship creates a positive feed-forward loop in control of breast cancer cell proliferation. Furthermore, the co-dependent association between S6K1 and ERα may be exploited in the development of targeted breast cancer therapies.

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

    PubMed

    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.

  10. Homologous desensitization of human histamine H₃ receptors expressed in CHO-K1 cells.

    PubMed

    Osorio-Espinoza, Angélica; Escamilla-Sánchez, Juan; Aquino-Jarquin, Guillermo; Arias-Montaño, José-Antonio

    2014-02-01

    Histamine H₃ receptors (H₃Rs) modulate the function of the nervous system at the pre- and post-synaptic levels. In this work we aimed to determine whether, as other G protein-coupled receptors (GPCRs), H₃Rs desensitize in response to agonist exposure. By using CHO-K1 cells stably transfected with the human H₃R (hH3R) we show that functional responses (inhibition of forskolin-induced cAMP accumulation in intact cells and stimulation of [(35)S]-GTPγS binding to cell membranes) were markedly reduced after agonist exposure. For cAMP accumulation assays the effect was significant at 60 min with a maximum at 90 min. Agonist exposure resulted in decreased binding sites for the radioligand [(3)H]-N-methyl-histamine ([(3)H]-NMHA) to intact cells and modified the sub-cellular distribution of H₃Rs, as detected by sucrose density gradients and [(3)H]-NMHA binding to cell membranes, suggesting receptor internalization. The reduction in the inhibition of forskolin-stimulated cAMP formation observed after agonist pre-incubation was prevented by incubation in hypertonic medium or in ice-cold medium. Agonist-induced loss in binding sites was also prevented by hypertonic medium or incubation at 4 °C, but not by filipin III, indicating clathrin-dependent endocytosis. Immunodetection showed that CHO-K1 cells express GPCR kinases (GRKs) 2/3, and both the GRK general inhibitor ZnCl₂ and a small interfering RNA against GRK-2 reduced receptor desensitization. Taken together these results indicate that hH₃Rs experience homologous desensitization upon prolonged exposure to agonists, and that this process involves the action of GRK-2 and internalization via clathrin-coated vesicles.

  11. PDGF induces SphK1 expression via Egr-1 to promote pulmonary artery smooth muscle cell proliferation.

    PubMed

    Sysol, Justin R; Natarajan, Viswanathan; Machado, Roberto F

    2016-06-01

    Pulmonary arterial hypertension (PAH) is a progressive, life-threatening disease for which there is currently no curative treatment available. Pathologic changes in this disease involve remodeling of the pulmonary vasculature, including marked proliferation of pulmonary artery smooth muscle cells (PASMCs). Recently, the bioactive lipid sphingosine-1-phosphate (S1P) and its activating kinase, sphingosine kinase 1 (SphK1), have been shown to be upregulated in PAH and promote PASMC proliferation. The mechanisms regulating the transcriptional upregulation of SphK1 in PASMCs are unknown. In this study, we investigated the role of platelet-derived growth factor (PDGF), a PAH-relevant stimuli associated with enhanced PASMC proliferation, on SphK1 expression regulation. In human PASMCs (hPASMCs), PDGF significantly increased SphK1 mRNA and protein expression and induced cell proliferation. Selective inhibition of SphK1 attenuated PDGF-induced hPASMC proliferation. In silico promoter analysis for SphK1 identified several binding sites for early growth response protein 1 (Egr-1), a PDGF-associated transcription factor. Luciferase assays demonstrated that PDGF activates the SphK1 promoter in hPASMCs, and truncation of the 5'-promoter reduced PDGF-induced SphK1 expression. Stimulation of hPASMCs with PDGF induced Egr-1 protein expression, and direct binding of Egr-1 to the SphK1 promoter was confirmed by chromatin immunoprecipitation analysis. Inhibition of ERK signaling prevented induction of Egr-1 by PDGF. Silencing of Egr-1 attenuated PDGF-induced SphK1 expression and hPASMC proliferation. These studies demonstrate that SphK1 is regulated by PDGF in hPASMCs via the transcription factor Egr-1, promoting cell proliferation. This novel mechanism of SphK1 regulation may be a therapeutic target in pulmonary vascular remodeling in PAH. PMID:27099350

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

    PubMed

    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.

  13. Differential protein expression following low temperature culture of suspension CHO-K1 cells

    PubMed Central

    Kumar, Niraj; Gammell, Patrick; Meleady, Paula; Henry, Michael; Clynes, Martin

    2008-01-01

    Background To ensure maximal productivity of recombinant proteins (rP) during production culture it is typical to encourage an initial phase of rapid cell proliferation to achieve high biomass followed by a stationary phase where cellular energies are directed towards production of rP. During many such biphasic cultures, the initial phase of rapid cell growth at 37°C is followed by a growth arrest phase induced through reduction of the culture temperature. Low temperature induced growth arrest is associated with many positive phenotypes including increased productivity, sustained viability and an extended production phase, although the mechanisms regulating these phenotypes during mild hypothermia are poorly understood. Results In this study differential protein expression in suspension CHO-K1 cells was investigated following a reduction of the culture temperature from 37°C to 31°C in comparison to standard batch culture maintained at 37°C using 2D-DIGE (Fluorescence 2-D Difference Gel Electrophoresis) and mass spectrometry (MS). There is only limited proteomic analysis of suspension-grown CHO cells describing a direct comparison of temperature shifted versus non-temperature shifted cultures using 2D-DIGE. This investigation has enabled the identification of temperature-dependent as well as temperature-independent proteomic changes. 201 proteins were observed as differentially expressed following temperature shift, of which 118 were up regulated. Of the 53 proteins identified by MALDI-ToF MS, 23 were specifically differentially expressed upon reduction of the culture temperature and were found related to a variety of cellular functions such as regulation of growth (HNRPC), cap-independent translation (EIF4A), apoptosis (importin-α), the cytoskeleton (vimentin) and glycoprotein quality control (alpha glucosidase 2). Conclusion These results indicate the extent of the temperature response in CHO-K1 cells and suggest a number of key regulatory proteins and

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

    PubMed

    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. PMID:27465603

  15. Genotoxic effects of environmental estrogen-like compounds in CHO-K1 cells.

    PubMed

    Tayama, Sumiko; Nakagawa, Yoshio; Tayama, Kuniaki

    2008-01-01

    Some environmental estrogen-like compounds, such as bisphenol A (BPA), 4-nonylphenol (NP), 4-octylphenol (OP), propyl p-hydroxybenzoate (P-PHBA), and butyl p-hydroxybenzoate (B-PHBA), synthetic estrogen, diethylstilbestrol (DES), and natural estrogen, 17beta-estradiol (E2), were studied for their genotoxicity in CHO-K1 cells using sister-chromatid exchange (SCE), chromosome aberration (CA), and DNA strand break (comet) assays. Six of the chemicals, excluding E2, caused DNA migration in the comet assay and induced SCEs at one or more of the highest doses. Among the chemicals, OP produced an especially high incidence of SCEs. Structural CA was induced by five of the chemicals, excluding OP and NP, and BPA, E2, and DES also induced aneuploid cells. E2 and DES particularly increased the rate of polyploidy at high doses. The incidence of colchicine-mitosis-like (c-mitotic) figures suggesting spindle disrupting effects was also detected with five of the chemicals, excluding OP and NP, and six of the chemicals, excluding E2, caused endoreduplication (ERD), a form of nuclear polyploidization induced by block of cell cycle at G2 phase, at one or more high doses. Our present results suggest that OP and NP cause repairable DNA damage, including SCEs, and do not result in CA, while the damage caused by DES, BPA, P-PHBA, and B-PHBA results in the induction of CAs together with SCEs probably because of imperfect repair. We are unable to explain the observation that the DNA damage caused by E2 resulted in CA induction but not DNA migration or SCE induction, except for speculating that the DNA damage is different from that caused by DES and the estrogen-like chemicals. Our findings also suggest that E2, DES and BPA have aneuploidogenic properties, and that the former two of chemicals also are polyploidy-inducing agents. PMID:17913570

  16. Cytochalasin J affects chromosome congression and spindle microtubule organization in PtK1 cells.

    PubMed

    Snyder, J A; Cohen, L

    1995-01-01

    PtK1 cells were treated with 10 micrograms/ml cytochalasin J (CJ) for 15 min at various stages of mitosis. When applied at nuclear envelope breakdown (NEB) chromosome congression was blocked or substantially slowed, and chromosomes failed to show organization patterns typical of prometaphase. Spindle microtubule (MT) numbers appeared unaffected as judged by the pattern of birefringent retardation. However, ultrastructural analysis showed MTs to be reorganized within the spindle domain with some exhibiting fragmentation and others failing to interact with poorly defined kinetochore laminae. The spindle domain took on a curved, almost banana-like shape, as related to the position of the centrosomes and lack of orientation of chromosomes. Serial section analysis of kinetochore regions showed reduced contour length and maturation of the kinetochore plate with few MTs associated with this structure. Cells similarly treated with 10 micrograms/ml CJ at NEB for 15 min and then released into conditioned medium for 15 min showed the most chromosomes resumed congression to the metaphase plate. Ultrastructural analysis revealed a more normal organization of spindle MTs, but kinetochore structure remained affected. CJ treatment of cells in prometaphase slightly affected chromosomes congression with most chromosomes aligning at the metaphase plate after 10-15 min of treatment. Ultrastructural analysis showed that astral MTs were disrupted and spindle MTs were fragmented; few MTs coursed from kinetochore to pole. Kinetochore structure was also affected with only small numbers of short MTs seen associated with kinetochores. Application of CJ at anaphase onset had little effect on anaphase A and B, but cytokinesis failed to occur. Anti-tubulin staining of a monolayer of cells treated with 10 micrograms/ml CJ for 15 min showed that over 60% of mitotic figures exhibited changes in MT organization. Cells showing the greatest effect of treatment had several foci of bundles of MTs, as

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

    PubMed

    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.

  18. Cytogenetic effects of piperonyl butoxide and safrole in CHO-K1 cells.

    PubMed

    Tayama, S

    1996-07-01

    Recently, hepatocarcinogenicity in rats and mice was reported with regard to the methylenedioxyphenyl compound, piperonyl butoxide (PB), which is used as a synergist for pyrethrins and related insecticides. Induction of sister-chromatid exchanges (SCEs) and chromosomal aberrations (CAs) due to PB were investigated using CHO-K1 cells with or without rat liver S9 fraction (S9); at the same time, the effects of safrole (SF), a methylenedioxyphenyl compound and a weak hepatocarcinogen, were also examined. PB (0.25 and 0.3 mM) and SF (0.8 mM) caused a slight but significant increase in SCEs followed by a cell-cycle delay in the 3-h treatment without S9. In the presence of S9 (4.5%), the cytotoxicity of PB or SF was weakened greatly or slightly, the top dose capable of cell division was raised to 0.6 mM (2-fold) or 1 mM, respectively. PB with S9 induced SCE at doses of 0.4 and 0.5 mM, and caused endoreduplications (ERDs, 7%) at a dose of 0.6 mM, while SF caused a dose-related significant increase in SCE at all doses used (0.4-1 mM) with S9. Genotoxicity of the metabolites of PB or SF was cleared by changing the dose of S9 (1.5-9%) while holding the dose of each chemical constant. In the case of SF (0.6 mM), induction of SCE, ERD and cell-cycle delay intensified almost in a dose-effect relationship, and CAs and a high level of ERD (14%) were caused by a 9% dose of S9. The concentration of unchanged SF in the incubated medium was certainly in inverse proportion to the dose of S9. This strongly suggests that the metabolites of SF are genotoxic. In the case of PB (0.3 mM), no positive responses were produced in the cultures, even with a high level of S9, though the amount of unchanged PB left in the incubated medium was very slight. This indicates that the metabolites of PB may not be genotoxic. In conclusion, PB and SF are possible to somewhat induce SCE at high dose(s) in the absence of S9, and the genotoxic effects of SF are more intensified in the presence of S9 than in

  19. MAP3K1

    PubMed Central

    Pham, Trang T.; Angus, Steven P.

    2013-01-01

    MAP3K1 is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family of serine/threonine kinases. MAP3K1 regulates JNK activation and is unique among human kinases in that it also encodes an E3 ligase domain that ubiquitylates c-Jun and ERK1/2. Full length MAP3K1 regulates cell migration and contributes to pro-survival signaling while its caspase 3-mediated cleavage generates a C-terminal kinase domain that promotes apoptosis. The critical function of MAP3K1 in cell fate decisions suggests that it may be a target for deregulation in cancer. Recent large-scale genomic studies have revealed that MAP3K1 copy number loss and somatic missense or nonsense mutations are observed in a significant number of different cancers, being most prominent in luminal breast cancer. The alteration of MAP3K1 in diverse cancer types demonstrates the importance of defining phenotypes for possible therapeutic targeting of tumor cell vulnerabilities created when MAP3K1 function is lost or gained. PMID:24386504

  20. Evidence that hematopoietic stem cell function is preserved during aging in long-lived S6K1 mutant mice

    PubMed Central

    Selman, Colin; Sinclair, Amy; Pedroni, Silvia M.A.; Irvine, Elaine E.; Michie, Alison M.; Withers, Dominic J.

    2016-01-01

    The mechanistic target of rapamycin (mTOR) signalling pathway plays a highly conserved role in aging; mice lacking ribosomal protein S6 kinase 1 (S6K1−/−) have extended lifespan and healthspan relative to wild type (WT) controls. Exactly how reduced mTOR signalling induces such effects is unclear, although preservation of stem cell function may be important. We show, using gene expression analyses, that there was a reduction in expression of cell cycle genes in young (12 week) and aged (80 week) S6K1−/− BM-derived c-Kit+ cells when compared to age-matched WT mice, suggesting that these cells are more quiescent in S6K1−/− mice. In addition, we investigated hematopoietic stem cell (HSC) frequency and function in young and aged S6K1−/− and WT mice. Young, but not aged, S6K1−/− mice had more LSK (lineage−, c-Kit+, Sca-1+) cells (% of bone marrow (BM)), including the most primitive long-term repopulating HSCs (LT-HSC) relative to WT controls. Donor-derived engraftment of LT-HSCs in recipient mice was unaffected by genotype in young mice, but was enhanced in transplants using LT-HSCs derived from aged S6K1−/− mice. Our results are the first to provide evidence that age-associated HSC functional decline is ameliorated in a long-lived mTOR mutant mouse. PMID:27083004

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

    PubMed

    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. PMID:26921102

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

    PubMed

    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.

  3. Arsenite induces cell transformation by reactive oxygen species, AKT, ERK1/2, and p70S6K1

    SciTech Connect

    Carpenter, Richard L.; Jiang, Yue; Jing, Yi; He, Jun; Rojanasakul, Yon; Liu, Ling-Zhi; Jiang, Bing-Hua

    2011-10-28

    Highlights: Black-Right-Pointing-Pointer Chronic exposure to arsenite induces cell proliferation and transformation. Black-Right-Pointing-Pointer Arsenite-induced transformation increases ROS production and downstream signalings. Black-Right-Pointing-Pointer Inhibition of ROS levels via catalase reduces arsenite-induced cell transformation. Black-Right-Pointing-Pointer Interruption of AKT, ERK, or p70S6K1 inhibits arsenite-induced cell transformation. -- Abstract: Arsenic is naturally occurring element that exists in both organic and inorganic formulations. The inorganic form arsenite has a positive association with development of multiple cancer types. There are significant populations throughout the world with high exposure to arsenite via drinking water. Thus, human exposure to arsenic has become a significant public health problem. Recent evidence suggests that reactive oxygen species (ROS) mediate multiple changes to cell behavior after acute arsenic exposure, including activation of proliferative signaling and angiogenesis. However, the role of ROS in mediating cell transformation by chronic arsenic exposure is unknown. We found that cells chronically exposed to sodium arsenite increased proliferation and gained anchorage-independent growth. This cell transformation phenotype required constitutive activation of AKT, ERK1/2, mTOR, and p70S6K1. We also observed these cells constitutively produce ROS, which was required for the constitutive activation of AKT, ERK1/2, mTOR, and p70S6K1. Suppression of ROS levels by forced expression of catalase also reduced cell proliferation and anchorage-independent growth. These results indicate cell transformation induced by chronic arsenic exposure is mediated by increased cellular levels of ROS, which mediates activation of AKT, ERK1/2, and p70S6K1.

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

    PubMed

    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. PMID:17418577

  5. Downregulation of LSD1 suppresses the proliferation, tumorigenicity and invasion of papillary thyroid carcinoma K1 cells

    PubMed Central

    KONG, LING-LING; MAN, DONG-MEI; WANG, TIAN; ZHANG, GUO-AN; CUI, WEN

    2016-01-01

    The present study aimed to evaluate the effects of lysine-specific demethylase 1 (LSD1) downregulation, induced by small interfering RNA (siRNA) transfection, on the proliferation, colony formation, migration and invasion of the papillary thyroid carcinoma K1 cell line. The siRNA targeting LSD1 and scrambled non-targeting siRNA were each transfected into papillary thyroid carcinoma K1 cells. Downregulation of LSD1 mRNA and protein level was evaluated by reverse transcription-quantitative polymerase chain reaction, and immunocytochemical (ICC) analysis and western blotting, respectively. A Cell Counting kit-8 assay was applied to estimate the effect of LSD1-siRNA on cell growth. Migration and invasion abilities were estimated by Transwell chamber assay. A soft agar colony formation assay was performed to estimate the effect of LSD1-siRNA on tumorigenicity in vitro. ICC data showed that LSD1 protein was strongly expressed in the blank and control K1 cells compared with the LSD1-siRNA cells (F=15.192, P<0.01). Compared with the control cells, cells transfected with siRNA targeting LSD1 exhibited significant downregulation of LSD1 mRNA (t=6.845, P<0.01) and protein (F=53.764, P<0.01) levels. siRNA targeting LSD1 also downregulated cell proliferation following transfection for 24, 48 and 72 h (t=4.777, P<0.001; t=3.302, P=0.003; and t=3.017, P=0.006, respectively). Compared with the control group, the amount of cell invasion was gradually reduced in the LSD1-siRNA group (t=12.301, P<0.01). The number of migrating cells was significantly higher in the negative control group compared with the LSD1-siRNA group (t=7.911, P<0.01), and the ability of colony formation in the LSD1-siRNA cells was notably reduced in the soft agar formation assay (t=3.612, P=0.005). siRNA targeting LSD1 efficiently inhibits the proliferation, colony formation, migration and invasion of papillary thyroid carcinoma K1 cells. PMID:27073501

  6. High prevalence of MAP2K1 mutations in variant and IGHV4-34-expressing hairy-cell leukemias.

    PubMed

    Waterfall, Joshua J; Arons, Evgeny; Walker, Robert L; Pineda, Marbin; Roth, Laura; Killian, J Keith; Abaan, Ogan D; Davis, Sean R; Kreitman, Robert J; Meltzer, Paul S

    2014-01-01

    To understand the genetic mechanisms driving variant and IGHV4-34-expressing hairy-cell leukemias, we performed whole-exome sequencing of leukemia samples from ten affected individuals, including six with matched normal samples. We identified activating mutations in the MAP2K1 gene (encoding MEK1) in 5 of these 10 samples and in 10 of 21 samples in a validation set (overall frequency of 15/31), suggesting potential new strategies for treating individuals with these diseases.

  7. The inhibition of CHO-K1-BH4 cell proliferation and induction of chromosomal aberrations by brevetoxins in vitro

    PubMed Central

    Sayer, A.N.; Hu, Q.; Bourdelais, A.J.; Baden, D.G.; Gibson, J.E.

    2009-01-01

    Brevetoxins (PbTxs) are highly potent trans-syn polyether neurotoxins produced during blooms of several species of marine dinoflagellates, most notably Karenia brevis. These neurotoxins act on voltage-sensitive sodium channels prolonging the active state. During red tides, the commercial fishing and tourism industries experience millions of dollars of lost revenue. Human consumption of shellfish contaminated with PbTxs results in neurotoxic shellfish poisoning (NSP). Additionally, blooms of K. brevis are potentially responsible for adverse human health effects such as respiratory irritation and airway constriction in coastal residents. There is little information regarding the full range of potential toxic effects caused by PbTxs. Recent evidence suggests that PbTxs are genotoxic substances. The purpose of this study was to determine if PbTxs could induce chromosomal aberrations and inhibit cellular proliferation in CHO-K1-BH4 cells, and if so, could the damage be negated or reduced by the PbTx antagonist brevenal. Results from the chromosomal aberrations assay demonstrated that PbTxs are potent inducers of CHO-K1-BH4 chromosome damage. Results from the inhibition of cellular proliferation assays demonstrated that PbTxs inhibit the ability of CHO-K1-BH4 cells to proliferate, an effect which can be reduced with brevenal. PMID:16487644

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

    PubMed

    Hanot, Camille C; Choi, Young Suk; Anani, Tareq B; Soundarrajan, Dharsan; David, Allan E

    2016-01-01

    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. PMID:26729108

  9. Carbamates: A study on genotoxic, cytotoxic, and apoptotic effects induced in Chinese hamster ovary (CHO-K1) cells.

    PubMed

    Soloneski, Sonia; Kujawski, Maciej; Scuto, Anna; Larramendy, Marcelo L

    2015-08-01

    In vitro effects of the carbamates pirimicarb and zineb and their formulations Aficida® (50% pirimicarb) and Azzurro® (70% zineb), respectively, were evaluated in Chinese hamster ovary (CHO-K1) cells. Whereas the cytokinesis-blocked micronucleus cytome assay was employed to test for genotoxicity, MTT, neutral red (NR), and apoptosis evaluation were used as tests for estimating cell viability and succinic dehydrogenase activity, respectively. Concentrations tested were 10-300 μg/ml for pirimicarb and Aficida®, and 1-50 μg/ml for zineb and Azzurro®. All compounds were able to increase the frequency of micronuclei. A marked reduction in the nuclear division index was observed after treatment with 5 μg/ml of zineb and Azzurro® and 10 μg/ml of Azzurro®. Alterations in the cellular morphology not allowing the recognition of binucleated cells exposed to 300 μg/ml pirimicarb and Aficida® as well as 10-50 μg/ml zineb and Azzurro®. All four compounds induced inhibition of both cell viability and succinic dehydrogenase activity and trigger apoptosis in CHO-K1 cells, at least when exposed for 24 h. The data herein demonstrate the genotoxic and cytotoxic effects exerted by these carbamates and reveal the potential risk factor of these pesticides, still extensively used worldwide, for both human health and the environment.

  10. Evolutionary story of a satellite DNA from Phodopus sungorus (Rodentia, Cricetidae).

    PubMed

    Paço, Ana; Adega, Filomena; Meštrović, Nevenka; Plohl, Miroslav; Chaves, Raquel

    2014-10-21

    With the goal to contribute for the understanding of satellite DNA evolution and its genomic involvement, in this work it was isolated and characterized the first satellite DNA (PSUcentSat) from Phodopus sungorus (Cricetidae). Physical mapping of this sequence in P. sungorus showed large PSUcentSat arrays located at the heterochromatic (peri)centromeric region of five autosomal pairs and Y-chromosome. The presence of orthologous PSUcentSat sequences in the genomes of other Cricetidae and Muridae rodents was also verified, presenting however, an interspersed chromosomal distribution. This distribution pattern suggests a PSUcentSat-scattered location in an ancestor of Muridae/Cricetidae families, that assumed afterwards, in the descendant genome of P. sungorus a restricted localization to few chromosomes in the (peri)centromeric region. We believe that after the divergence of the studied species, PSUcentSat was most probably highly amplified in the (peri)centromeric region of some chromosome pairs of this hamster by recombinational mechanisms. The bouquet chromosome configuration (prophase I) possibly displays an important role in this selective amplification, providing physical proximity of centromeric regions between chromosomes with similar size and/or morphology. This seems particularly evident for the acrocentric chromosomes of P. sungorus (including the Y-chromosome), all presenting large PSUcentSat arrays at the (peri)centromeric region. The conservation of this sequence in the studied genomes and its (peri)centromeric amplification in P. sungorus strongly suggests functional significance, possibly displaying this satellite family different functions in the different genomes. The verification of PSUcentSat transcriptional activity in normal proliferative cells suggests that its transcription is not stage-limited, as described for some other satellites.

  11. Evolutionary Story of a Satellite DNA from Phodopus sungorus (Rodentia, Cricetidae)

    PubMed Central

    Paço, Ana; Adega, Filomena; Meštrović, Nevenka; Plohl, Miroslav; Chaves, Raquel

    2014-01-01

    With the goal to contribute for the understanding of satellite DNA evolution and its genomic involvement, in this work it was isolated and characterized the first satellite DNA (PSUcentSat) from Phodopus sungorus (Cricetidae). Physical mapping of this sequence in P. sungorus showed large PSUcentSat arrays located at the heterochromatic (peri)centromeric region of five autosomal pairs and Y-chromosome. The presence of orthologous PSUcentSat sequences in the genomes of other Cricetidae and Muridae rodents was also verified, presenting however, an interspersed chromosomal distribution. This distribution pattern suggests a PSUcentSat-scattered location in an ancestor of Muridae/Cricetidae families, that assumed afterwards, in the descendant genome of P. sungorus a restricted localization to few chromosomes in the (peri)centromeric region. We believe that after the divergence of the studied species, PSUcentSat was most probably highly amplified in the (peri)centromeric region of some chromosome pairs of this hamster by recombinational mechanisms. The bouquet chromosome configuration (prophase I) possibly displays an important role in this selective amplification, providing physical proximity of centromeric regions between chromosomes with similar size and/or morphology. This seems particularly evident for the acrocentric chromosomes of P. sungorus (including the Y-chromosome), all presenting large PSUcentSat arrays at the (peri)centromeric region. The conservation of this sequence in the studied genomes and its (peri)centromeric amplification in P. sungorus strongly suggests functional significance, possibly displaying this satellite family different functions in the different genomes. The verification of PSUcentSat transcriptional activity in normal proliferative cells suggests that its transcription is not stage-limited, as described for some other satellites. PMID:25336681

  12. C-terminal region of GADD34 regulates eIF2α dephosphorylation and cell proliferation in CHO-K1 cells.

    PubMed

    Otsuka, Ryo; Harada, Nagakatsu; Aoki, Shouhei; Shirai, Kanna; Nishitsuji, Kazuchika; Nozaki, Ayane; Hatakeyama, Adzumi; Shono, Masayuki; Mizusawa, Noriko; Yoshimoto, Katsuhiko; Nakaya, Yutaka; Kitahata, Hiroshi; Sakaue, Hiroshi

    2016-01-01

    GADD34 is a member of a growth arrest and DNA damage (GADD)-inducible gene family. Here, we established a novel Chinese hamster ovary (CHO)-K1-derived cell line, CHO-K1-G34M, which carries a nonsense mutation (termed the Q525X mutation) in the GADD34 gene. The Q525X mutant protein lacks the C-terminal 66 amino acids required for GADD34 to bind to and activate protein phosphatase 1 (PP1). We investigated the effects of GADD34 with or without the Q525X mutation on the phosphorylation status of PP1 target proteins, including the α subunit of eukaryotic initiation factor 2 (eIF2α) and glycogen synthase kinase 3β (GSK3β). CHO-K1-G34M cells had higher levels of eIF2α phosphorylation compared to the control CHO-K1-normal cells both in the presence and absence of endoplasmic reticulum stress. Overexpression of the wild-type GADD34 protein in CHO-K1-normal cells largely reduced eIF2α phosphorylation, while overexpression of the Q525X mutant did not produce similar reductions. Meanwhile, neither wild type nor Q525X mutation of GADD34 affected the GSK3β phosphorylation status. GADD34 also did not affect the canonical Wnt signaling pathway downstream of GSK3β. Cell proliferation rates were higher, while expression levels of the cyclin-dependent kinase inhibitor p21 were lower in CHO-K1-G34M cells compared to the CHO-K1-normal cells. The GADD34 Q525X mutant had a reduced ability to inhibit cell proliferation and enhance p21 expression of the CHO-K1-normal cells compared to the wild-type GADD34 protein. These results suggest that the GADD34 protein C-terminal plays important roles in regulating not only eIF2α dephosphorylation but also cell proliferation in CHO-K1 cells.

  13. High prevalence of MAP2K1 mutations in variant and IGHV4-34-expressing hairy-cell leukemias.

    PubMed

    Waterfall, Joshua J; Arons, Evgeny; Walker, Robert L; Pineda, Marbin; Roth, Laura; Killian, J Keith; Abaan, Ogan D; Davis, Sean R; Kreitman, Robert J; Meltzer, Paul S

    2014-01-01

    To understand the genetic mechanisms driving variant and IGHV4-34-expressing hairy-cell leukemias, we performed whole-exome sequencing of leukemia samples from ten affected individuals, including six with matched normal samples. We identified activating mutations in the MAP2K1 gene (encoding MEK1) in 5 of these 10 samples and in 10 of 21 samples in a validation set (overall frequency of 15/31), suggesting potential new strategies for treating individuals with these diseases. PMID:24241536

  14. Phenotypic and functional characterization of a new human macrophage cell line K1m demonstrating immunophagocytic activity and signalling through HLA class II.

    PubMed Central

    Dialynas, D P; Lee, M J; Shao, L E; Tan, P C; Yu, J

    1997-01-01

    A human macrophage line, designated K1m, has been established from peripheral blood. K1m expresses a number of lineage-specific markers as well as a broad array of intercellular adhesion molecules. In particular, K1m expresses high levels of human leucocyte antigen (HLA) class I and class II. In response to ligation of HLA class II (HLA-DR), but not in response to ligation of HLA class 1, K1m forms tighter homotypic aggregates and develops a striking 'stellate' culture phenotype. K1m also expresses Fc receptors for immunoglobulin G (IgG) (CD64, CD32, and CD16) and can be shown to phagocytose polystyrene latex beads, as well as neuroblastoma cells in the presence of tumour-specific monoclonal antibody (mAb). The K1m cell line should therefore prove useful for studying both signalling through macrophage HLA class II and immunophagocytosis. Images Figure 5 Figure 6 PMID:9176097

  15. Increases in intracellular calcium via activation of an endogenous P2-purinoceptor in cultured CHO-K1 cells.

    PubMed Central

    Iredale, P. A.; Hill, S. J.

    1993-01-01

    1. Increases in intracellular calcium ([Ca2+]i) were measured in chinese hamster cultured ovary cells (clone, CHO-K1), by use of the fluorescent, calcium-sensitive dye, fura-2. 2. Addition of both ATP and UTP elicited rapid increases in [Ca2+]i due to mobilization from intracellular stores and calcium entry across the plasma membrane. 3. Omission of calcium from the extracellular medium and pre-incubation with the inorganic calcium channel blocker, nickel (Ni2+) prevented the calcium entry components of the responses. 4. Investigation of the concentration-response relationships of various analogues of ATP suggests the presence of a purinoceptor which cannot be characterized as P2X or P2Y. In addition, there appears to be a sub-population of P2Y-purinoceptors which do not cross-react with the 'nucleotide' receptor population. 5. Cross-desensitization and additivity experiments suggest that both ATP and UTP activate the same receptor. 6. Pre-incubation with the tumour-promoting agent, beta-phorbol-12,13 dibutyrate (PDBu), caused a reduction in the increases in [Ca2+]i, suggesting a role for protein kinase C in feedback inhibition of purinoceptor responses in this cell line. 7. In summary, we present evidence for the existence of an endogenous P2U-purinoceptor (or 'nucleotide receptor') which is linked to increases in [Ca2+]i in CHO-K1 cells. PMID:8306069

  16. T-Cell-Specific Deletion of Map3k1 Reveals the Critical Role for Mekk1 and Jnks in Cdkn1b-Dependent Proliferative Expansion

    PubMed Central

    Suddason, Tesha; Anwar, Saba; Charlaftis, Nikolaos; Gallagher, Ewen

    2016-01-01

    Summary MAPK signaling is important for T lymphocyte development, homeostasis, and effector responses. To better understand the role of Mekk1 (encoded by Map3k1) in T cells, we conditionally deleted Map3k1 in LckCre/+Map3k1f/f mice, and these display larger iNKT cell populations within the liver, spleen, and bone marrow. Mekk1 signaling controls splenic and liver iNKT cell expansion in response to glycolipid antigen. LckCre/+Map3k1f/f mice have enhanced liver damage in response to glycolipid antigen. Mekk1 regulates Jnk activation in iNKT cells and binds and transfers Lys63-linked poly-ubiquitin onto Carma1. Map3k1 is critical for the regulation of p27Kip1 (encoded by Cdkn1b). PMID:26774476

  17. In vitro infection by Ehrlichia ruminantium of baby hamster kidney (BHK), Chinese hamster ovary (CHO-K1) and Madin Darby bovine kidney (MDBK) cells.

    PubMed

    Zweygarth, E; Josemans, A I

    2003-06-01

    The Welgevonden stock of Ehrlichia ruminantium, aetiological agent of heartwater, was propagated in baby hamster kidney (BHK) cells, Chinese hamster ovary (CHO-K1) cells and Madin Darby bovine kidney (MDBK) cells. The cultures required supplementation of the medium with cycloheximide for reliable growth of E. ruminantium. Growth of the Welgevonden stock in BHK and CHO-K1 cells could lead to the development of suspension cultures suitable for the mass production of E. ruminantium for an inactivated elementary body vaccine.

  18. Heterologous, PKC-Mediated Desensitization of Human Histamine H3 Receptors Expressed in CHO-K1 Cells.

    PubMed

    Montejo-López, Wilber; Rivera-Ramírez, Nayeli; Escamilla-Sánchez, Juan; García-Hernández, Ubaldo; Arias-Montaño, José-Antonio

    2016-09-01

    Desensitization is a major mechanism to regulate the functional response of G protein-coupled receptors. In this work we studied whether the human histamine H3 receptor of 445 amino acids (hH3R445) experiences heterologous desensitization mediated by PKC activation. Bioinformatic analysis indicated the presence of Serine and Threonine residues susceptible of PKC-mediated phosphorylation on the third intracellular loop and the carboxyl terminus of the hH3R445. In CHO-K1 cells stably transfected with the hH3R445 direct PKC activation by phorbol 12-myristate 13-acetate (TPA, 200 nM) abolished H3R-mediated inhibition of forskolin-stimulated cAMP accumulation. Activation of endogenous purinergic receptors by ATP (adenosine 5'-triphosphate, 10 μM) increased the free calcium intracellular concentration ([Ca(2+)]i) confirming their coupling to phospholipase C stimulation. Incubation with ATP also abolished H3R-mediated inhibition of forskolin-induced cAMP accumulation, and this effect was prevented by the PKC inhibitors Ro-31-8220 and Gö-6976. Pre-incubation with TPA or ATP reduced H3R-mediated stimulation of [(35)S]-GTPγS binding to membranes from CHO-K1-hH3R445 cells by 39.7 and 54.2 %, respectively, with no change in the agonist potency, and the effect was prevented by either Ro-31-8220 or Gö-6976. Exposure to ATP or TPA also resulted in the loss of cell surface H3Rs (-30.4 and -45.1 %) as evaluated by [(3)H]-NMHA binding to intact cells. These results indicate that the hH3R445 undergoes heterologous desensitization upon activation of receptors coupled to PKC stimulation. PMID:27350581

  19. The goya mouse mutant reveals distinct newly identified roles for MAP3K1 in the development and survival of cochlear sensory hair cells.

    PubMed

    Parker, Andrew; Cross, Sally H; Jackson, Ian J; Hardisty-Hughes, Rachel; Morse, Susan; Nicholson, George; Coghill, Emma; Bowl, Michael R; Brown, Steve D M

    2015-12-01

    Mitogen-activated protein kinase, MAP3K1, plays an important role in a number of cellular processes, including epithelial migration during eye organogenesis. In addition, studies in keratinocytes indicate that MAP3K1 signalling through JNK is important for actin stress fibre formation and cell migration. However, MAP3K1 can also act independently of JNK in the regulation of cell proliferation and apoptosis. We have identified a mouse mutant, goya, which exhibits the eyes-open-at-birth and microphthalmia phenotypes. In addition, these mice also have hearing loss. The goya mice carry a splice site mutation in the Map3k1 gene. We show that goya and kinase-deficient Map3k1 homozygotes initially develop supernumerary cochlear outer hair cells (OHCs) that subsequently degenerate, and a progressive profound hearing loss is observed by 9 weeks of age. Heterozygote mice also develop supernumerary OHCs, but no cellular degeneration or hearing loss is observed. MAP3K1 is expressed in a number of inner-ear cell types, including outer and inner hair cells, stria vascularis and spiral ganglion. Investigation of targets downstream of MAP3K1 identified an increase in p38 phosphorylation (Thr180/Tyr182) in multiple cochlear tissues. We also show that the extra OHCs do not arise from aberrant control of proliferation via p27KIP1. The identification of the goya mutant reveals a signalling molecule involved with hair-cell development and survival. Mammalian hair cells do not have the ability to regenerate after damage, which can lead to irreversible sensorineural hearing loss. Given the observed goya phenotype, and the many diverse cellular processes that MAP3K1 is known to act upon, further investigation of this model might help to elaborate upon the mechanisms underlying sensory hair cell specification, and pathways important for their survival. In addition, MAP3K1 is revealed as a new candidate gene for human sensorineural hearing loss.

  20. The goya mouse mutant reveals distinct newly identified roles for MAP3K1 in the development and survival of cochlear sensory hair cells

    PubMed Central

    Parker, Andrew; Cross, Sally H.; Jackson, Ian J.; Hardisty-Hughes, Rachel; Morse, Susan; Nicholson, George; Coghill, Emma; Bowl, Michael R.; Brown, Steve D. M.

    2015-01-01

    ABSTRACT Mitogen-activated protein kinase, MAP3K1, plays an important role in a number of cellular processes, including epithelial migration during eye organogenesis. In addition, studies in keratinocytes indicate that MAP3K1 signalling through JNK is important for actin stress fibre formation and cell migration. However, MAP3K1 can also act independently of JNK in the regulation of cell proliferation and apoptosis. We have identified a mouse mutant, goya, which exhibits the eyes-open-at-birth and microphthalmia phenotypes. In addition, these mice also have hearing loss. The goya mice carry a splice site mutation in the Map3k1 gene. We show that goya and kinase-deficient Map3k1 homozygotes initially develop supernumerary cochlear outer hair cells (OHCs) that subsequently degenerate, and a progressive profound hearing loss is observed by 9 weeks of age. Heterozygote mice also develop supernumerary OHCs, but no cellular degeneration or hearing loss is observed. MAP3K1 is expressed in a number of inner-ear cell types, including outer and inner hair cells, stria vascularis and spiral ganglion. Investigation of targets downstream of MAP3K1 identified an increase in p38 phosphorylation (Thr180/Tyr182) in multiple cochlear tissues. We also show that the extra OHCs do not arise from aberrant control of proliferation via p27KIP1. The identification of the goya mutant reveals a signalling molecule involved with hair-cell development and survival. Mammalian hair cells do not have the ability to regenerate after damage, which can lead to irreversible sensorineural hearing loss. Given the observed goya phenotype, and the many diverse cellular processes that MAP3K1 is known to act upon, further investigation of this model might help to elaborate upon the mechanisms underlying sensory hair cell specification, and pathways important for their survival. In addition, MAP3K1 is revealed as a new candidate gene for human sensorineural hearing loss. PMID:26542706

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

    PubMed

    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

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

    PubMed

    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

  3. Inhibition of topoisomerase II{alpha} activity in CHO K1 cells by 2-[(aminopropyl)amino]ethanethiol (WR-1065)

    SciTech Connect

    Grdina, D.J. |; Constantinou, A.; Shigematsu, N.; Murley, J.S.

    1993-06-01

    The aminothiol 2-[(aminopropyl)amino]ethanethiol (WR-1065) is the active thiol of the clinically studied radioprotective agent S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721). WR-1065 is an effective radiation protector and antimutagenic agent when it is administered 30 min prior to radiation exposure to Chinese hamster ovary Kl cells at a concentration of 4 mM. Under these exposure conditions, topoisomerase (topo) I and II activities and associated protein contents were measured in the K1 cell line using the DNA relaxation assay, the P4 unknotting assay, and immunoblotting, respectively. WR-1065 was ineffective in modifying topo I activity, but it did reduce topo IIa activity by an average of 50 percent. The magnitude of topo IIa protein content, however, was not affected by these exposure conditions. Cell cycle effects were monitored by the method of flow cytometry. Exposure of cells to 4 mM WR-1065 for a period of up to 6 h resulted in a buildup of cells in the G2 compartment. However, in contrast to topo II inhibitors used in chemotherapy, WR-1065 is an effective radioprotector agent capable of protecting against both radiation-induced cell lethality and mutagenesis. One of several mechanisms of radiation protection attributed to aminothiol compounds such as WR-1065 has been their ability to affect endogenous enzymatic reactions involved in DNA synthesis, repair, and cell cycle progression. These results are consistent with such a proposed mechanism and demonstrate in particular a modifying effect by 2-[(aminopropyl)amino]ethanethiol on type II topoisomerase, which is involved in DNA synthesis.

  4. Investigation of superparamagnetic (Fe3O4) nanoparticles and magnetic field exposures on CHO-K1 cell line

    NASA Astrophysics Data System (ADS)

    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.

  5. Surface modification by argon plasma treatment improves antioxidant defense ability of CHO-k1 cells on titanium surfaces.

    PubMed

    de Queiroz, Jana Dara Freires; Leal, Angélica Maria de Sousa; Terada, Maysa; Agnez-Lima, Lucymara Fassarela; Costa, Isolda; Pinto, Nadja Cristhina de Souza; de Medeiros, Silvia Regina Batistuzzo

    2014-04-01

    Titanium is one of the most used materials in implants and changes in its surface can modify the cellular functional response to better implant fixation. An argon plasma treatment generates a surface with improved mechanical proprieties without modifying its chemical composition. Oxidative stress induced by biomaterials is considered one of the major causes of implant failure and studies in this field are fundamental to evaluate the biocompatibility of a new material. Therefore, in this work, induction of oxidative stress by titanium surfaces subjected to plasma treatment (PTTS) was evaluated. The viability of CHO-k1 cells was higher on PTTS discs. Cells grown on titanium surfaces are subjected to intracellular oxidative stress. Titanium discs subjected to the plasma treatment induced less oxidative stress than the untreated ones, which resulted in improved cellular survival. These were associated with improved cellular antioxidant response in Plasma Treated Titanium Surface (PTTS). Furthermore, a decrease in protein and DNA oxidative damage was observed on cells grown on the roughed surface when compared to the smooth one. In conclusion, our data suggest that the treatment of titanium with argon plasma may improve its biocompatible, thus improving its performance as implants or as a scaffold in tissue engineering.

  6. MAP3K1 function is essential for cytoarchitecture of the mouse organ of Corti and survival of auditory hair cells

    PubMed Central

    Yousaf, Rizwan; Meng, Qinghang; Hufnagel, Robert B.; Xia, Ying; Puligilla, Chandrakala; Ahmed, Zubair M.; Riazuddin, Saima

    2015-01-01

    ABSTRACT 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. PMID:26496772

  7. MAP3K1 function is essential for cytoarchitecture of the mouse organ of Corti and survival of auditory hair cells.

    PubMed

    Yousaf, Rizwan; Meng, Qinghang; Hufnagel, Robert B; Xia, Ying; Puligilla, Chandrakala; Ahmed, Zubair M; Riazuddin, Saima

    2015-12-01

    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 Map3k1(tm1Yxia) mutant mice was open eyelids at birth. Here, we report that homozygous Map3k1(tm1Yxia) 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, Map3k1(tm1Yxia) 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.

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

    SciTech Connect

    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. - 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.

  9. Cytochalasin J treatment significantly alters mitotic spindle microtubule organization and kinetochore structure in PtK1 cells.

    PubMed

    Wrench, G A; Snyder, J A

    1997-01-01

    It has previously been demonstrated that treatment of mitotic PtK1 cells with 10-20 microg/ml cytochalasin J (CJ) blocks or slows chromosome motion and has a significant effect on spindle architecture [Snyder and Cohen, 1995: Cell Motil. Cytoskeleton 32:245-257]. Spindle microtubules (MTs) were shown to reorganize within the spindle domain, with kinetochore MTs (kMTs) reduced in number and non-kinetochore MTs (nkMTs) shown to splay outside the original spindle domain. In some cases, bundles of MTs were shown to be refocused away from the original spindle poles, creating the appearance of a multi-polar spindle. In this paper we use serial section electron microscopy, coupled with computer-assisted reconstruction techniques, to determine the rearrangement of spindle MTs and chromosome position following brief treatments of mitotic cells with 10-20 microg/ml CJ at various stages of mitosis. CJ treatment of prometaphase cells reduces the number of kMTs and the size and organization of the kinetochore lamina. Instead of kinetochore bundles of MTs aligned parallel to one another and running from kinetochore to pole, this class of MTs is highly fragmented. Non-kinetochore MTs are also highly fragmented, usually less than 2 microm long, and remain relatively straight over short distances, with some MTs arranged at an oblique angle to the longitudinal spindle axis. In approximately 30% of cells treated with CJ, the failure of a small number of chromosomes to attach to spindle fibers can be documented. These chromosomes show a significant change in the organization of the kinetochore laminae. Light microscopic analysis of cells treated with CJ reveals loss of chromosome congression, with chromsomes usually located at the periphery of the spindle and some completely detached from the spindle. Cells treated with 10 microg/ml CJ for 10 min and released into tissue culture medium show a resumption of chromosome motion within a few minutes, both during congression and anaphase

  10. Characterization of CHO-K1 cells stably expressing PDE-IV enzymes. Whole-cell cAMP determinations vs broken-cell enzymatic assays.

    PubMed

    Pon, D J; Plant, M; Tkach, J; Boulet, L; Muise, E; Allen, R A; Rodger, I W

    1998-01-01

    A CHO-K1 cell line stably expressing a recombinant full-length human PDE-IVa (HSPDE4A4B) enzyme was established under hygromycin B selection. Full-length expression of the protein was determined by Western blot analysis, which revealed the presence of a 125-kDa immunoreactive band using rabbit anti-PDE-IVa antibodies. The potency of inhibitor compounds was examined by their ability to increase cAMP in the whole-cell, and by their ability to inhibit cAMP hydrolysis in a 100,000 g supernatant (soluble enzyme preparation) obtained from the same cell line. Inhibition of the expressed PDE-IVa activity by selective PDE-IV inhibitors--(R) and (S)-rolipram, RS 14203, and CDP 840--at 100 nM substrate demonstrated that RS 14203 and CDP 840 were the most potent with IC50 = 9 nM, followed by (R)-rolipram (IC50 = 110 nM) and (S)-rolipram (IC50 = 420 nM). The rank order of potencies of the inhibitors in elevating cAMP in the whole-cell assay was quite different from that on the soluble enzyme. RS 14203 was still the most potent compound in elevating cAMP. Moreover, the relative rank order of potencies between CDP 840 and (R)-rolipram changed dramatically, such that (R)-rolipram was more potent than CDP 840 = (S)-rolipram. An apparent 30-fold stereoselectivity between (R)- and (S)-rolipram was also noted. The whole-cell rank order of potencies was also maintained when PKA activity ratios were measured in place of cAMP levels. The ability of the compounds to elevate cAMP in the stable CHO-K1 cells appeared to track better with the potency of the compounds against the high-affinity (Sr) conformer of the enzyme rather than the low-affinity catalytic state.

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

    NASA Technical Reports Server (NTRS)

    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

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

    PubMed Central

    Baisón-Olmo, Fernando; Galindo-Moreno, María; Ramos-Morales, Francisco

    2015-01-01

    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. PMID:25972862

  13. Inhibition of p70S6K1 Activation by Pdcd4 Overcomes the Resistance to an IGF-1R/IR Inhibitor in Colon Carcinoma Cells.

    PubMed

    Zhang, Yan; Wang, Qing; Chen, Li; Yang, Hsin-Sheng

    2015-03-01

    Agents targeting insulin-like growth factor 1 receptor (IGF-1R) are being actively examined in clinical trials. Although there has been some initial success of single-agent targeting IGF-1R, attempts in later studies failed because of resistance. This study aimed to understand the effects of programmed cell death 4 (Pdcd4) on the chemosensitivity of the IGF-1R inhibitor OSI-906 in colorectal cancer cells and the mechanism underlying this impact. Using OSI-906-resistant and -sensitive colorectal cancer cells, we found that the Pdcd4 level directly correlates with cell chemosensitivity to OSI-906. In addition, tumors derived from Pdcd4 knockdown cells resist the growth inhibitory effect of OSI-906 in a colorectal cancer xenograft mouse model. Moreover, Pdcd4 enhances the antiproliferative effect of OSI-906 in resistant cells through suppression of p70S6K1 activation. Knockdown of p70S6K1, but not p70S6K2, significantly increases the chemosensitivity of OSI-906 in cultured colorectal cancer cells. Furthermore, the combination of OSI-906 and PF-4708671, a p70S6K1 inhibitor, efficiently suppresses the growth of OSI-906-resistant colon tumor cells in vitro and in vivo. Taken together, activation of p70S6K1 that is inhibited by Pdcd4 is essential for resistance to the IGF-1R inhibitor in colon tumor cells, and the combinational treatment of OSI-906 and PF-4708671 results in enhanced antiproliferation effects in colorectal cancer cells in vitro and in vivo, providing a novel venue to overcome the resistance to the IGF-1R inhibitor in treating colorectal cancer. PMID:25573956

  14. Okadaic acid induces DNA fragmentation via caspase-3-dependent and caspase-3-independent pathways in Chinese hamster ovary (CHO)-K1 cells.

    PubMed

    Kitazumi, Ikuko; Maseki, Yoko; Nomura, Yoshiko; Shimanuki, Akiko; Sugita, Yumi; Tsukahara, Masayoshi

    2010-01-01

    DNA fragmentation is a hallmark of apoptosis that occurs in a variety of cell types; however, it remains unclear whether caspase-3 is required for its induction. To investigate this, we produced caspase-3 knockout Chinese hamster ovary (CHO)-K1 cells and examined the effects of gene knockout and treatment with caspase-3 inhibitors. Okadaic acid (OA) is a potent inhibitor of the serine/threonine protein phosphatases (PPs) PP1 and PP2A, which induce apoptotic cellular reactions. Treatment of caspase-3(-/-) cells with OA induced DNA fragmentation, indicating that caspase-3 is not an essential requirement. However, in the presence of benzyloxycarbonyl-Asp-Glu-Val-Asp (OMe) fluoromethylketone (z-DEVD-fmk), DNA fragmentation occurred in CHO-K1 cells but not in caspase-3(-/-) cells, suggesting that caspase-3 is involved in OA-induced DNA fragmentation that does not utilize DEVDase activity. In the absence of caspase-3, DEVDase activity may play an important role. In addition, OA-induced DNA fragmentation was reduced but not blocked in CHO-K1 cells, suggesting that caspase-3 is involved in caspase-independent OA-induced DNA fragmentation. Furthermore, OA-induced cleavage of caspase-3 and DNA fragmentation were blocked by pretreatment with the wide-ranging serine protease inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride. These results suggest that serine proteases regulate DNA fragmentation upstream of caspase-3.

  15. MiR-145 is downregulated in human ovarian cancer and modulates cell growth and invasion by targeting p70S6K1 and MUC1

    SciTech Connect

    Wu, Huijuan; Xiao, ZhengHua; Wang, Ke; Liu, Wenxin; Hao, Quan

    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.

  16. Atorvastatin partially inhibits the epithelial-mesenchymal transition in A549 cells induced by TGF-β1 by attenuating the upregulation of SphK1.

    PubMed

    Fan, Zhiqiang; Jiang, Handong; Wang, Zili; Qu, Jieming

    2016-08-01

    Statins are the most effective drugs used in the reduction of intracellular synthesis of cholesterol. Numerous studies have confirmed that statins reduce the risk of multiple types of cancers. Statin use in cancer patients is associated with reduced cancer-related mortality. Epithelial-to-mesenchymal transition (EMT), a complicated process programmed by multiple genes, is an important mechanism of cancer metastasis. We explored the effect and mechanism of atorvastatin on the EMT process in A549 cells by establishing an EMT model in vitro induced by TGF-β1, and evaluated the effects of atorvastatin on the lower signaling pathway of TGF-β1 stimulation. Our results showed that atorvastatin partially inhibited the EMT process, and inhibited cell migration and actin filament remodeling. Transcriptional upregulation of ZEB1 and protein sphingosine kinase 1 (SphK1) induced by TGF-β1 was also suppressed. SphK1 plasmid transient transfection strengthened the EMT process induced by TGF-β1 in the presence of atorvastatin. Our experiments confirmed that atorvastatin can partially inhibit the EMT process of non-small cell lung cancer cells induced by TGF-β1 by attenuating the upregulation of SphK1. PMID:27349500

  17. SSCA1-K1

    SciTech Connect

    Baker, Matthew

    2015-11-06

    SSCA1-K1 is a parallel implementation of kernel 1 of the SSCA1 benchmark suite released by the DARPA HPCS program. This kernel is able to run in parallel on a distributed shared memory system at extreme scales using OpenSHMEM.

  18. A novel regulatory element (E77) isolated from CHO‐K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells

    PubMed Central

    Kang, Shin‐Young; Kim, Yeon‐Gu; Kang, Seunghee; Lee, Hong Weon

    2016-01-01

    Abstract 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. PMID:26762773

  19. Induction of chromosomal damage in CHO-K1 cells and their repair-deficient mutant XRS5 by x-ray and particle irradiation

    NASA Astrophysics Data System (ADS)

    Nasonova, E.; Ritter, S.; Fomenkova, T.; Kraft, G.

    The cytogenetic effects of X-rays and Au ions were investigated in repair-proficient CHO-K1 cells and their radiosensitive mutant strain xrs5, which shows a defect in the rejoining of DNA double-strand breaks. Both cell lines were synchronized by mitotic shake off, irradiated in G_1-phase with either 250 kV X-rays or 780 MeV/u Au ions (LET: 1150 keV/mum) and chromosome aberrations were analyzed in first post-irradiation metaphases. Isoeffective doses of X-rays for the induction of aberrant cells and aberrations per cell were about 14 times lower for xrs5 than for CHO-K1 cells. After high LET radiation the difference in the cytogenetic response of both cell lines was drastically diminished. Furthermore, the analysis of the aberration types induced by sparsely and densely ionizing radiation showed for both cell lines specific changes in the spectrum of aberration types as LET increases. The experimental results are discussed with respect to the different types of lesions induced by sparsely and densely ionizing radiation.

  20. Interactions of Neuropathogenic Escherichia coli K1 (RS218) and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

    PubMed Central

    Yousuf, Farzana Abubakar; Yousuf, Zuhair; Iqbal, Junaid; Siddiqui, Ruqaiyyah; Khan, Hafsa; Khan, Naveed Ahmed

    2014-01-01

    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. PMID:24818136

  1. Macrophage receptor with collagenous structure (MARCO) is a dynamic adhesive molecule that enhances uptake of carbon nanotubes by CHO-K1 Cells

    SciTech Connect

    Hirano, Seishiro; Fujitani, Yuji; Furuyama, Akiko; Kanno, Sanae

    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.

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

    PubMed

    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.

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

    PubMed

    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. PMID:27035094

  4. Functional properties of alpha7 nicotinic acetylcholine receptors co-expressed with RIC-3 in a stable recombinant CHO-K1 cell line.

    PubMed

    Roncarati, Renza; Seredenina, Tamara; Jow, Brian; Jow, Flora; Papini, Silvia; Kramer, Angela; Bothmann, Hendrick; Dunlop, John; Terstappen, Georg C

    2008-04-01

    Heterologous functional expression of alpha7 nicotinic acetylcholine receptors (nAChRs) is difficult to achieve in mammalian cell lines, and the reasons have been associated with a lack of expression of the putative chaperone factor RIC-3. Here, we describe the generation and functional and pharmacological characterization of a Chinese hamster ovary (CHO)-K1 cell line co-expressing the human alpha7 nAChR and RIC-3. Stable recombinant cells expressing alpha7 nAChR on the plasma membrane were selected by binding of fluorochrome-conjugated alpha-bungarotoxin and fluorescence-activated cell sorting. The presence of functional alpha7 channels was demonstrated by whole cell patch clamp recordings. Nicotine and acetylcholine induced rapid desensitizing currents with 50% effective concentration values of 14 and 37 microM, respectively, with agonist-evoked currents detected in approximately 75% of the cell population. Surprisingly, when tested in a FLIPR (Molecular Devices, Sunnyvale, CA) Ca(2+) assay, activation of alpha7 nAChRs was measured only when nicotinic agonists were applied either in the presence of the positive allosteric modulator (PAM) PNU-120596 or after pretreatment of cells with the tyrosine kinase inhibitor genistein. No Ca(2+) influx was measured upon addition of agonists alone or together with allosteric potentiators such as 5-hydroxyindole that predominantly increase the apparent peak amplitude without robustly affecting the current desensitization rate, as exemplified by PNU-120596. These results show that functional alpha7 nAChRs can stably be expressed in the non-neuronal CHO-K1 cell line. This recombinant cell system is useful for characterization of alpha7 nAChRs and to study the mechanism of action of chemical modulators, in particular the detection of PAMs capable of slowing receptor desensitization kinetics.

  5. Analysis of possible genotoxicity of the herbicide flurochloridone and its commercial formulations: Endo III and Fpg alkaline comet assays in Chinese hamster ovary (CHO-K1) cells.

    PubMed

    Soloneski, Sonia; Nikoloff, Noelia; Larramendy, Marcelo L

    2016-02-01

    Cytotoxic and genotoxic effects of flurochloridone (FLC) and its formulations Twin Pack Gold(®) and Rainbow(®) were evaluated in CHO-K1 cells. Using the alkaline single-cell gel electrophoresis (SCGE) assay, we observed that FLC (15 μg/ml), Twin Pack Gold(®) or Rainbow(®) induced primary DNA damage, increasing the frequency of damaged nucleoids. Vitamin E pretreatment did not modify the effect. Decreased cell viability was observed only in Twin Pack Gold(®)-treated cultures and was significantly ameliorated by vitamin E. Post-treatment of herbicide-damaged CHO-K1 cells with the enzymes Endo III or Fpg did not increase FLC-, Twin Pack Gold(®)-, or Rainbow(®)-induced DNA damage. These results demonstrate that neither FLC nor FLC-based formulations induce DNA damage through hydroxyl radical or lipid alkoxyl radical production, and that the induced DNA lesions were not related to oxidative damage at the purine/pyrimidine level. Our observations strongly suggest that the cytotoxic effects observed after Twin Pack Gold(®) exposure are due to the excipients contained within the technical formulation rather than FLC itself.

  6. Analysis of possible genotoxicity of the herbicide flurochloridone and its commercial formulations: Endo III and Fpg alkaline comet assays in Chinese hamster ovary (CHO-K1) cells.

    PubMed

    Soloneski, Sonia; Nikoloff, Noelia; Larramendy, Marcelo L

    2016-02-01

    Cytotoxic and genotoxic effects of flurochloridone (FLC) and its formulations Twin Pack Gold(®) and Rainbow(®) were evaluated in CHO-K1 cells. Using the alkaline single-cell gel electrophoresis (SCGE) assay, we observed that FLC (15 μg/ml), Twin Pack Gold(®) or Rainbow(®) induced primary DNA damage, increasing the frequency of damaged nucleoids. Vitamin E pretreatment did not modify the effect. Decreased cell viability was observed only in Twin Pack Gold(®)-treated cultures and was significantly ameliorated by vitamin E. Post-treatment of herbicide-damaged CHO-K1 cells with the enzymes Endo III or Fpg did not increase FLC-, Twin Pack Gold(®)-, or Rainbow(®)-induced DNA damage. These results demonstrate that neither FLC nor FLC-based formulations induce DNA damage through hydroxyl radical or lipid alkoxyl radical production, and that the induced DNA lesions were not related to oxidative damage at the purine/pyrimidine level. Our observations strongly suggest that the cytotoxic effects observed after Twin Pack Gold(®) exposure are due to the excipients contained within the technical formulation rather than FLC itself. PMID:26921020

  7. A new karyotype for Rhipidomys (Rodentia, Cricetidae) from Southeastern Brazil

    PubMed Central

    de Carvalho, Ana Heloisa; Lopes, Maria Olímpia Garcia; Svartman, Marta

    2012-01-01

    Abstract In this work we present a new karyotype for Rhipidomys Tschudi, 1845 (Cricetidae, Rodentia) from Brazil. Our chromosome analyses included GTG- and CBG-banding patterns, the localization of the nucleolus organizer regions after silver staining (Ag-NORs) and fluorescence in situ hybridization (FISH) with a telomere probe. The new karyotype is composed of 44 chromosomes and has a fundamental number (number of autosomal arms) of 48. Most Rhipidomys species already karyotyped presented similar complements with 2n=44, but their fundamental numbers varied from FN=46 to 80, a variation that has been mainly attributed to pericentric inversions. The comparison of this new karyotype to those of other Rhipidomys already reported allowed us to conclude that it is a distinctive chromosome complement, which can be of great use as a tool for the very complicated taxonomic identification in this genus. PMID:24260664

  8. Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media.

    PubMed

    Mohmad-Saberi, Salfarina Ezrina; Hashim, Yumi Zuhanis Has-Yun; Mel, Maizirwan; Amid, Azura; Ahmad-Raus, Raha; Packeer-Mohamed, Vasila

    2013-08-01

    An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system. In this study, global metabolite (metabolomics) analysis approach was used to try and understand the relationships between types of media used, culture growth behavior and productivity. CHO-KI cells producing IGF-1 were obtained from ATCC and grown in T-flask (37 °C, 5 % CO2) until 70-80 % confluent in RPMI 1640 and Ham's F12, respectively. Samples were taken at 8-hourly intervals for routine cell counting, biochemical responses, insulin like growth factor-1 (IGF-1) protein concentration and global metabolite analysis (gas chromatography mass spectrometry, GCMS). Conditioned media from each time point were spun down before injection into GCMS. Data from GCMS were then transferred to SIMCA-P + Version 12 for chemometric evaluation using principal component analysis. The results showed that while routine analysis gave only subtle differences between the media, global metabolite analysis was able to clearly separate the culture based on growth media with growth phases as confounding factor. Different types of media also appeared to affect IGF-1 production. Asparagine was found to be indicative of healthiness of cells and production of high IGF-1. Meanwhile identification of ornithine and lysine in death phase was found to be associated with apoptosis and oversupplied nutrient respectively. Using the biomarkers revealed in the study, several bioprocessing strategies including medium improvement and in-time downstream processing can be potentially implemented to achieve efficient CHO culture system.

  9. Coupling of a transfected human brain A1 adenosine receptor in CHO-K1 cells to calcium mobilisation via a pertussis toxin-sensitive mechanism.

    PubMed Central

    Iredale, P. A.; Alexander, S. P.; Hill, S. J.

    1994-01-01

    1. The presence of A1 adenosine receptors in CHO-K1 cells transfected with the human brain A1 sequence was confirmed by ligand binding studies using 8-cyclopentyl-[3H] 1,3-dipropylxanthine ([3H]-DPCPX). 2. Alterations in intracellular calcium ([Ca2+]i) were measured with the calcium-sensitive dye, fura-2. 3. N6-cyclopentyladenosine (CPA), the selective A1 agonist, and 5'-N-ethylcarboxaminoadenosine (NECA), a relatively non-selective adenosine receptor agonist, elicited rapid, biphasic increases in [Ca2+]i which involved both mobilisation from intracellular stores and calcium entry. 4. The calcium response to CPA was significantly inhibited by the selective A1 antagonist DPCPX. The non-selective adenosine receptor, xanthine amino congener (XAC), was less potent. 5. The calcium response to CPA was completely prevented by pretreatment of the cells with pertussis toxin implicating the involvement of Gi in the receptor-mediated response. 6. In summary, we present evidence for the coupling of transfected human brain A1 adenosine receptors in CHO-K1 cells to mobilisation of [Ca2+]i via a pertussis toxin-sensitive G protein. PMID:8032613

  10. Inclusion of S-sepharose beads in the culture medium significantly improves recovery of secreted rBPI(21) from transfected CHO-K1 cells.

    PubMed

    Horwitz, A H; Carroll, S F; Williams, R E; Liu, P S

    2000-02-01

    rBPI(23), a recombinant N-terminal fragment of human bactericidal/permeability-increasing protein (BPI), kills gram-negative bacteria and binds endotoxin. rBPI(21), a variant, in which cysteine 132 is changed to alanine, retains the activities of rBPI(23). Initial attempts using conventional ion-exchange chromatography to purify rBPI(23) from culture supernatants of transfected CHO-K1 cells resulted in lower than expected yields. Also, ELISA of supernatants from CHO-K1 transfectants expressing rBPI(23) or rBPI(21) yielded variable signals. Results from pulse-chase experiments using [(35)S]methionine had indicated that rBPI(23) could not be detected in the culture medium by 7 h of chase, suggesting that these proteins were degraded and/or bound to cells, media components, or vessel surfaces. To address these issues, we developed a novel process whereby sterile S-Sepharose beads were added directly to the cell culture medium. For attached cells, the beads were added to confluent cultures with serum-free medium for the expression phase, while for suspension-adapted cells, beads were added at the beginning of culture growth. The S-Sepharose was then separated from cells and media and washed, and BPI was eluted with high-salt buffer. This approach yielded up to a 50-fold improvement in recovery of rBPI(23) and rBPI(21) from roller bottles, shake flasks, and 2-liter fermenters. It also resulted in improved detection and quantitation of secreted rBPI(23) and rBPI(21) by ELISA. Results of competition binding studies with iodinated rBPI(21) in conjunction with unlabeled rBPI(21) and rBPI(23) or with heparin demonstrated that these proteins bound specifically and with high affinity to heparan-containing sites on the surface of the CHO-K1 cells. We conclude that the S-Sepharose included in the culture medium captures the BPI protein products as they are secreted and protects them from degradation and/or irreversible binding to cell surfaces. This method has been scaled up to

  11. 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.

    PubMed

    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.

  12. 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

    PubMed Central

    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

  13. Relevance of kinetochore size and microtubule-binding capacity for stable chromosome attachment during mitosis in PtK1 cells.

    PubMed

    McEwen, B F; Ding, Y; Heagle, A B

    1998-02-01

    Chromosomes attach to the mitotic spindle via their kinetochores. The average number of spindle microtubules binding to each kinetochore varies with species, the stage of mitosis, and the length of time that the kinetochore has been attached to the spindle. In this report, we investigate how kinetochore microtubule number varies with kinetochore size and chromosome size in PtK1 cells. From an analysis of serial-section electron micrographs, we determined that the average surface area of metaphase, taxol-treated metaphase, and anaphase kinetochores is 0.16 +/- 0.05 microm2 (N = 181). Surprisingly, kinetochore microtubules are packed more densely on the smaller kinetochores, as seen by a reduction in the average spacing between kinetochore microtubules from 89 nm to 59 nm. Our interpretation of this result is that PtK1 cells require a minimum kinetochore microtubule-binding capacity for survival during repeated rounds of mitotic division. We estimate the lower limit to be 23 kinetochore microtubules and suggest that this capacity is required to ensure stable attachment during the dynamic and highly stochastic process of kinetochore fiber formation. There is a modest but statistically significant increase in kinetochore microtubule number with chromosome size, indicating that chromosome size is a minor determinant of kinetochore microtubule number.

  14. Landfill leachate sludge use as soil additive prior and after electrocoagulation treatment: A cytological assessment using CHO-k1 cells.

    PubMed

    Morozesk, M; Bonomo, M M; Rocha, L D; Duarte, I D; Zanezi, E R L; Jesus, H C; Fernandes, M N; Matsumoto, S T

    2016-09-01

    Electrocoagulation has recently attracted attention as a potential technique for treating toxic effluents due to its versatility and environmental compatibility, generating a residue chemically suitable to be used as a soil additive. In the present study, landfill leachate sludge hazardous effects were investigated prior and after electrocoagulation process using in vitro assays with the mammalian cells CHO-k1. An integrated strategy for risk assessment was used to correctly estimate the possible adverse landfill leachate sludge effects on human health and ecosystem. Electrocoagulation process proved to be an effective treatment due to possibility to improve effluent adverse characteristics and produce sludge with potential to be used as soil additive. Despite low cytoxicity, the residue presented genotoxic and mutagenic effects, indicating a capacity to induce genetic damages, probably due to induction of polyploidization process in cells. The observed effects demand an improvement of waste management methods for reduce negative risks of landfill leachate sludge application.

  15. Landfill leachate sludge use as soil additive prior and after electrocoagulation treatment: A cytological assessment using CHO-k1 cells.

    PubMed

    Morozesk, M; Bonomo, M M; Rocha, L D; Duarte, I D; Zanezi, E R L; Jesus, H C; Fernandes, M N; Matsumoto, S T

    2016-09-01

    Electrocoagulation has recently attracted attention as a potential technique for treating toxic effluents due to its versatility and environmental compatibility, generating a residue chemically suitable to be used as a soil additive. In the present study, landfill leachate sludge hazardous effects were investigated prior and after electrocoagulation process using in vitro assays with the mammalian cells CHO-k1. An integrated strategy for risk assessment was used to correctly estimate the possible adverse landfill leachate sludge effects on human health and ecosystem. Electrocoagulation process proved to be an effective treatment due to possibility to improve effluent adverse characteristics and produce sludge with potential to be used as soil additive. Despite low cytoxicity, the residue presented genotoxic and mutagenic effects, indicating a capacity to induce genetic damages, probably due to induction of polyploidization process in cells. The observed effects demand an improvement of waste management methods for reduce negative risks of landfill leachate sludge application. PMID:27243586

  16. Effect of Brazilian propolis (AF-08) on genotoxicity, cytotoxicity and clonogenic death of Chinese hamster ovary (CHO-K1) cells irradiated with (60)Co gamma-radiation.

    PubMed

    Santos, Geyza Spigoti; Tsutsumi, Shigetoshi; Vieira, Daniel Perez; Bartolini, Paolo; Okazaki, Kayo

    2014-03-01

    The present study was conducted in order to evaluate the effect of Brazilian propolis (AF-08; 5, 10, 15, 30, 50, 100, and 200μg/mL) in protecting CHO-K1 cells against genotoxic and cytotoxic damage and clonogenic death induced by (60)Co gamma-radiation (1.0, 2.0, 4.0, and 6.0Gy). For this purpose, three interlinked endpoints were analyzed: induction of DNA damage by use of the micronucleus (MN) test (genotoxic damage), cell viability by means of the MTS assay, and differential staining (cytotoxic damage) and clonogenic death via the colony-formation test (cytotoxic damage). The MN test revealed that propolis alone (5-100μg/mL) was not genotoxic up to 100μg/mL and that 30μg/mL of propolis reduced the radiation-induced DNA damage (∼56% reduction, p<0.05), exhibiting a radio-protective effect on irradiated CHO-K1 cells. On the other hand, analysis of cytotoxicity showed that a concentration of 50μg/mL presented a significant proliferative effect (p<0.001) when associated with radiation, decreasing the percentage of necrotic cells (p<0.01). No mediated cytotoxic effect was found, but the concentration of 200μg/mL was toxic when analyzed at 24 and 48h via the differential staining technique, but not at 72h after irradiation, analyzed with the MTS assay. Differential staining also showed that necrosis was the main death modality in irradiated cells and that apoptosis was induced only at the toxic concentration of propolis (200μg/mL). Concerning the clonogenic capacity, a concentration of 50μg/mL also exhibited a significant stimulating effect on cell proliferation (p<0.001), in agreement with the data from differential staining. Taken together, these data suggest that the use of propolis AF-08 for the prevention of the adverse effects of ionizing radiation is promising. Nevertheless, additional investigations are necessary for a better understanding of potential applications of propolis to improve human health.

  17. TGFβ-Mediated induction of SphK1 as a potential determinant in human MDA-MB-231 breast cancer cell bone metastasis

    PubMed Central

    Stayrook, Keith R; Mack, Justin K; Cerabona, Donna; Edwards, Daniel F; Bui, Hai H; Niewolna, Maria; Fournier, Pierrick GJ; Mohammad, Khalid S; Waning, David L; Guise, Theresa A

    2015-01-01

    Mechanistic understanding of the preferential homing of circulating tumor cells to bone and their perturbation on bone metabolism within the tumor–bone microenvironment remains poorly understood. Alteration in both transforming growth factor β (TGFβ) signaling and sphingolipid metabolism results in the promotion of tumor growth and metastasis. Previous studies using MDA-MB-231 human breast cancer-derived cell lines of variable metastatic potential were queried for changes in sphingolipid metabolism genes to explore correlations between TGFβ dependence and bone metastatic behavior. Of these genes, only sphingosine kinase-1 (SPHK1) was identified to be significantly increased following TGFβ treatment. Induction of SPHK1 expression correlated to the degree of metastatic capacity in these MDA-MB-231-derived cell lines. We demonstrate that TGFβ mediates the regulation of SPHK1 gene expression, protein kinase activity and is critical to MDA-MB-231 cell viability. Furthermore, a bioinformatic analysis of human breast cancer gene expression supports SPHK1 as a hallmark TGFβ target gene that also bears the genetic fingerprint of the basal-like/triple-negative breast cancer molecular subtype. These data suggest a potential new signaling axis between TGFβ/SphK1 that may have a role in the development, prognosis or the clinical phenotype associated with tumor-bone metastasis. PMID:26157579

  18. Phylogeography of Trichuris populations isolated from different Cricetidae rodents.

    PubMed

    Callejón, Rocío; De Rojas, Manuel; Feliú, Carlos; Balao, Francisco; Marrugal, Angela; Henttonen, Heikki; Guevara, Diego; Cutillas, Cristina

    2012-11-01

    The phylogeography of Trichuris populations (Nematoda) collected from Cricetidae rodents (Muroidea) from different geographical regions was studied. Ribosomal DNA (Internal Transcribed Spacers 1 and 2, and mitochondrial DNA (cytochrome c- oxidase subunit 1 partial gene) have been used as molecular markers. The nuclear internal transcribed spacers (ITSs) 1 and 2 showed 2 clear-cut geographical and genetic lineages: one of the Nearctic region (Oregon), although the second was widespread throughout the Palaearctic region and appeared as a star-like structure in the minimum spanning network. The mitochondrial results revealed that T. arvicolae populations from the Palaearctic region were separated into 3 clear-cut geographical and genetic lineages: populations from Northern Europe, populations from Southern (Spain) and Eastern Europe (Croatia, Belarus, Kazahstan), and populations from Italy and France (Eastern Pyrénean Mountains). Phylogenetic analysis obtained on the basis of ITS1-5·8S-ITS2 rDNA sequences did not show a differential geographical structure; however, these markers suggest a new Trichuris species parasitizing Chionomys roberti and Cricetulus barabensis. The mitochondrial results revealed that Trichuris populations from arvicolinae rodents show signals of a post-glacial northward population expansion starting from the Pyrenees and Italy. Apparently, the Pyrenees and the Alps were not barriers to the dispersal of Trichuris populations.

  19. N- and 6-O-Sulfated Heparan Sulfates Mediate Internalization of Coxsackievirus B3 Variant PD into CHO-K1 Cells

    PubMed Central

    Zautner, Andreas E.; Jahn, Birgit; Hammerschmidt, Elke; Wutzler, Peter; Schmidtke, Michaela

    2006-01-01

    Recently, it was demonstrated that the coxsackievirus B3 variant PD (CVB3 PD) is able to infect coxsackievirus-adenovirus receptor (CAR)-lacking cells by using heparan sulfates (HS) as additional receptors (A. E. Zautner, U. Korner, A. Henke, C. Badorff, and M. Schmidtke, J. Virol. 77:10071-10077, 2003). For this study, competition experiments with growth factors binding to known HS sequences as well as with specifically desulfated heparins were performed with Chinese hamster ovary cells (CHO-K1) to determine the structural requirements of HS for interaction with CVB3. Hepatocyte growth factor interacting with HS sequences containing [IdUA-GlcNSO3(6OSO3)]n, but not basic fibroblast growth factor binding to [HexUA-GlcNSO3-HexUA-GlcNSO3-IdUA(2OSO3)]n, was shown to compete effectively with CVB3 PD for cell surface HS. Whereas unmodified heparin and 2-O-desulfated heparin strongly inhibited the CVB3 PD-induced cytopathic effect, the antiviral activity was markedly reduced after N-, O- and 6-O-desulfation of heparin. Taken together, these results indicate that 6-O- and N-sulfation of GlcNAc of HS is crucial for HS interaction with CVB3 PD and that the disaccharide [IdUA-GlcNSO3(6OSO3)]n is involved in viral binding. Results from experiments with various inhibitors of endocytic pathways suggest that HS-mediated virus internalization is pH dependent. Despite the fact that CVB3 PD initiates infection about four times slower by making use of HS as a receptor than by using CAR, the time required for a complete viral life cycle in Chinese hamster ovary cells was independent of the utilized receptor. PMID:16775350

  20. The surface protease ompT serves as Escherichia coli K1 adhesin in binding to human brain micro vascular endothelial cells.

    PubMed

    Wan, Lei; Guo, Yan; Hui, Chang-Ye; Liu, Xiao-Lu; Zhang, Wen-Bing; Cao, Hong; Cao, Hong

    2014-05-01

    Escherichia coli (E. coli) K1 is the most common bacteria that cause meningitis in the neonatal period. But it's not entirely clear about how E. coli crosses the blood-brain barrier. The features of the ompT deletion in meningitic E. coli infection were texted in vitro. In comparison with the parent strain, the isogenic ompT deletion mutant was significantly less adhesive to human brain microvascular endothelial cells (HBMEC). The adhesion-deficient phenotype of the mutant was restored to the level of the wild-type by complementing with low-level OmpT expression plasmid. Interestingly, the adhesion was enhanced by point mutation at the OmpT proposed catalytic residue D85. Compared with the poor adhesive activity of bovine serum albumin-coated fluorescent beads, recombinant OmpT or catalytically inactive variant of OmpT-coated beads bound to HBMEC monolayer effectively. Our study suggests that OmpT is important for bacterial adhesion while entering into central nervous system, and the adhesion does not involve in the proteolytic activity of OmpT.

  1. Kinetochore fiber maturation in PtK1 cells and its implications for the mechanisms of chromosome congression and anaphase onset.

    PubMed

    McEwen, B F; Heagle, A B; Cassels, G O; Buttle, K F; Rieder, C L

    1997-06-30

    Kinetochore microtubules (kMts) are a subset of spindle microtubules that bind directly to the kinetochore to form the kinetochore fiber (K-fiber). The K-fiber in turn interacts with the kinetochore to produce chromosome motion toward the attached spindle pole. We have examined K-fiber maturation in PtK1 cells using same-cell video light microscopy/serial section EM. During congression, the kinetochore moving away from its spindle pole (i.e., the trailing kinetochore) and its leading, poleward moving sister both have variable numbers of kMts, but the trailing kinetochore always has at least twice as many kMts as the leading kinetochore. A comparison of Mt numbers on sister kinetochores of congressing chromosomes with their direction of motion, as well as distance from their associated spindle poles, reveals that the direction of motion is not determined by kMt number or total kMt length. The same result was observed for oscillating metaphase chromosomes. These data demonstrate that the tendency of a kinetochore to move poleward is not positively correlated with the kMt number. At late prometaphase, the average number of Mts on fully congressed kinetochores is 19.7 +/- 6.7 (n = 94), at late metaphase 24.3 +/- 4.9 (n = 62), and at early anaphase 27.8 +/- 6.3 (n = 65). Differences between these distributions are statistically significant. The increased kMt number during early anaphase, relative to late metaphase, reflects the increased kMt stability at anaphase onset. Treatment of late metaphase cells with 1 microM taxol inhibits anaphase onset, but produces the same kMt distribution as in early anaphase: 28.7 +/- 7. 4 (n = 54). Thus, a full complement of kMts is not sufficient to induce anaphase onset. We also measured the time course for kMt acquisition and determined an initial rate of 1.9 kMts/min. This rate accelerates up to 10-fold during the course of K-fiber maturation, suggesting an increased concentration of Mt plus ends in the vicinity of the kinetochore at

  2. Selective Modulation of Protein Kinase C α over Protein Kinase C ε by Curcumin and Its Derivatives in CHO-K1 Cells.

    PubMed

    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. Selective Modulation of Protein Kinase C α over Protein Kinase C ε by Curcumin and Its Derivatives in CHO-K1 Cells.

    PubMed

    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. PMID

  4. Polydatin attenuates AGEs-induced upregulation of fibronectin and ICAM-1 in rat glomerular mesangial cells and db/db diabetic mice kidneys by inhibiting the activation of the SphK1-S1P signaling pathway.

    PubMed

    Chen, Cheng; Huang, Kaipeng; Hao, Jie; Huang, Junying; Yang, Zhiying; Xiong, Fengxiao; Liu, Peiqing; Huang, Heqing

    2016-05-15

    We previously demonstrated that activation of sphingosine kinase 1 (SphK1)- sphingosine 1- phosphate (S1P) signaling pathway by high glucose (HG) plays a pivotal role in increasing the expression of fibronectin (FN), an important fibrotic component, by promoting the DNA-binding activity of transcription factor activator protein 1 (AP-1) in glomerular mesangial cells (GMCs) under diabetic conditions. As a multi-target anti-oxidative drug, polydatin (PD) has been shown to have renoprotective effects on experimental diabetes. However, whether PD could resist diabetic nephropathy (DN) by regulating SphK1-S1P signaling pathway needs further investigation. Here, we found that PD significantly reversed the upregulated FN and ICAM-1 expression in GMCs exposed to AGEs. Simultaneously, PD dose-dependently inhibited SphK1 levels at the protein expression and kinase activity and attenuated S1P production under AGEs treatment conditions. In addition, PD reduced SphK activity in GMCs transfected with wild-type SphK(WT) plasmid and significantly suppressed SphK1-mediated increase of FN and ICAM-1 levels under normal conditions. Furthermore, we found that the AGEs-induced upregulation of phosphorylation of c-Jun at Ser63 and Ser73 and c-Fos at Ser32, DNA-binding activity and transcriptional activity of AP-1 were blocked by PD. In comparison with db/db model group, PD treatment suppressed SphK1 levels (mRNA, protein expression, and activity) and S1P production, reversed the upregulation of FN, ICAM-1, c-Jun, and c-Fos in the kidney tissues of diabetic mice, and finally ameliorated renal injury in db/db mice. These findings suggested that the downregulation of SphK1-S1P signaling pathway is probably a novel mechanism by which PD suppressed AGEs-induced FN and ICAM-1 expression and improved renal dysfunction of diabetic models. PMID:26948947

  5. Elevated Protein Kinase D3 (PKD3) Expression Supports Proliferation of Triple-negative Breast Cancer Cells and Contributes to mTORC1-S6K1 Pathway Activation*

    PubMed Central

    Huck, Bettina; Duss, Stephan; Hausser, Angelika; Olayioye, Monilola A.

    2014-01-01

    Here, we show that the expression of the Golgi-localized serine-threonine kinase protein kinase D3 (PKD3) is elevated in triple-negative breast cancer (TNBC). Using an antibody array, we identified PKD3 to trigger the activation of S6 kinase 1 (S6K1), a main downstream target of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. Accordingly, PKD3 knockdown in TNBC cells led to reduced S6K1 phosphorylation, which was associated with impaired activation of mTORC1 at endolysosomal membranes, the accumulation of the mannose 6-phosphate receptor in and the recruitment of the autophagy marker light chain 3 to enlarged acidic vesicles. We further show that PKD3 depletion strongly inhibited cell spreading and proliferation of TNBC cells, identifying this kinase as a potential novel molecular therapeutic target in TNBC. Together, our data suggest that PKD3 in TNBC cells provides a molecular connection between the Golgi and endolysosomal compartments to enhance proliferative mTORC1-S6K1 signaling. PMID:24337579

  6. Pterygodermatites (Paucipectines) baiomydis n. sp. (Nematoda: Rictulariidae), a parasite of Baiomys taylori (Cricetidae)

    PubMed Central

    Lynggaard, Christina; García-Prieto, Luis; Guzmán-Cornejo, Carmen; Osorio-Sarabia, David

    2014-01-01

    Pterygodermatites (Paucipectines) baiomydis n. sp., an intestinal parasite of the northern pygmy mouse, Baiomys taylori (Cricetidae), collected in La Yerbabuena, Colima, Mexico, is described herein. Specimens were studied using light and scanning electronic microscopy. This is the 19th species of the subgenus Paucipectines described worldwide and the fourth collected in Mexico. It is differentiated from the remaining species in the subgenus by having 25 perioral denticles, arranged in a triangle (seven on each lateroventral margin, and eleven on the dorsal margin), and 10 pairs of caudal papillae. PMID:25375029

  7. A new genus and species of lungworm (Nemata: Metastrongyloidea) from Akodon mollis Thomas, 1894 (Rodentia: Cricetidae) in Peru.

    PubMed

    Morales, Maria Elizabeth; Ubelaker, John E; Gardner, Scott L

    2012-06-01

    Akodonema luzsarmientae n.g., n.sp. (Nemata: Metastrongyloidea) is described from the pulmonary arteries and heart from several individuals of "soft grass mouse," Akodon mollis (Rodentia: Cricetidae), collected in the region of Ancash, Peru. The new genus and species is distinguished by a reduction of the dorsal ray to 2 small widely separated papillae. PMID:22746394

  8. Pterygodermatites (Paucipectines) baiomydis n. sp. (Nematoda: Rictulariidae), a parasite of Baiomys taylori (Cricetidae).

    PubMed

    Lynggaard, Christina; García-Prieto, Luis; Guzmán-Cornejo, Carmen; Osorio-Sarabia, David

    2014-01-01

    Pterygodermatites (Paucipectines) baiomydis n. sp., an intestinal parasite of the northern pygmy mouse, Baiomys taylori (Cricetidae), collected in La Yerbabuena, Colima, Mexico, is described herein. Specimens were studied using light and scanning electronic microscopy. This is the 19th species of the subgenus Paucipectines described worldwide and the fourth collected in Mexico. It is differentiated from the remaining species in the subgenus by having 25 perioral denticles, arranged in a triangle (seven on each lateroventral margin, and eleven on the dorsal margin), and 10 pairs of caudal papillae.

  9. Antidepressants activate the lysophosphatidic acid receptor LPA(1) to induce insulin-like growth factor-I receptor transactivation, stimulation of ERK1/2 signaling and cell proliferation in CHO-K1 fibroblasts.

    PubMed

    Olianas, Maria C; Dedoni, Simona; Onali, Pierluigi

    2015-06-15

    Different lines of evidence indicate that the lysophosphatidic acid (LPA) receptor LPA1 is involved in neurogenesis, synaptic plasticity and anxiety-related behavior, but little is known on whether this receptor can be targeted by neuropsychopharmacological agents. The present study investigated the effects of different antidepressants on LPA1 signaling. We found that in Chinese hamster ovary (CHO)-K1 fibroblasts expressing endogenous LPA1 tricyclic and tetracyclic antidepressants and fluoxetine induced the phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2) and CREB. This response was antagonized by either LPA1 blockade with Ki16425 and AM966 or knocking down LPA1 with siRNA. Antidepressants induced ERK1/2 phosphorylation in human embryonic kidney (HEK)-293 cells overexpressing LPA1, but not in wild-type cells. In PathHunter™ assay measuring receptor-β-arrestin interaction, amitriptyline, mianserin and fluoxetine failed to induce activation of LPA2 and LPA3 stably expressed in CHO-K1 cells. ERK1/2 stimulation by antidepressants and LPA was suppressed by pertussis toxin and inhibition of Src, phosphatidylinositol-3 kinase and insulin-like growth factor-I receptor (IGF-IR) activities. Antidepressants and LPA induced tyrosine phosphorylation of IGF-IR and insulin receptor-substrate-1 through LPA1 and Src. Prolonged exposure of CHO-K1 fibroblasts to either mianserin, mirtazapine or LPA enhanced cell proliferation as indicated by increased [(3)H]-thymidine incorporation and Ki-67 immunofluorescence. This effect was inhibited by blockade of LPA1- and ERK1/2 activity. These data provide evidence that different antidepressants induce LPA1 activation, leading to receptor tyrosine kinase transactivation, stimulation of ERK1/2 signaling and enhanced cell proliferation.

  10. Inhibition of topoisomerase II activity in repair-proficient CHO K1 cells by 2-[(aminopropyl)amino]ethanethiol (WR-1065)

    SciTech Connect

    Grdina, D.J.; Constantinou, A.; Shigematsu, N.

    1992-09-01

    The aminothiol 2-[(aminopropyl)amino]ethanethiol (WR-1065) is the active thiol of the clinically studied radioprotective agent S-2-(3-aminopropylamino) ethylphosphorothioic acid (WR-2721). WR-1065 is an effective radiation protector under in vitro conditions when it is administered 30 min prior to radiation exposure at a concentration of 4 mM to repair-proficient Chinese hamster ovary Kl cells (i.e., a dose modification factor of 1.4). In contrast, the DNA double-strand break, repair-deficient Chinese hamster ovary xrs-5 cell line is not protected under these conditions (i.e., a dose modification factor of 1.0). Topoisomerase (topo) I and II activities and protein contents were measured in both Kl and xrs-5 cell lines and were found to be similar in magnitude. Neither exposure to radiation, to WR-1065, or to both affected these variables in xrs-5 cells. WR 1065 was effective, however, in reducing topo 11 activity by a factor of 2 in the repair-proficient Kl cell line. Topo II protein content, however, was not affected by these exposure conditions. One of several mechanisms of radiation protection attributed to aminothiol compounds has been their ability to affect enzymatic reactions involved in DNA synthesis, repair, and cell cycle progression. These results demonstrate a modifying effect by 2-[(aminopropyl)amino]ethanethiol on a specific nuclear enzyme (i.e., type H topoisomerase), which is involved in DNA synthesis. These results also suggest that differences do exist between the topo 11 enzymes isolated from the parent repair-proficient Kl and the DNA double-strand break, repair-deficient xrs-5 mutant cell lines.

  11. Decontamination of hot cells K-1, K-3, M-1, M-3, and A-1, M-Wing, Building 200: Project final report Argonne National Laboratory-East

    SciTech Connect

    Cheever, C.L.; Rose, R.W.

    1996-09-01

    The purpose of this project was to remove radioactively contaminated materials and equipment from the hot cells, to decontaminate the hot cells, and to dispose of the radioactive waste. The goal was to reduce stack releases of Rn-220 and to place the hot cells in an emptied, decontaminated condition with less than 10 {micro}Sv/h (1 mrem/h) general radiation background. The following actions were needed: organize and mobilize a decontamination team; prepare decontamination plans and procedures; perform safety analyses to ensure protection of the workers, public, and environment; remotely size-reduce, package, and remove radioactive materials and equipment for waste disposal; remotely decontaminate surfaces to reduce hot cell radiation background levels to allow personnel entries using supplied air and full protective suits; disassemble and package the remaining radioactive materials and equipment using hands-on techniques; decontaminate hot cell surfaces to remove loose radioactive contaminants and to attain a less than 10 {micro}Sv/h (1 mrem/h) general background level; document and dispose of the radioactive and mixed waste; and conduct a final radiological survey.

  12. Src mediates endocytosis of TWIK-related acid-sensitive K+ 1 channels in PC12 cells in response to nerve growth factor.

    PubMed

    Matsuoka, Hidetada; Inoue, Masumi

    2015-08-15

    TWIK-related acid-sensitive K(+) (TASK) channels produce background K(+) currents. We elucidated that TASK1 channels in rat adrenal medullary cells and PC12 cells are internalized in a clathrin-dependent manner in response to nerve growth factor (NGF). Here, the molecular mechanism for this internalization in PC12 cells was explored. The combination of enzyme inhibitors with tropomyosin receptor kinase A mutants revealed that the internalization was mediated by both phospholipase C and phosphatidylinositol 3-kinase pathways that converge on protein kinase C with the consequent activation of Src, a nonreceptor tyrosine kinase. The NGF-induced endocytosis of TASK1 channels did not occur in the presence of the Src inhibitor or with the expression of a kinase-dead Src mutant. Additionally, NGF induced a transient colocalization of Src with the TASK1 channel, but not the TASK1 mutant, in which tyrosine at 370 was replaced with phenylalanine. This TASK1 mutant showed no increase in tyrosine phosphorylation and markedly diminished internalization in response to NGF. We concluded that NGF induces endocytosis of TASK1 channels via tyrosine phosphorylation in its carboxyl terminus.

  13. Development of Organometallic S6K1 Inhibitors

    PubMed Central

    2015-01-01

    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. PMID:25356520

  14. Vaccinia virus A25 and A26 proteins are fusion suppressors for mature virions and determine strain-specific virus entry pathways into HeLa, CHO-K1, and L cells.

    PubMed

    Chang, Shu-Jung; Chang, Yu-Xun; Izmailyan, Roza; Tang, Yin-Liang; Chang, Wen

    2010-09-01

    Mature vaccinia virus enters cells through either fluid-phase endocytosis/macropinocytosis or plasma membrane fusion. This may explain the wide range of host cell susceptibilities to vaccinia virus entry; however, it is not known how vaccinia virus chooses between these two pathways and which viral envelope proteins determine such processes. By screening several recombinant viruses and different strains, we found that mature virions containing the vaccinia virus A25 and A26 proteins entered HeLa cells preferentially through a bafilomycin-sensitive entry pathway, whereas virions lacking these two proteins entered through a bafilomycin-resistant pathway. To investigate whether the A25 and A26 proteins contribute to entry pathway specificity, two mutant vaccinia viruses, WRDeltaA25L and WRDeltaA26L, were subsequently generated from the wild-type WR strain. In contrast to the WR strain, both the WRDeltaA25L and WRDeltaA26L viruses became resistant to bafilomycin, suggesting that the removal of the A25 and A26 proteins bypassed the low-pH endosomal requirement for mature virion entry. Indeed, WRDeltaA25L and WRDeltaA26L virus infections of HeLa, CHO-K1, and L cells immediately triggered cell-to-cell fusion at a neutral pH at 1 to 2 h postinfection (p.i.), providing direct evidence that viral fusion machinery is readily activated after the removal of the A25 and A26 proteins to allow virus entry through the plasma membrane. In summary, our data support a model that on vaccinia mature virions, the viral A25 and A26 proteins are low-pH-sensitive fusion suppressors whose inactivation during the endocytic route results in viral and cell membrane fusion. Our results also suggest that during virion morphogenesis, the incorporation of the A25 and A26 proteins into mature virions may help restrain viral fusion activity until the time of infections.

  15. Epizootiology of Tacaribe serocomplex viruses (Arenaviridae) associated with neotomine rodents (Cricetidae, Neotominae) in southern California.

    PubMed

    Milazzo, Mary Louise; Cajimat, Maria N B; Mauldin, Matthew R; Bennett, Stephen G; Hess, Barry D; Rood, Michael P; Conlan, Christopher A; Nguyen, Kiet; Wekesa, J Wakoli; Ramos, Ronald D; Bradley, Robert D; Fulhorst, Charles F

    2015-02-01

    The objective of this study was to advance our knowledge of the epizootiology of Bear Canyon virus and other Tacaribe serocomplex viruses (Arenaviridae) associated with wild rodents in California. Antibody (immunoglobulin G [IgG]) to a Tacaribe serocomplex virus was found in 145 (3.6%) of 3977 neotomine rodents (Cricetidae: Neotominae) captured in six counties in southern California. The majority (122 or 84.1%) of the 145 antibody-positive rodents were big-eared woodrats (Neotoma macrotis) or California mice (Peromyscus californicus). The 23 other antibody-positive rodents included a white-throated woodrat (N. albigula), desert woodrat (N. lepida), Bryant's woodrats (N. bryanti), brush mice (P. boylii), cactus mice (P. eremicus), and deer mice (P. maniculatus). Analyses of viral nucleocapsid protein gene sequence data indicated that Bear Canyon virus is associated with N. macrotis and/or P. californicus in Santa Barbara County, Los Angeles County, Orange County, and western Riverside County. Together, analyses of field data and antibody prevalence data indicated that N. macrotis is the principal host of Bear Canyon virus. Last, the analyses of viral nucleocapsid protein gene sequence data suggested that the Tacaribe serocomplex virus associated with N. albigula and N. lepida in eastern Riverside County represents a novel species (tentatively named "Palo Verde virus") in the genus Arenavirus. PMID:25700047

  16. Climatic niche conservatism and ecological opportunity in the explosive radiation of arvicoline rodents (Arvicolinae, Cricetidae).

    PubMed

    Lv, Xue; Xia, Lin; Ge, Deyan; Wu, Yongjie; Yang, Qisen

    2016-05-01

    Climatic niche conservatism shapes patterns of diversity in many taxonomic groups, while ecological opportunity (EO) can trigger rapid speciation that is less constrained by the amount of time a lineage has occupied a given habitat. These two processes are well studied, but limited research has considered their joint and relative roles in shaping diversity patterns. We characterized climatic and biogeographic variables for 102 species of arvicoline rodents (Arvicolinae, Cricetidae), testing the effects of climatic niche conservatism and EO on arvicoline diversification as lineages transitioned between biogeographic regions. We found that the amount of time a lineage has occupied a precipitation niche is positively correlated with diversity along a precipitation gradient, suggesting climatic niche conservatism. In contrast, shift in diversification rate explained diversity patterns along a temperature gradient. Our results suggest that an indirect relationship exists between temperature and diversification that is associated with EO as arvicoline rodents colonized warm Palearctic environments. Climatic niche conservatism alone did not fully explain diversity patterns under density-dependence, highlighting the additional importance of EO-related processes in promoting the explosive radiation in arvicoline rodents and shaping diversity pattern among biogeographic regions and along climatic gradients.

  17. Epizootiology of Tacaribe Serocomplex Viruses (Arenaviridae) Associated with Neotomine Rodents (Cricetidae, Neotominae) in Southern California

    PubMed Central

    Milazzo, Mary Louise; Cajimat, Maria N. B.; Mauldin, Matthew R.; Bennett, Stephen G.; Hess, Barry D.; Rood, Michael P.; Conlan, Christopher A.; Nguyen, Kiet; Wekesa, J. Wakoli; Ramos, Ronald D.; Bradley, Robert D.

    2015-01-01

    Abstract The objective of this study was to advance our knowledge of the epizootiology of Bear Canyon virus and other Tacaribe serocomplex viruses (Arenaviridae) associated with wild rodents in California. Antibody (immunoglobulin G [IgG]) to a Tacaribe serocomplex virus was found in 145 (3.6%) of 3977 neotomine rodents (Cricetidae: Neotominae) captured in six counties in southern California. The majority (122 or 84.1%) of the 145 antibody-positive rodents were big-eared woodrats (Neotoma macrotis) or California mice (Peromyscus californicus). The 23 other antibody-positive rodents included a white-throated woodrat (N. albigula), desert woodrat (N. lepida), Bryant's woodrats (N. bryanti), brush mice (P. boylii), cactus mice (P. eremicus), and deer mice (P. maniculatus). Analyses of viral nucleocapsid protein gene sequence data indicated that Bear Canyon virus is associated with N. macrotis and/or P. californicus in Santa Barbara County, Los Angeles County, Orange County, and western Riverside County. Together, analyses of field data and antibody prevalence data indicated that N. macrotis is the principal host of Bear Canyon virus. Last, the analyses of viral nucleocapsid protein gene sequence data suggested that the Tacaribe serocomplex virus associated with N. albigula and N. lepida in eastern Riverside County represents a novel species (tentatively named “Palo Verde virus”) in the genus Arenavirus. PMID:25700047

  18. Climatic niche conservatism and ecological opportunity in the explosive radiation of arvicoline rodents (Arvicolinae, Cricetidae).

    PubMed

    Lv, Xue; Xia, Lin; Ge, Deyan; Wu, Yongjie; Yang, Qisen

    2016-05-01

    Climatic niche conservatism shapes patterns of diversity in many taxonomic groups, while ecological opportunity (EO) can trigger rapid speciation that is less constrained by the amount of time a lineage has occupied a given habitat. These two processes are well studied, but limited research has considered their joint and relative roles in shaping diversity patterns. We characterized climatic and biogeographic variables for 102 species of arvicoline rodents (Arvicolinae, Cricetidae), testing the effects of climatic niche conservatism and EO on arvicoline diversification as lineages transitioned between biogeographic regions. We found that the amount of time a lineage has occupied a precipitation niche is positively correlated with diversity along a precipitation gradient, suggesting climatic niche conservatism. In contrast, shift in diversification rate explained diversity patterns along a temperature gradient. Our results suggest that an indirect relationship exists between temperature and diversification that is associated with EO as arvicoline rodents colonized warm Palearctic environments. Climatic niche conservatism alone did not fully explain diversity patterns under density-dependence, highlighting the additional importance of EO-related processes in promoting the explosive radiation in arvicoline rodents and shaping diversity pattern among biogeographic regions and along climatic gradients. PMID:27061935

  19. A single-point mutation (Ala280Val) in the third intracellular loop alters the signalling properties of the human histamine H3 receptor stably expressed in CHO-K1 cells

    PubMed Central

    Flores-Clemente, Cecilia; Osorio-Espinoza, Angélica; Escamilla-Sánchez, Juan; Leurs, Rob; Arias, Juan-Manuel; Arias-Montaño, José-Antonio

    2013-01-01

    Background and Purpose An alanine to valine exchange at amino acid position 280 (A280V) in the third intracellular loop of the human histamine H3 receptor was first identified in a patient suffering from Shy–Drager syndrome and later reported as a risk factor for migraine. Here, we have compared the pharmacological and signalling properties of wild-type (hH3RWT) and A280V mutant (hH3RA280V) receptors stably expressed in CHO-K1 cells. Experimental Approach The hH3RA280V cDNA was created by overlapping extension PCR amplification. Receptor expression and affinity were assessed by radioligand (N-α-[methyl-3H]-histamine) binding to cell membranes, and receptor function by the inhibition of forskolin-induced cAMP accumulation and stimulation of ERK1/2 phosphorylation in intact cells, as well as stimulation of [35S]-GTPγS binding to cell membranes. Key Results Both receptors were expressed at similar levels with no significant differences in their affinities for H3 receptor ligands. Upon activation the hH3RWT was significantly more efficacious to inhibit forskolin-induced cAMP accumulation and to stimulate [35S]-GTPγS binding, with no difference in pEC50 estimates. The hH3RWT was also more efficacious to stimulate ERK1/2 phosphorylation, but this difference was not significant. The inverse agonist ciproxifan was more efficacious at hH3RWT to reduce [35S]-GTPγS binding but, for both receptors, failed to enhance forskolin-induced cAMP accumulation. Conclusions and Implications The A280V mutation reduces the signalling efficacy of the human H3 receptor. This effect may be relevant to the pathophysiology of disorders associated with the mutation. Linked Articles This article is part of a themed issue on Histamine Pharmacology Update. To view the other articles in this issue visit http://dx.doi.org/10.1111/bph.2013.170.issue-1 PMID:23713487

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

    SciTech Connect

    Bian, Chuan-Xiu; Shi, Zhumei; Meng, Qiao; Jiang, Yue; Liu, Ling-Zhi; Jiang, Bing-Hua

    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.

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

    PubMed

    Bian, Chuan-Xiu; Shi, Zhumei; Meng, Qiao; Jiang, Yue; Liu, Ling-Zhi; Jiang, Bing-Hua

    2010-07-30

    The 70kDa 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-1alpha binding site at its enhancer region. The expression of p70S6K1 siRNA specifically inhibited HIF-1alpha, but not HIF-1beta 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-1alpha expression in tumor tissues. Our results suggest that p70S6K1 is required for tumor growth and angiogenesis through HIF-1alpha and VEGF expression, providing a molecular mechanism of human ovarian cancer mediated by p70S6K1 signaling.

  2. K1 and K3 capsular antigens of Klebsiella induce tumor necrosis factor activities.

    PubMed

    Choy, Y M; Tsang, S F; Kong, S K; Leung, K N; Parolis, H; Lee, C Y; Fung, K P

    1996-01-01

    Capsular polysaccharide antigens isolated from Klebsiella pneumoniae sero-type 1 (K1) and sero-type 3 (K3) could induce tumor necrosis factor-alpha in ICR mice. K1 and K3 capsular antigens were found to be non-toxic by brine shrimp bioassay. When injected into Ehrlich ascites tumor-bearing mice, both K1 and K3 capsular antigens exhibited significant suppression in the growth of tumor cells. The significance of these observations is discussed.

  3. The Severe Adverse Reaction to Vitamin K1 Injection Is Anaphylactoid Reaction but Not Anaphylaxis

    PubMed Central

    Mi, Yan-Ni; Ping, Na-Na; Xiao, Xue; Zhu, Yan-Bing; Liu, Jing; Cao, Yong-Xiao

    2014-01-01

    The severe adverse reaction to vitamin K1 injection is always remarkable and is thought to result from anaphylaxis. Paradoxically, however, some patients administered vitamin K1 injection for the first time have adverse reactions. Using beagle dogs, the present study tested the hypothesis that the response to vitamin K1 is an anaphylactoid reaction. The results showed that serious anaphylaxis-like symptoms appeared in beagle dogs after the administration of vitamin K1 injection for the first time. The plasma histamine concentration increased, and blood pressure decreased sharply. After sensitization, dogs were challenged with vitamin K1 injection and displayed the same degree of symptoms as prior to sensitization. However, when the vitamin K1 injection-sensitized dogs were challenged with a vitamin K1-fat emulsion without solubilizers such asTween-80, the abnormal reactions did not occur. Furthermore, there was no significant change in the plasma immunoglobulin E concentration after vitamin K1 challenge. Following treatment with vitamin K1 injection, the release of histamine and β-hexosaminidase by rat basophilic leukemia-2H3 cells as well as the rate of apoptosis increased. The Tween-80 group displayed results similar to those observed following vitamin K1 injection in vivo. However, the dogs in the vitamin K1-fat emulsion group did not display any abnormal behavior or significant change in plasma histamine. Additionally, degranulation and apoptosis did not occur in rat basophilic leukemia-2H3 cells. Our results indicate that the adverse reaction induced by vitamin K1 injection is an anaphylactoid reaction, not anaphylaxis. Vitamin K1 injection induces the release of inflammatory factors via a non-IgE-mediated immune pathway, for which the trigger may be the solubilizer. PMID:24594861

  4. Amphimerus bragai n. sp. (Digenea: Opisthorchiidae), a parasite of the rodent Nectomys squamipes (Cricetidae) from Minas Gerais, Brazil.

    PubMed

    de Moraes Neto, A H; Thatcher, V E; Lanfredi, R M

    1998-01-01

    Amphimerus bragai n.sp. (Digenea, Opisthorchiidae) from the bile ducts of a rodent from the State of Minas Gerais, Brazil, Nectomys squamipes (Cricetidae), is described. The new species as studied by both light and scanning electron microscopy. A table is presented comparing the measurements of the new species with those of A. lancea (Diesing, 1850) and A. vallecaucensis Thatcher; 1970, parasites of dolphins and marsupials, respectively. The new species is similar in size and body form to A. vallecaucensis from which it differs in having a vitellarium that extends to the acetabulum while that of the former species are limited to the posterior one-third of the body. Additionally, the new species is from a rodent.

  5. An introduction to the systematics of Akodon orophilus Osgood, 1913 (Rodentia: Cricetidae) with the description of a new species.

    PubMed

    Jiménez, Carlos F; Pacheco, Víctor; Vivas, Dan

    2013-01-01

    The genus Akodon is one of the most species-rich rodent lineages in South America. In Peru, this genus contains 14 species subdivided in two groups: aerosus and boliviensis. Akodon orophilus Osgood, 1913 (Rodentia: Cricetidae) is a member of the Akodon aerosus group that inhabits the northern Peruvian montane forest, but is poorly characterized and its distribution is vaguely known. We review the status of the species based on morphology, morphometric and karyology, and compared with all members of the Akodon aerosus clade from Peruvian montane forests. As a result, we provide a complete redescription of A. orophilus, present new information on its natural history, restrict its distribution to the east of Río Marai6n, and describe a new species of Akodon from Huánuco Department, central Peru, a population previously assigned to A. orophilus. PMID:26312340

  6. S6K1 determines the metabolic requirements for BCR-ABL survival.

    PubMed

    Barger, J F; Gallo, C A; Tandon, P; Liu, H; Sullivan, A; Grimes, H L; Plas, D R

    2013-01-24

    In chronic myelogenous leukemia, the constitutive activation of the BCR-ABL kinase transforms cells to an addicted state that requires glucose metabolism for survival. We investigated S6K1, a protein kinase that drives glycolysis in leukemia cells, as a target for counteracting glucose-dependent survival induced by BCR-ABL. BCR-ABL potently activated S6K1-dependent signaling and glycolysis. Although S6K1 knockdown or rapamycin treatment suppressed glycolysis in BCR-ABL-transformed cells, these treatments did not induce cell death. Instead, loss of S6K1 triggered compensatory activation of fatty-acid oxidation, a metabolic program that can support glucose-independent cell survival. Fatty-acid oxidation in response to S6K1 inactivation required the expression of the fatty-acid transporter carnitine palmitoyl transferase 1c, which was recently linked to rapamycin resistance in cancer. Finally, addition of an inhibitor of fatty-acid oxidation significantly enhanced cytotoxicity in response to S6K1 inactivation. These data indicate that S6K1 dictates the metabolic requirements mediating BCR-ABL survival and provide a rationale for combining targeted inhibitors of signal transduction, with strategies to interrupt oncogene-induced metabolism. PMID:22391570

  7. Deciphering Gene Expression Program of MAP3K1 in Mouse Eyelid Morphogenesis

    PubMed Central

    Jin, Chang; Chen, Jing; Meng, Qinghang; Carreira, Vinicius; Tam, Neville N. C.; Geh, Esmond; Karyala, Saikumar; Ho, Shuk-Mei; Zhou, Xiangtian; Medvedovic, Mario; Xia, Ying

    2012-01-01

    Embryonic eyelid closure involves forward movement and ultimate fusion of the upper and lower eyelids, an essential step of mammalian ocular surface development. Although its underlying mechanism of action is not fully understood, a functional mitogen-activated protein kinase kinase kinase 1 (MAP3K1) is required for eyelid closure. Here we investigate the molecular signatures of MAP3K1 in eyelid morphogenesis. At mouse gestational day E15.5, the developmental stage immediately prior to eyelid closure, MAP3K1 expression is predominant in the eyelid leading edge (LE) and the inner eyelid (IE) epithelium. We used Laser Capture Microdissection (LCM) to obtain highly enriched LE and IE cells from wild type and MAP3K1-deficient fetuses and analyzed genome-wide expression profiles. The gene expression data led to the identification of three distinct developmental features of MAP3K1. First, MAP3K1 modulated Wnt and Sonic hedgehog signals, actin reorganization, and proliferation only in LE but not in IE epithelium, illustrating the temporal-spatial specificity of MAP3K1 in embryogenesis. Second, MAP3K1 potentiated AP-2α expression and SRF and AP-1 activity, but its target genes were enriched for binding motifs of AP-2α and SRF, and not AP-1, suggesting the existence of novel MAP3K1-AP-2α/SRF modules in gene regulation. Third, MAP3K1 displayed variable effects on expression of lineage specific genes in the LE and IE epithelium, revealing potential roles of MAP3K1 in differentiation and lineage specification. Using LCM and expression array, our studies have uncovered novel molecular signatures of MAP3K1 in embryonic eyelid closure. PMID:23201579

  8. S6K1 alternative splicing modulates its oncogenic activity and regulates mTORC1

    PubMed Central

    Ben-Hur, Vered; Denichenko, Polina; Siegfried, Zahava; Maimon, Avi; Krainer, Adrian; Davidson, Ben; Karni, Rotem

    2016-01-01

    Ribosomal S6 Kinase 1 (S6K1) is a major mTOR downstream signaling molecule which regulates cell size and translation efficiency. Here we report that short isoforms of S6K1 are over-produced in breast cancer cell lines and tumors. Overexpression of S6K1 short isoforms induces transformation of human breast epithelial cells. The long S6K1 variant (Iso-1) induced opposite effects: It inhibits Ras-induced transformation and tumor formation, while its knockdown or knockout induced transformation, suggesting that Iso-1 has a tumor suppressor activity. We further found that S6K1 short isoforms bind and activate mTORC1, elevating 4E-BP1 phosphorylation, cap-dependent translation and Mcl-1 protein levels. Both a phosphorylation-defective 4E-BP1 mutant and the mTORC1 inhibitor rapamycin partially blocked the oncogenic effects of S6K1 short isoforms, suggesting that these are mediated by mTORC1 and 4E-BP1. Thus, alternative splicing of S6K1 acts as a molecular switch in breast cancer cells elevating oncogenic isoforms that activate mTORC1. PMID:23273915

  9. 26 CFR 31.3306(k)-1 - Agricultural labor.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Agricultural labor. 31.3306(k)-1 Section 31.3306(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(k)-1 Agricultural...

  10. 26 CFR 31.3306(k)-1 - Agricultural labor.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 15 2011-04-01 2011-04-01 false Agricultural labor. 31.3306(k)-1 Section 31.3306(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(k)-1 Agricultural...

  11. 26 CFR 31.3306(k)-1 - Agricultural labor.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 15 2012-04-01 2012-04-01 false Agricultural labor. 31.3306(k)-1 Section 31.3306(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(k)-1 Agricultural...

  12. 26 CFR 31.3306(k)-1 - Agricultural labor.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 15 2013-04-01 2013-04-01 false Agricultural labor. 31.3306(k)-1 Section 31.3306(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(k)-1 Agricultural...

  13. 26 CFR 31.3306(k)-1 - Agricultural labor.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 15 2014-04-01 2014-04-01 false Agricultural labor. 31.3306(k)-1 Section 31.3306(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3306(k)-1 Agricultural...

  14. Influence of growth temperature of Escherichia coli on K1 capsular antigen production and resistance to opsonization.

    PubMed Central

    Bortolussi, R; Ferrieri, P; Quie, P G

    1983-01-01

    When Escherichia coli strains that produce K1 capsular polysaccharide antigen at 37 degrees C were grown at 22 degrees C, K1 antigen was not detected in the supernatant or washed-cell fraction of broth cultures. Significant amounts of K1 polysaccharide were detected only when the organism was grown at temperatures of 30 degrees C or higher. Rabbits immunized with an E. coli K1 strain (serotype O18ac:K1:H7) grown at 37 degrees C produced agglutinating antibody to somatic antigen and precipitating and agglutinating antibody to capsular K1 antigen; those immunized with this strain grown at 22 degrees C produced antibody to somatic antigen, but not to K1 antigen. Antibody to somatic antigen was markedly reduced by adsorption with the organism grown at 22 degrees C, while antibody to capsular antigen was not. E. coli K1 strains grown at 37 degrees C (K1 present) resisted phagocytosis and killing if they were opsonized solely by the alternative complement pathway (ACP) using magnesium ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid-chelated serum. When these strains were grown at 22 degrees C (K1 absent), they were opsonized efficiently by the ACP (28 versus 94% killing, respectively; P less than 0.001). In addition, a non-K1 mutant of an E. coli K1 strain was opsonized efficiently by the ACP although its encapsulated K1 parent was not. Sensitivity of E. coli strains to the bactericidal activity of serum was observed in strains with and without K1 capsular antigen. These studies demonstrated that production of K1 polysaccharide antigen was regulated by environmental temperature and that K1 capsule plays an essential role in rendering the organism resistant to opsonization by the ACP. PMID:6341228

  15. Comparison of protein patterns of xrs-5, a radiosensitive Chinese hamster ovary cell line, and CHO-K1, its radioresistant parent, using two-dimensional gel-electrophoresis

    SciTech Connect

    Kramer, J.M. . Dept. of Zoology)

    1991-01-01

    X-ray sensitive strains of Chinese hamster ovary cell lines have been used to analyze radiation repair mechanisms. One cell line, xrs-5, has been shown to be very sensitive to ionizing radiation and radical forming chemical mutagens. This sensitivity is thought to be a result a mutation in the DNA double strand break (DSB) repair mechanism, and its characterization has been a goal of several repair mechanism studies. Using two-dimensional gel electrophoresis, we have detected a protein (MW approximately 55KD) in the DNA/Nuclear Matrix (nucleoid) cell fraction of CHO-Kl cells that is absent in the nucleoid fraction of xrs-5. This protein is present, however, in both CHO-Kl and xrs-5 whole cell protein maps. To determine whether the 55KD protein is responsible for the radiosensitive and defective DSB repair phenotype of xrs-5 cells, studies are now underway to analyze revertants of xrs-5 that are proficient in DSB repair. Furthermore, an effort to sequence the protein in question is planned. 23 refs., 2 figs.

  16. Taxonomic position of Eothenomys wardi (Arvicolinae: Cricetidae) based on morphological and molecular analyses with a detailed description of the species.

    PubMed

    Zeng, Tao; Jin, Wei; Sun, Zhi Yu; Liu, Yang; Murphy, Robert W; Fu, Jian Rong; Wang, Xin; Hou, Quan Fen; Tu, Fei Yun; Liao, Rui; Liu, Shao Ying; Yue, Bi Song

    2013-01-01

    Ward's Red-backed Vole (Eothenomys wardi) is a rodent from the family Cricetidae. This endemic species occurs only in extreme northwestern Yunnan province, China in the Mekong and Salween river divide. It occupies steep cliffs at 2,800 to 4,250 m above sea level on the remote Qinghai-Tibetan plateau. The validity of E. wardi is controversial and no specimens exist apart from the nominal series. In 2010, we collected 38 topotypes of E. wardi from Meri Snow Mountain. The results of our phylogenetic analyses based on nucleotide sequences of the mitochondrial genes cytochrome b (cytb) and cytochrome c oxidase subunit one (COI) suggest that E. wardi is the sister group of E. custos, against its previously presumed sister species or conspecific species E. chinensis. In addition, seven out of 34 morphological characters differentiate E. wardi from other members of the genus Eothenomys. Therefore, we consider E. wardi to be a valid species and we provide its detailed morphological description.

  17. Metaphase chromosome and nucleoid differences between CHO-K1 and its radiosensitive derivative xrs-5

    SciTech Connect

    Schwartz, J.L. |; Cowan, J.M.; Moan, E.; Sedita, B.A.; Stephens, J.; Vaughan, A.T.M.

    1992-05-01

    The Chinese hamster ovary (CHO) cell line xrs-5 is a radiation-sensitive mutant isolated from CHO-K1 cells. The radiation sensitivity is associated with a defect in DNA double-strand break rejoining. Chromatin structure also appears altered in xrs-5 cells compared to the parental CHO-K1 cells. Metaphase chromosomes from xrs-5 are more condensed in appearance than CHO-K1 chromosomes. The overcondensed look is not the result of colcemid sensitivity. Electron microscopy studies suggest that xrs-5 metaphase chromosomes have larger loops of chromatin extending out from the chromosome core. There are also differences between CHO-K1 and xrs-5 cells in the size and fluorescence pattern of ethidium bromide-stained nucleoid preparations. These results suggest that there is a fundamental difference between CHO-K1 and xrs-5 in either the organization of the supercoiled loops of DNA attached to the nuclear matrix or in the nature of the proteins that attach the DNA to the matrix. These alterations in chromosome structure may underlie, in part, the radiation sensitivity of xrs-5 cells.

  18. The Cricetidae (Rodentia, Mammalia) from the Ulantatal area (Inner Mongolia, China): New data concerning the evolution of Asian cricetids during the Oligocene

    NASA Astrophysics Data System (ADS)

    Gomes Rodrigues, Helder; Marivaux, Laurent; Vianey-Liaud, Monique

    2012-08-01

    The Oligocene fossil deposits of Ulantatal in Inner Mongolia show an amazing faunal richness, comparable to the highly diversified contemporaneous faunas from the Valley of Lakes in Central Mongolia. To date, only a few taxa have been described. The present study consists of the description of 13 species of cricetid rodents from seven localities ranging in age from the late Early Oligocene to the Late Oligocene epoch. Most of them are new and belong to Eucricetodontinae, the dominating cricetid group at Ulantatal. These taxa give new information regarding the evolution of the Cricetidae in Central and Eastern Asia during the Oligocene. Four new species, Eucricetodon jilantaiensis nov. sp., Eucricetodon bagus nov. sp., Bagacricetodon tongi nov. gen., nov. sp. and Plesiodipus wangae nov. sp., show noticeable evolutionary trends. These species display more derived dental characters than their European contemporaries, in which they are much more comparable to Miocene forms. This observation reinforces the assumed early diversification of cricetids in this part of Asia. A striking case of sympatric evolution is indicated by the similarity of size and dental morphology of two sibling species, Eucricetodon asiaticus and Eucricetodon jilantaiensis nov. sp. Other taxa such as Witenia yolua nov. sp., Pseudocricetops matthewi nov. gen., nov. sp. and a primitive Tachyoryctoidinae, are scarcely represented and present unusual morphologies. The Cricetidae from Ulantatal also provide evidence suggesting faunal exchanges between Asia and Europe through the Paratethyan pathway during the second half of the Paleogene.

  19. Modulation of nociceptive dural input to the trigeminocervical complex through GluK1 kainate receptors.

    PubMed

    Andreou, Anna P; Holland, Philip R; Lasalandra, Michele P; Goadsby, Peter J

    2015-03-01

    Migraine is a common and disabling neurologic disorder, with important psychiatric comorbidities. Its pathophysiology involves activation of neurons in the trigeminocervical complex (TCC). Kainate receptors carrying the glutamate receptor subunit 5 (GluK1) are present in key brain areas involved in migraine pathophysiology. To study the influence of kainate receptors on trigeminovascular neurotransmission, we determined the presence of GluK1 receptors within the trigeminal ganglion and TCC with immunohistochemistry. We performed in vivo electrophysiologic recordings from TCC neurons and investigated whether local or systemic application of GluK1 receptor antagonists modulated trigeminovascular transmission. Microiontophoretic application of a selective GluK1 receptor antagonist, but not of a nonspecific ionotropic glutamate receptor antagonist, markedly attenuated cell firing in a subpopulation of neurons activated in response to dural stimulation, consistent with selective inhibition of postsynaptic GluK1 receptor-evoked firing seen in all recorded neurons. In contrast, trigeminovascular activation was significantly facilitated in a different neuronal population. The clinically active kainate receptor antagonist LY466195 attenuated trigeminovascular activation in all neurons. In addition, LY466195 demonstrated an N-methyl-d-aspartate receptor-mediated effect. This study demonstrates a differential role of GluK1 receptors in the TCC, antagonism of which can inhibit trigeminovascular activation through postsynaptic mechanisms. Furthermore, the data suggest a novel, possibly presynaptic, modulatory role of trigeminocervical kainate receptors in vivo. Differential activation of kainate receptors suggests unique roles for this receptor in pro- and antinociceptive mechanisms in migraine pathophysiology. PMID:25679470

  20. Development of nanoemulsions for topical delivery of vitamin K1.

    PubMed

    Campani, Virginia; Biondi, Marco; Mayol, Laura; Cilurzo, Francesco; Pitaro, Michele; De Rosa, Giuseppe

    2016-09-10

    Vitamin K1 (VK1) is a natural and lipophilic compound currently used in dermatological formulations. In this work, nanoemulsions containing VK1 have been proposed to overcome some issues associated to semisolid VK1-incorporating formulations. The study has been focused on the design of a lipid-free aqueous formulation, easy to prepare and with low cost of production. Thus, a simply protocol, using a low-energy method, has been used to spontaneously form the nanoemulsions. The nanoemulsion composition has been optimized to improve its physical stability during storage in different conditions. Then, the possibility to administer VK1-containing nanoemulsions by nebulization without significant alteration of the formulation was tested. Moreover, the VK1 accumulation into the skin layers have been evaluated through permeation experiments on Franz cells, ATR-FITR analysis, confocal laser scanning microscopy (CLSM) observations, and high performance liquid chromatography (HPLC) analysis. The study demonstrated that NEs represent an interesting option for the commercial development of an aqueous spray formulation for the topical delivery of VK1.

  1. Skeletal myocyte hypertrophy requires mTOR kinase activity and S6K1

    SciTech Connect

    Park, In-Hyun . E-mail: ihpark@uiuc.edu; Erbay, Ebru; Nuzzi, Paul; Chen Jie

    2005-09-10

    The protein kinase mammalian target of rapamycin (mTOR) is a central regulator of cell proliferation and growth, with the ribosomal subunit S6 kinase 1 (S6K1) as one of the key downstream signaling effectors. A critical role of mTOR signaling in skeletal muscle differentiation has been identified recently, and an unusual regulatory mechanism independent of mTOR kinase activity and S6K1 is revealed. An mTOR pathway has also been reported to regulate skeletal muscle hypertrophy, but the regulatory mechanism is not completely understood. Here, we report the investigation of mTOR's function in insulin growth factor I (IGF-I)-induced C2C12 myotube hypertrophy. Added at a later stage when rapamycin no longer had any effect on normal myocyte differentiation, rapamycin completely blocked myocyte hypertrophy as measured by myotube diameter. Importantly, a concerted increase of average myonuclei per myotube was observed in IGF-I-stimulated myotubes, which was also inhibited by rapamycin added at a time when it no longer affected normal differentiation. The mTOR protein level, its catalytic activity, its phosphorylation on Ser2448, and the activity of S6K1 were all found increased in IGF-I-stimulated myotubes compared to unstimulated myotubes. Using C2C12 cells stably expressing rapamycin-resistant forms of mTOR and S6K1, we provide genetic evidence for the requirement of mTOR and its downstream effector S6K1 in the regulation of myotube hypertrophy. Our results suggest distinct mTOR signaling mechanisms in different stages of skeletal muscle development: While mTOR regulates the initial myoblast differentiation in a kinase-independent and S6K1-independent manner, the hypertrophic function of mTOR requires its kinase activity and employs S6K1 as a downstream effector.

  2. Variability in pK'1 of human plasma.

    PubMed

    Tibi, L; Bhattacharya, S S; Flear, C T

    1982-05-01

    (1) Results were presented of an investigation of the relationship between ionic strength and PK'1 (negative logarithmic form of the apparent overall first dissociation constant of carbonic acid in plasma) in separated human plasma at constant PCO2. (2) Ionic strength was varied by adding dry NaCl to diluted aliquots of plasma from six healthy people and dry NaCl plus dry NaHCO3 to diluted aliquots of plasma from six other healthy people. pK'1 was determined from simultaneous measurements of pH and PCO2, and measurements of TCO2. Values for solubility factor for CO2 (s) were corrected for differences in plasma water and in Na concentration. All plasmas were equilibrated at 37 degrees C in a tonometer with a constant gas mixture (5% CO2, 12% O2, 83% N2). Precision of pK'1 determinations averaged 0.003. pK'1 was also determined on fresh undiluted healthy plasma, similarly tonometered. (3) We report (a) considerable variability in pK'1 of fresh undiluted healthy plasma (from 6.0197 to 6.1217); and also in extrapolated values for pK'1 at a notional zero ionic strength (6.179 to 6.325); (b) that variation in plasma ionic strength alters pK'1; (c) that change in plasma bicarbonate [HCO3]p can also change pK'1; (d) that change in pK'1, changes measured pH. (4) Implications are discussed. PMID:6805986

  3. CTX-M β-Lactamase Production and Virulence of Escherichia coli K1

    PubMed Central

    Dubois, Damien; Prasadarao, Nemani V.; Mittal, Rahul; Bret, Laurent; Roujou-Gris, Marie

    2009-01-01

    We report a patient with neonatal meningitis caused by a CTX-M-1–producing Escherichia coli K1 strain. The influence of CTX-M production on virulence was investigated in cell culture and a newborn mouse model of meningitis. CTX-M production had no influence on virulence but was a major factor in clinical outcome. PMID:19961682

  4. 0.33-k1 ArF lithography for 100-nm DRAM

    NASA Astrophysics Data System (ADS)

    Bok, Cheol-Kyu; Kim, Seok-Kyun; Kim, Hee-Bom; Oh, Jin-Sung; Ahn, Chang-Nam; Shin, Ki-Soo

    2002-07-01

    We have evaluated 0.33k1 ArF lithography using 0.63NA scanner to develop 100 nm DRAM. ArF resist problems were resist pattern shrinkage during CD SEM measurement, resist pattern collapse during wet development and poor etch resistance. Off-Site Measurement (OSM) method has been developed for decreasing pattern shrinkage. With OSM method, 8nm of CD shrinkage was down to 2nm for 100nm L/S patterns. We have found a proper BARC material that prevents resist patterns falling down. Lack of etch resistance was compensated by hard mask. With W/SiN hard mask, acrylate- type resist patterns were transferred well into W/poly-Si gate patterns. We have simulated process window of critical DRAM cell patterns (isolation, gate, bit line contact, storage node) in the simple off-axis illumination (OAI) and optical proximity correction (OPC) conditions based on single exposure. Simulation results were verified by lithography tests and it turned out that 0.33k1 process was possible with exposure latitude of above 10% and focus latitude of more than 0.4 micrometers . 0.33k1 ArF lithography was successfully implemented into 100 nm DRAM with CD uniformity of 10nm (3 (sigma) ) and overlay accuracy of 30 nm (mean +3 (sigma) ). We have also evaluated double exposure technique using dipole illumination targeting 90 nm in order to see the possibility of 0.29k1 process. 0.29k1 process was also likely to be possible, although some specific improvements were recommended for the wider process window. From the simulation and resist patterning results, we believe that 0.85 NA lens will be able to extend ArF lithography into 75 nm by single exposure technology using crosspole illumination (0.33k1 process) and 65 nm by double exposure technology using dipole and crosspole illumination (0.29k1 process).

  5. Sam68 Regulates S6K1 Alternative Splicing during Adipogenesis

    PubMed Central

    Song, Jingwen

    2015-01-01

    The requirement for alternative splicing during adipogenesis is poorly understood. The Sam68 RNA binding protein is a known regulator of alternative splicing, and mice deficient for Sam68 exhibit adipogenesis defects due to defective mTOR signaling. Sam68 null preadipocytes were monitored for alternative splicing imbalances in components of the mTOR signaling pathway. Herein, we report that Sam68 regulates isoform expression of the ribosomal S6 kinase gene (Rps6kb1). Sam68-deficient adipocytes express Rps6kb1-002 and its encoded p31S6K1 protein, in contrast to wild-type adipocytes that do not express this isoform. Sam68 binds an RNA sequence encoded by Rps6kb1 intron 6 and prevents serine/arginine-rich splicing factor 1 (SRSF1)-mediated alternative splicing of Rps6kb1-002, as assessed by cross-linking and immunoprecipitation (CLIP) and minigene assays. Depletion of p31S6K1 with small interfering RNAs (siRNAs) partially restored adipogenesis of Sam68-deficient preadipocytes. The ectopic expression of p31S6K1 in wild-type 3T3-L1 cells resulted in adipogenesis differentiation defects, showing that p31S6K1 is an inhibitor of adipogenesis. Our findings indicate that Sam68 is required to prevent the expression of p31S6K1 in adipocytes for adipogenesis to occur. PMID:25776557

  6. MAP3K1 functionally interacts with Axin1 in the canonical Wnt signalling pathway.

    PubMed

    Sue Ng, Ser; Mahmoudi, Tokameh; Li, Vivian S W; Hatzis, Pantelis; Boersema, Paul J; Mohammed, Shabaz; Heck, Albert J; Clevers, Hans

    2010-01-01

    A central point of regulation in the Wnt/beta-catenin signalling pathway is the formation of the beta-catenin destruction complex. Axin1, an essential negative regulator of Wnt signalling, serves as a scaffold within this complex and is critical for rapid turnover of beta-catenin. To examine the mechanism by which Wnt signalling disables the destruction complex, we used an immunoprecipitation-coupled proteomics approach to identify novel endogenous binding partners of Axin1. We found mitogen-activated protein kinase kinase kinase 1 (MAP3K1) as an Axin1 interactor in Ls174T colorectal cancer (CRC) cells. Importantly, confirmation of this interaction in HEK293T cells indicated that the Axin1-MAP3K1 interaction is induced and modulated by Wnt stimulation. siRNA depletion of MAP3K1 specifically abrogated TCF/LEF-driven transcription and Wnt3A-driven endogenous gene expression in both HEK293T as well as DLD-1 CRC. Expression of ubiquitin ligase mutants of MAP3K1 abrogated TCF/LEF transcription, whereas kinase mutants had no effect in TCF-driven activity, highlighting the essential role of the MAP3K1 E3 ubiquitin ligase activity in regulation of the Wnt/beta-catenin pathway. These results suggest that MAP3K1, previously reported as an Axin1 inter-actor in c-Jun NH(2)-terminal kinase pathway, is also involved in the canonical Wnt signalling pathway and positively regulates expression of Wnt target genes. PMID:20128690

  7. Enantiomers of warfarin and vitamin K1 metabolism.

    PubMed Central

    Choonara, I A; Haynes, B P; Cholerton, S; Breckenridge, A M; Park, B K

    1986-01-01

    The effect of the individual enantiomers of warfarin at steady state (1 mg daily) was investigated in five healthy volunteers. Both enantiomers produced a significant increase in prothrombin time, but the increase with S warfarin (1.8 +/- 0.8 s, mean +/- s.d.) was greater than with R warfarin (1.0 +/- 0.3 s), despite lower steady state plasma concentrations of S warfarin, due to its more rapid clearance. Following the administration of vitamin K1, the maximum plasma concentration and area under the plasma concentration time curve values for the metabolite vitamin K1 2,3-epoxide were greater after S warfarin than after R warfarin. The greater anticoagulant potency of S warfarin is reflected by a greater degree of inhibition of vitamin K1 epoxide reductase. PMID:3567019

  8. A Supplementary Program for Environmental Education, Mathematics, Grade K-1.

    ERIC Educational Resources Information Center

    Warpinski, Robert

    Presented in these teacher's guides for grades K-1 are lesson plans and ideas for integrating mathematics and environmental education. Each lesson originates with a fundamental concept pertaining to the environment and states, in addition, its discipline area, subject area, and problem orientation. Following this, behavioral objectives and…

  9. Quantum Analysis of k=-1 Robertson-Walker Universe

    NASA Astrophysics Data System (ADS)

    Dariescu, Ciprian; Dariescu, Marina-Aura

    2015-11-01

    The (k=-1)-Robertson-Walker spacetime is under investigation. With the derived Hamilton operator, we are solving the Wheeler-De Witt Equation and its Schrödinger-like extension, for physically important forms of the effective potential. The closed form solutions, expressed in terms of Heun's functions, allow us to comment on the occurrence of Universe from highly probable quantum states.

  10. Space Access for Small Satellites on the K-1

    NASA Astrophysics Data System (ADS)

    Faktor, L.

    Affordable access to space remains a major obstacle to realizing the increasing potential of small satellites systems. On a per kilogram basis, small launch vehicles are simply too expensive for the budgets of many small satellite programs. Opportunities for rideshare with larger payloads on larger launch vehicles are still rare, given the complications associated with coordinating delivery schedules and deployment orbits. Existing contractual mechanisms are also often inadequate to facilitate the launch of multiple payload customers on the same flight. Kistler Aerospace Corporation is committed to lowering the price and enhancing the availability of space access for small satellite programs through the fully-reusable K-1 launch vehicle. Kistler has been working with a number of entities, including Astrium Ltd., AeroAstro, and NASA, to develop innovative approaches to small satellite missions. The K-1 has been selected by NASA as a Flight Demonstration Vehicle for the Space Launch Initiative. NASA has purchased the flight results during the first four K-1 launches on the performance of 13 advanced launch vehicle technologies embedded in the K-1 vehicle. On K-1 flights #2-#4, opportunities exist for small satellites to rideshare to low-earth orbit for a low-launch price. Kistler's flight demonstration contract with NASA also includes options to fly Add-on Technology Experiment flights. Opportunities exist for rideshare payloads on these flights as well. Both commercial and government customers may take advantage of the rideshare pricing. Kistler is investigating the feasibility of flying dedicated, multiple small payload missions. Such a mission would launch multiple small payloads from a single customer or small payloads from different customers. The orbit would be selected to be compatible with the requirements of as many small payload customers as possible, and make use of reusable hardware, standard interfaces (such as the existing MPAS) and verification plans

  11. A tale of tails: Sialidase is key to success in a model of phage therapy against K1-capsulated Escherichia coli

    SciTech Connect

    Bull, J.J.; Vimr, E.R.; Molineux, I.J.

    2010-03-01

    Prior studies treating mice infected with Escherichia coli O18:K1:H7 observed that phages requiring the K1 capsule for infection (K1-dep) were superior to capsule-independent (K1-ind) phages. We show that three K1-ind phages all have low fitness when grown on cells in serum whereas fitnesses of four K1-dep phages were high. The difference is serum-specific, as fitnesses in broth overlapped. Sialidase activity was associated with all K1-dep virions tested but no K1-ind virions, a phenotype supported by sequence analyses. Adding endosialidase to cells infected with K1-ind phage increased fitness in serum by enhancing productive infection after adsorption. We propose that virion sialidase activity is the primary determinant of high fitness on cells grown in serum, and thus in a mammalian host. Although the benefit of sialidase is specific to K1-capsulated bacteria, this study may provide a scientific rationale for selecting phages for therapeutic use in many systemic infections.

  12. Patterning strategy for low-K1 lithography

    NASA Astrophysics Data System (ADS)

    Hwang, David H.; Cheng, Wen-Hao

    2004-08-01

    Moore's law has been guiding the semiconductor industry for four decades. Lithography is the key enabler to keep the industry on the technology treadmill. Lithographers have been facing unprecedented challenges during last five years to keep the technology on the technology treadmill by developing various kinds of resolution enhancement techniques (RETs). In low K1 regime, co-optimization of design, layout mask, OPC, lithography and etching is the primary strategy to deliver a production-worthy patterning solution. Optical shrink is not a trivial task anymore. Intel always pursues parallel patterning techniques based on the dual exposure wavelength patterning strategy. While EUVL is the preferred patterning solution for 32nm node, 193nm immersion lithography with super high NA illumination is one of the parallel patterning strategies. The effects of polarization at super high NA illumination on mask technology, such as lens reduction ratio, blank absorber thickness and image imbalance correction, and restriction on design layout are addressed in this paper. Contact patterning is extremely challenging at low K1. Contact shape factor (circularity) which impacts the design rule will be discussed in this paper. Explosion of data file size and mask write time, stringent mask CD control and mask defect disposition are direct consequences of low-K1/high-MEEF (Mask Error Enhancement Factor) lithography. Mask makers alone cannot resolve the challenges in a cost effective manner. A seamless integration solution is a must.

  13. S6K1 Negatively Regulates TAK1 Activity in the Toll-Like Receptor Signaling Pathway

    PubMed Central

    Kim, So Yong; Baik, Kyung-Hwa; Baek, Kwan-Hyuck; Chah, Kyong-Hwa; Kim, Kyung Ah; Moon, Gyuyoung; Jung, Eunyu; Kim, Seong-Tae; Shim, Jae-Hyuck; Greenblatt, Matthew B.; Chun, Eunyoung

    2014-01-01

    Transforming growth factor β (TGF-β)-activated kinase 1 (TAK1) is a key regulator in the signals transduced by proinflammatory cytokines and Toll-like receptors (TLRs). The regulatory mechanism of TAK1 in response to various tissue types and stimuli remains incompletely understood. Here, we show that ribosomal S6 kinase 1 (S6K1) negatively regulates TLR-mediated signals by inhibiting TAK1 activity. S6K1 overexpression causes a marked reduction in NF-κB and AP-1 activity induced by stimulation of TLR2 or TLR4. In contrast, S6K1−/− and S6K1 knockdown cells display enhanced production of inflammatory cytokines. Moreover, S6K1−/− mice exhibit decreased survival in response to challenge with lipopolysaccharide (LPS). We found that S6K1 inhibits TAK1 kinase activity by interfering with the interaction between TAK1 and TAB1, which is a key regulator protein for TAK1 catalytic function. Upon stimulation with TLR ligands, S6K1 deficiency causes a marked increase in TAK1 kinase activity that in turn induces a substantial enhancement of NF-κB-dependent gene expression, indicating that S6K1 is negatively involved in the TLR signaling pathway by the inhibition of TAK1 activity. Our findings contribute to understanding the molecular pathogenesis of the impaired immune responses seen in type 2 diabetes, where S6K1 plays a key role both in driving insulin resistance and modulating TLR signaling. PMID:24277938

  14. Loop quantum cosmology of k=1 FRW models

    SciTech Connect

    Ashtekar, Abhay; Pawlowski, Tomasz; Singh, Parampreet; Vandersloot, Kevin

    2007-01-15

    The closed, k=1, FRW model coupled to a massless scalar field is investigated in the framework of loop quantum cosmology using analytical and numerical methods. As in the k=0 case, the scalar field can be again used as emergent time to construct the physical Hilbert space and introduce Dirac observables. The resulting framework is then used to address a major challenge of quantum cosmology: resolving the big-bang singularity while retaining agreement with general relativity at large scales. It is shown that the framework fulfills this task. In particular, for states which are semiclassical at some late time, the big bang is replaced by a quantum bounce and a recollapse occurs at the value of the scale factor predicted by classical general relativity. Thus, the 'difficulties' pointed out by Green and Unruh in the k=1 case do not arise in a more systematic treatment. As in k=0 models, quantum dynamics is deterministic across the deep Planck regime. However, because it also retains the classical recollapse, in contrast to the k=0 case one is now led to a cyclic model. Finally, we clarify some issues raised by Laguna's recent work addressed to computational physicists.

  15. A new species of Reithrodontomys, subgenus Aporodon (Cricetidae: Neotominae), from the highlands of Costa Rica, with comments on Costa Rican and Panamanian Reithrodontomys

    USGS Publications Warehouse

    Gardner, Alfred L.; Carleton, Michael D.

    2009-01-01

    A new species of the rodent genus Reithrodontomys (Cricetidae: Neotominae) is described from Cerro Asuncion in the western Cordillera de Talamanca, Costa Rica. The long tail, elongate rostrum, bulbous braincase, and complex molars of the new species associate it with members of the subgenus Aporodon, tenuirostris species group. In its diminutive size and aspects of cranial shape, the new species (Reithrodontomys musseri, sp. nov.) most closely resembles R. microdon, a form known from highlands in Guatemala and Chiapas, Mexico. In the course of differentially diagnosing the new species, we necessarily reviewed the Costa Rican and Panamanian subspecies of R. mexicanus based on morphological comparisons, study of paratypes and vouchers used in recent molecular studies, and morphometric analyses. We recognize Reithrodontomys cherrii (Allen, 1891) and R. garichensis finders and Pearson, 1940, as valid species, and allocate R. mexicanus potrerograndei Goodwin, 1945, as a subjective synonym of R. brevirostris Goodwin, 1943. Critical review of museum specimens collected subsequent to Hooper's (1952) revision is needed and would do much to improve understanding of Reithrodontomys taxonomy and distribution in Middle America.

  16. Rediscovery and New Morphological Data on Two Hassalstrongylus (Nematoda: Heligmonellidae) Coparasitic in the Marsh Rat Holochilus chacarius (Rodentia: Cricetidae) from Argentina.

    PubMed

    Digiani, María Celina; Notarnicola, Juliana; Navone, Graciela T

    2015-10-01

    Two species of Hassalstrongylus Durette-Desset, 1971, coparasitic in Holochilus chacarius Thomas (Rodentia, Cricetidae) and not recorded since their original description in 1937, were newly found in their type host and locality. Hassalstrongylus mazzai (Freitas, Lent and Almeida, 1937) and Hassalstrongylus argentinus (Freitas, Lent and Almeida, 1937) were obtained from Ho. chacarius from 2 different populations: one from Salta Province (northwest Argentina) and another from Chaco Province (northeast Argentina). The species described as Heligmonoides mazzai Freitas, Lent and Almeida, 1937 had been transferred to Hassalstrongylus even though its synlophe had never been studied. We provide the first descriptions and illustrations of the synlophe of males and females of Hassalstrongylus mazzai and the female of H. argentinus and account for morphological and metrical variability. We confirm, through the study of the synlophe, the placement of Hassalstrongylus mazzai in the genus Hassalstrongylus and designate neotypes for the species because the type material deposited by the authors could not be found. Females of both species were morphologically very similar, and a principal components analysis (PCA) performed on some morphometrical characters showed that the body length, uterus length, and an unexpected character as the number of eggs were useful characters in the discrimination of both species.

  17. Rediscovery and New Morphological Data on Two Hassalstrongylus (Nematoda: Heligmonellidae) Coparasitic in the Marsh Rat Holochilus chacarius (Rodentia: Cricetidae) from Argentina.

    PubMed

    Digiani, María Celina; Notarnicola, Juliana; Navone, Graciela T

    2015-10-01

    Two species of Hassalstrongylus Durette-Desset, 1971, coparasitic in Holochilus chacarius Thomas (Rodentia, Cricetidae) and not recorded since their original description in 1937, were newly found in their type host and locality. Hassalstrongylus mazzai (Freitas, Lent and Almeida, 1937) and Hassalstrongylus argentinus (Freitas, Lent and Almeida, 1937) were obtained from Ho. chacarius from 2 different populations: one from Salta Province (northwest Argentina) and another from Chaco Province (northeast Argentina). The species described as Heligmonoides mazzai Freitas, Lent and Almeida, 1937 had been transferred to Hassalstrongylus even though its synlophe had never been studied. We provide the first descriptions and illustrations of the synlophe of males and females of Hassalstrongylus mazzai and the female of H. argentinus and account for morphological and metrical variability. We confirm, through the study of the synlophe, the placement of Hassalstrongylus mazzai in the genus Hassalstrongylus and designate neotypes for the species because the type material deposited by the authors could not be found. Females of both species were morphologically very similar, and a principal components analysis (PCA) performed on some morphometrical characters showed that the body length, uterus length, and an unexpected character as the number of eggs were useful characters in the discrimination of both species. PMID:26193068

  18. Genetic analysis of chromosomal mutations in the polysialic acid gene cluster of Escherichia coli K1.

    PubMed Central

    Vimr, E R; Aaronson, W; Silver, R P

    1989-01-01

    The kps gene cluster of Escherichia coli K1 encodes functions for sialic acid synthesis, activation, polymerization, and possibly translocation of polymer to the cell surface. The size and complexity of this membrane polysaccharide biosynthetic cluster have hindered genetic mapping and functional descriptions of the kps genes. To begin a detailed investigation of the polysialic acid synthetic mechanism, acapsular mutants were characterized to determine their probable defects in polymer synthesis. The mutants were tested for complementation with kps fragments subcloned from two separately isolated, functionally intact kps gene clusters. Complementation was assayed by immunological and biochemical methods and by sensitivity to the K1-specific bacteriophage K1F. The kps cluster consisted of a central 5.8-kilobase region that contained at least two genes coding for sialic acid synthetic enzymes, a gene encoding the sialic acid-activating enzyme, and a gene encoding the sialic acid polymerase. This biosynthetic region is flanked on one side by an approximately 2.8-kilobase region that contains a potential regulatory locus and at least one structural gene for a polypeptide that appears to function in polysialic acid assembly. Flanking the biosynthetic region on the opposite side is a 6- to 8.4-kilobase region that codes for at least three proteins which may also function in polymer assembly and possibly in translocating polymer to the outer cell surface. Results of transduction crosses supported these conclusions and indicated that some of the kps genes flanking the central biosynthetic region may not function directly in transporting polymer to the cell surface. The results also demonstrate that the map position and probable function of most of the kps cluster genes have been identified. Images PMID:2644224

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

    SciTech Connect

    Aubert, Bernard; Bona, M.; Karyotakis, Y.; Lees, J.P.; Poireau, V.; Prencipe, E.; Prudent, X.; Tisserand, V.; Garra Tico, J.; Grauges, E.; Lopez, L.; Palano, Antimo; Pappagallo, M.; Eigen, G.; Stugu, Bjarne; Sun, L.; 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.

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

    PubMed

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

    2015-01-01

    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. PMID:25821933

  1. Penguin-dominated B →ϕ K1(1270) and ϕ K1(1400 ) decays in the perturbative QCD approach

    NASA Astrophysics Data System (ADS)

    Liu, Xin; Zou, Zhi-Tian; Xiao, Zhen-Jun

    2014-11-01

    We investigate the C P -averaged branching ratios, the polarization fractions, the relative phases, and the C P -violating asymmetries of the penguin-dominated B →ϕ K1(1270 ) and ϕ K1(1400 ) decays in the perturbative QCD (pQCD) approach, where K1(1270 ) and K1(1400 ) are believed to be the mixtures of two distinct types of axial-vector K1 A(P1 3 ) and K1 B(P1 1 ) states with different behavior, however, their mixing angle θK1is still a hot and controversial topic presently. By numerical evaluations with two different mixing angles θK 1˜3 3∘ and 58° and phenomenological analysis, we find that: (i) the pQCD predictions for the branching ratio, the longitudinal polarization fraction and the direct C P violation of B±→ϕ K1(1270 )±decay with the smaller angle 33° are in good agreement with the currently available data; (ii) though the central values significantly exceed the available upper limit, both pQCD predictions of Br (B±→ϕ K1(1400 )±) with two different mixing angles are consistent with that obtained in QCD factorization and with the preliminary data in 2 σ errors. These results and other relevant predictions for the considered decays will be further tested by the LHCb and the forthcoming Super-B experiments; (iii) the weak annihilation contributions can play an important role in B →ϕ K1(1270 ) and ϕ K1(1400 ) decays; (d) these pQCD predictions combined with the future precision measurements can examine the reliability of the factorization approach employed here, but also explore the complicated QCD dynamics and mixing angle θK 1 of the axial-vector K1(1270 ) and K1(1400 ) system.

  2. FimH adhesin of Escherichia coli K1 type 1 fimbriae activates BV-2 microglia

    SciTech Connect

    Lee, Jongseok; Shin, Sooan; Teng, C.-H.; Hong, Suk Jin; Kim, Kwang Sik . E-mail: kwangkim@jhmi.edu

    2005-09-02

    The generation of intense inflammation in the subarachnoid space in response to meningitis-causing bacteria contributes to brain dysfunction and neuronal injury in bacterial meningitis. Microglia, the major immune effector cells in the central nervous system (CNS), become activated by bacterial components to produce proinflammatory immune mediators. In this study, we showed that FimH adhesin, a tip component of type 1 fimbriae of meningitis-causing Escherichia coli K1, activated the murine microglial cell line, BV-2, which resulted in the production of nitric oxide and the release of tumor necrosis factor-{alpha}. Mitogen-activated protein kinases, ERK and p-38, and nuclear factor-{kappa}B were involved in FimH adhesin-mediated microglial activation. These findings suggest that FimH adhesin contributes to the CNS inflammatory response by virtue of activating microglia in E. coli meningitis.

  3. [Antimicrobial sensitivity of Escherichia coli strains with K1 antigen isolated from pregnant women and newborns].

    PubMed

    Kaczmarek, Agnieszka; Budzyńska, Anna; Gospodarek, Eugenia

    2011-01-01

    The aim of this study was comparison of the susceptibility to antibiotics of E. coli strains with K1 antigen (E. coli K1+) and non-K1 E. coli strains (E. coli K1-). This study included 67 of E. coli K1+ and 67 of E. coli K1- strains isolated in the time period from June to September of 2008 from pregnant women and newborns hospitalized at dr. J. Biziel University Hospital number 2 L. Rydygier Collegium Medicum in Bydgoszcz Nicolaus Copernicus University in Toruń. Antimicrobial susceptibility of E. coli strains was tested by the disc-diffusion method, on the Mueller Hinton 2 Agar (Becton Dickinson). It was found that 64,2% of E. coli K1+ strains and 53,7% of E. coli K1-strains were susceptible to all tested antibiotics and chemioterapeutics. E. coli K1- strains were more often than E. coli K1+ nonsusceptible to at least one antimicrobial agent. The obtained results indicate that E. coli K1+ strains significant differed in the susceptibility to ampicillin/sulbactam (85,1% versus 95,5%) (p=0,041), cephalothin (70,1% versus 85,1%) (p=0,038) and tetracycline (91,0% versus 74,6%) (p=0,012) from E. coli K1-strains. All tested E. coli K1+ and K1-strains were sensitive to piperacillin/tazobactam, cefoperazone/sulbactam, cefotaxime, ceftazidime, cefepime, imipenem, amikacin, netilmicin and tigecycline. There weren't the ESBL-producing strains among tested E. coli K1+ and K1- rods.

  4. The role of PI3K/AKT-related PIP5K1α and the discovery of its selective inhibitor for treatment of advanced prostate cancer

    PubMed Central

    Semenas, Julius; Hedblom, Andreas; Miftakhova, Regina R.; Sarwar, Martuza; Larsson, Rikard; Shcherbina, Liliya; Johansson, Martin E.; Härkönen, Pirkko; Sterner, Olov; Persson, Jenny L.

    2014-01-01

    Nitrogen-containing heterocyclic compounds are an important class of molecules that are commonly used for the synthesis of candidate drugs. Phosphatidylinositol-4-phosphate 5-kinase-α (PIP5Kα) is a lipid kinase, similar to PI3K. However, the role of PIP5K1α in oncogenic processes and the development of inhibitors that selectively target PIP5K1α have not been reported. In the present study we report that overexpression of PIP5K1α is associated with poor prognosis in prostate cancer and correlates with an elevated level of the androgen receptor. Overexpression of PIP5K1α in PNT1A nonmalignant cells results in an increased AKT activity and an increased survival, as well as invasive malignant phenotype, whereas siRNA-mediated knockdown of PIP5K1α in aggressive PC-3 cells leads to a reduced AKT activity and an inhibition in tumor growth in xenograft mice. We further report a previously unidentified role for PIP5K1α as a druggable target for our newly developed compound ISA-2011B using a high-throughput KINOMEscan platform. ISA-2011B was discovered during our synthetic studies of C-1 indol-3-yl substituted 1,2,3,4-tetrahydroisoquinolines via a Pictet-Spengler approach. ISA-2011B significantly inhibits growth of tumor cells in xenograft mice, and we show that this is mediated by targeting PIP5K1α-associated PI3K/AKT and the downstream survival, proliferation, and invasion pathways. Further, siRNA-mediated knockdown of PIP5K1α exerts similar effects on PC3 cells as ISA-2011B treatment, significantly inhibiting AKT activity, increasing apoptosis and reducing invasion. Thus, PIP5K1α has high potential as a drug target, and compound ISA-2011B is interesting for further development of targeted cancer therapy. PMID:25071204

  5. The role of PI3K/AKT-related PIP5K1α and the discovery of its selective inhibitor for treatment of advanced prostate cancer.

    PubMed

    Semenas, Julius; Hedblom, Andreas; Miftakhova, Regina R; Sarwar, Martuza; Larsson, Rikard; Shcherbina, Liliya; Johansson, Martin E; Härkönen, Pirkko; Sterner, Olov; Persson, Jenny L

    2014-09-01

    Nitrogen-containing heterocyclic compounds are an important class of molecules that are commonly used for the synthesis of candidate drugs. Phosphatidylinositol-4-phosphate 5-kinase-α (PIP5Kα) is a lipid kinase, similar to PI3K. However, the role of PIP5K1α in oncogenic processes and the development of inhibitors that selectively target PIP5K1α have not been reported. In the present study we report that overexpression of PIP5K1α is associated with poor prognosis in prostate cancer and correlates with an elevated level of the androgen receptor. Overexpression of PIP5K1α in PNT1A nonmalignant cells results in an increased AKT activity and an increased survival, as well as invasive malignant phenotype, whereas siRNA-mediated knockdown of PIP5K1α in aggressive PC-3 cells leads to a reduced AKT activity and an inhibition in tumor growth in xenograft mice. We further report a previously unidentified role for PIP5K1α as a druggable target for our newly developed compound ISA-2011B using a high-throughput KINOMEscan platform. ISA-2011B was discovered during our synthetic studies of C-1 indol-3-yl substituted 1,2,3,4-tetrahydroisoquinolines via a Pictet-Spengler approach. ISA-2011B significantly inhibits growth of tumor cells in xenograft mice, and we show that this is mediated by targeting PIP5K1α-associated PI3K/AKT and the downstream survival, proliferation, and invasion pathways. Further, siRNA-mediated knockdown of PIP5K1α exerts similar effects on PC3 cells as ISA-2011B treatment, significantly inhibiting AKT activity, increasing apoptosis and reducing invasion. Thus, PIP5K1α has high potential as a drug target, and compound ISA-2011B is interesting for further development of targeted cancer therapy.

  6. The role of PI3K/AKT-related PIP5K1α and the discovery of its selective inhibitor for treatment of advanced prostate cancer.

    PubMed

    Semenas, Julius; Hedblom, Andreas; Miftakhova, Regina R; Sarwar, Martuza; Larsson, Rikard; Shcherbina, Liliya; Johansson, Martin E; Härkönen, Pirkko; Sterner, Olov; Persson, Jenny L

    2014-09-01

    Nitrogen-containing heterocyclic compounds are an important class of molecules that are commonly used for the synthesis of candidate drugs. Phosphatidylinositol-4-phosphate 5-kinase-α (PIP5Kα) is a lipid kinase, similar to PI3K. However, the role of PIP5K1α in oncogenic processes and the development of inhibitors that selectively target PIP5K1α have not been reported. In the present study we report that overexpression of PIP5K1α is associated with poor prognosis in prostate cancer and correlates with an elevated level of the androgen receptor. Overexpression of PIP5K1α in PNT1A nonmalignant cells results in an increased AKT activity and an increased survival, as well as invasive malignant phenotype, whereas siRNA-mediated knockdown of PIP5K1α in aggressive PC-3 cells leads to a reduced AKT activity and an inhibition in tumor growth in xenograft mice. We further report a previously unidentified role for PIP5K1α as a druggable target for our newly developed compound ISA-2011B using a high-throughput KINOMEscan platform. ISA-2011B was discovered during our synthetic studies of C-1 indol-3-yl substituted 1,2,3,4-tetrahydroisoquinolines via a Pictet-Spengler approach. ISA-2011B significantly inhibits growth of tumor cells in xenograft mice, and we show that this is mediated by targeting PIP5K1α-associated PI3K/AKT and the downstream survival, proliferation, and invasion pathways. Further, siRNA-mediated knockdown of PIP5K1α exerts similar effects on PC3 cells as ISA-2011B treatment, significantly inhibiting AKT activity, increasing apoptosis and reducing invasion. Thus, PIP5K1α has high potential as a drug target, and compound ISA-2011B is interesting for further development of targeted cancer therapy. PMID:25071204

  7. 17 CFR 201.550 - Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... to Exchange Act Section 12(k)(1)(A). 201.550 Section 201.550 Commodity and Securities Exchanges... Suspensions § 201.550 Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A). (a) Petition for termination of suspension. Any person adversely affected by a suspension pursuant to Section 12(k)(1)(A)...

  8. 17 CFR 201.550 - Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... to Exchange Act Section 12(k)(1)(A). 201.550 Section 201.550 Commodity and Securities Exchanges... Suspensions § 201.550 Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A). (a) Petition for termination of suspension. Any person adversely affected by a suspension pursuant to Section 12(k)(1)(A)...

  9. 17 CFR 201.550 - Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... to Exchange Act Section 12(k)(1)(A). 201.550 Section 201.550 Commodity and Securities Exchanges... Suspensions § 201.550 Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A). (a) Petition for termination of suspension. Any person adversely affected by a suspension pursuant to Section 12(k)(1)(A)...

  10. 17 CFR 201.550 - Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... to Exchange Act Section 12(k)(1)(A). 201.550 Section 201.550 Commodity and Securities Exchanges... Suspensions § 201.550 Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A). (a) Petition for termination of suspension. Any person adversely affected by a suspension pursuant to Section 12(k)(1)(A)...

  11. 17 CFR 201.550 - Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... to Exchange Act Section 12(k)(1)(A). 201.550 Section 201.550 Commodity and Securities Exchanges... Suspensions § 201.550 Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A). (a) Petition for termination of suspension. Any person adversely affected by a suspension pursuant to Section 12(k)(1)(A)...

  12. 26 CFR 31.3402(k)-1 - Special rule for tips.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 15 2013-04-01 2013-04-01 false Special rule for tips. 31.3402(k)-1 Section 31.3402(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Collection of Income Tax at Source § 31.3402(k)-1 Special rule for tips. (a) Withholding of income tax...

  13. 26 CFR 1.1031(k)-1 - Treatment of deferred exchanges.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Treatment of deferred exchanges. 1.1031(k)-1 Section 1.1031(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Common Nontaxable Exchanges § 1.1031(k)-1 Treatment of...

  14. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 7 2013-04-01 2013-04-01 false Communist-controlled organizations. 1.501(k)-1 Section 1.501(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1...

  15. 26 CFR 31.3402(k)-1 - Special rule for tips.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 15 2011-04-01 2011-04-01 false Special rule for tips. 31.3402(k)-1 Section 31.3402(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Collection of Income Tax at Source § 31.3402(k)-1 Special rule for tips. (a) Withholding of income tax...

  16. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 7 2011-04-01 2009-04-01 true Communist-controlled organizations. 1.501(k)-1 Section 1.501(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1...

  17. 26 CFR 31.3402(k)-1 - Special rule for tips.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 15 2012-04-01 2012-04-01 false Special rule for tips. 31.3402(k)-1 Section 31.3402(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Collection of Income Tax at Source § 31.3402(k)-1 Special rule for tips. (a) Withholding of income tax...

  18. 26 CFR 1.1031(k)-1 - Treatment of deferred exchanges.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 11 2014-04-01 2014-04-01 false Treatment of deferred exchanges. 1.1031(k)-1 Section 1.1031(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Common Nontaxable Exchanges § 1.1031(k)-1 Treatment...

  19. 26 CFR 1.162(k)-1 - Disallowance of deduction for reacquisition payments.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... payments. 1.162(k)-1 Section 1.162(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Corporations § 1.162(k)-1 Disallowance of deduction for reacquisition payments. (a) In general... corporation to reacquire its stock from an ESOP that are used in a manner described in section...

  20. 26 CFR 1.1031(k)-1 - Treatment of deferred exchanges.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 11 2013-04-01 2013-04-01 false Treatment of deferred exchanges. 1.1031(k)-1 Section 1.1031(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Common Nontaxable Exchanges § 1.1031(k)-1 Treatment...

  1. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 7 2012-04-01 2012-04-01 false Communist-controlled organizations. 1.501(k)-1 Section 1.501(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1...

  2. 26 CFR 1.1031(k)-1 - Treatment of deferred exchanges.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 11 2012-04-01 2012-04-01 false Treatment of deferred exchanges. 1.1031(k)-1 Section 1.1031(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Common Nontaxable Exchanges § 1.1031(k)-1 Treatment...

  3. 26 CFR 31.3402(k)-1 - Special rule for tips.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 15 2014-04-01 2014-04-01 false Special rule for tips. 31.3402(k)-1 Section 31.3402(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Collection of Income Tax at Source § 31.3402(k)-1 Special rule for tips. (a) Withholding of income tax...

  4. 26 CFR 1.162(k)-1 - Disallowance of deduction for reacquisition payments.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... payments. 1.162(k)-1 Section 1.162(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Corporations § 1.162(k)-1 Disallowance of deduction for reacquisition payments. (a) In general... corporation to reacquire its stock from an ESOP that are used in a manner described in section...

  5. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 7 2014-04-01 2013-04-01 true Communist-controlled organizations. 1.501(k)-1 Section 1.501(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1...

  6. 26 CFR 31.3402(k)-1 - Special rule for tips.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Special rule for tips. 31.3402(k)-1 Section 31.3402(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Collection of Income Tax at Source § 31.3402(k)-1 Special rule for tips. (a) Withholding of income tax...

  7. 26 CFR 1.162(k)-1 - Disallowance of deduction for reacquisition payments.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... payments. 1.162(k)-1 Section 1.162(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Corporations § 1.162(k)-1 Disallowance of deduction for reacquisition payments. (a) In general... corporation to reacquire its stock from an ESOP that are used in a manner described in section...

  8. 26 CFR 1.162(k)-1 - Disallowance of deduction for reacquisition payments.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... payments. 1.162(k)-1 Section 1.162(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Corporations § 1.162(k)-1 Disallowance of deduction for reacquisition payments. (a) In general... corporation to reacquire its stock from an ESOP that are used in a manner described in section...

  9. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Communist-controlled organizations. 1.501(k)-1 Section 1.501(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1...

  10. 26 CFR 1.162(k)-1 - Disallowance of deduction for reacquisition payments.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... payments. 1.162(k)-1 Section 1.162(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Corporations § 1.162(k)-1 Disallowance of deduction for reacquisition payments. (a) In general... corporation to reacquire its stock from an ESOP that are used in a manner described in section...

  11. 26 CFR 1.1031(k)-1 - Treatment of deferred exchanges.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 11 2011-04-01 2011-04-01 false Treatment of deferred exchanges. 1.1031(k)-1 Section 1.1031(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Common Nontaxable Exchanges § 1.1031(k)-1 Treatment...

  12. Involvement of NarK1 and NarK2 Proteins in Transport of Nitrate and Nitrite in the Denitrifying Bacterium Pseudomonas aeruginosa PAO1

    PubMed Central

    Sharma, Vandana; Noriega, Chris E.; Rowe, John J.

    2006-01-01

    Two transmembrane proteins were tentatively classified as NarK1 and NarK2 in the Pseudomonas genome project and hypothesized to play an important physiological role in nitrate/nitrite transport in Pseudomonas aeruginosa. The narK1 and narK2 genes are located in a cluster along with the structural genes for the nitrate reductase complex. Our studies indicate that the transcription of all these genes is initiated from a single promoter and that the gene complex narK1K2GHJI constitutes an operon. Utilizing an isogenic narK1 mutant, a narK2 mutant, and a narK1K2 double mutant, we explored their effect on growth under denitrifying conditions. While the ΔnarK1::Gm mutant was only slightly affected in its ability to grow under denitrification conditions, both the ΔnarK2::Gm and ΔnarK1K2::Gm mutants were found to be severely restricted in nitrate-dependent, anaerobic growth. All three strains demonstrated wild-type levels of nitrate reductase activity. Nitrate uptake by whole-cell suspensions demonstrated both the ΔnarK2::Gm and ΔnarK1K2::Gm mutants to have very low yet different nitrate uptake rates, while the ΔnarK1::Gm mutant exhibited wild-type levels of nitrate uptake. Finally, Escherichia coli narK rescued both the ΔnarK2::Gm and ΔnarK1K2::Gm mutants with respect to anaerobic respiratory growth. Our results indicate that only the NarK2 protein is required as a nitrate/nitrite transporter by Pseudomonas aeruginosa under denitrifying conditions. PMID:16391109

  13. Ferredoxin Is Involved in Secretion of Cytotoxic Necrotizing Factor 1 across the Cytoplasmic Membrane in Escherichia coli K1

    PubMed Central

    Yu, Hao; Kim, Kwang Sik

    2010-01-01

    We previously showed that cytotoxic necrotizing factor 1 (CNF1) contributes to Escherichia coli K1 invasion of human brain microvascular endothelial cells (HBMEC) and interacts with the receptor on the surface of HBMEC. CNF1 is the cytoplasmic protein, and it remains incompletely understood how CNF1 is secreted across the inner and outer membranes in E. coli K1. In order to investigate the genetic determinants for secretion of CNF1 in E. coli K1, we performed Tn5 mutagenesis screening by applying β-lactamase as a reporter to monitor secretion of CNF1. We identified a Tn5 mutant that exhibited no β-lactamase activity in the culture supernatant and in which the mutated gene encodes a ferredoxin gene (fdx). In the fdx deletion mutant, there was no evidence of translocation of CNF1 into HBMEC. Western blot analysis of the fdx deletion mutant revealed that ferredoxin is involved in translocation of CNF1 across the cytoplasmic membrane. The fdx mutant exhibited significantly decreased invasion of HBMEC, similar to the decreased HBMEC invasion observed with the CNF1 mutant. The failures to secrete CNF1 and invade HBMEC of the fdx mutant were restored to the levels of the parent strain by complementation with fdx. These findings demonstrate for the first time that ferredoxin is involved in secretion of CNF1 across the inner membrane in meningitis-causing E. coli K1. PMID:19917710

  14. Contribution of S6K1/MAPK Signaling Pathways in the Response to Oxidative Stress: Activation of RSK and MSK by Hydrogen Peroxide

    PubMed Central

    Schneider, Taiane; Bonan, Carla Denise; Bartrons, Ramon; Ventura, Francesc; Rodrigues de Oliveira, Jarbas; Rosa, Jose Luis

    2013-01-01

    Cells respond to different kind of stress through the coordinated activation of signaling pathways such as MAPK or p53. To find which molecular mechanisms are involved, we need to understand their cell adaptation. The ribosomal protein, S6 kinase 1 (S6K1), is a common downstream target of signaling by hormonal or nutritional stress. Here, we investigated the initial contribution of S6K1/MAPK signaling pathways in the cell response to oxidative stress produced by hydrogen peroxide (H2O2). To analyze S6K1 activation, we used the commercial anti-phospho-Thr389-S6K1 antibody most frequently mentioned in the bibliography. We found that this antibody detected an 80-90 kDa protein that was rapidly phosphorylated in response to H2O2 in several human cells. Unexpectedly, this phosphorylation was insensitive to both mTOR and PI3K inhibitors, and knock-down experiments showed that this protein was not S6K1. RSK and MSK proteins were candidate targets of this phosphorylation. We demonstrated that H2O2 stimulated phosphorylation of RSK and MSK kinases at residues that are homologous to Thr389 in S6K1. This phosphorylation required the activity of either p38 or ERK MAP kinases. Kinase assays showed activation of RSK and MSK by H2O2. Experiments with mouse embryonic fibroblasts from p38 animals’ knockout confirmed these observations. Altogether, these findings show that the S6K1 signaling pathway is not activated under these conditions, clarify previous observations probably misinterpreted by non-specific detection of proteins RSK and MSK by the anti-phospho-Thr389-S6K1 antibody, and demonstrate the specific activation of MAPK signaling pathways through ERK/p38/RSK/MSK by H2O2. PMID:24058693

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

    NASA Astrophysics Data System (ADS)

    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.

  16. Broadening diversity in the Arostrilepis horrida complex: Arostrilepis kontrimavichusi n. sp. (Cyclophyllidea: Hymenolepididae) in the western red-backed vole Myodes californicus (Merriam) (Cricetidae: Arvicolinae) from temperate latitudes of the Pacific Northwest, North America.

    PubMed

    Makarikov, Arseny A; Hoberg, Eric P

    2016-06-01

    Specimens originally identified provisionally as Hymenolepis horrida (Linstow, 1901) [later Arostrilepis horrida (Linstow, 1901)] in Myodes californicus (Merriam) from near the Pacific coastal zone of southern Oregon are revised. Specimens in western red-backed voles represent an undescribed species of Arostrilepis Mas Coma & Tenora, 1997, contributing to recognition and resolution of a broadening complex encompassing cryptic diversity for these hymenolepidid tapeworms distributed across the Holarctic region. Consistent with recent studies defining diversity in the genus, the form, dimensions, and spination (pattern, shape and size) of the cirrus are diagnostic. Among 12 nominal congeners, specimens of A. kontrimavichusi n. sp. are further distinguished by the relative position and length of the cirrus-sac, arrangement of the testes and relative size of the external seminal vesicle and seminal receptacle. Specimens from Oregon voles represent the fifth endemic hymenolepidid in this genus from the Nearctic. Host range for the North American assemblage of species includes Cricetidae (Arvicolinae and Neotominae), Heteromyidae, Geomyidae, and rarely Sciuridae.

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

    PubMed

    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.

  18. Rapid Detection of K1 Hypervirulent Klebsiella pneumoniae by MALDI-TOF MS

    PubMed Central

    Huang, Yonglu; Li, Jiaping; Gu, Danxia; Fang, Ying; Chan, Edward W.; Chen, Sheng; Zhang, Rong

    2015-01-01

    Hypervirulent strains of Klebsiella pneumoniae (hvKP) are genetic variants of K. pneumoniae which can cause life-threatening community-acquired infection in healthy individuals. Currently, methods for efficient differentiation between classic K. pneumoniae (cKP) and hvKP strains are not available, often causing delay in diagnosis and treatment of hvKP infections. To address this issue, we devised a Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) approach for rapid identification of K1 hvKP strains. Four standard algorithms, genetic algorithm (GA), support vector machine (SVM), supervised neural network (SNN), and quick classifier (QC), were tested for their power to differentiate between K1 and non-K1 strains, among which SVM was the most reliable algorithm. Analysis of the receiver operating characteristic curves of the interest peaks generated by the SVM model was found to confer highly accurate detection sensitivity and specificity, consistently producing distinguishable profiles for K1 hvKP and non-K1 strains. Of the 43 K. pneumoniae modeling strains tested by this approach, all were correctly identified as K1 hvKP and non-K1 capsule type. Of the 20 non-K1 and 17 K1 hvKP validation isolates, the accuracy of K1 hvKP and non-K1 identification was 94.1 and 90.0%, respectively, according to the SVM model. In summary, the MALDI-TOF MS approach can be applied alongside the conventional genotyping techniques to provide rapid and accurate diagnosis, and hence prompt treatment of infections caused by hvKP. PMID:26733976

  19. Role of GluK1 Kainate Receptors in Seizures, Epileptic Discharges, and Epileptogenesis

    PubMed Central

    Fritsch, Brita; Reis, Janine; Gasior, Maciej; Kaminski, Rafal M.

    2014-01-01

    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. PMID:24760837

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

    PubMed

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

    2016-10-01

    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 p150(Glued) subunit of dynactin. IC-p150(Glued) 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

  1. 26 CFR 1.401(k)-1 - Certain cash or deferred arrangements.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... determined in accordance with regulations and rulings issued by the Department of Labor. See 29 CFR 2510.3... 26 Internal Revenue 5 2013-04-01 2013-04-01 false Certain cash or deferred arrangements. 1.401(k)-1 Section 1.401(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE...

  2. 26 CFR 1.401(k)-1 - Certain cash or deferred arrangements.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... determined in accordance with regulations and rulings issued by the Department of Labor. See 29 CFR 2510.3... 26 Internal Revenue 5 2014-04-01 2014-04-01 false Certain cash or deferred arrangements. 1.401(k)-1 Section 1.401(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE...

  3. 26 CFR 1.401(k)-1 - Certain cash or deferred arrangements.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... determined in accordance with regulations and rulings issued by the Department of Labor. See 29 CFR 2510.3... 26 Internal Revenue 5 2011-04-01 2011-04-01 false Certain cash or deferred arrangements. 1.401(k)-1 Section 1.401(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE...

  4. 26 CFR 1.401(k)-1 - Certain cash or deferred arrangements.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... determined in accordance with regulations and rulings issued by the Department of Labor. See 29 CFR 2510.3... 26 Internal Revenue 5 2012-04-01 2011-04-01 true Certain cash or deferred arrangements. 1.401(k)-1 Section 1.401(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  5. 26 CFR 1.401(k)-1 - Certain cash or deferred arrangements.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... rulings issued by the Department of Labor. See 29 CFR 2510.3-102. (3) Rules applicable to cash or deferred... 26 Internal Revenue 5 2010-04-01 2010-04-01 false Certain cash or deferred arrangements. 1.401(k)-1 Section 1.401(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE...

  6. Combination drug scheduling defines a "window of opportunity" for chemopotentiation of gemcitabine by an orally bioavailable, selective ChK1 inhibitor, GNE-900.

    PubMed

    Blackwood, Elizabeth; Epler, Jennifer; Yen, Ivana; Flagella, Michael; O'Brien, Tom; Evangelista, Marie; Schmidt, Stephen; Xiao, Yang; Choi, Jonathan; Kowanetz, Kaska; Ramiscal, Judi; Wong, Kenton; Jakubiak, Diana; Yee, Sharon; Cain, Gary; Gazzard, Lewis; Williams, Karen; Halladay, Jason; Jackson, Peter K; Malek, Shiva

    2013-10-01

    Checkpoint kinase 1 (ChK1) is a serine/threonine kinase that functions as a central mediator of the intra-S and G2-M cell-cycle checkpoints. Following DNA damage or replication stress, ChK1-mediated phosphorylation of downstream effectors delays cell-cycle progression so that the damaged genome can be repaired. As a therapeutic strategy, inhibition of ChK1 should potentiate the antitumor effect of chemotherapeutic agents by inactivating the postreplication checkpoint, causing premature entry into mitosis with damaged DNA resulting in mitotic catastrophe. Here, we describe the characterization of GNE-900, an ATP-competitive, selective, and orally bioavailable ChK1 inhibitor. In combination with chemotherapeutic agents, GNE-900 sustains ATR/ATM signaling, enhances DNA damage, and induces apoptotic cell death. The kinetics of checkpoint abrogation seems to be more rapid in p53-mutant cells, resulting in premature mitotic entry and/or accelerated cell death. Importantly, we show that GNE-900 has little single-agent activity in the absence of chemotherapy and does not grossly potentiate the cytotoxicity of gemcitabine in normal bone marrow cells. In vivo scheduling studies show that optimal administration of the ChK1 inhibitor requires a defined lag between gemcitabine and GNE-900 administration. On the refined combination treatment schedule, gemcitabine's antitumor activity against chemotolerant xenografts is significantly enhanced and dose-dependent exacerbation of DNA damage correlates with extent of tumor growth inhibition. In summary, we show that in vivo potentiation of gemcitabine activity is mechanism based, with optimal efficacy observed when S-phase arrest and release is followed by checkpoint abrogation with a ChK1 inhibitor.

  7. Revealing Different Roles of the mTOR-Targets S6K1 and S6K2 in Breast Cancer by Expression Profiling and Structural Analysis

    PubMed Central

    Karlsson, Elin; Magić, Ivana; Bostner, Josefine; Dyrager, Christine; Lysholm, Fredrik; Hallbeck, Anna-Lotta; Stål, Olle; Lundström, Patrik

    2015-01-01

    Background The AKT/mTORC1/S6K pathway is frequently overstimulated in breast cancer, constituting a promising therapeutic target. The benefit from mTOR inhibitors varies, likely as a consequence of tumour heterogeneity, and upregulation of several compensatory feed-back mechanisms. The mTORC1 downstream effectors S6K1, S6K2, and 4EBP1 are amplified and overexpressed in breast cancer, associated with a poor outcome and divergent endocrine treatment benefit. S6K1 and S6K2 share high sequence homology, but evidence of partly distinct biological functions is emerging. The aim of this work was to explore possible different roles and treatment target potentials of S6K1 and S6K2 in breast cancer. Materials and methods Whole-genome expression profiles were compared for breast tumours expressing high levels of S6K1, S6K2 or 4EBP1, using public datasets, as well as after in vitro siRNA downregulation of S6K1 and/or S6K2 in ZR751 breast cancer cells. In silico homology modelling of the S6K2 kinase domain was used to evaluate its possible structural divergences to S6K1. Results Genome expression profiles were highly different in S6K1 and S6K2 high tumours, whereas S6K2 and 4EBP1 profiles showed significant overlaps, both correlated to genes involved in cell cycle progression, among these the master regulator E2F1. S6K2 and 4EBP1 were inversely associated with IGF1 levels, and their prognostic value was shown to be restricted to tumours positive for IGFR and/or HER2. In vitro, S6K1 and S6K2 silencing resulted in upregulation of genes in the mTORC1 and mTORC2 complexes. Isoform-specific silencing also showed distinct patterns, e.g. S6K2 downregulation lead to upregulation of several cell cycle associated genes. Structural analyses of the S6K2 kinase domain showed unique structure patterns, deviating from those of S6K1, facilitating the development of isoform-specific inhibitors. Our data support emerging proposals of distinct biological features of S6K1 and S6K2, suggesting

  8. Binding mode of an α-amino acid-linked quinoxaline-2,3-dione analogue at glutamate receptor subtype GluK1.

    PubMed

    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.

  9. B→K1π(K) decays in the perturbative QCD approach

    NASA Astrophysics Data System (ADS)

    Zhang, Zhi-Qing; Hou, Zhi-Wei; Yang, Yueling; Sun, Junfeng

    2014-10-01

    Within the framework of the perturbative QCD approach, we study the two-body charmless decays B→K1(1270)(K1(1400))π(K). We find the following results: (i) The decays B¯0→K1(1270)+π-, K1(1400)+π- are incompatible with the present experimental data. There exists a similar situation for the decays B¯0→a1(1260)+K-, b1(1235)+K-, which are usually considered that the nonperturbative contributions are needed to explain the data. But the difference is that the nonperturbative contributions seem to play opposite roles in these two groups of decays. (ii) The pure annihilation type decays B¯0→K1±(1270)K∓, K1±(1400)K∓ are good channels to test whether an approach can be used to calculate correctly the strength of the penguin-annihilation amplitudes. Their branching ratios are predicted at 10-7 order, which are larger than the QCDF results. (iii) The dependence of the direct CP-violating asymmetries of these decays on the mixing angle θK_1 are also considered.

  10. Contribution of vitamin K1 to the electron spin polarization in spinach photosystem I

    SciTech Connect

    Rustandi, R.R.; Snyder, S.W.; Feezel, L.L.; Michalski, T.J.; Norris, J.R.; Thurnauer, M.C.; Biggins, J. )

    1990-09-04

    The electron spin polarized (ESP) electron paramagnetic resonance (EPR) signal observed in spinach photosystem I (PSI) particles was examined in preparations depleted of vitamin K1 by solvent extraction and following biological reconstitution by the quinone. The ESP EPR signal was not detected in the solvent-extracted PSI sample but was restored upon reconstitution with either protonated or deuterated vitamin K1 under conditions that also restored electron transfer to the terminal PSI acceptors. Reconstitution using deuterated vitamin K1 resulted in a line narrowing of the ESP EPR signal, supporting the conclusion that the ESP EPR signals in the reconstituted samples arise from a radical pair consisting of the oxidized PSI primary donor, P700+, and reduced vitamin K1.

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

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    Mueller, George E.; Kohrs, Dick; Bailey, Richard; Lai, Gary

    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 3 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.

  13. Vitamin K1 metabolism in relation to pharmacodynamic response in anticoagulated patients.

    PubMed Central

    Choonara, I A; Scott, A K; Haynes, B P; Cholerton, S; Breckenridge, A M; Park, B K

    1985-01-01

    The disposition of, and pharmacological response to, a single intravenous dose of vitamin K1 (10 mg) was studied in eleven patients on daily warfarin therapy. The pharmacokinetics of vitamin K1 in patients were similar to those reported previously in healthy volunteers, terminal half-life 1.7 h. All patients had been taking warfarin for at least 3 months. Steady state warfarin plasma concentrations ranged from 0.5 to 1.4 micrograms ml-1. Prothrombin complex activity ranged from 15 to 28.5%. There was considerable inter-individual variation in pharmacodynamic response as expressed by prothrombin complex activity (PCA) and Factor VII. The maximum values for PCA and Factor VII were reached at 24-96 h and 24-48 h, respectively, after the administration of vitamin K1. Vitamin K1 (10 mg) has a long duration of action (greater than 168 h) in terms of clotting factor synthesis in patients on steady state warfarin. All the patients on warfarin had measurable levels (CPmax 0.3-1.2 micrograms ml-1) of vitamin K1 2, 3-epoxide. There was a significant correlation between the pharmacodynamic response as expressed by change in % PCA and the AUC for vitamin K1 2,3-epoxide (P less than 0.05). PMID:4091996

  14. Essential role for SphK1/S1P signaling to regulate hypoxia-inducible factor 2α expression and activity in cancer

    PubMed Central

    Bouquerel, P; Gstalder, C; Müller, D; Laurent, J; Brizuela, L; Sabbadini, R A; Malavaud, B; Pyronnet, S; Martineau, Y; Ader, I; Cuvillier, O

    2016-01-01

    The sphingosine kinase-1/sphingosine 1-phosphate (SphK1/S1P) signaling pathway has been reported to modulate the expression of the canonical transcription factor hypoxia-inducible HIF-1α in multiple cell lineages. HIF-2α is also frequently overexpressed in solid tumors but its role has been mostly studied in clear cell renal cell carcinoma (ccRCC), the most common form of kidney cancer, where HIF-2α has been established as a driver of a more aggressive disease. In this study, the role of SphK1/S1P signaling with regard to HIF-2α was investigated in various cancer cell models including ccRCC cells. Under hypoxic conditions or in ccRCC lacking a functional von Hippel-Lindau (VHL) gene and expressing high levels of HIF-2α, SphK1 activity controls HIF-2α expression and transcriptional activity through a phospholipase D (PLD)-driven mechanism. SphK1 silencing promotes a VHL-independent HIF-2α loss of expression and activity and reduces cell proliferation in ccRCC. Importantly, downregulation of SphK1 is associated with impaired Akt and mTOR signaling in ccRCC. Taking advantage of a monoclonal antibody neutralizing extracellular S1P, we show that inhibition of S1P extracellular signaling blocks HIF-2α accumulation in ccRCC cell lines, an effect mimicked when the S1P transporter Spns2 or the S1P receptor 1 (S1P1) is silenced. Here, we report the first evidence that the SphK1/S1P signaling pathway regulates the transcription factor hypoxia-inducible HIF-2α in diverse cancer cell lineages notably ccRCC, where HIF-2α has been established as a driver of a more aggressive disease. These findings demonstrate that SphK1/S1P signaling may act as a canonical regulator of HIF-2α expression in ccRCC, giving support to its inhibition as a therapeutic strategy that could contribute to reduce HIF-2 activity in ccRCC. PMID:26974204

  15. Atrial proarrhythmia due to increased inward rectifier current (I(K1)) arising from KCNJ2 mutation--a simulation study.

    PubMed

    Kharche, Sanjay; Garratt, Clifford J; Boyett, Mark R; Inada, Shin; Holden, Arun V; Hancox, Jules C; Zhang, Henggui

    2008-01-01

    Atrial fibrillation (AF) has been linked to increased inward rectifier potassium current, I(K1), either due to AF-induced electrical remodelling, or from functional changes due to the Kir2.1 V93I mutation. The aim of this simulation study was to identify at cell and tissue levels' mechanisms by which increased I(K1) facilitates and perpetuates AF. The Courtemanche et al. human atrial cell action potential (AP) model was modified to incorporate reported changes in I(K1) induced by the Kir2.1 V93I mutation in both heterozygous (Het) and homozygous (Hom) mutant forms. The modified models for wild type (WT), Het and Hom conditions were incorporated into homogeneous 1D, 2D and 3D tissue models. Restitution curves of AP duration (APD), effective refractory period (ERP) and conduction velocity (CV) were computed and both the temporal and the spatial vulnerability of atrial tissue to re-entry were measured. The lifespan and tip meandering pattern of re-entry were also characterised. For comparison, parallel simulations were performed by incorporating into the Courtmanche et al. model a linear increase in maximal I(K1) conductance. It was found that the gain-in-function of V93I 'mutant'I(K1) led to abbreviated atrial APs and flattened APD, ERP and CV restitution curves. It also hyperpolarised atrial resting membrane potential and slowed down intra-atrial conduction. V93I 'mutant'I(K1) reduced the tissue's temporal vulnerability but increased spatial vulnerability to initiate and sustain re-entry, resulting in an increased overall susceptibility of atrial tissue to arrhythmogenesis. In the 2D model, spiral waves self-terminated for WT (lifespan < 3.3 s) tissue, but persisted in Het and Hom tissues for the whole simulation period (lifespan > 10 s). The tip of the spiral wave meandered more in WT tissue than in Het and Hom tissues. Increased I(K1) due to augmented maximal conductance produced similar results to those of Het and Hom Kir2.1 V93I mutant conditions. In the 3D

  16. An investigation of the pharmacological response to vitamin K1 in the rabbit.

    PubMed Central

    Winn, M. J.; Cholerton, S.; Park, B. K.

    1988-01-01

    1. The relationship between pharmacological response and disposition of a dose of vitamin K1 (10 mgkg-1, i.v.) in normal rabbits and in rabbits treated with the coumarin anticoagulant brodifacoum, has been studied. 2. High performance liquid chromatography (h.p.l.c.) with electrochemical detection (EC) was used to determine concentrations of vitamin K1 in plasma, whole liver homogenate, and liver microsomes. 3. After intravenous administration of vitamin K1, plasma concentrations of the vitamin declined in a tri-exponential fashion. There were no differences between the two groups over the first 24 h of the experiment. However, between 24 h and the end of the study, plasma concentrations of vitamin K1 in the presence of brodifacoum were significantly (P less than or equal to 0.05) below those of vehicle-treated rabbits. 4. Seventy-two hours after administration of vitamin K1, plasma concentrations of the vitamin were not different from normal. 5. Three hours after administration of vitamin K1, the concentrations of the vitamin in whole liver were 46.6 +/- 4.3 micrograms g-1 in the presence of brodifacoum, and 32.8 +/- 6.4 micrograms g-1 in the absence of brodifacoum; and were significantly (P less than or equal to 0.05) greater than normal (127.7 +/- 44.3 ng g-1). Likewise, microsomal concentrations of vitamin K1 (4.00 +/- 2.38 micrograms mg-1 protein, and 2.65 +/- 1.01 micrograms mg-1 protein, in the presence and absence of brodifacoum, respectively) were significantly (P less than or equal to 0.01) greater than normal (16.0 +/- 3.5 ng mg-1 protein).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3207975

  17. K1 and K15 of Kaposi's Sarcoma-Associated Herpesvirus Are Partial Functional Homologues of Latent Membrane Protein 2A of Epstein-Barr Virus

    PubMed Central

    Steinbrück, Lisa; Gustems, Montse; Medele, Stephanie; Schulz, Thomas F.; Lutter, Dominik

    2015-01-01

    ABSTRACT The human herpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are associated with Hodgkin's lymphoma (HL) and Primary effusion lymphomas (PEL), respectively, which are B cell malignancies that originate from germinal center B cells. PEL cells but also a quarter of EBV-positive HL tumor cells do not express the genuine B cell receptor (BCR), a situation incompatible with survival of normal B cells. EBV encodes LMP2A, one of EBV's viral latent membrane proteins, which likely replaces the BCR's survival signaling in HL. Whether KSHV encodes a viral BCR mimic that contributes to oncogenesis is not known because an experimental model of KSHV-mediated B cell transformation is lacking. We addressed this uncertainty with mutant EBVs encoding the KSHV genes K1 or K15 in lieu of LMP2A and infected primary BCR-negative (BCR−) human B cells with them. We confirmed that the survival of BCR– B cells and their proliferation depended on an active LMP2A signal. Like LMP2A, the expression of K1 and K15 led to the survival of BCR− B cells prone to apoptosis, supported their proliferation, and regulated a similar set of cellular target genes. K1 and K15 encoded proteins appear to have noncomplementing, redundant functions in this model, but our findings suggest that both KSHV proteins can replace LMP2A's key activities contributing to the survival, activation and proliferation of BCR– PEL cells in vivo. IMPORTANCE Several herpesviruses encode oncogenes that are receptor-like proteins. Often, they are constitutively active providing important functions to the latently infected cells. LMP2A of Epstein-Barr virus (EBV) is such a receptor that mimics an activated B cell receptor, BCR. K1 and K15, related receptors of Kaposi's sarcoma-associated herpesvirus (KSHV) expressed in virus-associated tumors, have less obvious functions. We found in infection experiments that both viral receptors of KSHV can replace LMP2A and deliver functions

  18. S6K1 Phosphorylation of H2B Mediates EZH2 Trimethylation of H3: A Determinant of Early Adipogenesis.

    PubMed

    Yi, Sang Ah; Um, Sung Hee; Lee, Jaecheol; Yoo, Ji Hee; Bang, So Young; Park, Eun Kyung; Lee, Min Gyu; Nam, Ki Hong; Jeon, Ye Ji; Park, Jong Woo; You, Jueng Soo; Lee, Sang-Jin; Bae, Gyu-Un; Rhie, Jong Won; Kozma, Sara C; Thomas, George; Han, Jeung-Whan

    2016-05-01

    S6K1 has been implicated in a number of key metabolic responses, which contribute to obesity. Critical among these is the control of a transcriptional program required for the commitment of mesenchymal stem cells to the adipocytic lineage. However, in contrast to its role in the cytosol, the functions and targets of nuclear S6K1 are unknown. Here, we show that adipogenic stimuli trigger nuclear translocation of S6K1, leading to H2BS36 phosphorylation and recruitment of EZH2 to H3, which mediates H3K27 trimethylation. This blocks Wnt gene expression, inducing the upregulation of PPARγ and Cebpa and driving increased adipogenesis. Consistent with this finding, white adipose tissue from S6K1-deficient mice exhibits no detectable H2BS36 phosphorylation or H3K27 trimethylation, whereas both responses are highly elevated in obese humans or in mice fed a high-fat diet. These findings define an S6K1-dependent mechanism in early adipogenesis, contributing to the promotion of obesity. PMID:27151441

  19. S6K1 Phosphorylation of H2B Mediates EZH2 Trimethylation of H3: A Determinant of Early Adipogenesis.

    PubMed

    Yi, Sang Ah; Um, Sung Hee; Lee, Jaecheol; Yoo, Ji Hee; Bang, So Young; Park, Eun Kyung; Lee, Min Gyu; Nam, Ki Hong; Jeon, Ye Ji; Park, Jong Woo; You, Jueng Soo; Lee, Sang-Jin; Bae, Gyu-Un; Rhie, Jong Won; Kozma, Sara C; Thomas, George; Han, Jeung-Whan

    2016-05-01

    S6K1 has been implicated in a number of key metabolic responses, which contribute to obesity. Critical among these is the control of a transcriptional program required for the commitment of mesenchymal stem cells to the adipocytic lineage. However, in contrast to its role in the cytosol, the functions and targets of nuclear S6K1 are unknown. Here, we show that adipogenic stimuli trigger nuclear translocation of S6K1, leading to H2BS36 phosphorylation and recruitment of EZH2 to H3, which mediates H3K27 trimethylation. This blocks Wnt gene expression, inducing the upregulation of PPARγ and Cebpa and driving increased adipogenesis. Consistent with this finding, white adipose tissue from S6K1-deficient mice exhibits no detectable H2BS36 phosphorylation or H3K27 trimethylation, whereas both responses are highly elevated in obese humans or in mice fed a high-fat diet. These findings define an S6K1-dependent mechanism in early adipogenesis, contributing to the promotion of obesity.

  20. Fcγ Receptor I Alpha Chain (CD64) Expression in Macrophages Is Critical for the Onset of Meningitis by Escherichia coli K1

    PubMed Central

    Selvaraj, Suresh K.; Wooster, David G.; Babu, M. Madan; Schreiber, Alan D.; Verbeek, J. Sjef; Prasadarao, Nemani V.

    2010-01-01

    Neonatal meningitis due to Escherichia coli K1 is a serious illness with unchanged morbidity and mortality rates for the last few decades. The lack of a comprehensive understanding of the mechanisms involved in the development of meningitis contributes to this poor outcome. Here, we demonstrate that depletion of macrophages in newborn mice renders the animals resistant to E. coli K1 induced meningitis. The entry of E. coli K1 into macrophages requires the interaction of outer membrane protein A (OmpA) of E. coli K1 with the alpha chain of Fcγ receptor I (FcγRIa, CD64) for which IgG opsonization is not necessary. Overexpression of full-length but not C-terminal truncated FcγRIa in COS-1 cells permits E. coli K1 to enter the cells. Moreover, OmpA binding to FcγRIa prevents the recruitment of the γ-chain and induces a different pattern of tyrosine phosphorylation of macrophage proteins compared to IgG2a induced phosphorylation. Of note, FcγRIa−/− mice are resistant to E. coli infection due to accelerated clearance of bacteria from circulation, which in turn was the result of increased expression of CR3 on macrophages. Reintroduction of human FcγRIa in mouse FcγRIa−/− macrophages in vitro increased bacterial survival by suppressing the expression of CR3. Adoptive transfer of wild type macrophages into FcγRIa−/− mice restored susceptibility to E. coli infection. Together, these results show that the interaction of FcγRI alpha chain with OmpA plays a key role in the development of neonatal meningitis by E. coli K1. PMID:21124939

  1. Deletion of C7L and K1L genes leads to significantly decreased virulence of recombinant vaccinia virus TianTan.

    PubMed

    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.

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

    USGS Publications Warehouse

    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.

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

    PubMed Central

    Matin, Abdul

    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. 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. PMID:22355201

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

    PubMed

    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.

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

    PubMed

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

    2016-01-01

    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. PMID:26574010

  6. Final report of the APMP water flow key comparison: APMP.M.FF-K1

    NASA Astrophysics Data System (ADS)

    Lee, Kwang-Bock; Chun, Sejong; Terao, Yoshiya; Thai, Nguyen Hong; Tsair Yang, Cheng; Tao, Meng; Gutkin, Mikhail B.

    2011-01-01

    The key comparison, APMP.M.FF-K1, was undertaken by APMP/TCFF, the Technical Committee for Fluid Flow (TCFF) under the Asia Pacific Metrology Program (APMP). One objective of the key comparison was to demonstrate the degree of equivalence among six participating laboratories (KRISS, NMIJ, VMI, CMS, NIM and VNIIM) in water flow rate metrology by comparing the results with the key comparison reference value (KCRV) determined from the CCM.FF-K1 key comparison. The other objective of this key comparison was to provide supporting evidence for the calibration and measurement capabilities (CMCs), which had been declared by the participating laboratories during this key comparison. The Transfer Standard Package (TSP) was a Coriolis mass flowmeter, which had been used in the CCM.FF-K1 key comparison. Because the K-factors in the APMP.M.FF-K1 key comparison were slightly lower than the K-factors of the CCM.FF-K1 key comparison due to long-term drifts of the TSP, a correction value D was introduced. The value of D was given by a weighted sum between two link laboratories (NMIJ and KRISS), which participated in both the CCM.FF-K1 and the APMP.M.FF-K1 key comparisons. By this correction, the K-factors were laid between 12.004 and 12.017 at either low (Re = 254 000) or high (Re = 561 000) flow rates. Most of the calibration data were within expected uncertainty bounds. However, some data showed undulations, which gave large fluctuations of the metering factor at Re = 561 000. Calculation of degrees of equivalence showed that all the participating laboratories had deviations between -0.009 and 0.007 pulses/kg from the CCM.FF-K1 KCRV at either the low or the high flow rates. In case of En calculation, all the participating laboratories showed values less than 1, indicating that the corrected K-factors of all the laboratories were equivalent with the KCRV at both Re = 254 000 and 561 000. When the corrected K-factors from two participating laboratories were compared, all the

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

    PubMed

    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. Broadening diversity in the Arostrilepis horrida complex: Arostrilepis kontrimavichusi n. sp. (Cyclophyllidea: Hymenolepididae) in the western red-backed vole Myodes californicus (Merriam) (Cricetidae: Arvicolinae) from temperate latitudes of the Pacific Northwest, North America.

    PubMed

    Makarikov, Arseny A; Hoberg, Eric P

    2016-06-01

    Specimens originally identified provisionally as Hymenolepis horrida (Linstow, 1901) [later Arostrilepis horrida (Linstow, 1901)] in Myodes californicus (Merriam) from near the Pacific coastal zone of southern Oregon are revised. Specimens in western red-backed voles represent an undescribed species of Arostrilepis Mas Coma & Tenora, 1997, contributing to recognition and resolution of a broadening complex encompassing cryptic diversity for these hymenolepidid tapeworms distributed across the Holarctic region. Consistent with recent studies defining diversity in the genus, the form, dimensions, and spination (pattern, shape and size) of the cirrus are diagnostic. Among 12 nominal congeners, specimens of A. kontrimavichusi n. sp. are further distinguished by the relative position and length of the cirrus-sac, arrangement of the testes and relative size of the external seminal vesicle and seminal receptacle. Specimens from Oregon voles represent the fifth endemic hymenolepidid in this genus from the Nearctic. Host range for the North American assemblage of species includes Cricetidae (Arvicolinae and Neotominae), Heteromyidae, Geomyidae, and rarely Sciuridae. PMID:27221000

  9. 76 FR 7847 - Glenn A. Baxter, Application To Renew License for Amateur Radio Service Station K1MAN

    Federal Register 2010, 2011, 2012, 2013, 2014

    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...

  10. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... deductibility of certain dividend distributions. (Temporary) 1.404(k)-1T Section 1.404(k)-1T Internal Revenue... (CONTINUED) 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)...

  11. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... deductibility of certain dividend distributions. (Temporary) 1.404(k)-1T Section 1.404(k)-1T Internal Revenue... (CONTINUED) 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)...

  12. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false Electric Arc Furnace (EAF) CH4 Emission Factors K Table K-1 to Subpart K of Part 98 Protection of Environment ENVIRONMENTAL PROTECTION.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4...

  13. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... deductibility of certain dividend distributions. (Temporary) 1.404(k)-1T Section 1.404(k)-1T Internal Revenue... (CONTINUED) 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)...

  14. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false Electric Arc Furnace (EAF) CH4 Emission Factors K Table K-1 to Subpart K of Part 98 Protection of Environment ENVIRONMENTAL PROTECTION.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4...

  15. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... deductibility of certain dividend distributions. (Temporary) 1.404(k)-1T Section 1.404(k)-1T Internal Revenue... (CONTINUED) 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)...

  16. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false Electric Arc Furnace (EAF) CH4 Emission Factors K Table K-1 to Subpart K of Part 98 Protection of Environment ENVIRONMENTAL PROTECTION.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4...

  17. 77 FR 64848 - Proposed Collection; Comment Request for Form 1120S, Schedule D, Schedule K-1, and Schedule M-3

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-23

    ... Internal Revenue Service Proposed Collection; Comment Request for Form 1120S, Schedule D, Schedule K-1, and... With Total Assets of $10 Million or More, and Schedule K-1 (Form 1120S), Shareholder's Share of Income... Losses and Built-in Gains, Schedule K-1 (Form 1120S), Shareholder's Share of Income, Credits,...

  18. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Code of Federal Regulations, 2010 CFR

    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:...

  19. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false Electric Arc Furnace (EAF) CH4 Emission Factors K Table K-1 to Subpart K of Part 98 Protection of Environment ENVIRONMENTAL PROTECTION.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4...

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

    PubMed

    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.

  1. Energy: Multidisciplinary Activities for the Classroom. Top Hit Energy Lesson Plans, K-1, 2-6.

    ERIC Educational Resources Information Center

    National Energy Foundation, Salt Lake City, UT.

    This six-volume set of multidisciplinary instructional materials developed by the National Energy Foundation (NEF) presents energy activities for grades K-1, 2-6. The instructional materials are teacher-developed, teacher-tested, and multi-disciplinary. The lesson plans and activities are organized around seven goal areas of a NEF developed…

  2. The Greatest Educational Change America Has Ever Seen, 2002: Lesson Plans for Grades K-1.

    ERIC Educational Resources Information Center

    United States Mint (Dept. of Treasury), Washington, DC.

    This teacher's guide on the 50 state quarters emitted by the United States Mint includes 6 "teacher-friendly" lesson plans that fit easily into the curriculum of grades K-1; reproducible student worksheets that coincide with each lesson; "fun" state facts and information on the new quarter designs; and USA map template with state outlines. These…

  3. The Greatest Educational Change America Has Ever Seen. [Teaching Guide]. Grades K-1.

    ERIC Educational Resources Information Center

    United States Mint (Dept. of Treasury), Washington, DC.

    This teaching guide for grades K-1 focuses on the 1999-2000 United States Mint 50 State Quarters Program, which includes new quarter designs for the following states: Delaware, Pennsylvania, New Jersey, Georgia, Connecticut, Massachusetts, Maryland, South Carolina, New Hampshire, and Virginia. The guide includes six lesson plans that fit easily…

  4. The Greatest Educational Change America Has Ever Seen, 2001: Lesson Plans for Grades K-1.

    ERIC Educational Resources Information Center

    United States Mint (Dept. of Treasury), Washington, DC.

    This teacher's guide about the 50 state quarters produced by the United States Mint includes 6 lesson plans that fit easily into the social studies curriculum for grades K-1. The lesson plans include reproducible student work pages that coincide with each lesson; state facts and information about the new 2001 state quarter designs (New York, North…

  5. easyCBM® Reading Criterion Related Validity Evidence: Grades K-1. Technical Report #1309

    ERIC Educational Resources Information Center

    Lai, Cheng-Fei; Alonzo, Julie; Tindal, Gerald

    2013-01-01

    In this technical report, we present the results of a study to gather criterion-related evidence for Grade K-1 easyCBM® reading measures. We used correlations to examine the relation between the easyCBM® measures and other published measures with known reliability and validity evidence, including the Dynamic Indicators of Basic Early Literacy…

  6. Two Genome Sequences of Klebsiella pneumoniae Strains with Sequence Type 23 and Capsular Serotype K1

    PubMed Central

    Lin, Hsi-Hsu; Chen, Yao-Shen; Hsiao, Hao-Wen; Hsueh, Pei-Tan; Ni, Wei-Fen

    2016-01-01

    Here, we report the whole-genome sequences of Klebsiella pneumoniae ED2 and ED23, isolated, respectively, from bacteremic patients with liver abscesses (ED2) and patients with primary liver abscess and metastatic meningitis (ED23). Both strains were of multilocus sequence type 23 with capsule serotype K1. PMID:27795261

  7. Isolation and Characterization of the K1 (L) Antigen of Escherichia coli1

    PubMed Central

    Bolaños, Roger; DeWitt, Charles W.

    1966-01-01

    Bolaños, Roger (Tulane University, New Orleans, La.), and Charles W. DeWitt. Isolation and characterization of the K1 (L) antigen of Escherichia coli. J. Bacteriol. 91:987–996. 1966.—An acidic polysaccharide with the serological properties of the K1 (L) antigen has been isolated from Escherichia coli O2:K1:H4 by means of phenol-water extraction, fractionation with hexadecyltrimethylammonium bromide (Cetavlon), filtration through Sephadex G-200, and chromatography on anion-exchange cellulose columns. Nucleic acid and protein content were reduced to a nondetectable level. There is no contamination with O antigen. The active material appears in two different positions in the Cetavlon fractionation, each with a slightly different serological specificity, as followed by the inhibition of passive hemagglutination. By paper chromatography, the polysaccharide moiety of the O antigenic fraction is composed of glucose, galactose, hexosamine and rhamnose. The absence of colanic acid in either K1 fraction was not proven, although its participation in our assay system, as well as the participation of the common or cross-reacting antigen, was ruled out by serological means. PMID:5326105

  8. Antinociceptive effects of MSVIII-19, a functional antagonist of the GluK1 kainate receptor.

    PubMed

    Qiu, Chang-Shen; Lash-Van Wyhe, Leanne; Sasaki, Makoto; Sakai, Ryuichi; Swanson, Geoffrey T; Gereau, Robert W

    2011-05-01

    The ionotropic glutamate receptor subunit, GluK1 (GluR5), is expressed in many regions of the nervous system related to sensory transmission. Recently, a selective ligand for the GluK1 receptor, MSVIII-19 (8,9-dideoxy-neodysiherbaine), was synthesized as a derivative of dysiherbaine, a toxin isolated from the marine sponge Lendenfeldia chondrodes. MSVIII-19 potently desensitizes GluK1 receptors without channel activation, rendering it useful as a functional antagonist. Given the high selectivity for GluK1 and the proposed role for this glutamate receptor in nociception, we sought to test the analgesic potential of MSVIII-19 in a series of models of inflammatory, neuropathic, and visceral pain in mice. MSVIII-19 delivered intrathecally dose-dependently reduced formalin-induced spontaneous behaviors and reduced thermal hypersensitivity 3 hours after formalin injection and 24 hours after complete Freund's adjuvant-induced inflammation, but had no effect on mechanical sensitivity in the same models. Intrathecal MSVIII-19 significantly reduced both thermal hyperalgesia and mechanical hypersensitivity in the chronic constriction injury model of neuropathic pain, but had no effect in the acetic acid model of visceral pain. Peripheral administration of MSVIII-19 had no analgesic efficacy in any of these models. Finally, intrathecal MSVIII-19 did not alter responses in Tail-flick tests or performance on the accelerating RotaRod. These data suggest that spinal administration of MSVIII-19 reverses hypersensitivity in several models of pain in mice, supporting the clinical potential of GluK1 antagonists for the management of pain. PMID:21324591

  9. Loss of Plastoglobule Kinases ABC1K1 and ABC1K3 Causes Conditional Degreening, Modified Prenyl-Lipids, and Recruitment of the Jasmonic Acid Pathway[W

    PubMed Central

    Lundquist, Peter K.; Poliakov, Anton; Giacomelli, Lisa; Friso, Giulia; Appel, Mason; McQuinn, Ryan P.; Krasnoff, Stuart B.; Rowland, Elden; Ponnala, Lalit; Sun, Qi; van Wijk, Klaas J.

    2013-01-01

    Plastoglobules (PGs) are plastid lipid-protein particles. This study examines the function of PG-localized kinases ABC1K1 and ABC1K3 in Arabidopsis thaliana. Several lines of evidence suggested that ABC1K1 and ABC1K3 form a protein complex. Null mutants for both genes (abc1k1 and abc1k3) and the double mutant (k1 k3) displayed rapid chlorosis upon high light stress. Also, k1 k3 showed a slower, but irreversible, senescence-like phenotype during moderate light stress that was phenocopied by drought and nitrogen limitation, but not cold stress. This senescence-like phenotype involved degradation of the photosystem II core and upregulation of chlorophyll degradation. The senescence-like phenotype was independent of the EXECUTER pathway that mediates genetically controlled cell death from the chloroplast and correlated with increased levels of the singlet oxygen–derived carotenoid β-cyclocitral, a retrograde plastid signal. Total PG volume increased during light stress in wild type and k1 k3 plants, but with different size distributions. Isolated PGs from k1 k3 showed a modified prenyl-lipid composition, suggesting reduced activity of PG-localized tocopherol cyclase (VTE1), and was consistent with loss of carotenoid cleavage dioxygenase 4. Plastid jasmonate biosynthesis enzymes were recruited to the k1 k3 PGs but not wild-type PGs, while pheophytinase, which is involved in chlorophyll degradation, was induced in k1 k3 and not wild-type plants and was localized to PGs. Thus, the ABC1K1/3 complex contributes to PG function in prenyl-lipid metabolism, stress response, and thylakoid remodeling. PMID:23673981

  10. Mutations in MAP3K1 Cause 46,XY Disorders of Sex Development and Implicate a Common Signal Transduction Pathway in Human Testis Determination

    PubMed Central

    Pearlman, Alexander; Loke, Johnny; Le Caignec, Cedric; White, Stefan; Chin, Lisa; Friedman, Andrew; Warr, Nicholas; Willan, John; Brauer, David; Farmer, Charles; Brooks, Eric; Oddoux, Carole; Riley, Bridget; Shajahan, Shahin; Camerino, Giovanna; Homfray, Tessa; Crosby, Andrew H.; Couper, Jenny; David, Albert; Greenfield, Andy; Sinclair, Andrew; Ostrer, Harry

    2010-01-01

    Investigations of humans with disorders of sex development (DSDs) resulted in the discovery of many of the now-known mammalian sex-determining genes, including SRY, RSPO1, SOX9, NR5A1, WT1, NR0B1, and WNT4. Here, the locus for an autosomal sex-determining gene was mapped via linkage analysis in two families with 46,XY DSD to the long arm of chromosome 5 with a combined, multipoint parametric LOD score of 6.21. A splice-acceptor mutation (c.634-8T>A) in MAP3K1 segregated with the phenotype in the first family and disrupted RNA splicing. Mutations were demonstrated in the second family (p.Gly616Arg) and in two of 11 sporadic cases (p.Leu189Pro, p.Leu189Arg)—18% prevalence in this cohort of sporadic cases. In cultured primary lymphoblastoid cells from family 1 and the two sporadic cases, these mutations altered the phosphorylation of the downstream targets, p38 and ERK1/2, and enhanced binding of RHOA to the MAP3K1 complex. Map3k1 within the syntenic region was expressed in the embryonic mouse gonad prior to, and after, sex determination. Thus, mutations in MAP3K1 that result in 46,XY DSD with partial or complete gonadal dysgenesis implicate this pathway in normal human sex determination. PMID:21129722

  11. Capsular polysaccharide vaccine for Group B Neisseria meningitidis, Escherichia coli K1, and Pasteurella haemolytica A2

    PubMed Central

    Robbins, John B.; Schneerson, Rachel; Xie, Guilin; Hanson, Lars Å.; Miller, Mark A.

    2011-01-01

    We reviewed the literature that is the basis for our proposal that (2→8)-α-Neu5Ac conjugates will be safe and effective vaccines for Group B meningococci (GBMs), Escherichia coli K1, and Pasteurella haemolytica A2. Although (2→8)-α-Neu5Ac is a virulence factor and a protective antigen of these three pathogens, it is also a component of normal tissues (neural cell adhesion molecule). Natural, anti–(2→8)-α-Neu5Ac present in most adults, vaccine-induced antibodies, and even high levels of spontaneously appearing monoclonal anti–(2→8)-α-Neu5Ac did not cause autoimmunity. Although it is not possible to prove a null hypothesis, there are no epidemiologic, serologic, immunologic, or clinical data to indicate that (2→8)-α-Neu5Ac antibodies will induce pathology or an autoimmune disease. No increased pathology caused by these antibodies was found, even in neonates and infants of mothers recovered from GBM meningitis. The lack of pathology mediated by anti–(2→8)-α-Neu5Ac may be explained by different presentations of (2→8)-α-Neu5Ac on bacterial and mammalian cells and by the unusual physicochemical properties of anti–(2→8)-α-Neu5Ac. Based on clinical and experimental data collected over 30 y and because (2→8)-α-Neu5Ac is an essential virulence factor and a protective antigen for GBM, E. coli K1, and P. haemolytica A2, protein conjugates of it are easy to prepare using inexpensive and plentiful ingredients, and they would be compatible with routinely administered infant vaccines, clinical studies of these conjugates should proceed. PMID:22025709

  12. Photogenerated radical intermediates of vitamin K 1: a time-resolved resonance Raman study

    NASA Astrophysics Data System (ADS)

    Balakrishnan, G.; Umapathy, S.

    1999-01-01

    Quinones play a vital role in the process of electron transfer in bacterial photosynthetic reaction centers. It is of interest to investigate the photochemical reactions involving quinones with a view to elucidating the structure-function relationships in the biological processes. Resonance Raman spectra of radical anions and the time-resolved resonance Raman spectra of vitamin K 1 (model compound for Q A in Rhodopseudomonas viridis, a bacterial photosynthetic reception center) are presented. The photochemical intermediates of vitamin K 1, viz. radical anion, ketyl radical and o-quinone methide have been identified. The vibrational assignments of all these intermediates are made on the basis of comparison with our earlier TR3 studies on radical anions of naphthoquinone and menaquinone.

  13. Cortical GluK1 kainate receptors modulate scratching in adult mice.

    PubMed

    Descalzi, Giannina; Chen, Tao; Koga, Kohei; Li, Xiang-Yao; Yamada, Kaori; Zhuo, Min

    2013-09-01

    Recent investigations into the mechanisms mediating itch transmission have focused on spinal mechanisms, whereas few studies have investigated the role of the cerebral cortex in itch-related behaviors. Human imaging studies show that several cortical regions are active in correspondence with itch, including the anterior cingulate cortex (ACC). We present here evidence of cortical modulation of pruritogen-induced scratching behavior. We combine pharmacological, genetic, and electrophysiological approaches to show that cortical GluK1-containing kainate (KA) receptors are involved in scratching induced by histamine and non-histamine-dependent itching stimuli. We further show that scratching corresponds with enhanced excitatory transmission in the ACC through KA receptor modulation of inhibitory circuitry. In addition, we found that inhibiting GluK1-containing KA receptors in the ACC also reduced behavioral nociceptive responses induced by formalin. Our results reveal a new role of the cortex in pruritogen-induced scratching. PMID:23786569

  14. Particle motion in generalized Dirac's monopoles of dimension 2k + 1

    NASA Astrophysics Data System (ADS)

    Bai, Zhanqiang

    2016-08-01

    By using Meng's idea in his generalization of the classical MICZ-Kepler problem, we obtained the equations of motion of a charged particle in the field of generalized Dirac monopole in odd dimensional Euclidean spaces. The main result is that for every particle trajectory r : I → ℝ2k+1∖{0}, there is a 2-dimensional cone with vertex at the origin on which r is a geodesic.

  15. Final report on RMO Vickers key comparison COOMET M.H-K1

    NASA Astrophysics Data System (ADS)

    Aslanyan, E.; Menelao, F.; Herrmann, K.; Aslanyan, A.; Pivovarov, V.; Galat, E.; Dovzhenko, Y.; Zhamanbalin, M.

    2013-01-01

    This report describes a COOMET key comparison on Vickers hardness scales involving five National Metrology Institutes: PTB (Germany), BelGIM (Belarus), NSC IM (Ukraine), KazInMetr (Kazakhstan) and VNIIFTRI (Russia). The pilot laboratory was VNIIFTRI, and PTB acted as the linking institute to key comparisons CCM.H-K1.b and CCM.H-K1.c conducted for the Vickers hardness scales HV1 and HV30, respectively. The comparison was also conducted for the HV5 Vickers hardness scale, since this scale is most frequently used in practice in Russia and CIS countries that work according to GOST standards. In the key comparison, two sets of hardness reference blocks for the Vickers hardness scales HV1, HV5 and HV30 consisting each of three hardness reference blocks with hardness levels of 450 HV and 750 HV were used. The measurement results and uncertainty assessments for HV1 and HV30 hardness scales, as announced by BelGIM, NSC IM, KazInMetr and VNIIFTRI, are in good agreement with the key comparison reference values of CCM.H-K1.b and CCM.H-K1.c. The comparison results for the HV5 hardness scale are viewed as additional information, since up to today no CCM key comparisons on this scale have yet been carried out. 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 (CIPM MRA).

  16. Volatile Composition of Comet C/2012 K1 (PanSTARRS)

    NASA Astrophysics Data System (ADS)

    Roth, Nathan; Gibb, Erika; Bonev, Boncho P.; DiSanti, Michael A.; Villanueva, Geronimo L.; Paganini, Lucas; Mumma, Michael J.

    2015-11-01

    On 2014 May 22 and 24 we characterized the volatile composition of the dynamically new Oort Cloud comet C/2012 K1 (PanSTARRS) using the long-slit, high resolution (λ/Δλ ≈ 25,000) infared echelle spectrograph (NIRSPEC) at the 10m Keck 2 telescope on Maunakea, HI. We detected fluorescent emission from six primary species (H2O, HCN, CH4, C2H6, CH3OH, and CO) and prompt emission from one product species (OH* - a directory proxy for H­2O). Upper limits were derived for C2H2 and H2CO. We report rotational temperatures, production rates, and mixing ratios (relative to water). Based on the inventory of comets characterized to date, mixing ratios of trace gases in C/2012 K1 (PanSTARRS) are about normal - CH3OH and C2H6 are slightly enriched, CO, CH4, HCN, and H2CO are average, and C­2H2 is depleted. I will discuss C/2012 K1 (PanSTARRS) in the context of an emerging taxonomy for comets based on volatile composition.This work is supported through the NASA Missouri Space Grant Consortium and the National Science Foundation (NSF 1211362), the NASA Astrobiology Institute through a grant to the Goddard Center for Astrobiology (811073.02.12.03.91), and the NASA Planetary Astronomy Program (811073.02.03.03.43).

  17. Plasma superwarfarin levels and vitamin K1 treatment in dogs with anticoagulant rodenticide poisoning.

    PubMed

    Robben, J H; Kuijpers, E A; Mout, H C

    1998-01-01

    The plasma concentration, plasma half-life (t1/2), and mean residence time (MRT) of rodenticide anticoagulants were determined in 21 dogs in which a preliminary diagnosis of anticoagulant rodenticide poisoning had been made. Brodifacoum, difethialone, and difenacoum were detected by high-performance liquid chromatography (HPLC) in the plasma of 13, 3, and 2 dogs, respectively. At presentation the plasma concentration ranged from below the detection limit (10 ng/L) to 851 ng/L. Toxin could not be detected in 3 dogs, despite these animals showing characteristic coagulation disturbances and a positive response to therapy with vitamin K1. In 7 dogs the estimated t1/2 of brodifacoum ranged from 0.9 to 4.7 (median 2.4) days with a MRT of 1.9 to 3.7 (median 2.8) days. In 2 dogs the individual t1/2 of difethialone was 2.2 and 3.2 days and the MRT was 2.3 and 2.8 days, respectively. Two dogs died during emergency treatment. Treatment in the remaining 19 dogs consisted of the administration of vitamin K1 and supportive therapy. The dose of vitamin K1 was reduced in a stepwise manner as long as the prothrombin time remained within physiological limits. The variation in initial plasma concentrations of the anticoagulants combined with the results of treatment support the idea that an individual therapeutic approach is warranted. PMID:9477532

  18. [Role of mitochondrial alternative oxidase (AOX) pathway in photoprotection in Rumex K-1 leaves].

    PubMed

    Meng, Xiang-Long; Zhang, Li-Tao; Zhang, Zi-Shan; Gao, Hui-Yuan; Meng, Qing-Wei

    2012-07-01

    Taking Rumex K-1 leaves as test materials, this paper studied the role of mitochondrial alternative oxidase (AOX) pathway in photoprotection under different light intensities. Under low light intensity (200 micromol x m(-2) x s(-1)), and after treated with salicylhydroxamic acid to inhibit the AOX pathway, the leaf actual photochemical efficiency of PS II, linear electron transport rate of photosynthesis, and photosynthetic O2 evolution rate all decreased significantly while the non-Q(B) reducing reaction center had a significant increase, indicating that under low light, the photoinhibition was aggravated while the scavenging enzymes of reactive oxygen species (ROS) increased, which avoided the over-accumulation of ROS and partially alleviated the photoinhibition of Rumex K-1 leaves. Under high light intensity (800 micromol x m(-2) x s(-1)), the inhibition of AOX pathway caused more severe photoinhibition, and the increased activities of ROS scavenging enzymes were insufficient to prevent the over-accumulation of ROS. This study demonstrated that AOX pathway played an important role in the photoprotection in Rumex K-1 leaves under both high and low light intensities, and the role of AOX pathway in photoprotection under high light could be irreplaceable by the other photoprotection pathways in chloroplast.

  19. Piezoelectricity in K1-xNaxNbO3: First-principles calculation

    NASA Astrophysics Data System (ADS)

    Li, Qiang; Rui, Zhang; Lv, Tian-Quan; Zheng, Li-Mei

    2015-05-01

    The piezoelectric properties of K1-xNaxNbO3 are studied by using first-principles calculations within virtual crystal approximation. To understand the critical factors for the high piezoelectric response in K1-xNaxNbO3, the total energy, piezoelectric coefficient, elastic property, density of state, Born effective charge, and energy barrier on polarization rotation paths are systematically investigated. The morphotropic phase boundary in K1-xNaxNbO3 is predicted to occur at x = 0.521, which is in good agreement with the available experimental data. At the morphotropic phase boundary, the longitudinal piezoelectric coefficient d33 of orthorhombic K0.5Na0.5NbO3 reaches a maximum value. The rotated maximum of is found to be along the 50° direction away from the spontaneous polarization (close to the [001] direction). The moderate bulk and shear modulus are conducive to improving the piezoelectric response. By analyzing the energy barrier on polarization rotation paths, it is found that the polarization rotation of orthorhombic K0.5Na0.5NbO3 becomes easier compared with orthorhombic KNbO3, which proves that the high piezoelectric response is attributed to the flattening of the free energy at compositions close to the morphotropic phase boundary. Project supported by the National Basic Research Program of China (Grant No. 2013CB632900).

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

    NASA Astrophysics Data System (ADS)

    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.

  1. RNA polymerase activity in PtK1 micronuclei containing individual chromosomes: an in vitro and in situ study

    SciTech Connect

    Labidi, B.; Gregoire, M.; Frackowiak, S.; Hernandez-Verdun, D.; Bouteille, M.

    1987-03-01

    Micronuclei have been induced by colchicine in rat kangaroo (Potorous tridactylis) PtK1 cells. The synthesis of RNA was investigated both in isolated micronuclei by quantifying RNA polymerase activities at different ionic strengths with or without inhibitors, and in micronucleated cells by radioautography after (/sup 3/H)uridine pulse labeling. In vitro transcription shows that isolated micronuclei are able to take up (/sup 3/H)UTP. The rate curves of incorporation are close to those of isolated diploid nuclei, though the level of incorporation was relatively lower (65-70%) than control nuclei. This indicates that micronuclei react to the ionic environment and to inhibitors in the same manner as described for many species of isolated diploid nuclei. The labelling distributions plotted from radioautographs show that micronuclei were able to efficiently incorporate the hot precursor. Furthermore, for short pulses there is no homogeneity in the labelling density among the different micronuclei and there is no correlation between the labelling intensity and the size of micronuclei. After 60-min pulse time, there is an enhanced uptake of (/sup 3/H)uridine and all the micronuclei exhibit considerable labelling, although less than control cells. Thus, the micronuclei exhibit some characteristic RNA transcriptional activity in situ as well as after isolation. This material should be a particular interesting model with which to study the physiological activity and the role of each individual interphasic chromosome.

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

    PubMed

    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.

  3. EUV resolution enhancement techniques (RETs) for k1 0.4 and below

    NASA Astrophysics Data System (ADS)

    Hsu, Stephen; Howell, Rafael; Jia, Jianjun; Liu, Hua-Yu; Gronlund, Keith; Hansen, Steve; Zimmermann, Jörg

    2015-03-01

    Due to the exponential growth of mobile wireless devices, low-power logic chips continue to drive device scaling. To enable sub-10 nm device scaling at an affordable cost, there is a strong need for single exposure advanced lithography. Extreme ultraviolet lithography (EUVL) is one of the most promising candidates to support the design rules for sub-10 nm. The aggressive mobile device design rules continue to push the critical dimension (CD) and pitch and put very stringent demands on the lithography performance such as pattern placement control, image contrast, critical dimension uniformity (CDU), and line width roughness (LWR). In this paper we report the latest advances in resolution enhancement techniques to address low k1 challenges in EUV lithography, specifically: minimizing the pattern placement error, enhancing the through-focus contrast, and reducing the impact of stochastic effects. We have developed an innovative source-mask optimization (SMO) method to significantly reduce edge placement errors (EPE) [1] [2]. Aggressive design rules using the state-of-the-art NA of 0.33 of the NXE:3300B and its successor tools can have imaging below k1 = 0.4, which can extend the current process capabilities for single exposure high volume manufacturing (HVM). Burkhardt et al. reported in a previous study that inserting a sub-resolution assist feature (SRAF) within semi-isolated features introduces strong Bossung tilts and best focus shifts, and a general solution for random pitches is not apparent [3]. Kang observed the same issues and proposed to introduce spherical aberrations to correct these effects while having a global impact on the full-chip [4]. In this work we introduce a new methodology to apply SRAFs to improve contrast, reduce best focus shift, and improve process window. Finally, the lower number of photons of EUV and the small feature size brings serious issue of the stochastic effect that causes the line-edge-roughness (LER) and local CD uniformity

  4. Adenoviral delivery of truncated MMP-8 fused with the hepatocyte growth factor mutant 1K1 ameliorates liver cirrhosis and promotes hepatocyte proliferation

    PubMed Central

    Liu, Jinghua; Li, Jianbo; Fu, Weiwei; Tang, Jiacheng; Feng, Xu; Chen, Jiang; Liang, Yuelong; Jin, Ren’an; Xie, Anyong; Cai, Xiujun

    2015-01-01

    Liver cirrhosis is a chronic liver disease caused by chronic liver injury, which activates hepatic stellate cells (HSCs) and the secretion of extracellular matrix (ECM). Cirrhosis accounts for an extensive level of morbidity and mortality worldwide, largely due to lack of effective treatment options. In this study, we have constructed a fusion protein containing matrix metal-loproteinase 8 (MMP-8) and the human growth factor mutant 1K1 (designated cMMP8-1K1) and delivered it into hepatocytes and in vivo and in cell culture via intravenous injection of fusion protein-harboring adenovirus. In doing so, we found that the cMMP8-1K1 fusion protein promotes the proliferation of hepatocytes, likely resulting from the combined inhibition of type I collagen secretion and the degradation of the ECM in the HSCs. This fusion protein was also observed to ameliorate liver cirrhosis in our mouse model. These changes appear to be linked to changes in downstream gene expression. Taken together, these results suggest a possible strategy for the treatment of liver cirrhosis and additional work is warranted. PMID:26527860

  5. Escherichia coli K1 Modulates Peroxisome Proliferator-Activated Receptor γ and Glucose Transporter 1 at the Blood-Brain Barrier in Neonatal Meningitis.

    PubMed

    Krishnan, Subramanian; Chang, Alexander C; Stoltz, Brian M; Prasadarao, Nemani V

    2016-10-01

    Escherichia coli K1 meningitis continues to be a major threat to neonatal health. Previous studies demonstrated that outer membrane protein A (OmpA) of E. coli K1 interacts with endothelial cell glycoprotein 96 (Ecgp96) in the blood-brain barrier to enter the central nervous system. Here we show that the interaction between OmpA and Ecgp96 downregulates peroxisome proliferator-activated receptor γ (PPAR-γ) and glucose transporter 1 (GLUT-1) levels in human brain microvascular endothelial cells, causing disruption of barrier integrity and inhibition of glucose uptake. The suppression of PPAR-γ and GLUT-1 by the bacteria in the brain microvessels of newborn mice causes extensive pathophysiology owing to interleukin 6 production. Pretreatment with partial or selective PPAR-γ agonists ameliorate the pathological outcomes of infection by suppressing interleukin 6 production in the brain. Thus, inhibition of PPAR-γ and GLUT-1 by E. coli K1 is a novel pathogenic mechanism in meningitis, and pharmacological upregulation of PPAR-γ and GLUT-1 levels may provide novel therapeutic avenues. PMID:27456707

  6. Adenoviral delivery of truncated MMP-8 fused with the hepatocyte growth factor mutant 1K1 ameliorates liver cirrhosis and promotes hepatocyte proliferation.

    PubMed

    Liu, Jinghua; Li, Jianbo; Fu, Weiwei; Tang, Jiacheng; Feng, Xu; Chen, Jiang; Liang, Yuelong; Jin, Ren'an; Xie, Anyong; Cai, Xiujun

    2015-01-01

    Liver cirrhosis is a chronic liver disease caused by chronic liver injury, which activates hepatic stellate cells (HSCs) and the secretion of extracellular matrix (ECM). Cirrhosis accounts for an extensive level of morbidity and mortality worldwide, largely due to lack of effective treatment options. In this study, we have constructed a fusion protein containing matrix metal-loproteinase 8 (MMP-8) and the human growth factor mutant 1K1 (designated cMMP8-1K1) and delivered it into hepatocytes and in vivo and in cell culture via intravenous injection of fusion protein-harboring adenovirus. In doing so, we found that the cMMP8-1K1 fusion protein promotes the proliferation of hepatocytes, likely resulting from the combined inhibition of type I collagen secretion and the degradation of the ECM in the HSCs. This fusion protein was also observed to ameliorate liver cirrhosis in our mouse model. These changes appear to be linked to changes in downstream gene expression. Taken together, these results suggest a possible strategy for the treatment of liver cirrhosis and additional work is warranted.

  7. Phenotypic Heterogeneity in Expression of the K1 Polysaccharide Capsule of Uropathogenic Escherichia coli and Downregulation of the Capsule Genes during Growth in Urine.

    PubMed

    King, Jane E; Aal Owaif, Hasan A; Jia, Jia; Roberts, Ian S

    2015-07-01

    Uropathogenic Escherichia coli (UPEC) is the major causative agent of uncomplicated urinary tract infections (UTI). The K1 capsule on the surface of UPEC strains is a key virulence factor, and its expression may be important in the onset and progression of UTI. In order to understand capsule expression in more detail, we analyzed its expression in the UPEC strain UTI89 during growth in rich medium (LB medium) and urine and during infection of a bladder epithelial cell line. Comparison of capsule gene transcription using a chromosomal gfp reporter fusion showed a significant reduction in transcription during growth in urine compared to that during growth in LB medium. When examined at the single-cell level, following growth in both media, capsule gene expression appears to be heterogeneous, with two distinct green fluorescent protein (GFP)-expressing populations. Using anti-K1 antibody, we showed that this heterogeneity in gene expression results in two populations of encapsulated and unencapsulated cells. We demonstrated that the capsule hinders attachment to and invasion of epithelial cells and that the unencapsulated cells within the population preferentially adhere to and invade bladder epithelial cells. We found that once internalized, UTI89 starts to produce capsule to aid in its intracellular survival and spread. We propose that this observed phenotypic diversity in capsule expression is a fitness strategy used by the bacterium to deal with the constantly changing environment of the urinary tract.

  8. Phenotypic Heterogeneity in Expression of the K1 Polysaccharide Capsule of Uropathogenic Escherichia coli and Downregulation of the Capsule Genes during Growth in Urine

    PubMed Central

    King, Jane E.; Aal Owaif, Hasan A.; Jia, Jia

    2015-01-01

    Uropathogenic Escherichia coli (UPEC) is the major causative agent of uncomplicated urinary tract infections (UTI). The K1 capsule on the surface of UPEC strains is a key virulence factor, and its expression may be important in the onset and progression of UTI. In order to understand capsule expression in more detail, we analyzed its expression in the UPEC strain UTI89 during growth in rich medium (LB medium) and urine and during infection of a bladder epithelial cell line. Comparison of capsule gene transcription using a chromosomal gfp reporter fusion showed a significant reduction in transcription during growth in urine compared to that during growth in LB medium. When examined at the single-cell level, following growth in both media, capsule gene expression appears to be heterogeneous, with two distinct green fluorescent protein (GFP)-expressing populations. Using anti-K1 antibody, we showed that this heterogeneity in gene expression results in two populations of encapsulated and unencapsulated cells. We demonstrated that the capsule hinders attachment to and invasion of epithelial cells and that the unencapsulated cells within the population preferentially adhere to and invade bladder epithelial cells. We found that once internalized, UTI89 starts to produce capsule to aid in its intracellular survival and spread. We propose that this observed phenotypic diversity in capsule expression is a fitness strategy used by the bacterium to deal with the constantly changing environment of the urinary tract. PMID:25870229

  9. Draft Genome Sequence of Thermus scotoductus Strain K1, Isolated from a Geothermal Spring in Karvachar, Nagorno Karabakh.

    PubMed

    Saghatelyan, Ani; Poghosyan, Lianna; Panosyan, Hovik; Birkeland, Nils-Kåre

    2015-01-01

    The 2,379,636-bp draft genome sequence of Thermus scotoductus strain K1, isolated from geothermal spring outlet located in the Karvachar region in Nagorno Karabakh is presented. Strain K1 shares about 80% genome sequence similarity with T. scotoductus strain SA-01, recovered from a deep gold mine in South Africa. PMID:26564055

  10. Draft Genome Sequence of Thermus scotoductus Strain K1, Isolated from a Geothermal Spring in Karvachar, Nagorno Karabakh

    PubMed Central

    Saghatelyan, Ani; Poghosyan, Lianna

    2015-01-01

    The 2,379,636-bp draft genome sequence of Thermus scotoductus strain K1, isolated from geothermal spring outlet located in the Karvachar region in Nagorno Karabakh is presented. Strain K1 shares about 80% genome sequence similarity with T. scotoductus strain SA-01, recovered from a deep gold mine in South Africa. PMID:26564055

  11. 75 FR 10018 - Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and K-1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-04

    ... Internal Revenue Service Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and... Form 1041 and related Schedules D, J, and K-1, U.S. Income Tax Return for Estates and Trusts. DATES... Trusts (Schedule J), and Beneficiary's Share of Income, Deductions, Credits, etc. (Schedule K-1)....

  12. OPC model calibration for CPL patterning at extreme low k1

    NASA Astrophysics Data System (ADS)

    Shi, Xuelong; Laidig, Tom; Chen, J. Fung; Van Den Broeke, Douglas; Hsu, Stephen; Hsu, Michael; Wampler, Kurt E.; Hollerbach, Uwe; Park, Jung Chul; Yu, Linda

    2004-12-01

    Model based optical proximity correction (OPC) to enhance image fidelity and process robustness has become one of the most critical components that enable the low k1 optical lithography. To meet the challenges imposed by the previously unthinkable low k1 for manufacturing with most stringent dimension control requirements, a capable OPC model to meet such an aggressive lithography challenges has been urgently called upon. In addition to providing better accuracy for the currently implemented process technologies, the new OPC model must work well with Chromeless Phase Lithography (CPL) in which the topography on the mask is rather significant, and Double Dipole Lithography (DDL) in which two masks and two exposures are needed. It must also be able to intelligently take into account the effect from the more aggressive illuminations, usch as customer designed illuminator and experimental measured illuminator profile from the scanners. The physical and mathematical foundation of the model must be well thought of to meet the requirements for the above-mentioned applications. We have extended our Eigen Decomposition Model (EDM) for model OPC treatment into the high NA regime, in which the vector characteristics of light and thin film stack are taken into account. For CPL calibration, it has been found that 3D mask topography effect cannot be ignored in order to achieve satisfactory model accuracy.

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

    PubMed

    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. PMID:18999657

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

    PubMed

    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.

  15. Effects of lunar and mars dust simulants on HaCaT keratinocytes and CHO-K1 fibroblasts

    NASA Astrophysics Data System (ADS)

    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

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

    PubMed

    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.

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

    PubMed

    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.

  18. 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

    NASA Astrophysics Data System (ADS)

    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.

  19. 75 FR 42831 - Proposed Collection; Comment Request for Form 1065, Schedule C, Schedule D, Schedule K-1...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-22

    ... Items), Schedule L (Balance Sheets per Books), Schedule M-1 (Reconciliation of Income (Loss) per Books.... (Schedule K-1), Balance Sheets per Books (Schedule L), Reconciliation of Income (Loss) per Books With...

  20. Role of K1 capsule antigen in cirrhotic patients with Escherichia coli spontaneous bacterial peritonitis in southern Taiwan.

    PubMed

    Wang, M C; Lin, W H; Tseng, C C; Wu, A B; Teng, C H; Yan, J J; Wu, J J

    2013-03-01

    Spontaneous bacterial peritonitis (SBP) is one of the most serious complications in patients with cirrhosis. This study aimed to investigate the prevalence of SBP caused by Escherichia coli isolates with or without the K1 capsule antigen in cirrhotic patients and the outcome. From January 2004 to January 2012, a total of 54 and 41 E. coli strains derived from patients with SBP and intestinal perforation (IP), respectively, were included for comparison in this study. Bacterial characteristics including phylogenetic groups, K1 capsule antigen, and 14 virulence factor genetic determinants, as well as data regarding patient characteristics, clinical manifestations, and in-hospital deaths, were collected and analyzed. The prevalence of the K1 capsule antigen gene neuA was more common in SBP isolates compared to IP isolates (28 % vs. 10 %, p = 0.0385). Phylogenetic groups B2 and group D were dominant in E. coli isolates with and without the K1 capsule antigen, respectively. The prevalence of virulence factors genes papG II, ompT, and usp was higher in E. coli K1 strains. There were 26 deaths (48 %) during hospitalization. Presence of the K1 capsule antigen in E. coli isolates was significantly associated with in-hospital death in cirrhotic patients with SBP (42 % vs. 14 %, p = 0.0331). This study demonstrates a higher prevalence of the K1 capsule antigen in E. coli SBP compared to E. coli peritonitis caused by IP. There were significant associations between the K1 capsule antigen and in-hospital mortality and bacterial virulence in cirrhotic patients with E. coli SBP.

  1. Molecular characterization and expression profile of MAP2K1ip1/MP1 gene from tiger shrimp, Penaeus monodon.

    PubMed

    Yang, Lishi; Liu, Xianjun; Huang, Jianhua; Yang, Qibin; Qiu, Lihua; Liu, Wenjing; Jiang, Shigui

    2012-05-01

    MAPK kinase 1 interacting protein 1 (MAP2K1ip1) is an important scaffold proteins of the mitogen-activated protein kinase (MAPK) pathway that form an active signaling module and enhance the specificity and spatiality of MAPK signaling. In the present study, we identified and characterized a MAP2K1ip1 cDNA from tiger shrimp Penaeus monodon (designated as PmMAP2K1ip1). The open reading frame of PmMAP2K1ip1 is 372 bp encoding 123 amino-acid residues with a MAPK interaction domain. The predicted PmMAP2Kip1 protein is 13.6 KDa with the theoretical isoelectric point of 6.3. PmMAP2K1ip1 shared the highest amino acid with Nasonia vitripennis and Strongylocentrotus purpuratus, at 48% and 47.5%, respectively. Phylogenic analysis shows PmMAP2Kip1 is clustering with SpMAP2Kip1, and close to the group of MAP2Kip1s from insect. Furthermore, semiquantitative RT-PCR revealed PmMAP2Kip1 is widely distributed in most examined tissues except nerve, and high expressed in ovary, hemocyte, intestines and hepatopancreas. Meanwhile, PmMAP2k1ip1 is expressed ubiquitously during larval and sex gland development, and keep a high level at the initial development stage. Quantitative real time RT-PCR revealed PmMAP2K1ip1 were up-regulated by lipopolysaccharide and peptidoglycan (PGN) in haemocyte. These data reveal MAP2K1ip1 is a multifunction protein that involved development and immune response. It is benefit to characterize other MAPK signal genes and elucidate the molecular regulation mechanism of MAPK signaling in tiger shrimp. PMID:22209950

  2. Molecular characterization and expression profile of MAP2K1ip1/MP1 gene from tiger shrimp, Penaeus monodon.

    PubMed

    Yang, Lishi; Liu, Xianjun; Huang, Jianhua; Yang, Qibin; Qiu, Lihua; Liu, Wenjing; Jiang, Shigui

    2012-05-01

    MAPK kinase 1 interacting protein 1 (MAP2K1ip1) is an important scaffold proteins of the mitogen-activated protein kinase (MAPK) pathway that form an active signaling module and enhance the specificity and spatiality of MAPK signaling. In the present study, we identified and characterized a MAP2K1ip1 cDNA from tiger shrimp Penaeus monodon (designated as PmMAP2K1ip1). The open reading frame of PmMAP2K1ip1 is 372 bp encoding 123 amino-acid residues with a MAPK interaction domain. The predicted PmMAP2Kip1 protein is 13.6 KDa with the theoretical isoelectric point of 6.3. PmMAP2K1ip1 shared the highest amino acid with Nasonia vitripennis and Strongylocentrotus purpuratus, at 48% and 47.5%, respectively. Phylogenic analysis shows PmMAP2Kip1 is clustering with SpMAP2Kip1, and close to the group of MAP2Kip1s from insect. Furthermore, semiquantitative RT-PCR revealed PmMAP2Kip1 is widely distributed in most examined tissues except nerve, and high expressed in ovary, hemocyte, intestines and hepatopancreas. Meanwhile, PmMAP2k1ip1 is expressed ubiquitously during larval and sex gland development, and keep a high level at the initial development stage. Quantitative real time RT-PCR revealed PmMAP2K1ip1 were up-regulated by lipopolysaccharide and peptidoglycan (PGN) in haemocyte. These data reveal MAP2K1ip1 is a multifunction protein that involved development and immune response. It is benefit to characterize other MAPK signal genes and elucidate the molecular regulation mechanism of MAPK signaling in tiger shrimp.

  3. How to obtain accurate resist simulations in very low-k1 era?

    NASA Astrophysics Data System (ADS)

    Chiou, Tsann-Bim; Park, Chan-Ha; Choi, Jae-Seung; Min, Young-Hong; Hansen, Steve; Tseng, Shih-En; Chen, Alek C.; Yim, Donggyu

    2006-03-01

    A procedure for calibrating a resist model iteratively adjusts appropriate parameters until the simulations of the model match the experimental data. The tunable parameters may include the shape of the illuminator, the geometry and transmittance/phase of the mask, light source and scanner-related parameters that affect imaging quality, resist process control and most importantly the physical/chemical factors in the resist model. The resist model can be accurately calibrated by measuring critical dimensions (CD) of a focus-exposure matrix (FEM) and the technique has been demonstrated to be very successful in predicting lithographic performance. However, resist model calibration is more challenging in the low k1 (<0.3) regime because numerous uncertainties, such as mask and resist CD metrology errors, are becoming too large to be ignored. This study demonstrates a resist model calibration procedure for a 0.29 k1 process using a 6% halftone mask containing 2D brickwall patterns. The influence of different scanning electron microscopes (SEM) and their wafer metrology signal analysis algorithms on the accuracy of the resist model is evaluated. As an example of the metrology issue of the resist pattern, the treatment of a sidewall angle is demonstrated for the resist line ends where the contrast is relatively low. Additionally, the mask optical proximity correction (OPC) and corner rounding are considered in the calibration procedure that is based on captured SEM images. Accordingly, the average root-mean-square (RMS) error, which is the difference between simulated and experimental CDs, can be improved by considering the metrological issues. Moreover, a weighting method and a measured CD tolerance are proposed to handle the different CD variations of the various edge points of the wafer resist pattern. After the weighting method is implemented and the CD selection criteria applied, the RMS error can be further suppressed. Therefore, the resist CD and process window can

  4. The mitochondrial alternative oxidase pathway protects the photosynthetic apparatus against photodamage in Rumex K-1 leaves

    PubMed Central

    2012-01-01

    Background It is known that excess reducing equivalents in the form of NADPH in chloroplasts can be transported via shuttle machineries, such as the malate-oxaloacetate (OAA) shuttle, into the mitochondria, where they are efficiently oxidised by the mitochondrial alternative oxidase (AOX) respiratory pathway. Therefore, it has been speculated that the AOX pathway may protect plants from photoinhibition, but the mechanism by which this protection occurs remains to be elucidated. Results The observation that the malate-OAA shuttle activity and the AOX pathway capacity increased markedly after intense light treatment in Rumex K-1 leaves indicates that excess NADPH was transported from the chloroplasts and oxidised by the AOX pathway. The inhibition of the AOX pathway by salicylhydroxamic acid (SHAM) caused the over-reduction of the photosystem I (PSI) acceptor side, as indicated by the increases in the extent of reduction of P700+. Furthermore, the photosynthetic linear electron flow was restricted, which was indicated by the decreases in the PSII electron transport rate (ETR) and the photosynthetic O2 evolution rate. The restriction of the photosynthetic linear electron flow, which generates the thylakoid ΔpH, inevitably decreased the de-epoxidation of the xanthophyll cycle (ΔPRI). Therefore, the induction of non-photochemical quenching (NPQ) was suppressed when the AOX pathway was inhibited. The effect of the inhibition of the AOX pathway on NPQ induction was less at 20 mM NaHCO3 than at 1 mM NaHCO3. The suppression of NPQ induction by the inhibition of the AOX pathway was also observed during the induction phase of photosynthesis. In addition, the inhibition of the AOX pathway increased the accumulation of hydrogen peroxide (H2O2), suggesting that the AOX pathway functions as an antioxidant mechanism. Conclusions The inhibition of the AOX pathway resulted in the rapid accumulation of NADPH in the chloroplasts, which caused the over-reduction of the PSI acceptor

  5. Resolution of structure of PIP5K1A reveals molecular mechanism for its regulation by dimerization and dishevelled

    PubMed Central

    Hu, Jian; Yuan, Qianying; Kang, Xue; Qin, Yuanbo; Li, Lin; Ha, Ya; Wu, Dianqing

    2015-01-01

    Type I phosphatidylinositol phosphate kinase (PIP5K1) phosphorylates the head group of phosphatidylinositol 4-phosphate (PtdIns4P) to generate PtdIns4,5P2, which plays important roles in a wide range of cellular functions including Wnt signalling. However, the lack of its structural information has hindered the understanding of its regulation. Here we report the crystal structure of the catalytic domain of zebrafish PIP5K1A at 3.3 Å resolution. This molecule forms a side-to-side dimer. Mutagenesis study of PIP5K1A reveals two adjacent interfaces for the dimerization and interaction with the DIX domain of the Wnt signalling molecule dishevelled. Although these interfaces are located distally to the catalytic/substrate-binding site, binding to these interfaces either through dimerization or the interaction with DIX stimulates PIP5K1 catalytic activity. DIX binding additionally enhances PIP5K1 substrate binding. Thus, this study elucidates regulatory mechanisms for this lipid kinase and provides a paradigm for the understanding of PIP5K1 regulation by their interacting molecules. PMID:26365782

  6. Structure Function Studies of Vaccinia Virus Host Range Protein K1 Reveal a Novel Functional Surface for Ankyrin Repeat Proteins

    SciTech Connect

    Li, Yongchao; Meng, Xiangzhi; Xiang, Yan; Deng, Junpeng

    2010-06-15

    Poxvirus host tropism at the cellular level is regulated by virus-encoded host range proteins acting downstream of virus entry. The functioning mechanisms of most host range proteins are unclear, but many contain multiple ankyrin (ANK) repeats, a motif that is known for ligand interaction through a concave surface. We report here the crystal structure of one of the ANK repeat-containing host range proteins, the vaccinia virus K1 protein. The structure, at a resolution of 2.3 {angstrom}, showed that K1 consists entirely of ANK repeats, including seven complete ones and two incomplete ones, one each at the N and C terminus. Interestingly, Phe82 and Ser83, which were previously shown to be critical for K1's function, are solvent exposed and located on a convex surface, opposite the consensus ANK interaction surface. The importance of this convex surface was further supported by our additional mutagenesis studies. We found that K1's host range function was negatively affected by substitution of either Asn51 or Cys47 and completely abolished by substitution of both residues. Cys47 and Asn51 are also exposed on the convex surface, spatially adjacent to Phe82 and Ser83. Altogether, our data showed that K1 residues on a continuous convex ANK repeat surface are critical for the host range function, suggesting that K1 functions through ligand interaction and does so with a novel ANK interaction surface.

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

    NASA Astrophysics Data System (ADS)

    Woodward, Charles E.; Kelley, Michael S. P.; 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.

    2015-08-01

    We present pre-perihelion infrared 8-31 μm 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 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 μm 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, apeak = 0.6 μm, large. The sub-micron coma grains are dominated by amorphous carbon, with a silicate-to-carbon ratio of {0.80}-0.20+0.25. The silicate crystalline mass fraction is {0.20}-0.10+0.30, similar to with other dynamically new comets exhibiting weak 10 μm silicate features. The bolometric dust albedo of the coma dust is 0.14 ± 0.01 at a phase angle of 34.°76, and the average dust production rate, corrected to zero phase, at the epoch of our observations was Afρ ≃ 5340 cm.

  8. Role of F225 in O-phosphoserine sulfhydrylase from Aeropyrum pernix K1.

    PubMed

    Takeda, Emi; Kunimoto, Kohei; Kawai, Yoshito; Kataoka, Misumi; Ishikawa, Kazuhiko; Nakamura, Takashi

    2016-09-01

    O-Phosphoserine sulfhydrylase (OPSS) synthesizes cysteine from O-phospho-L-serine (OPS) and sulfide. We have determined the three-dimensional structures of OPSS from hyperthermophilic archaeon Aeropyrum pernix K1 (ApOPSS) in complex with aminoacrylate intermediate (AA) formed from pyridoxal 5'-phosphate with OPS or in complex with cysteine and compared them with that of ApOPSS. We found an orientational change of F225 at the active-site entrance and constructed an F225A mutant to examine its activities and AA stability and clarify the role of F225 in ApOPSS. The OPS and O-acetyl-L-serine (OAS) sulfhydrylase activities of the F225A mutant decreased by 4.2- and 15-fold compared to those of the wild-type (wt) ApOPSS, respectively. The ability of OPS and OAS to form AA also decreased by 12- and 27-fold, respectively. AA was less stable in the F225A mutant than in the wt ApOPSS. Simulated docking showed that leaving groups, such as phosphate and acetate, were oriented to the inside of the active site in the F225A mutant, whereas they were oriented to the entrance in the wt ApOPSS. These results suggest that F225 in ApOPSS plays important roles in maintaining the hydrophobic environment of AA from solvent water and in controlling the orientation of leaving groups. PMID:27377295

  9. Temperature dependence of kinetic parameters for hyperthermophilic glutamate dehydrogenase from Aeropyrum pernix K1.

    PubMed

    Bhuiya, Mohammad W; Sakuraba, Haruhiko; Ohshima, Toshihisa

    2002-04-01

    The temperature dependence of the steady-state kinetic parameters for a glutamate dehydrogenase from Aeropyrum pernix K1 was investigated. The enzyme showed a biphasic kinetic characteristic for L-glutamate and a monophasic one for NADP at 50-90 degrees C. At low concentrations of L-glutamate the Km decreased from 2.02 to 0.56 mM and the catalytic efficiency (Vmax/Km) markedly increased (4-150 micromol x mg(-1) x mM(-1)) along with the increase of temperature from 50 to 90 degrees C. At high concentrations of the substrate the Km was fairly high and approximately constant (around 225 mM), and the catalytic efficiency was low and its temperature-dependent change was small. The Km (0.039 mM) for NADP did not change with the increase of temperature. In the reductive amination, the Kms for 2-oxoglutarate (1.81 and 9.37 mM at low and high levels of ammonia, respectively) were independent on temperature, but the Kms for ammonia and NADPH rose from 86 to 185 mM and 0.050 to 0.175 mM, respectively.

  10. Scanner performance predictor and optimizer in further low-k1 lithography

    NASA Astrophysics Data System (ADS)

    Aoyama, Hajime; Nakashima, Toshiharu; Ogata, Taro; Kudo, Shintaro; Kita, Naonori; Ikeda, Junji; Matsui, Ryota; Yamamoto, Hajime; Sukegawa, Ayako; Makino, Katsushi; Murayama, Masayuki; Masaki, Kazuo; Matsuyama, Tomoyuki

    2014-03-01

    Due to the importance of errors in lithography scanners, masks, and computational lithography in low-k1 lithography, application software is used to simultaneously reduce them. We have developed "Masters" application software, which is all-inclusive term of critical dimension uniformity (CDU), optical proximity effect (OPE), overlay (OVL), lens control (LNS), tool maintenance (MNT) and source optimization for wide process window (SO), for compensation of the issues on imaging and overlay. In this paper, we describe the more accurate and comprehensive solution of OPE-Master, LNS-Master and SO-Master with functions of analysis, prediction and optimization. Since OPE-Master employed a rigorous simulation, a root cause of error in OPE matching was found out. From the analysis, we had developed an additional knob and evaluated a proof-of- concept for the improvement. Influence of thermal issues on projection optics is evaluated with a heating prediction, and an optimization with scanner knobs on an optimized source taken into account mask 3D effect for obtaining usable process window. Furthermore, we discuss a possibility of correction for reticle expansion by heating comparing calculation and measurement.

  11. Plasma disposition of vitamin K1 in relation to anticoagulant poisoning.

    PubMed Central

    Park, B K; Scott, A K; Wilson, A C; Haynes, B P; Breckenridge, A M

    1984-01-01

    The disposition of vitamin K1, after intravenous (10 mg) and oral doses (10 mg and 50 mg) was studied in six healthy male subjects. After intravenous administration, the plasma concentration-time profile was adequately fitted with an average terminal half-life of 1.7 h. After oral administration (10 mg and 50 mg) the availability of vitamin K showed marked inter-individual variation (10-63%). With the higher dose intra-individual variation was also observed. Experiments in brodifacoum-anticoagulated rabbits demonstrate that the duration of action of a pharmacological dose (10 mg/kg) is short (9 h) and that high plasma concentrations (ca 1 microgram/ml) of the vitamin are required to drive clotting factor synthesis during maximum coumarin anticoagulation. Taken collectively, these data indicate that the short duration of action of vitamin K, frequently observed in cases of coumarin poisoning, is a consequence of requirements for high vitamin K concentrations and rapid clearance of the vitamin. PMID:6508974

  12. Role of F225 in O-phosphoserine sulfhydrylase from Aeropyrum pernix K1.

    PubMed

    Takeda, Emi; Kunimoto, Kohei; Kawai, Yoshito; Kataoka, Misumi; Ishikawa, Kazuhiko; Nakamura, Takashi

    2016-09-01

    O-Phosphoserine sulfhydrylase (OPSS) synthesizes cysteine from O-phospho-L-serine (OPS) and sulfide. We have determined the three-dimensional structures of OPSS from hyperthermophilic archaeon Aeropyrum pernix K1 (ApOPSS) in complex with aminoacrylate intermediate (AA) formed from pyridoxal 5'-phosphate with OPS or in complex with cysteine and compared them with that of ApOPSS. We found an orientational change of F225 at the active-site entrance and constructed an F225A mutant to examine its activities and AA stability and clarify the role of F225 in ApOPSS. The OPS and O-acetyl-L-serine (OAS) sulfhydrylase activities of the F225A mutant decreased by 4.2- and 15-fold compared to those of the wild-type (wt) ApOPSS, respectively. The ability of OPS and OAS to form AA also decreased by 12- and 27-fold, respectively. AA was less stable in the F225A mutant than in the wt ApOPSS. Simulated docking showed that leaving groups, such as phosphate and acetate, were oriented to the inside of the active site in the F225A mutant, whereas they were oriented to the entrance in the wt ApOPSS. These results suggest that F225 in ApOPSS plays important roles in maintaining the hydrophobic environment of AA from solvent water and in controlling the orientation of leaving groups.

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

    NASA Astrophysics Data System (ADS)

    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.

  14. Could S6K1 immunopositivity be used to distinguish early and advanced stages of endometrioid endometrial adenocarcinoma?

    PubMed Central

    Gün, İsmet; Özdamar, Özkan; Küçükodacı, Zafer; Muhçu, Murat; Demirel, Dilaver

    2016-01-01

    Objective To assess whether the immunopositivity of S6K1, a crucial effector of the mTOR signaling pathway, varies between early-stage low-grade and advanced-stage high-grade endometrial endometrioid adenocarcinoma (EEA) as well as to discuss its prognostic significance. Material and Methods A total of 22 normal endometrial tissue samples (Control group) and 41 EEA specimens (Study group) were enrolled in the study, and all the samples underwent immunohistochemical staining for S6 kinase alpha (S6K1). The study group was further evaluated in two subgroups; stage 1A, grade 1 (Group 1) and stage ≥1A, grade 2 or 3 (Group 2). Group 2 patients were considered as a poor prognosis for EEA. The samples were examined by two independent pathologists. Statistical analyses were performed using the Student’s t-test for continuous variables, the Chi-square test for categorical variables, and one-way analysis of variance for the comparison of multiple variables. Results The immunopositivity rate for all the included EEA patients was 56.1%, whereas none of the 22 normal endometrial tissue samples revealed immunoreactivity for S6K1. The immunopositivity rates were significantly different between Groups 1 and 2 [38.1% (8/21) and 75.0% (15/20), respectively, p=0.039]. When S6K1 positivity was used as a criterion of poor prognosis in EEA, the sensitivity, specificity, positive predictive value, and negative predictive value were calculated to be 62%, 75%, 72%, and 65%, respectively (OR: 4.9 and 95% CI: 1.3–18.7). Conclusion S6K1 was positive in the majority of EEAs and malignancies at an advanced stage. Higher grade disease had a significantly higher rate of S6K1 positivity. S6K1 immunopositivity appears to be a promising method to predict poor prognosis in EEA.

  15. Final report on the EURAMET.PR-K1.a-2009 comparison of spectral irradiance 250 nm—2500 nm

    NASA Astrophysics Data System (ADS)

    Goodman, Teresa; Servantes, William; Woolliams, Emma; Sperfeld, Peter; Simionescu, Mihai; Blattner, Peter; Källberg, Stefan; Khlevnoy, Boris; Dekker, Paul

    2015-01-01

    This report gives the results of the EURAMET.PR-K1.a-2009 comparison of spectral irradiance over the wavelength range 250 nm—2500 nm. Seven laboratories took part, including the pilot. In general the results are consistent, with a few exceptions as explained in the report. The EURAMET.PR-K1.a key comparison detailed in this report was carried out to establish the degrees of equivalence for the participating European laboratories with respect to the Key Comparison Reference Value (KCRV) of the CCPR-K1.a key comparison. The EURAMET.PR-K1.a key comparison was piloted by the National Physical Laboratory (NPL), who also acted as pilot for the CCPR-K1.a key comparison; a further linkage to the KCRV of the CCPR-K1.a key comparison was provided through the participation of the Physikalisch-Technische Bundesanstalt (PTB) in both comparisons. The other participants were: National Institute of Metrology of Romania (INM-RO), Federal Office of Metrology (METAS), VSL Dutch Metrology Institute (VSL), SP Technical Research Institute of Sweden (SP) and All Russian Institute for Optical and Physical Measurements (VNIIOFI). Measurements were made by each laboratory at 44 designated wavelengths, or a subset of these wavelengths. The link laboratories made measurements at all 44 wavelengths. For the purposes of analysis each wavelength has been treated independently, as for the CCPR K1.a comparison. 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 CCPR, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  16. Could S6K1 immunopositivity be used to distinguish early and advanced stages of endometrioid endometrial adenocarcinoma?

    PubMed Central

    Gün, İsmet; Özdamar, Özkan; Küçükodacı, Zafer; Muhçu, Murat; Demirel, Dilaver

    2016-01-01

    Objective To assess whether the immunopositivity of S6K1, a crucial effector of the mTOR signaling pathway, varies between early-stage low-grade and advanced-stage high-grade endometrial endometrioid adenocarcinoma (EEA) as well as to discuss its prognostic significance. Material and Methods A total of 22 normal endometrial tissue samples (Control group) and 41 EEA specimens (Study group) were enrolled in the study, and all the samples underwent immunohistochemical staining for S6 kinase alpha (S6K1). The study group was further evaluated in two subgroups; stage 1A, grade 1 (Group 1) and stage ≥1A, grade 2 or 3 (Group 2). Group 2 patients were considered as a poor prognosis for EEA. The samples were examined by two independent pathologists. Statistical analyses were performed using the Student’s t-test for continuous variables, the Chi-square test for categorical variables, and one-way analysis of variance for the comparison of multiple variables. Results The immunopositivity rate for all the included EEA patients was 56.1%, whereas none of the 22 normal endometrial tissue samples revealed immunoreactivity for S6K1. The immunopositivity rates were significantly different between Groups 1 and 2 [38.1% (8/21) and 75.0% (15/20), respectively, p=0.039]. When S6K1 positivity was used as a criterion of poor prognosis in EEA, the sensitivity, specificity, positive predictive value, and negative predictive value were calculated to be 62%, 75%, 72%, and 65%, respectively (OR: 4.9 and 95% CI: 1.3–18.7). Conclusion S6K1 was positive in the majority of EEAs and malignancies at an advanced stage. Higher grade disease had a significantly higher rate of S6K1 positivity. S6K1 immunopositivity appears to be a promising method to predict poor prognosis in EEA. PMID:27651726

  17. Study of antihypertensive mechanism of Tribulus terrestris in 2K1C hypertensive rats: role of tissue ACE activity.

    PubMed

    Sharifi, Ali M; Darabi, Radbod; Akbarloo, Nasrin

    2003-10-24

    Tribulus terrestris is a natural herb used for treating many diseases including hypertension. According to previous reports, aqueous extract of tribulus fruits may have some antihypertensive effect with an unknown mechanism. The present study investigated the antihypertensive mechanism of tribulus in 2K1C hypertensive rats by measurement of circulatory and local ACE activity in aorta, heart, kidney and lung. Four groups of rats were selected; control, sham, operated or hypertensive and tribulus treated hypertensive group. Hypertension was induced using silver clip on renal artery by surgery. Four weeks after surgery, a single daily dose of 10 mg/kg of lyophilized aqueous extract of tribulus fruit were given orally to 2K1C rats for four weeks. ACE activity was determined by high performance liquid chromatography (HPLC). Blood pressure was measured by the tail-cuff method. The systolic blood pressure (SBP) was significantly increased in 2K1C rats compared to control rats. The SBP of tribulus fed hypertensive rats was significantly decreased compared to hypertensive rats. The ACE activity in all tissues of 2K1C rats including: aorta, heart, kidney, lung as well as serum were significantly increased compared to normal rats. The ACE activity in all tissues of tribulus fed hypertensive rats was significantly lower than that of hypertensive rats, which was more pronounced in kidney. These results indicated that there is a negative correlation between consumption of tribulus and ACE activity in serum and different tissues in 2K1C rats.

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

    PubMed Central

    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

  19. Heads or Tails: Genotyping of Hepatitis C Virus Concerning the 2k/1b Circulating Recombinant Form.

    PubMed

    Schuermans, Wim; Orlent, Hans; Desombere, Isabelle; Descheemaeker, Patrick; Van Vlierberghe, Hans; Geerts, Anja; Verhelst, Xavier; Reynders, Marijke; Padalko, Elizaveta

    2016-01-01

    As different hepatitis C virus (HCV) genotypes respond differently to initiated therapy, correct HCV genotyping is essential. A potential risk for misclassification of the intergenotypic HCV circulating recombinant form (CRF) 2k/1b strains exists, depending on the genotyping method used. The aim was to investigate the differences in HCV genotyping methods with regard to CRF 2k/1b and to gain insight in the prevalence of the CRF 2k/1b. Genotyping results by Versant HCV Genotype Assay were compared with nonstructural protein 5B (NS5B) sequencing. In total, from November 2001 until March 2015, 3296 serum samples were analyzed by Versant HCV Genotype Assay. As misclassified CRF is harbored among HCV genotype 2, we further focused our search on 142 (4.3%) samples positive for HCV genotype 2. On 116 (81.7%) retrieved samples, the NS5B sequencing was performed. Twelve out of the 116 retrieved samples (10.3%) were classified as CRF 2k/1b by sequencing of the NS5B region. Ten of these 12 samples were originally misclassified as genotype 2a or 2c, while 2 of them were misclassified as genotype 2. Our results show that the current prevalence of CRF 2k/1b is underestimated. The importance of correct HCV genotyping is emphasized, considering the tailored choice of treatment regimen and overall prognosis. PMID:27563879

  20. Jack Polynomials as Fractional Quantum Hall States and the Betti Numbers of the ( k + 1)-Equals Ideal

    NASA Astrophysics Data System (ADS)

    Zamaere, Christine Berkesch; Griffeth, Stephen; Sam, Steven V.

    2014-08-01

    We show that for Jack parameter α = -( k + 1)/( r - 1), certain Jack polynomials studied by Feigin-Jimbo-Miwa-Mukhin vanish to order r when k + 1 of the coordinates coincide. This result was conjectured by Bernevig and Haldane, who proposed that these Jack polynomials are model wavefunctions for fractional quantum Hall states. Special cases of these Jack polynomials include the wavefunctions of Laughlin and Read-Rezayi. In fact, along these lines we prove several vanishing theorems known as clustering properties for Jack polynomials in the mathematical physics literature, special cases of which had previously been conjectured by Bernevig and Haldane. Motivated by the method of proof, which in the case r = 2 identifies the span of the relevant Jack polynomials with the S n -invariant part of a unitary representation of the rational Cherednik algebra, we conjecture that unitary representations of the type A Cherednik algebra have graded minimal free resolutions of Bernstein-Gelfand-Gelfand type; we prove this for the ideal of the ( k + 1)-equals arrangement in the case when the number of coordinates n is at most 2 k + 1. In general, our conjecture predicts the graded S n -equivariant Betti numbers of the ideal of the ( k + 1)-equals arrangement with no restriction on the number of ambient dimensions.

  1. Heads or Tails: Genotyping of Hepatitis C Virus Concerning the 2k/1b Circulating Recombinant Form

    PubMed Central

    Schuermans, Wim; Orlent, Hans; Desombere, Isabelle; Descheemaeker, Patrick; Van Vlierberghe, Hans; Geerts, Anja; Verhelst, Xavier; Reynders, Marijke; Padalko, Elizaveta

    2016-01-01

    As different hepatitis C virus (HCV) genotypes respond differently to initiated therapy, correct HCV genotyping is essential. A potential risk for misclassification of the intergenotypic HCV circulating recombinant form (CRF) 2k/1b strains exists, depending on the genotyping method used. The aim was to investigate the differences in HCV genotyping methods with regard to CRF 2k/1b and to gain insight in the prevalence of the CRF 2k/1b. Genotyping results by Versant HCV Genotype Assay were compared with nonstructural protein 5B (NS5B) sequencing. In total, from November 2001 until March 2015, 3296 serum samples were analyzed by Versant HCV Genotype Assay. As misclassified CRF is harbored among HCV genotype 2, we further focused our search on 142 (4.3%) samples positive for HCV genotype 2. On 116 (81.7%) retrieved samples, the NS5B sequencing was performed. Twelve out of the 116 retrieved samples (10.3%) were classified as CRF 2k/1b by sequencing of the NS5B region. Ten of these 12 samples were originally misclassified as genotype 2a or 2c, while 2 of them were misclassified as genotype 2. Our results show that the current prevalence of CRF 2k/1b is underestimated. The importance of correct HCV genotyping is emphasized, considering the tailored choice of treatment regimen and overall prognosis. PMID:27563879

  2. Aerobic growth of Anoxybacillus pushchinoensis K1(T): emended descriptions of A. pushchinoensis and the genus Anoxybacillus

    NASA Technical Reports Server (NTRS)

    Pikuta, Elena; Cleland, David; Tang, Jane

    2003-01-01

    In this work, corrections are made to the descriptions of the species Anoxybacillus pushchinoensis corrig. and the genus ANOXYBACILLUS: Experiments to determine the relationship of A. pushchinoensis K1(T) to oxygen showed that it was capable of aerobic growth, but preferred to grow anaerobically. During aerobic growth, the redox indicator resazurin was reduced as a result of hydrogen gas production. The facultatively anaerobic nature of K1(T) was ascertained by cultivation in aerobic liquid medium, where growth began at the bottom of the tube. The anaerobic nature of K1(T) was also indicated by a negative catalase reaction. This work is submitted to correct the description of the species A. pushchinoensis from obligate anaerobe to aerotolerant anaerobe and to emend the description of the genus Anoxybacillus from obligate anaerobes or facultative anaerobes to aerotolerant anaerobes or facultative anaerobes.

  3. The bioavailability of a mixed micellar preparation of vitamin K1, and its procoagulant effect in anticoagulated rabbits.

    PubMed

    Winn, M J; White, P M; Scott, A K; Pratt, S K; Park, B K

    1989-04-01

    We have investigated the pharmacokinetics and procoagulant activity of a new, mixed-micellar preparation of vitamin K1 (MM-K) in male New Zealand White rabbits. Oral administration of MM-K alone caused a significant (P less than 0.01) increase in the plasma concentrations of vitamin K1 as measured by normal-phase high-performance liquid chromatography (HPLC). Maximum plasma concentrations of vitamin K1 (450 ng mL-1, range 133-824 ng mL-1) were recorded at 3.3 h (range 3-5 h), and were significantly (P less than 0.05) greater than those seen after administration of an existing polyethoxylated castor oil preparation (PE-K; Konakion), which were 260 ng mL-1, range 198-390 ng mL-1 (tmax 0.8 h, range 0.4-1.2 h). AUC after MM-K (4.6 micrograms mL-1 h-1, range 2.1-6.3 micrograms ML-1 h-1) was also significantly (P less than 0.05) greater than after PE-K (1.6 micrograms mL-1 h-1, range 1.0-2.1 micrograms ML-1 h-1). However, the bioavailability of vitamin K1 after administration of MM-K was poor (9.4%), and there was considerable intra-individual variability between the concentrations of vitamin K1 recorded in the plasma samples. Both preparations of vitamin K1 stimulated clotting factor synthesis in rabbits anticoagulated with the potent and long-acting coumarin, brodifacoum. Maximum stimulation of clotting factor synthesis by vitamin K1 after MM-K was 87%, range 44-124% (%PCA). The maximum was seen later (tmax 12 h) than after PE-K (PCA 82%, range 47-125%; tmax 5 h). However, there was considerable intra-individual variability in response to both MM-K and PE-K.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2568466

  4. GluK1 antagonists from 6-(tetrazolyl)phenyl decahydroisoquinoline derivatives: in vitro profile and in vivo analgesic efficacy.

    PubMed

    Martinez-Perez, Jose A; Iyengar, Smriti; Shannon, Harlan E; Bleakman, David; Alt, Andrew; Clawson, David K; Arnold, Brian M; Bell, Michael G; Bleisch, Thomas J; Castaño, Ana M; Del Prado, Miriam; Dominguez, Esteban; Escribano, Ana M; Filla, Sandra A; Ho, Ken H; Hudziak, Kevin J; Jones, Carrie K; Mateo, Ana; Mathes, Brian M; Mattiuz, Edward L; Ogden, Ann Marie L; Simmons, Rosa Maria A; Stack, Douglas R; Stratford, Robert E; Winter, Mark A; Wu, Zhipei; Ornstein, Paul L

    2013-12-01

    We have explored the decahydroisoquinoline scaffold, bearing a phenyl tetrazole, as GluK1 antagonists with potential as oral analgesics. We have established the optimal linker atom between decahydroisoquinoline and phenyl rings and demonstrated an improvement of both the affinity for the GluK1 receptor and the selectivity against the related GluA2 receptor with proper phenyl substitution. In this Letter, we also disclose in vivo data that led to the discovery of LY545694·HCl, a compound with oral efficacy in two persistent pain models.

  5. 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.

    PubMed

    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

  6. Fine-scale mapping of the 5q11.2 breast cancer locus reveals at least three independent risk variants regulating MAP3K1.

    PubMed

    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

  7. Fine-Scale Mapping of the 5q11.2 Breast Cancer Locus Reveals at Least Three Independent Risk Variants Regulating MAP3K1

    PubMed Central

    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

  8. Fine-scale mapping of the 5q11.2 breast cancer locus reveals at least three independent risk variants regulating MAP3K1.

    PubMed

    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.

  9. Structure of a putative trans-editing enzyme for prolyl-tRNA synthetase from Aeropyrum pernix K1 at 1.7 Å resolution

    SciTech Connect

    Murayama, Kazutaka; Kato-Murayama, Miyuki; Katsura, Kazushige; Uchikubo-Kamo, Tomomi; Yamaguchi-Hirafuji, Machiko; Kawazoe, Masahito; Akasaka, Ryogo; Hanawa-Suetsugu, Kyoko; Hori-Takemoto, Chie; Terada, Takaho; Shirouzu, Mikako; Yokoyama, Shigeyuki

    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%)

  10. Genetic mutations in the K1 and K10 genes of patients with epidermolytic hyperkeratosis. Correlation between location and disease severity.

    PubMed Central

    Syder, A J; Yu, Q C; Paller, A S; Giudice, G; Pearson, R; Fuchs, E

    1994-01-01

    Epidermolytic hyperkeratosis (EH) is a skin disease caused by mutations in the genes encoding K1 and K10, the differentiation-specific keratins of epidermis. To explore the heterogeneity of mutations and to assess whether a correlation exists between disease severity and the extent to which a mutation interferes with keratin network formation, we determined the genetic bases of four severe incidences of EH and one unusually mild case. Two severe cases have the same mutation, K10-R156:C, at a conserved arginine that we previously showed was mutated to a histidine in two unrelated EH families. An additional severe case has a mutation six residues away, still within the amino end of the alpha-helical rod domain of K10. The other severe case has a mutation in the conserved carboxy end of the K1 rod. In contrast, affected members of the atypically mild family have a mutation just proximal to the conserved carboxy end of the K10 rod. By genetic engineering and gene transfection, we demonstrate that each mutation is functionally responsible for the keratin filament aberrations that are typical of keratinocytes cultured from these patients. Moreover, we show that the mild EH mutation less severely affects filament network formation. Taken together, our studies strengthen the link between filament perturbations, cell fragility, and degeneration. Images PMID:7512983

  11. 78 FR 23981 - Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and K-1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-23

    ... Internal Revenue Service Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and... Form 1041 and related Schedules D, J, and K-1, U.S. Income Tax Return for Estates and Trusts. DATES... Complex Trusts (Schedule J), and Beneficiary's Share of Income, Deductions, Credits, etc. (Schedule...

  12. Glucocorticoids limit acute lung inflammation in concert with inflammatory stimuli by induction of SphK1

    PubMed Central

    Vettorazzi, Sabine; Bode, Constantin; Dejager, Lien; Frappart, Lucien; Shelest, Ekaterina; Klaßen, Carina; Tasdogan, Alpaslan; Reichardt, Holger M.; Libert, Claude; Schneider, Marion; Weih, Falk; Henriette Uhlenhaut, N.; David, Jean-Pierre; Gräler, Markus; Kleiman, Anna; Tuckermann, Jan P.

    2015-01-01

    Acute lung injury (ALI) is a severe inflammatory disease for which no specific treatment exists. As glucocorticoids have potent immunosuppressive effects, their application in ALI is currently being tested in clinical trials. However, the benefits of this type of regimen remain unclear. Here we identify a mechanism of glucocorticoid action that challenges the long-standing dogma of cytokine repression by the glucocorticoid receptor. Contrarily, synergistic gene induction of sphingosine kinase 1 (SphK1) by glucocorticoids and pro-inflammatory stimuli via the glucocorticoid receptor in macrophages increases circulating sphingosine 1-phosphate levels, which proves essential for the inhibition of inflammation. Chemical or genetic inhibition of SphK1 abrogates the therapeutic effects of glucocorticoids. Inflammatory p38 MAPK- and mitogen- and stress-activated protein kinase 1 (MSK1)-dependent pathways cooperate with glucocorticoids to upregulate SphK1 expression. Our findings support a critical role for SphK1 induction in the suppression of lung inflammation by glucocorticoids, and therefore provide rationales for effective anti-inflammatory therapies. PMID:26183376

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

    PubMed Central

    Boonla, Orachorn; Kukongviriyapan, Upa; Pakdeechote, Poungrat; Kukongviriyapan, Veerapol; Pannangpetch, Patchareewan; Thawornchinsombut, Supawan

    2015-01-01

    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. PMID:26184305

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

    PubMed

    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. PMID:27617056

  15. Effects of the plant flavonoids silymarin and quercetin on arsenite-induced oxidative stress in CHO-K1 cells.

    PubMed

    Bongiovanni, G A; Soria, E A; Eynard, A R

    2007-06-01

    Chronic toxic effects of arsenic resulting from drinking water are a human health problem, especially in South-America and Asia. Arsenic is capable of influencing various cellular processes, causing adverse effects, including cancer. Although the exact mechanism of the action is not known, a correlation between oxidative stress, tumour promotion and arsenic exposure has been observed. We examined the effects of silymarin and quercetin, in counteracting oxidative stress produced by acute or sub-chronic sodium arsenite exposure. The stress responses to arsenite included an increase in the heat shock protein 70 kDa expression, lipid peroxidation assayed by conjugated dienes measure, and gamma-glutamyl-transpeptidase activity. We found that all these stress responses were eliminated by silymarin and quercetin in acute experiments. Both flavonoids diminished the conjugated dienes formation during sub-chronic cultures. Our results suggest that these antioxidant flavonoids, which may be easily incorporated into the diet, may afford a protective effect against arsenite-induced cytotoxicity.

  16. 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

    PubMed Central

    Pan, Zihao; Hu, Lin; Wang, Shaohui; Wang, Haojin; Leung, Frederick C.; Dai, Jianjun; Fan, Hongjie

    2014-01-01

    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. PMID:25397580

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

    PubMed

    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.

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

    PubMed

    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.

  19. Piperazine-2,3-dicarboxylic acid Derivatives as Dual Antagonists of NMDA and GluK1-Containing Kainate Receptors

    PubMed Central

    Irvine, Mark W.; Costa, Blaise M.; Dlaboga, Daniel; Culley, Georgia; Hulse, Richard; Scholefield, Caroline L.; Atlason, Palmi; Fang, Guangyu; Eaves, Richard; Morley, Richard; Mayo-Martin, Maria B.; Amici, Mascia; Bortolotto, Zuner A.; Donaldson, Lucy; Collingridge, Graham L.; Molnár, Elek; Monaghan, Daniel T.; Jane, David E.

    2011-01-01

    Competitive N-methyl-D-aspartate receptor (NMDAR) antagonists bind to the GluN2 subunit, of which there are four types (GluN2A-D). We report that some N1-substituted derivatives of cis-piperazine-2,3-dicarboxylic acid display improved relative affinity for GluN2C and GluN2D versus GluN2A and GluN2B. These derivatives also display subtype-selectivity among the more distantly related kainate receptor family. Compounds 18i and (−)-4 were the most potent kainate receptor antagonists and 18i was selective for GluK1 versus GluK2, GluK3 and AMPA receptors. Modeling studies revealed structural features required for activity at GluK1 subunits and suggested that S674 was vital for antagonist activity. Consistent with this hypothesis, replacing the equivalent residue in GluK3 (alanine) with a serine imparts 18i antagonist activity. Antagonists with dual GluN2D and GluK1 antagonist activity may have beneficial effects in various neurological disorders. Consistent with this idea, antagonist 18i (30 mg/Kg i.p.) showed antinociceptive effects in an animal model of mild nerve injury. PMID:22111545

  20. A global emerging disease of Klebsiella pneumoniae liver abscess: is serotype K1 an important factor for complicated endophthalmitis?

    PubMed Central

    Fung, C-P; Chang, F-Y; Lee, S-C; Hu, B-S; Kuo, B I-T; Liu, C-Y; Ho, M; Siu, L K

    2002-01-01

    Background and aims: Over the past two decades in Taiwan, pyogenic liver abscess has usually been caused by a single microorganism, Klebsiella pneumoniae, and is frequently associated with the serious complication of endophthalmitis, especially in diabetic patients. However, the relationship between the clinical presentation and bacterial factors remains unclear. The aim of this study was to investigate the clinical features of patients and the serotype and ribotype of K pneumoniae liver abscess. Methods: From July 1991 to June 1998, a total of 134 cases of K pneumoniae liver abscess with 248 K pneumoniae isolates from the same patients were collected from two large medical centres in northern Taiwan. Clinical data were collected from medical records. Serotyping and ribotyping were performed using the countercurrent immunoelectrophoresis method and automated Riboprinter. Results: Serotyping revealed that the most common serotypes were K1 (63.4%) and K2 (14.2%). K1 isolates occurred at a significantly higher frequency (p<0.01) than all other serotypes. Among 134 patients, 105 (78.4%) had suffered from diabetes mellitus for 3–15 years. Fourteen patients (10.4%) had metastatic infection to the eye causing septic endophthalmitis. Liver aspirates, and blood and vitreous pus cultures yielded the same serotype of K pneumoniae in all patients. Among patients with septic endophthalmitis, 92.3% (13/14) were diabetic, and 85.7% (12/14) of the isolates belonged to serotype K1. For molecular typing, different degrees of genetic polymorphism among isolates with the same K1 serotype suggested no particular prevalence of any one strain in K pneumoniae liver abscess. Conclusion: K pneumoniae serotype K1 was significantly associated with liver abscess and the complication of endophthalmitis, especially in diabetic patients. Physicians should request an immediate report of serotyping and susceptibility test results simultaneously if a diagnosis of pyogenic liver abscess has been

  1. Loss of plastoglobule kinases ABC1K1 and ABC1K3 causes conditional degreening, modified prenyl-lipids, and recruitment of the jasmonic acid pathway

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plastoglobules (PGs) are plastid lipid-protein particles. This study examines the function of PG-localized kinases ABC1K1 and ABC1K3 in Arabidopsis thaliana. Several lines of evidence suggested that ABC1K1 and ABC1K3 form a protein complex. Null mutants for both genes (abc1k1 and abc1k3) and the dou...

  2. Randomized, placebo-controlled trial of K1 acupoint acustimulation to prevent cisplatin- or oxaliplatin-induced nausea

    PubMed Central

    Shen, Yehua; Liu, Luming; Chiang, Joseph S.; Meng, Zhiqiang; Garcia, M. Kay; Chen, Zhen; Peng, Huiting; Bei, Wenying; Zhao, Qi; Spelman, Amy R.; Cohen, Lorenzo

    2014-01-01

    Background More than 70% of cancer patients experience chemotherapy-induced nausea and vomiting (CINV). We examined the effects of electrostimulation of the K1 acupoint located on the sole of the foot, as it is thought to have potential to control CINV. Methods In this trial, 103 patients diagnosed with primary or metastatic liver cancer were recruited before trans-catheter arterial infusion (TAI) of cisplatin (CDDP) or oxaliplatin (OXA) and randomized to group A (N=51; treated with the antiemetic tropisetron and acustimulation at the K1 acupoint for 20 minutes, 1-2 hours before TAI on the first day and then daily for the subsequent 5 days) or group B (N=53; treated with tropisetron and electrostimulation at a placebo point on the heel). The rate, intensity, and duration of nausea and vomiting were collected at baseline and then daily for 5 days after TAI. Quality of life was assessed daily using the MD Anderson Symptom Inventory (MDASI) and the EuroQoL scale. Results No differences were found between groups A and B in the incidence and degree of nausea or vomiting on day 1 or the consecutive 5 days. Patients in group A had better EuroQoL scores than did patients in group B (A: 72.83 versus B: 65.94, P = 0.04) on day 4 but not on the other days. No group differences were noted at any time point for MDASI scores. Conclusions Electrostimulation of K1 combined with antiemetics did not result in initial prevention of CDDP- or OXA-induced nausea or vomiting. PMID:25204437

  3. The pap Operon of Avian Pathogenic Escherichia coli Strain O1:K1 Is Located on a Novel Pathogenicity Island

    PubMed Central

    Kariyawasam, Subhashinie; Johnson, Timothy J.; Nolan, Lisa K.

    2006-01-01

    We have identified a 56-kb pathogenicity island (PAI) in avian pathogenic Escherichia coli strain O1:K1 (APEC-O1). This PAI, termed PAI IAPEC-O1, is integrated adjacent to the 3′ end of the pheV tRNA gene. It carries putative virulence genes of APEC (pap operon), other E. coli genes (tia and ireA), and a 1.5-kb region unique to APEC-O1. The kps gene cluster required for the biosynthesis of polysialic acid capsule was mapped to a location immediately downstream of this PAI. PMID:16369033

  4. Significant expression of a Chinese scorpion peptide, BmK1, in Escherichia coli through promoter engineering and gene dosage strategy.

    PubMed

    Wang, Jianfeng; Xiong, Zhiqiang; Yang, Yingying; Zhao, Na; Wang, Yong

    2014-01-01

    Heterologous expression is an efficient alternative to conventional extraction to produce a specific Buthus martensii Karsch (BmK) peptide. In this work, BmK1 was successfully expressed in Escherichia coli after genetic codon optimization, but BmK1 content was <6% of total cellular protein. To improve BmK1 expression, a trc promoter library with a wide relative strength was constructed, and three promoters, PpJF136 (0.55), PpJF325 (1.29), and PpJF288 (2.31), were selected to control BmK1 expression. A higher BmK1 expression (>13.9% of total protein) was obtained using a stronger promoter, PpJF325 . Furthermore, a maximum BmK1 content (>21.7% of total protein) was obtained by combining promoter PpJF325 and three copies of the BmK1 gene. The yield of the purified BmK1 achieved 196.74 mg L(-1) in E. coli BL21(DE3) pJF431, which was improved 2.09-fold compared with the control. This was the highest reported production of scorpion peptides in E. coli. PMID:24372571

  5. Boston Public Schools K1 and K2 Programs Needs Assessment. Internal Report to the Department of Early Childhood, Boston Public Schools

    ERIC Educational Resources Information Center

    Marshall, Nancy L.; Roberts, Joanne; Mills, Linda

    2006-01-01

    The Boston Public Schools (BPS) Department of Early Childhood commissioned a needs assessment of current kindergarten (K2) and preschool (K1) programs (1) to inform the BPS Department of Early Childhood about professional development needs to improve the quality of existing K1 and K2 programs; and (2) to inform the Department of additional…

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

    NASA Astrophysics Data System (ADS)

    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).

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

    PubMed

    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.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27647260

  9. Sulforaphane induces apoptosis in adipocytes via Akt/p70s6k1/Bad inhibition and ERK activation.

    PubMed

    Yao, Anjun; Shen, Yingzhuo; Wang, Anshi; Chen, Shiyong; Zhang, Huiqin; Chen, Fen; Chen, Zhongming; Wei, Hua; Zou, Zuquan; Shan, Yujuan; Zhang, Xiaohong

    2015-10-01

    Sulforaphane (SFN), an isothiocyanate isolated from cruciferous vegetables, possesses anti-oxidant and anti-cancer bioactivities. Moreover, SFN exerts its pro-apoptotic effects in some cancer lines. However, the effects and mechanisms of SFN on the regulation of apoptosis of adipocytes are still unknown. In this study, we found that SFN induced significant apoptosis in 3T3-L1 adipocytes and markedly decreased the cellular lipid content. Western blot demonstrated that SFN-induced apoptosis was mediated via the mitochondrial apoptosis pathway based on increased cleavage of poly-ADP-ribose-polymerase (PARP), release of cytochrome c into the cytoplasm, and activation of caspase-3, as well as decreased Bcl-2/Bax ratio. In addition, SFN markedly decreased phosphorylation of Akt and downstream proteins, p70s6k1 and Bad, and increased phosphorylation of ERK. Therefore, our findings clarified that SFN could induce 3T3-L1 adipocyte apoptosis via down-regulation of the Akt/p70s6k1/Bad pathway and up-regulation of the ERK pathway, suggesting SFN may be a promising agent for the treatment or prevention of obesity.

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

    PubMed

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

    2016-01-01

    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. PMID:27647260

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

    NASA Astrophysics Data System (ADS)

    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.

  12. Rotenone induction of hydrogen peroxide inhibits mTOR-mediated S6K1 and 4E-BP1/eIF4E pathways, leading to neuronal apoptosis.

    PubMed

    Zhou, Qian; Liu, Chunxiao; Liu, Wen; Zhang, Hai; Zhang, Ruijie; Liu, Jia; Zhang, Jinfei; Xu, Chong; Liu, Lei; Huang, Shile; Chen, Long

    2015-01-01

    Rotenone, a common pesticide and inhibitor of mitochondrial complex I, induces loss of dopaminergic neurons and consequential aspects of Parkinson's disease (PD). However, the exact mechanism of rotenone neurotoxicity is not fully elucidated. Here, we show that rotenone induced reactive oxygen species (ROS), leading to apoptotic cell death in PC12 cells and primary neurons. Pretreatment with catalase (CAT), a hydrogen peroxide-scavenging enzyme, attenuated rotenone-induced ROS and neuronal apoptosis, implying hydrogen peroxide (H₂O₂) involved, which was further verified by imaging intracellular H₂O₂ using a peroxide-selective probe H2DCFDA. Using thenoyltrifluoroacetone (TTFA), antimycin A, or Mito-TEMPO, we further demonstrated rotenone-induced mitochondrial H₂O₂-dependent neuronal apoptosis. Rotenone dramatically inhibited mTOR-mediated phosphorylation of S6K1 and 4E-BP1, which was also attenuated by CAT in the neuronal cells. Of interest, ectopic expression of wild-type mTOR or constitutively active S6K1, or downregulation of 4E-BP1 partially prevented rotenone-induced H₂O₂ and cell apoptosis. Furthermore, we noticed that rotenone-induced H₂O₂ was linked to the activation of caspase-3 pathway. This was evidenced by the finding that pretreatment with CAT partially blocked rotenone-induced cleavages of caspase-3 and poly (ADP-ribose) polymerase. Of note, zVAD-fmk, a pan caspase inhibitor, only partially prevented rotenone-induced apoptosis in PC12 cells and primary neurons. Expression of mTOR-wt, S6K1-ca, or silencing 4E-BP1 potentiated zVAD-fmk protection against rotenone-induced apoptosis in the cells. The results indicate that rotenone induction of H₂O₂ inhibits mTOR-mediated S6K1 and 4E-BP1/eIF4E pathways, resulting in caspase-dependent and -independent apoptosis in neuronal cells. Our findings suggest that rotenone-induced neuronal loss in PD may be prevented by activating mTOR signaling and/or administering antioxidants.

  13. Binding site and ligand flexibility revealed by high resolution crystal structures of GluK1 competitive antagonists

    PubMed Central

    Alushin, Gregory M.; Jane, David; Mayer, Mark L.

    2010-01-01

    The availability of crystal structures for the ligand binding domains of ionotropic glutamate receptors, combined with their key role in synaptic function in the normal and diseased brain, offers a unique selection of targets for pharmaceutical research compared to other drug targets for which the atomic structure of the ligand binding sites is not known. Currently only a few antagonist structures have been solved, and these reveal ligand specific conformational changes that hinder rational drug design. Here we report high resolution crystal structures for three kainate receptor GluK1 antagonist complexes which reveal new and unexpected modes of binding, highlighting the continued need for experimentally determined receptor-ligand complexes. PMID:20558186

  14. Final report, ongoing key comparison BIPM.QM-K1, ozone at ambient level, comparison with ISCIII (December 2014)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Sanchez, Carmen; Morillo Gomez, Pilar

    2015-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. 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 CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  15. Coexistence of the relaxor-like and ferroelectric behavior in K1-xLixTaO3

    NASA Astrophysics Data System (ADS)

    Trybuła, Z.; Dec, J.; Miga, S.; Łoś, Sz.; Trybuła, M.

    2016-08-01

    The results of low-temperature linear and nonlinear susceptibilities, polarization measurements and the dc electric field dependence of the dielectric properties of the lithium-doped potassium tantalate K1-xLixTaO3, x = 0.034 (KLT-3.4%Li) solid solution are presented. The coexistence of the relaxor-like and ferroelectric behavior and different mechanisms leading to either of them are discussed. The observed ferroelectric phase transition is of the first-order type with temperature hysteresis. This transition is due to the off-center motions of Ta ions in the octahedral environment of oxygen ions. Clusters of Li+ ions produce a relaxor-like behavior and random electric field. This field reduces the depolarization field and allows off-center motions of Ta ions and an appearance of spontaneous polarization.

  16. IL-10 administration reduces PGE-2 levels and promotes CR3-mediated clearance of Escherichia coli K1 by phagocytes in meningitis

    PubMed Central

    Mittal, Rahul; Gonzalez-Gomez, Ignacio; Panigrahy, Ashok; Goth, Kerstin; Bonnet, Richard

    2010-01-01

    Ineffectiveness of antibiotics in treating neonatal Escherichia coli K1 meningitis and the emergence of antibiotic-resistant strains evidently warrants new prevention strategies. We observed that administration of interleukin (IL)-10 during high-grade bacteremia clears antibiotic-sensitive and -resistant E. coli from blood of infected mice. Micro-CT studies of brains from infected animals displayed gross morphological changes similar to those observed in infected human neonates. In mice, IL-10, but not antibiotic or anti-TNF antibody treatment prevented brain damage caused by E. coli. IL-10 administration elevated CR3 expression in neutrophils and macrophages of infected mice, whereas infected and untreated mice displayed increased expression of FcγRI and TLR2. Neutrophils or macrophages pretreated with IL-10 ex vivo exhibited a significantly greater microbicidal activity against E. coli compared with cells isolated from wild-type or IL-10−/− mice. The protective effect of IL-10 was abrogated when CR3 was knocked-down in vivo by siRNA. The increased expression of CR3 in phagocytes was caused by inhibition of prostaglandin E-2 (PGE-2) levels, which were significantly increased in neutrophils and macrophages upon E. coli infection. These findings describe a novel modality of IL-10–mediated E. coli clearance by diverting the entry of bacteria via CR3 and preventing PGE-2 formation in neonatal meningitis. PMID:20498022

  17. IL-10 administration reduces PGE-2 levels and promotes CR3-mediated clearance of Escherichia coli K1 by phagocytes in meningitis.

    PubMed

    Mittal, Rahul; Gonzalez-Gomez, Ignacio; Panigrahy, Ashok; Goth, Kerstin; Bonnet, Richard; Prasadarao, Nemani V

    2010-06-01

    Ineffectiveness of antibiotics in treating neonatal Escherichia coli K1 meningitis and the emergence of antibiotic-resistant strains evidently warrants new prevention strategies. We observed that administration of interleukin (IL)-10 during high-grade bacteremia clears antibiotic-sensitive and -resistant E. coli from blood of infected mice. Micro-CT studies of brains from infected animals displayed gross morphological changes similar to those observed in infected human neonates. In mice, IL-10, but not antibiotic or anti-TNF antibody treatment prevented brain damage caused by E. coli. IL-10 administration elevated CR3 expression in neutrophils and macrophages of infected mice, whereas infected and untreated mice displayed increased expression of FcgammaRI and TLR2. Neutrophils or macrophages pretreated with IL-10 ex vivo exhibited a significantly greater microbicidal activity against E. coli compared with cells isolated from wild-type or IL-10-/- mice. The protective effect of IL-10 was abrogated when CR3 was knocked-down in vivo by siRNA. The increased expression of CR3 in phagocytes was caused by inhibition of prostaglandin E-2 (PGE-2) levels, which were significantly increased in neutrophils and macrophages upon E. coli infection. These findings describe a novel modality of IL-10-mediated E. coli clearance by diverting the entry of bacteria via CR3 and preventing PGE-2 formation in neonatal meningitis. PMID:20498022

  18. APMP key comparison for the measurement of air kerma for 60Co (APMP.RI(I)-K1.1)

    NASA Astrophysics Data System (ADS)

    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

  19. Fragmentation Origin of Major Sungrazing Comets C/1970 K1, C/1880 C1, and C/1843 D1

    NASA Astrophysics Data System (ADS)

    Sekanina, Zdenek; Chodas, Paul W.

    2002-12-01

    Following our recent successful modeling of the common origin of two of the brightest members of the Kreutz system of sungrazing comets, we now examine three other objects: C/1970 K1 (White-Ortiz-Bolelli), the most recent sungrazer discovered from Earth, C/1880 C1 (Great Southern Comet), and C/1843 D1 (Great March Comet). For White-Ortiz-Bolelli, five possible origin and orbit evolution scenarios are explored. We find that its parent was neither C/1965 S1 (Ikeya-Seki), nor C/1882 R1 (Great September Comet), nor the comet of 1106 (the presumed parent of Ikeya-Seki and the 1882 comet). The motion of C/1970 K1 is consistent with a scenario in which the parent was an unknown fragment that separated from the 1106 comet at the same time as, or shortly before, Ikeya-Seki and passed through perihelion in 1970 June-July, shortly after White-Ortiz-Bolelli. The separation of White-Ortiz-Bolelli from this fragment is found to have occurred around the mid-eighteenth century, at a heliocentric distance of about 150 AU, with a relative velocity of 3-5 m s-1 in the general direction of the Sun and to the north of the orbital plane. On the other hand, we conclude that the 1880 comet separated directly from C/1843 D1, the second brightest known sungrazer, some 100-150 days after the 1843 comet's previous perihelion passage in the eleventh century, at 2.5-3 AU from the Sun, with a relative velocity of slightly more than 7 m s-1 in the generally antisolar direction and to the south of the orbital plane. The pattern of fragmentation of the Kreutz system's members discovered from Earth begins to resemble the evolution of the system's minor fragments detected coronagraphically from aboard the SOHO spacecraft, and there is significant qualitative similarity with fragmentation of comet D/1993 F2 (Shoemaker-Levy 9).

  20. TOR and S6K1 promote translation reinitiation of uORF-containing mRNAs via phosphorylation of eIF3h.

    PubMed

    Schepetilnikov, Mikhail; Dimitrova, Maria; Mancera-Martínez, Eder; Geldreich, Angèle; Keller, Mario; Ryabova, Lyubov A

    2013-04-17

    Mammalian target-of-rapamycin (mTOR) triggers S6 kinase (S6K) activation to phosphorylate targets linked to translation in response to energy, nutrients, and hormones. Pathways of TOR activation in plants remain unknown. Here, we uncover the role of the phytohormone auxin in TOR signalling activation and reinitiation after upstream open reading frame (uORF) translation, which in plants is dependent on translation initiation factor eIF3h. We show that auxin triggers TOR activation followed by S6K1 phosphorylation at T449 and efficient loading of uORF-mRNAs onto polysomes in a manner sensitive to the TOR inhibitor Torin-1. Torin-1 mediates recruitment of inactive S6K1 to polysomes, while auxin triggers S6K1 dissociation and recruitment of activated TOR instead. A putative target of TOR/S6K1-eIF3h-is phosphorylated and detected in polysomes in response to auxin. In TOR-deficient plants, polysomes were prebound by inactive S6K1, and loading of uORF-mRNAs and eIF3h was impaired. Transient expression of eIF3h-S178D in plant protoplasts specifically upregulates uORF-mRNA translation. We propose that TOR functions in polysomes to maintain the active S6K1 (and thus eIF3h) phosphorylation status that is critical for translation reinitiation.

  1. Bioprocess optimization for production of thermoalkali-stable protease from Bacillus subtilis K-1 under solid-state fermentation.

    PubMed

    Singh, Satbir; Bajaj, Bijender Kumar

    2016-10-01

    Cost-effective production of proteases, which are robust enough to function under harsh process conditions, is always sought after due to their wide industrial application spectra. Solid-state production of enzymes using agro-industrial wastes as substrates is an environment-friendly approach, and it has several advantages such as high productivity, cost-effectiveness, being less labor-intensive, and less effluent production, among others. In the current study, different agro-wastes were employed for thermoalkali-stable protease production from Bacillus subtilis K-1 under solid-state fermentation. Agricultural residues such as cotton seed cake supported maximum protease production (728 U ml(-1)), which was followed by gram husk (714 U ml(-1)), mustard cake (680 U ml(-1)), and soybean meal (653 U ml(-1)). Plackett-Burman design of experiment showed that peptone, moisture content, temperature, phosphates, and inoculum size were the significant variables that influenced the protease production. Furthermore, statistical optimization of three variables, namely peptone, moisture content, and incubation temperature, by response surface methodology resulted in 40% enhanced protease production as compared to that under unoptimized conditions (from initial 728 to 1020 U ml(-1)). Thus, solid-state fermentation coupled with design of experiment tools represents a cost-effective strategy for production of industrial enzymes. PMID:26760481

  2. Probing the Molecular Complexity of Cometary Volatiles: The Case of C/2012 K1 (PanSTARRS)

    NASA Astrophysics Data System (ADS)

    Milam, Stefanie N.; Cordiner, Martin; Remijan, Anthony; Gicquel, Adeline; Charnley, Steven; Colom, Pierre; Crovisier, Jacques; Mumma, Michael; Boissier, Jeremie; Bockelee-Morvan, Dominique; Biver, Nicolas; Villanueva, Geronimo; Paganini, Lucas; Lis, Dariusz; Kuan, Yi-Jehng; Coulson, Iain

    2014-11-01

    Small bodies are considered to contain the most primitive material remaining from the formation of our solar system. Cometary ices trace the pristine volatile component and provide an important source of information regarding the physical and chemical conditions of the early Solar Nebula. However, not all observed species are native to the comet nucleus and some are likely formed in the coma (at least in part) with previous observations unable to ascertain the precise origin of fundamental species including H2CO, HCN, CO, CS, and HNC. Simultaneous, spatially and spectrally-resolved molecular emission maps of comets at mm and sub-mm wavelengths provide the key information required to probe the origin and nature of these volatiles. After months of monitoring cometary activity through OH observations at the NRAO GBT and the Nançay radio telescope, we observed comet C/2012 K1 (PanSTARRS) using ALMA Bands 6 and 7, sampling emission lines from HCN, CO, CS, HCO+, CH3OH, H2CO and HNC. In order to recover any extended flux resolved out by the interferometer, we simultaneously measured HCN and CH3OH with the APEX observatory. We will present full details of these unique observations, and an analysis of the observed spectra.

  3. Ligand-induced structural changes in the cyclic nucleotide-modulated potassium channel MloK1

    PubMed Central

    Kowal, Julia; Chami, Mohamed; Baumgartner, Paul; Arheit, Marcel; Chiu, Po-Lin; Rangl, Martina; Scheuring, Simon; Schröder, Gunnar F.; Nimigean, Crina M.; Stahlberg, Henning

    2014-01-01

    Cyclic nucleotide-modulated ion channels are important for signal transduction and pacemaking in eukaryotes. The molecular determinants of ligand gating in these channels are still unknown, mainly because of a lack of direct structural information. Here we report ligand-induced conformational changes in full-length MloK1, a cyclic nucleotide-modulated potassium channel from the bacterium Mesorhizobium loti, analysed by electron crystallography and atomic force microscopy. Upon cAMP binding, the cyclic nucleotide-binding domains move vertically towards the membrane, and directly contact the S1–S4 voltage sensor domains. This is accompanied by a significant shift and tilt of the voltage sensor domain helices. In both states, the inner pore-lining helices are in an ‘open’ conformation. We propose a mechanism in which ligand binding can favour pore opening via a direct interaction between the cyclic nucleotide-binding domains and voltage sensors. This offers a simple mechanistic hypothesis for the coupling between ligand gating and voltage sensing in eukaryotic HCN channels. PMID:24469021

  4. Thermostability and reactivity in organic solvent of O-phospho-L-serine sulfhydrylase from hyperthermophilic archaeon Aeropyrum pernix K1.

    PubMed

    Nakamura, Takashi; Asai, Shinji; Nakata, Kaori; Kunimoto, Kohei; Oguri, Masateru; Ishikawa, Kazuhiko

    2015-01-01

    O-phospho-l-serine sulfhydrylase (OPSS) from archaeon Aeropyrum pernix K1 is able to synthesize l-cysteine even at 80 °C. In this article, we compared thermal stability and reactivity in organic solvent of OPSS with those of O-acetyl-l-serine sulfhydrylase B (OASS-B) from Escherichia coli. As a result, the thermostability of OPSS was much higher than that of OASS-B. Moreover, the activity of OPSS increased in the reaction mixture containing the organic solvent, such as N, N'-dimethyl formamide and 1,4-dioxane, whereas that of OASS-B gradually decreased as the content of organic solvent increased. From the crystal structural analysis, the intramolecular electrostatic interactions of N-terminal domain in OPSS seemed to be correlated with the tolerance of OPSS to high temperature and organic solvent. These results indicate that OPSS is more superior to OASS-B for the industrial production of l-cysteine and unnatural amino acids that are useful pharmaceuticals in the presence of organic solvent.

  5. Interference harmonics and rigorous EM spectrum analysis method for low-k1 CD Bossung tilt correction

    NASA Astrophysics Data System (ADS)

    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.

  6. LDA'+DMFT investigation of electronic structure of K1 - x Fe2 - y Se2 superconductor

    NASA Astrophysics Data System (ADS)

    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.

  7. Systemic signalling in photosynthetic induction of Rumex K-1 (Rumex patientia × Rumex tianschaious) leaves.

    PubMed

    Hou, Fei; Jin, Li-Qiao; Zhang, Zi-Shan; Gao, Hui-Yuan

    2015-04-01

    The rapid induction of photosynthesis is critical for plants under light-fleck environment. Most previous studies about photosynthetic induction focused upon single leaf, but they did not consider the systemic integrity of plant. Here, we verified whether systemic signalling is involved in photosynthetic induction. Rumex K-1 (Rumex patientia × Rumex tianschaious) plants were grown under light-fleck condition. After whole night dark adaptation, different numbers of leaves (system leaf or SL) were pre-illuminated with light, and then the photosynthetic induction of other leaves (target leaf or TL) was investigated. This study showed that the pre-illumination of SL promoted photosynthetic induction in TL. This promotion was independent of the number of SL, the light intensity on SL and the distance between SL and TL, indicating that this systemic signalling is non-dose-dependent. More interestingly, the photosynthetic induction was promoted by only the pre-illumination of morphological upper leaf rather than the pre-illumination of morphological lower leaf, indicating that the transfer of this signal is directional. The results showed that the transfer of this systemic signalling depends upon the phloem. This systemic signalling helps plants to use light energy more efficiently under light flecks.

  8. Parallel down-regulation of chloride channel CLC-K1 and barttin mRNA in the thin ascending limb of the rat nephron by furosemide.

    PubMed

    Wolf, Konrad; Meier-Meitinger, Martina; Bergler, Tobias; Castrop, Hayo; Vitzthum, Helga; Riegger, Günter A J; Kurtz, Armin; Krämer, Bernhard K

    2003-09-01

    In the past few years the pivotal role of kidney Cl(-)channels (ClC-K) channels in maintaining salt and water homeostasis in the kidney has been established. The aim of the present study was to investigate the influence of the loop diuretic furosemide on the gene expression of the kidney chloride channel ClC-K1 and its recently described functional subunit barttin. Male Sprague Dawley rats received the loop diuretic furosemide (12 mg/kg/day) for 6 days. Rats had free access to 0.9% NaCl, 0.1%KCl solution to prevent volume depletion. Localisation and regulation of ClC-K1 and barttin mRNA was analysed by RNase protection and in situ hybridisation. Nephron-specific regulation was investigated by microdissection and real-time PCR quantification. In furosemide-treated rats ClC-K1 mRNA decreased to half in the inner medulla. In the renal cortex and outer medulla ClC-K1 mRNA levels were weak and did not change. Under furosemide treatment barttin mRNA was regulated in parallel with ClC-K1 mRNA. A significant mRNA decrease occurred after furosemide treatment in inner medulla (0.50 fold), whereas cortical and outer medulla levels remained unaffected. (35)S in situ hybridisation confirmed the regulation and distribution seen in the RNase protection assay experiments. Microdissection of the inner medullary collecting duct and thin limb of Henle's loop followed by real-time PCR revealed that CLC-K1 and barttin mRNA regulation in inner medulla was limited to the thin limb; mRNA levels in collecting ducts were not affected by furosemide treatment. Our findings imply that during furosemide treatment selective down-regulation of ClC-K1 and barttin mRNAs in thin limb plays a role in maintaining salt and water homeostasis. PMID:12759757

  9. The relationship between inhibition of vitamin K1 2,3-epoxide reductase and reduction of clotting factor activity with warfarin.

    PubMed Central

    Choonara, I A; Malia, R G; Haynes, B P; Hay, C R; Cholerton, S; Breckenridge, A M; Preston, F E; Park, B K

    1988-01-01

    1 The effect of low dose steady state warfarin (0.2 mg and 1 mg daily) on clotting factor activity and vitamin K1 metabolism was studied in seven healthy volunteers. 2 Steady state plasma warfarin concentrations were 41-99 ng ml-1 for the 0.2 mg dose and 157-292 ng ml-1 for the 1 mg dose. 3 There was a significant prolongation of the mean prothrombin time (0.9 s) after 1 mg warfarin daily, but no significant change in prothrombin time after 0.2 mg warfarin daily. There was no significant change in individual clotting factor activity (II, VII, IX or X) with either dose of warfarin. 4 Following the administration of a pharmacological dose of vitamin K1 (10 mg), all seven volunteers had detectable levels of vitamin K1 2,3-epoxide with both doses of warfarin (Cpmax 31-409 ng ml-1). 5 Both the Cpmax and the AUC for vitamin K1 2,3-epoxide were significantly greater on 1 mg of warfarin daily than 0.2 mg daily (P less than 0.01). 6 The apparent dissociation between inhibition of vitamin K1 2,3-epoxide reductase and reduction of clotting factor activity, produced by warfarin, may reflect the insensitivity of functional clotting factor assays to a small reduction in clotting factor concentration. PMID:3370190

  10. Inhibition of inducible nitric oxide controls pathogen load and brain damage by enhancing phagocytosis of Escherichia coli K1 in neonatal meningitis.

    PubMed

    Mittal, Rahul; Gonzalez-Gomez, Ignacio; Goth, Kerstin A; Prasadarao, Nemani V

    2010-03-01

    Escherichia coli K1 is a leading cause of neonatal meningitis in humans. In this study, we sought to determine the pathophysiologic relevance of inducible nitric oxide (iNOS) in experimental E. coli K1 meningitis. By using a newborn mouse model of meningitis, we demonstrate that E. coli infection triggered the expression of iNOS in the brains of mice. Additionally, iNOS-/- mice were resistant to E. coli K1 infection, displaying normal brain histology, no bacteremia, no disruption of the blood-brain barrier, and reduced inflammatory response. Treatment with an iNOS specific inhibitor, aminoguanidine (AG), of wild-type animals before infection prevented the development of bacteremia and the occurrence of meningitis. The infected animals treated with AG after the development of bacteremia also completely cleared the pathogen from circulation and prevented brain damage. Histopathological and micro-CT analysis of brains revealed significant damage in E. coli K1-infected mice, which was completely abrogated by AG administration. Peritoneal macrophages and polymorphonuclear leukocytes isolated from iNOS-/- mice or pretreated with AG demonstrated enhanced uptake and killing of the bacteria compared with macrophages and polymorphonuclear leukocytes from wild-type mice in which E. coli K1 survive and multiply. Thus, NO produced by iNOS may be beneficial for E. coli to survive inside the macrophages, and prevention of iNOS could be a therapeutic strategy to treat neonatal E. coli meningitis. PMID:20093483

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

    PubMed

    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.

  12. Phosphoproteomics Reveals Resveratrol-Dependent Inhibition of Akt/mTORC1/S6K1 Signaling

    PubMed Central

    2015-01-01

    Resveratrol, a plant-derived polyphenol, regulates many cellular processes, including cell proliferation, aging and autophagy. However, the molecular mechanisms of resveratrol action in cells are not completely understood. Intriguingly, resveratrol treatment of cells growing in nutrient-rich conditions induces autophagy, while acute resveratrol treatment of cells in a serum-deprived state inhibits autophagy. In this study, we performed a phosphoproteomic analysis after applying resveratrol to serum-starved cells with the goal of identifying the acute signaling events initiated by resveratrol in a serum-deprived state. We determined that resveratrol in serum-starved conditions reduces the phosphorylation of several proteins belonging to the mTORC1 signaling pathway, most significantly, PRAS40 at T246 and S183. Under these same conditions, we also found that resveratrol altered the phosphorylation of several proteins involved in various biological processes, most notably transcriptional modulators, represented by p53, FOXA1, and AATF. Together these data provide a more comprehensive view of both the spectrum of phosphoproteins upon which resveratrol acts as well as the potential mechanisms by which it inhibits autophagy in serum-deprived cells. PMID:25311616

  13. Phosphoproteomics reveals resveratrol-dependent inhibition of Akt/mTORC1/S6K1 signaling.

    PubMed

    Alayev, Anya; Doubleday, Peter F; Berger, Sara Malka; Ballif, Bryan A; Holz, Marina K

    2014-12-01

    Resveratrol, a plant-derived polyphenol, regulates many cellular processes, including cell proliferation, aging and autophagy. However, the molecular mechanisms of resveratrol action in cells are not completely understood. Intriguingly, resveratrol treatment of cells growing in nutrient-rich conditions induces autophagy, while acute resveratrol treatment of cells in a serum-deprived state inhibits autophagy. In this study, we performed a phosphoproteomic analysis after applying resveratrol to serum-starved cells with the goal of identifying the acute signaling events initiated by resveratrol in a serum-deprived state. We determined that resveratrol in serum-starved conditions reduces the phosphorylation of several proteins belonging to the mTORC1 signaling pathway, most significantly, PRAS40 at T246 and S183. Under these same conditions, we also found that resveratrol altered the phosphorylation of several proteins involved in various biological processes, most notably transcriptional modulators, represented by p53, FOXA1, and AATF. Together these data provide a more comprehensive view of both the spectrum of phosphoproteins upon which resveratrol acts as well as the potential mechanisms by which it inhibits autophagy in serum-deprived cells.

  14. Nucleotide sequence and genetic analysis of the neuD and neuB genes in region 2 of the polysialic acid gene cluster of Escherichia coli K1.

    PubMed Central

    Annunziato, P W; Wright, L F; Vann, W F; Silver, R P

    1995-01-01

    The K1 capsular polysaccharide, a polymer of sialic acid, is an important virulence determinant of extraintestinal pathogenic Escherichia coli. The genes responsible for the synthesis and expression of the polysialic acid capsule of E. coli K1 are located on the 17-kb kps gene cluster, which is functionally divided into three regions. Central region 2 encodes proteins necessary for the synthesis, activation, and polymerization of sialic acid, while flanking regions 1 and 3 are involved in polymer transport to the cell surface. In this study, we identified two genes at the proximal end of region 2, neuD and neuB, which encode proteins with predicted sizes of 22.7 and 38.7 kDa, respectively. Several observations suggest that the neuB gene encodes sialic acid synthase. EV24, a neuB chromosomal mutant that expresses a capsule when provided exogenous sialic acid, could be complemented in trans by the cloned neuB gene. In addition, NeuB has significant sequence similarity to the product of the cpsB gene of Neisseria meningitidis group B, which is postulated to encode sialic acid synthase. We also present data indicating that neuD has an essential role in K1 polymer production. Cells harboring pSR426, which contains all of region 2 but lacks region 1 and 3 genes, produce an intracellular polymer. In contrast, no polymer accumulated in cells carrying a derivative of pSR426 lacking a functional neuD gene. Unlike strains with mutations in neuB, however, neuD mutants are not complemented by exogenous sialic acid, suggesting that NeuD is not involved in sialic acid synthesis. Additionally, cells harboring a mutation in neuD accumulated sialic acid and CMP-sialic acid. We also found no significant differences between the endogenous and exogenous sialyltransferase activities of a neuD mutant and the wild-type organism. NeuD shows significant similarity to a family of bacterial acetyltransferases, leading to the theory that NeuD is an acetyltransferase which may exert its

  15. Mach's principle: Exact frame-dragging via gravitomagnetism in perturbed Friedmann-Robertson-Walker universes with K=({+-}1,0)

    SciTech Connect

    Schmid, Christoph

    2009-03-15

    We show that there is exact dragging of the axis directions of local inertial frames by a weighted average of the cosmological energy currents via gravitomagnetism for all linear perturbations of all Friedmann-Robertson-Walker (FRW) universes and of Einstein's static closed universe, and for all energy-momentum-stress tensors and in the presence of a cosmological constant. This includes FRW universes arbitrarily close to the Milne Universe and the de Sitter universe. Hence the postulate formulated by Ernst Mach about the physical cause for the time-evolution of inertial axes is shown to hold in general relativity for linear perturbations of FRW universes. - The time-evolution of local inertial axes (relative to given local fiducial axes) is given experimentally by the precession angular velocity {omega}-vector{sub gyro} of local gyroscopes, which in turn gives the operational definition of the gravitomagnetic field: B-vector{sub g}{identical_to}-2{omega}-vector{sub gyro}. The gravitomagnetic field is caused by energy currents J-vector{sub {epsilon}} via the momentum constraint, Einstein's G{sup 0-}circumflex{sub i-circumflex} equation, (-{delta}+{mu}{sup 2})A-vector{sub g}=-16{pi}G{sub N}J-vector{sub {epsilon}} with B-vector{sub g}=curl A-vector{sub g}. This equation is analogous to Ampere's law, but it holds for all time-dependent situations. {delta} is the de Rham-Hodge Laplacian, and {delta}=-curl curl for the vorticity sector in Riemannian 3-space. - In the solution for an open universe the 1/r{sup 2}-force of Ampere is replaced by a Yukawa force Y{sub {mu}}(r)=(-d/dr)[(1/R)exp(-{mu}r)], form-identical for FRW backgrounds with K=(-1,0). Here r is the measured geodesic distance from the gyroscope to the cosmological source, and 2{pi}R is the measured circumference of the sphere centered at the gyroscope and going through the source point. The scale of the exponential cutoff is the H-dot radius, where H is the Hubble rate, dot is the derivative with respect to

  16. KEY COMPARISON: Final report on CCPR K1-a: Spectral irradiance from 250 nm to 2500 nm

    NASA Astrophysics Data System (ADS)

    Woolliams, Emma R.; Fox, Nigel P.; Cox, Maurice G.; Harris, Peter M.; Harrison, Neil J.

    2006-01-01

    The CCPR K1-a key comparison of spectral irradiance (from 250 nm to 2500 nm) was carried out to meet the requirements of the Mutual Recognition Arrangement by 13 participating national metrology institutes (NMIs). Because of the fragile nature of the tungsten halogen lamps used as comparison artefacts, the comparison was arranged as a star comparison with three lamps per participant. NPL (United Kingdom) piloted the comparison and, by measuring all lamps, provided a link between participants' measurements. The other participants were BNM-INM (France), CENAM (Mexico), CSIRO (Australia), HUT (Finland), IFA-CSIC (Spain), MSL-IRL (New Zealand), NIM (China), NIST (United States of America), NMIJ (Japan), NRC (Canada), PTB (Germany) and VNIIOFI (Russian Federation). Before the analysis was completed and the results known, the pilot discussed with each participant which lamp measurements should be included as representative of their comparison. As a consequence of this check, at least one measurement was excluded from one third of the lamps because of changes due to transportations. The comparison thus highlighted the difficulty regarding the availability of suitable transfer standards for the dissemination of spectral irradiance. The use of multiple lamps and multiple measurements ensured sufficient redundancy that all participants were adequately represented. In addition, during this pre-draft A phase all participants had the opportunity to review the uncertainty budgets and methods of all other participants. This new process helped to ensure that all submitted results and their associated uncertainties were evaluated in a consistent manner. The comparison was analysed using a model-based method which regarded each lamp as having a stable spectral irradiance and the measurements made by an NMI as systematically influenced by a factor that applies to all that NMI's measurements. The aim of the analysis was to estimate the systematic factor for each NMI. Across the

  17. Collaboration of AMPK and PKC to induce phosphorylation of Ser413 on PIP5K1B resulting in decreased kinase activity and reduced PtdIns(4,5)P2 synthesis in response to oxidative stress and energy restriction.

    PubMed

    van den Bout, Iman; Jones, David R; Shah, Zahid H; Halstead, Jonathan R; Keune, Willem-Jan; Mohammed, Shabaz; D'Santos, Clive S; Divecha, Nullin

    2013-11-01

    The spatial and temporal regulation of the second messenger PtdIns(4,5)P2 has been shown to be crucial for regulating numerous processes in the cytoplasm and in the nucleus. Three isoforms of PIP5K1 (phosphatidylinositol 4-phosphate 5-kinase), A, B and C, are responsible for the regulation of the major pools of cellular PtdIns(4,5)P2. PIP5K1B is negatively regulated in response to oxidative stress although it remains unclear which pathways regulate its activity. In the present study, we have investigated the regulation of PIP5K1B by protein phosphorylation. Using MS analysis, we identified 12 phosphorylation sites on PIP5K1B. We developed a phospho-specific antibody against Ser413 and showed that its phosphorylation was increased in response to treatment of cells with phorbol ester, H2O2 or energy restriction. Using inhibitors, we define a stress-dependent pathway that requires the activity of the cellular energy sensor AMPK (AMP-activated protein kinase) and PKC (protein kinase C) to regulate Ser413 phosphorylation. Furthermore, we demonstrate that PKC can directly phosphorylate Ser413 in vitro. Mutation of Ser413 to aspartate to mimic serine phosphorylation decreased both PIP5K1B activity in vitro and PtdIns(4,5)P2 synthesis in vivo. Our studies show that collaboration between AMPK and PKC dictates the extent of Ser413 phosphorylation on PIP5K1B and regulates PtdIns(4,5)P2 synthesis.

  18. GluK1 antagonists from 6-(carboxy)phenyl decahydroisoquinoline derivatives. SAR and evaluation of a prodrug strategy for oral efficacy in pain models.

    PubMed

    Martinez-Perez, Jose A; Iyengar, Smriti; Shannon, Harlan E; Bleakman, David; Alt, Andrew; Arnold, Brian M; Bell, Michael G; Bleisch, Thomas J; Castaño, Ana M; Del Prado, Miriam; Dominguez, Esteban; Escribano, Ana M; Filla, Sandra A; Ho, Ken H; Hudziak, Kevin J; Jones, Carrie K; Mateo, Ana; Mathes, Brian M; Mattiuz, Edward L; Ogden, Ann Marie L; Simmons, Rosa Maria A; Stack, Douglas R; Stratford, Robert E; Winter, Mark A; Wu, Zhipei; Ornstein, Paul L

    2013-12-01

    The synthesis and structure-activity relationship of decahydroisoquinoline derivatives with various benzoic acid substitutions as GluK1 antagonists are described. Potent and selective antagonists were selected for a tailored prodrug approach in order to facilitate the evaluation of the new compounds in pain models after oral administration. Several diester prodrugs allowed for acceptable amino acid exposure and moderate efficacy in vivo.

  19. A Novel Saccharomyces cerevisiae Killer Strain Secreting the X Factor Related to Killer Activity and Inhibition of S. cerevisiae K1, K2 and K28 Killer Toxins.

    PubMed

    Melvydas, Vytautas; Bružauskaitė, Ieva; Gedminienė, Genovaitė; Šiekštelė, Rimantas

    2016-09-01

    It was determined that Kx strains secrete an X factor which can inhibit all known Saccharomyces cerevisiae killer toxins (K1, K2, K28) and some toxins of other yeast species-the phenomenon not yet described in the scientific literature. It was shown that Kx type yeast strains posess a killer phenotype producing small but clear lysis zones not only on the sensitive strain α'1 but also on the lawn of S. cerevisiae K1, K2 and K28 type killer strains at temperatures between 20 and 30 °C. The pH at which killer/antikiller effect of Kx strain reaches its maximum is about 5.0-5.2. The Kx yeast were identified as to belong to S. cerevisiae species. Another newly identified S. cerevisiae killer strain N1 has killer activity but shows no antikilller properties against standard K1, K2 and K28 killer toxins. The genetic basis for Kx killer/antikiller phenotype was associated with the presence of M-dsRNA which is bigger than M-dsRNA of standard S. cerevisiae K1, K2, K28 type killer strains. Killer and antikiller features should be encoded by dsRNA. The phenomenon of antikiller (inhibition) properties was observed against some killer toxins of other yeast species. The molecular weight of newly identified killer toxins which produces Kx type strains might be about 45 kDa.

  20. Time-course effects of aerobic exercise training on cardiovascular and renal parameters in 2K1C renovascular hypertensive rats.

    PubMed

    Maia, R C A; Sousa, L E; Santos, R A S; Silva, M E; Lima, W G; Campagnole-Santos, M J; Alzamora, A C

    2015-11-01

    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. PMID:26270472

  1. Bioluminescent Imaging Reveals Novel Patterns of Colonization and Invasion in Systemic Escherichia coli K1 Experimental Infection in the Neonatal Rat

    PubMed Central

    Witcomb, Luci A.; Collins, James W.; McCarthy, Alex J.; Frankel, Gadi

    2015-01-01

    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. PMID:26351276

  2. Structure–Activity Relationship Studies and in Vivo Activity of Guanidine-Based Sphingosine Kinase Inhibitors: Discovery of SphK1- and SphK2-Selective Inhibitors

    PubMed Central

    Kharel, Yugesh; Raje, Mithun R.; Gao, Ming; Tomsig, Jose L.; Lynch, Kevin R.; Santos, Webster L.

    2015-01-01

    Sphingosine 1-phosphate (S1P) is a pleiotropic signaling molecule that acts as a ligand for five G-protein coupled receptors (S1P1–5) whose downstream effects are implicated in a variety of important pathologies including sickle cell disease, cancer, inflammation, and fibrosis. The synthesis of S1P is catalyzed by sphingosine kinase (SphK) isoforms 1 and 2, and hence, inhibitors of this phosphorylation step are pivotal in understanding the physiological functions of SphKs. To date, SphK1 and 2 inhibitors with the potency, selectivity, and in vivo stability necessary to determine the potential of these kinases as therapeutic targets are lacking. Herein, we report the design, synthesis, and structure–activity relationship studies of guanidine-based SphK inhibitors bearing an oxadiazole ring in the scaffold. Our studies demonstrate that SLP120701, a SphK2-selective inhibitor (Ki = 1 μM), decreases S1P levels in histiocytic lymphoma (U937) cells. Surprisingly, homologation with a single methylene unit between the oxadiazole and heterocyclic ring afforded a SphK1-selective inhibitor in SLP7111228 (Ki = 48 nM), which also decreased S1P levels in cultured U937 cells. In vivo application of both compounds, however, resulted in contrasting effect in circulating levels of S1P. Administration of SLP7111228 depressed blood S1P levels while SLP120701 increased levels of S1P. Taken together, these compounds provide an in vivo chemical toolkit to interrogate the effect of increasing or decreasing S1P levels and whether such a maneuver can have implications in disease states. PMID:25643074

  3. Differential activities of bacteriophage depolymerase on bacterial polysaccharide: binding is essential but degradation is inhibitory in phage infection of K1-defective Escherichia coli.

    PubMed Central

    Pelkonen, S; Aalto, J; Finne, J

    1992-01-01

    Host range mutants were derived from bacteriophages PK1A and PK1E specific for the K1 polysialic acid capsule of Escherichia coli. The mutants were selected for their ability to infect E. coli bacteria with a low level of the K1 capsule. A specific loss of the cleaving activity of the phage endosialidase was observed in all the mutants, while the ability to bind specifically to the polysialic acid capsule was retained. The results indicate that the polysaccharide-binding activity of the bacteriophage enzyme is essential for the infection process. The cleaving activity, in contrast, is required for the penetration of the dense polysaccharide of wild-type bacteria but is inhibitory in the infection of bacteria with a sparse capsular polysaccharide. Images PMID:1447142

  4. Evaluation of a multiplex PCR for detection of serotypes K1, K2 and K5 in Klebsiella sp. and comparison of isolates within these serotypes.

    PubMed

    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.

  5. Combining Electron Crystallography and X-ray Crystallography to Study the MlotiK1 Cyclic Nucleotide-Regulated Potassium Channel

    SciTech Connect

    Clayton, G.; Aller, S; Wang, J; Unger, V; Morais-Cabral, J

    2009-01-01

    We have recently reported the X-ray structure of the cyclic nucleotide-regulated potassium channel, MlotiK1. Here we describe the application of both electron and X-ray crystallography to obtain high quality crystals. We suggest that the combined application of these techniques provides a useful strategy for membrane protein structure determination. We also present negative stain projection and cryo-data projection maps. These maps provide new insights about the properties of the MlotiK1 channel. In particular, a comparison of a 9 {angstrom} cryo-data projection with calculated model maps strongly suggests that there is a very weak interaction between the pore and the S1-S4 domains of this 6 TM tetrameric cation channel and that the S1-S4 domains can adopt multiple orientations relative to the pore.

  6. Investigation of PI3K/PKB/mTOR/S6K1 signaling pathway in relationship of type 2 diabetes and Alzheimer’s disease

    PubMed Central

    Ma, Yunqing; Wu, Dongke; Zhang, Wei; Liu, Jiankun; Chen, Siping; Hua, Binghong

    2015-01-01

    The aim of this study was to investigate the roles of PI3K/PKB/mTOR/S6K1 signaling pathway in the risk-increasing mechanisms of type 2 diabetes mellitus (T2DM) towards the Alzheimer’s disease (AD). Based on the high-sugar high-fat diet, the single intraperitoneal injection of streptozotocin was performed to induce the T2DM rat model; the immunohistochemistry and RT-PCR technique were then performed to detect the expression levels of mTOR, PI3K, PKB, S6K1 and phosphorylated Tau protein in the hippocampal tissues of each group. The related metabolic indicators of the T2DM group and the T2DM + AD group were significantly higher than the normal control group and the AD group (P<0.01); the Morris water maze test of the AD group and the learning and memory of the T2DM + AD group were than significantly decreased than the T2DM group (P<0.01); the T2DM + AD group exhibited significantly increased expression levels of mTOR, S6K1 and Tau protein in the hippocampal tissues than the AD group and the T2DM group (P<0.05), and while the expression levels of PI3K and PKB were decreased (P<0.05). Among the possible mechanisms through which T2DM increased the risk of AD, the dystransduction of insulin signaling pathway (PI3K/PKB/mTOR/S6K1) was the important cause of hyperphosphorylation of Tau protein, thus it prompted the AD occurrence. PMID:26770471

  7. Stability of fat-soluble vitamins A (retinol palmitate), E (tocopherol acetate), and K1 (phylloquinone) in total parenteral nutrition at home.

    PubMed

    Billion-Rey, F; Guillaumont, M; Frederich, A; Aulagner, G

    1993-01-01

    Our purpose was to extend previous studies of the stability of vitamins A (retinol palmitate), E (tocopherol acetate), and K1 (phylloquinone) to total parenteral nutrition at-home (TPNH) admixtures. First, stability over 20 days was tested. Experimental conditions included presence or absence of lipids, presence or absence of trace elements, and storage in a glass bottle or in a single or multi-layer plastic bag (ethylene vinyl acetate, polyvinyl chloride, Stedim 5, and Stedim 6). The 20-day storage studies were conducted at 4 degrees C or at ambient air temperature. The second part of the study consisted of exposing to natural light TPNH admixtures with or without lipids, but with trace elements, in the same containers (except polyvinyl chloride). Finally, a clinical situation of TPNH was simulated with a TPNH admixture prepared 11 days before the test in a Stedim 6 plastic bag and stored at 4 degrees C in total darkness. For vitamins A, E, and K1, we observed good stability for 20 days; the final concentrations ranged from 75% to 100% of initial concentrations whatever the conditions studied. It appears that there is no significant difference of action between all containers and that the presence or absence of lipids and trace elements in admixtures stored at 4 degrees C or ambient temperature makes no difference. With exposure to sunlight, vitamin losses were 100% at 3 hours for vitamin A and 50% for vitamin K1; vitamin E concentrations were unchanged after 12 hours of experiment.(ABSTRACT TRUNCATED AT 250 WORDS)

  8. CYP4F2 Is a Vitamin K1 Oxidase: An Explanation for Altered Warfarin Dose in Carriers of the V433M Variant

    PubMed Central

    McDonald, Matthew G.; Rieder, Mark J.; Nakano, Mariko; Hsia, Clara K.; Rettie, Allan E.

    2009-01-01

    Genetic polymorphisms in VKORC1 and CYP2C9, genes controlling vitamin K1 (VK1) epoxide reduction and (S)-warfarin metabolism, respectively, are major contributors to interindividual variability in warfarin dose. The V433M polymorphism (rs2108622) in CYP4F2 has also been associated with warfarin dose and speculatively linked to altered VK1 metabolism. Therefore, the purpose of the present study was to determine the role of CYP4F2 and the V433M polymorphism in the metabolism of VK1 by human liver. In vitro metabolic experiments with accompanying liquid chromatography-tandem mass spectrometry analysis demonstrated that recombinant CYP4F2 (Supersomes) and human liver microsomes supplemented with NADPH converted VK1 to a single product. A screen of all commercially available P450 Supersomes showed that only CYP4F2 was capable of metabolizing VK1 to this product. Steady-state kinetic analysis with recombinant CYP4F2 and with human liver microsomes revealed a substrate Km of 8 to 10 μM. Moreover, anti-CYP4F2 IgG, as well as several CYP4F2-selective chemical inhibitors, substantially attenuated the microsomal reaction. Finally, human liver microsomes genotyped for rs2108622 demonstrated reduced vitamin K1 oxidation and lower CYP4F2 protein concentrations in carriers of the 433M minor allele. These data demonstrate that CYP4F2 is a vitamin K1 oxidase and that carriers of the CYP4F2 V433M allele have a reduced capacity to metabolize VK1, secondary to an rs2108622-dependent decrease in steady-state hepatic concentrations of the enzyme. Therefore, patients with the rs2108622 polymorphism are likely to have elevated hepatic levels of VK1, necessitating a higher warfarin dose to elicit the same anticoagulant response. PMID:19297519

  9. Final report on the torque key komparison CCM.T-K1.2 measurand torque: 0 N.m, 500 N.m, 1000 N.m

    NASA Astrophysics Data System (ADS)

    Röske, Dirk

    2015-01-01

    The purpose of the CIPM subsequent bilateral comparison CCM.T-K1.2 was to link another participant, namely the National Institute of Metrology (Thailand), in short NIMT, to the CCM.T-K1 torque key comparison. The measuring capabilities up to 1000 N.m of dead-weight torque standard machines with supported lever were investigated. The pilot laboratory was the same in both comparisons—it was the Physikalisch-Technische Bundesanstalt (PTB, Braunschweig, Germany). The same two very stable torque transducers with well-known properties were used as travelling standards. The measurements at the participating laboratory were carried out between November 2007 and February 2008. According to the technical protocol, torque steps of 500 N.m and 1000 N.m had to be measured both in clockwise and anticlockwise directions. Corrections had to be applied to the results reported by the participants taking into account the use of different amplifiers, the creep (due to different loading times of the machines) and the environmental conditions in the laboratories (temperature and relative humidity of the ambient air). The results of the pilot laboratory in this bilateral comparison are in very good agreement with the same results obtained in the CCM.T-K1 comparison. For each of the transducers, the two torque steps and both senses of direction of the torque vector, the key comparison reference value of the CCM.T-K1 was taken, and the results of participant NIMT were calculated with respect to these values. The agreement between the results is very good. The smallest expanded (k = 2) relative uncertainty of the machine stated by the participant is 1 × 10-4. The results of the comparison support this uncertainty statement. 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 CCM, according

  10. Characterization of pII Family (GlnK1, GlnK2, GlnB) Protein Uridylylation in Response to Nitrogen Availability for Rhodopseudomonas palustris

    SciTech Connect

    Connelly, Heather M; Pelletier, Dale A; Lu, Tse-Yuan; Lankford, Patricia K; Hettich, Robert {Bob} L

    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 proteins

  11. Characterization of Escherichia coli K1 colominic acid-specific murine antibodies that are cross-protective against Neisseria meningitidis groups B, C, and Y.

    PubMed

    Park, In Ho; Lin, Jisheng; Choi, Ji Eun; Shin, Jeon-Soo

    2014-06-01

    The capsular polysaccharide (PS) of Neisseria meningitidis serogroup B (NMGB) is α(2-8)-linked N-acetylneuraminic acid (Neu5Ac), which is almost identical to the O-acetylated colominic acid (CA) of Escherichia coli K1 Although E. coli K1 has long been known to elicit cross-protective antibodies against NMGB, limited information on these highly cross-reactive antibodies is available. In the present study, six new monoclonal antibodies (mAbs) specific to both E. coli K1 CA and NMGB PS were produced by immunizing Balb/c mice with E. coli K1, and their serological and molecular properties were characterized, together with 12 previously reported hybridoma mAbs. Among the bactericidal mAbs against NMGB, both HmenB5 and HmenB18, which are genetically identical though of different mouse origins, were able to kill serogroup C and Y meningococci. Based on SPR sensograms, the binding affinity of HmenB18 for PS was suggested to be associated with at least two different binding forces: the polyanionicity of Neu5Ac and an interaction with the O-acetyl groups of Neu5Ac. Molecular analysis showed that similar to most mAbs presenting a few restricted V region germline genes, the V region genes of HmenB18 were 979% and 986% identical to the closest IGHV1-1401 and IGLV15-10301 germline gene alleles, respectively, and V-D-J editing in this mAb generated an unusually long VH-CDR3 sequence (17 amino acid residues), containing one basic arginine, two hydrophobic isoleucine residues and a 'YAMDY' motif. Models of the mAb combining sites demonstrate that most of the mAbs exhibited a wide, shallow groove with a high overall positive charge, as seen in mAb735, which is specific for a polyanionic helical epitope. In contrast, the combining site of HmenB18 was shown to be wide but to present a relatively weak positive charge, consistent with the extensive recognition by HmenB18 of the various structural epitopes formed with the Neu5Ac residue and its O-acetylation.

  12. Analysis of vitamin K1 in fruits and vegetables using accelerated solvent extraction and liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization.

    PubMed

    Jäpelt, Rie Bak; Jakobsen, Jette

    2016-02-01

    The objective of this study was to develop a rapid, sensitive, and specific analytical method to study vitamin K1 in fruits and vegetables. Accelerated solvent extraction and solid phase extraction was used for sample preparation. Quantification was done by liquid chromatography tandem mass 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. The repeatability was 5.2% and the internal reproducibility was 6.2%. Recovery was in the range 90-120%. No significant difference was observed between the results obtained by the present method and by a method using the same principle as the CEN-standard i.e. liquid-liquid extraction and post-column zinc reduction with fluorescence detection. Limit of quantification was estimated to 0.05 μg/100g fresh weight.

  13. Global ocean tides. Part IV. The diurnal luni-solar declination tide (K1), atlas of tidal charts and maps. Final report

    SciTech Connect

    Schwiderski, E.W.

    1981-05-15

    In Part I of this report (A060 913), a unique hydrodynamical interpolation technique was introduced, extensively tested, and evaluated in order to partial global ocean tides in great detail and with a high degree of accuracy. This novel method has been applied to construct the diurnal luni-solar declination (K1) ocean tide with a relative accuracy of better than 5 cm anywhere in the open oceans. The resulting tidal amplitudes and phases are tabulated on a 1 deg x 1 deg grid system in an atlas of 42 deg x 71 deg overlapping charts covering the whole oceanic globe. A corresponding atlas of global corange and cotidal maps is included to provide the reader with a quick general overview of the major tidal phenomena. The specifying hydrodynamical parameters of the model are listed along with quoted sources of empirical tide data, and significant tidal features are explained and discussed. The diurnal K1 ocean tide is found to resemble qualitatively the semidiurnal M2 and S2 tides presented in Parts II (AD-A084 694) and III (AD-A104 333) of this report. However, major shifts of the positions of the amphidromes are apparent.

  14. Analysis of vitamin K1 in fruits and vegetables using accelerated solvent extraction and liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization.

    PubMed

    Jäpelt, Rie Bak; Jakobsen, Jette

    2016-02-01

    The objective of this study was to develop a rapid, sensitive, and specific analytical method to study vitamin K1 in fruits and vegetables. Accelerated solvent extraction and solid phase extraction was used for sample preparation. Quantification was done by liquid chromatography tandem mass 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. The repeatability was 5.2% and the internal reproducibility was 6.2%. Recovery was in the range 90-120%. No significant difference was observed between the results obtained by the present method and by a method using the same principle as the CEN-standard i.e. liquid-liquid extraction and post-column zinc reduction with fluorescence detection. Limit of quantification was estimated to 0.05 μg/100g fresh weight. PMID:26304366

  15. SERENDIPITOUS DETECTION OF X-RAY EMISSION FROM THE HOT BORN-AGAIN CENTRAL STAR OF THE PLANETARY NEBULA K 1-16

    SciTech Connect

    Montez, Rodolfo Jr.; Kastner, Joel H. E-mail: jhk@cis.rit.edu

    2013-03-20

    We report the serendipitous detection of point-like X-ray emission from the hot, PG1159-type central star of the planetary nebula (CSPN) K 1-16 by the XMM-Newton and Chandra X-Ray Observatories. The CSPN lies superimposed on a galaxy cluster that includes an X-ray-bright quasar, but we have successfully isolated the CSPN X-ray emission from the strong diffuse background contributed by the quasar and intracluster gas. We have modeled the XMM-Newton and Chandra X-ray data, taking advantage of the contrasting detection efficiencies of the two observatories to better constrain the low-energy spectral response of Chandra's Advanced CCD Imaging Spectrometer. We find that the CSPN X-ray spectrum is well characterized by the combination of a non-local thermodynamic equilibrium model atmosphere with T{sub *} {approx} 135 kK and a carbon-rich, optically thin thermal plasma with T{sub X} {approx} 1 MK. These results for X-ray emission from the K 1-16 CSPN, combined with those obtained for other PG1159-type objects, lend support to the 'born-again' scenario for Wolf-Rayet and PG1159 CSPNe, wherein a late helium shell flash dredges up carbon-rich intershell material and ejects this material into the circumstellar environment.

  16. Vimentin, a Novel NF-κB Regulator, Is Required for Meningitic Escherichia coli K1-Induced Pathogen Invasion and PMN Transmigration across the Blood-Brain Barrier

    PubMed Central

    Zhang, Bao; Liu, Li-Qun; Wu, Xuedong; Mor-Vaknin, Nirit; Markovitz, David M.; Cao, Hong; Zhou, Yan-Hong

    2016-01-01

    Background NF-κB activation, pathogen invasion, polymorphonuclear leukocytes (PMN) transmigration (PMNT) across the blood-brain barrier (BBB) are the pathogenic triad hallmark features of bacterial meningitis, but the mechanisms underlying these events remain largely unknown. Vimentin, which is a novel NF-κB regulator, is the primary receptor for the major Escherichia coli K1 virulence factor IbeA that contributes to the pathogenesis of neonatal bacterial sepsis and meningitis (NSM). We have previously shown that IbeA-induced NF-κB signaling through its primary receptor vimentin as well as its co-receptor PTB-associated splicing factor (PSF) is required for pathogen penetration and leukocyte transmigration across the BBB. This is the first in vivo study to demonstrate how vimentin and related factors contributed to the pathogenic triad of bacterial meningitis. Methodology/Principal Findings The role of vimentin in IbeA+ E. coli K1-induced NF-κB activation, pathogen invasion, leukocyte transmigration across the BBB has now been demonstrated by using vimentin knockout (KO) mice. In the in vivo studies presented here, IbeA-induced NF-κB activation, E. coli K1 invasion and polymorphonuclear neutrophil (PMN) transmigration across the BBB were significantly reduced in Vim-/- mice. Decreased neuronal injury in the hippocampal dentate gyrus was observed in Vim-/- mice with meningitis. The major inflammatory regulator α7 nAChR and several signaling molecules contributing to NF-κB activation (p65 and p-CamKII) were significantly reduced in the brain tissues of the Vim-/- mice with E. coli meningitis. Furthermore, Vim KO resulted in significant reduction in neuronal injury and in α7 nAChR-mediated calcium signaling. Conclusion/Significance Vimentin, a novel NF-κB regulator, plays a detrimental role in the host defense against meningitic infection by modulating the NF-κB signaling pathway to increase pathogen invasion, PMN recruitment, BBB permeability and neuronal

  17. KEY COMPARISON: Final report, on-going key comparison BIPM.QM-K1: Ozone at ambient level, comparison with ISCIII, 2007

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Wielgosz, Robert; Morillo Gomez, Pilar; Sánchez Blaya, Carmen

    2009-01-01

    As part of the on-going key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone mole fraction range of 0 nmol/mol to 500 nmol/mol. 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).

  18. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with ISCIII (December 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Morillo Gomez, Pilar; Sánchez, Carmen

    2013-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. 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 (CIPM MRA).

  19. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with ISCIII (December 2010)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Morillo Gomez, Pilar; Sánchez, Carmen

    2011-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. 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).

  20. Synthesis, structural and vibrational studies on mixed alkali metal gadolinium double tungstate, K1-xNaxGd(WO4)2

    NASA Astrophysics Data System (ADS)

    Durairajan, A.; Thangaraju, D.; Moorthy Babu, S.

    2013-02-01

    Mixed alkali double tungstates K1-xNaxGd(WO4)2 (KNGW) (0 ⩽ x ⩽ 1) were synthesized by solid state reaction using sodium doped monoclinic KGd(WO4)2 (KGW). Synthesized KNGW powders were characterized using powder X-ray diffraction (XRD), differential thermal analysis (DTA), scanning electron microscopy (SEM) and Raman analysis. DTA analysis confirms that the melting point of the KGW matrix increases from 1063 °C to 1255 °C with increasing sodium content. The Powder XRD analyses reveal that mixed phases were observed up to 40 wt.% of Na in the KGW matrix above that percentage there is domination of scheelite structure in the synthesized powder. Polyhedral type, bi-pyramidal shape and spheroid shape morphology was observed for KGW, NKGW and NGW powders respectively. The Raman analysis was carried out to understand the vibrational characteristic changes with mixing of sodium ions in the KGW matrix.

  1. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with NMC, A*STAR, May 2013

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Wielgosz, Robert; Li, Hou; Hui, Wendy Liu; Norris, James E.; Guenther, Frank

    2014-01-01

    As part of the on-going key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Singapore maintained by the National Metrology Centre, A*STAR (NMC, A*STAR) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM), via a transfer standard maintained by the National Institute of Standards and Technology (NIST). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol 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 (CIPM MRA).

  2. KEY COMPARISON: BIPM comparison BIPM.RI(II)-K1.Cu-64 of the activity measurements of the radionuclide 64Cu

    NASA Astrophysics Data System (ADS)

    Michotte, C.; Courte, S.; Ratel, G.; Kossert, K.; Nähle, O. J.

    2009-01-01

    In 2009, the Physikalisch-Technische Bundesanstalt (PTB), Germany, submitted a sample of known activity of 64Cu to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM). The value of the activity submitted was about 9.3 MBq. The result of this new comparison has been approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), comparison identifier BIPM.RI(II)-K1.Cu-64. 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).

  3. Final report, on-going key comparison BIPM.QM-K1: ozone at ambient level, comparison with DMDM, July 2015

    NASA Astrophysics Data System (ADS)

    Viallon, J.; Moussay, P.; Wielgosz, R.; Bebic, J.; Norris, J. E.; Guenther, F.

    2016-01-01

    As part of the on-going key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Directorate of Measures and Precious Metals (DMDM) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM), via a transfer standard maintained by the National Institute of Standards and Technology (NIST). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol 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 (CIPM MRA).

  4. Investigation into the evolution of the structure of K1-x LixTa1-y NbyO3 single crystals under variations in temperature

    NASA Astrophysics Data System (ADS)

    Borisov, S. A.; Vakhrushev, S. B.; Koroleva, E. Yu.; Naberezhnov, A. A.; Syrnikov, P. P.; Simkin, V. G.; Kutnjak, Z.; Egami, T.; Dmowski, W.; Piekarz, P.

    2007-05-01

    The evolution of the structure of K1-x LixTa1-y NbyO3 single crystals with x = 0.001, y = 0.026, and 1900 ppm Cu (KLTN277) and with x = 0.0014 and y = 0.024 (KLTN123), which exhibit an extremely high permittivity (up to 4 × 105 in the quasi-static regime for the KLTN277 crystal), is investigated in the range from room temperature to 20 K. It is demonstrated that, upon cooling to the lowest temperatures, both crystals retain their cubic structure, but the lattice parameters pass through a minimum at the temperature of the observed anomalies of the dielectric response (˜50 K). In the neutron diffraction pattern of the KLTN123 sample, satellites appear in the vicinity of the (hhh) reflections at temperatures below ˜50 K. These satellites can be associated with the nucleation of the rhombohedral phase.

  5. Fe-vacancy ordering in superconducting K1–xFe2–ySe2: First-principles calculations and Monte Carlo simulations

    DOE PAGES

    Fang, Yong; Tai, Yuan -Yen; Deng, Junkai; Wu, Chao; Ding, Xiangdong; Sun, Jun; Salje, Ekhard K. H.

    2015-07-20

    Fe vacancies in the 33 K superconductor K1–xFe2–ySe2 show ordering schemes that may be correlated with its superconducting properties. First-principles calculations and kinetic Monte Carlo simulations lead to a very simple model for vacancy ordering. Repulsive dipolar interactions between Fe vacancies show three ground states: amore » $$\\sqrt{8}\\times \\sqrt{10}$$ rhombus-ordered structure for 12.5% vacancies, a $$\\sqrt{5}\\times \\sqrt{5}$$ squared lattice for 20% vacancies, and a $$\\sqrt{5}\\times \\sqrt{5}$$ rhombus-ordered structure for 25% vacancies. Other structural states are derived from these three ground states and may contain additional disordered spatial regions. As a result, the repulsive interaction between Fe vacancies arises from enhanced Fe–Se covalent bonds, which differs from the well-known attractive interaction of Fe vacancies in body-centered cubic Fe.« less

  6. Thermodynamic studies on the ferroelectric phase transition in neutron irradiated (Li(sub x)K(1-x))2SO4 crystals at high temperature

    NASA Astrophysics Data System (ADS)

    Kassem, M. E.; El-Khatib, A. M.; Ammar, E. A.; Denton, M. M.

    1989-05-01

    Thermodynamic studies of (Li(x)K(1-x))2SO4, LKS, mixed crystals have been made in the concentration range (x=0.1,0.2,..., x=0.5). The thermal behavior has been investigated by differential thermal analysis, DTA, and differential scanning calorimeter, DSC, in the vicinity of high temperature phases. Also, the effect of the mixed neutron field of fast and thermal neutrons (10 percent of the reactor neutron pile is fast neutrons) on the thermal properties of mixed crystals was studied. The results show a change in the transition temperature Tc, as well as the value of specific heat Cp at transition temperature, due to the change of the stoichiometric ratio and the radiation doses. The changes of enthalpy and entropy of mixed crystals have been estimated numerically. The obtained small values of Delta S/R is characteristic of incommensurate phase transition as previously confirmed by the results of neutron diffraction technique.

  7. Origin of giant piezoelectric effect in lead-free K1−xNaxTa1−yNbyO3 single crystals

    PubMed Central

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-01-01

    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. PMID:27160075

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

    PubMed

    Apró, William; Moberg, Marcus; Hamilton, D Lee; Ekblom, Björn; van Hall, Gerrit; Holmberg, Hans-Christer; Blomstrand, Eva

    2015-03-15

    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 assessed for glycogen content, kinase activity, protein phosphorylation, and gene expression. Mixed muscle fractional synthetic rate was measured at rest and during 3 h of recovery using the stable isotope technique. In ER, AMPK activity was elevated immediately after both endurance and resistance exercise (∼90%, P < 0.05) but was unchanged in R. Thr(389) phosphorylation of S6K1 was increased severalfold immediately after exercise (P < 0.05) in both trials and increased further throughout recovery. After 90 and 180 min recovery, S6K1 activity was elevated (∼55 and ∼110%, respectively, P < 0.05) and eukaryotic elongation factor 2 phosphorylation was reduced (∼55%, P < 0.05) with no difference between trials. In contrast, markers for protein catabolism were differently influenced by the two modes of exercise; ER induced a significant increase in gene and protein expression of MuRF1 (P < 0.05), which was not observed following R exercise only. In conclusion, cycling-induced elevation in AMPK activity does not inhibit mTOR complex 1 signaling after subsequent resistance exercise but may instead interfere with the hypertrophic response by influencing key components in protein breakdown.

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

    PubMed Central

    Miller, Steven L.; Aroniadou-Anderjaska, Vassiliki; Figueiredo, Taiza H.; Prager, Eric M.; Almeida-Suhett, Camila P.; Apland, James P.; Braga, Maria F.M.

    2015-01-01

    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 LD50 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 (1.2XLD50), 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. PMID:25689173

  10. Origin of giant piezoelectric effect in lead-free K1-xNaxTa1-yNbyO3 single crystals.

    PubMed

    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.

  11. Origin of giant piezoelectric effect in lead-free K1‑xNaxTa1‑yNbyO3 single crystals

    NASA Astrophysics Data System (ADS)

    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.

  12. Origin of giant piezoelectric effect in lead-free K1-xNaxTa1-yNbyO3 single crystals.

    PubMed

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-01-01

    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. PMID:27160075

  13. Sphingosine kinase-1 mediates androgen-induced osteoblast cell growth

    SciTech Connect

    Martin, Claire; Lafosse, Jean-Michel; Malavaud, Bernard; Cuvillier, Olivier

    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.

  14. Function and expression of CD1d and invariant natural killer T-cell receptor in the cotton rat (Sigmodon hispidus).

    PubMed

    Fichtner, Alina Suzann; Paletta, Daniel; Starick, Lisa; Schumann, Richard F; Niewiesk, Stefan; Herrmann, Thomas

    2015-12-01

    The cotton rat (Sigmodon hispidus) belongs to the rodent family of Cricetidae and provides a powerful model to study the pathogenesis of human respiratory viruses and measles virus. Recent studies in other rodent models have suggested a role for invariant natural killer T (iNKT) cells in antiviral immunity and vaccination against respiratory virus infections. Using new experimental tools, we provide the first evidence for a functional CD1d cell molecule (crCD1d) and iNKT T-cell receptor in cotton rats. The crCD1d cDNA sequence was identified and crCD1d transductants showed that monoclonal antibody WTH-2 stains crCD1d as efficiently as mouse or rat CD1d. The expression of crCD1d was clearly weaker for thymocytes and B cells, and higher for T cells, which is different to what is found in murine species. The antigen-presenting capacity of crCD1d was demonstrated with crCD1d-immunoglobulin dimers loaded with the glycolipid PBS57, which bound iNKT T-cell receptors. Evidence for functional cotton rat iNKT cells was provided by detection of interferon-γ and interleukin-4 in cultures of splenocytes stimulated with PBS57 and α-galactosylceramide and by specific staining of about 0·2% of splenocytes with PBS57-loaded crCD1d dimers. Canonical AV14/AJ18 rearrangements were identified and found to contain multiple members of the AV14 (AV11) family. One of them was expressed and found to bind CD1d dimers. In summary, these data provide the first evidence for functional CD1d molecules and iNKT T-cell receptors in cotton rats and provide the tools to analyse them both in the cotton rat model of infectious diseases.

  15. Efficient cleavage of single and clustered AP site lesions within mono-nucleosome templates by CHO-K1 nuclear extract contrasts with retardation of incision by purified APE1

    PubMed Central

    Eccles, Laura J.; Menoni, Hervé; Angelov, Dimitar; Lomax, Martine E.; O’Neill, Peter

    2015-01-01

    Clustered DNA damage is a unique characteristic of radiation-induced DNA damage and the formation of these sites poses a serious challenge to the cell’s repair machinery. Within a cell DNA is compacted, with nucleosomes being the first order of higher level structure. However, few data are reported on the efficiency of clustered-lesion processing within nucleosomal DNA templates. Here, we show retardation of cleavage of a single AP site by purified APE1 when contained in nucleosomal DNA, compared to cleavage of an AP site in non-nucleosomal DNA. This retardation seen in nucleosomal DNA was alleviated by incubation with CHO-K1 nuclear extract. When clustered DNA damage sites containing bistranded AP sites were present in nucleosomal DNA, efficient cleavage of the AP sites was observed after treatment with nuclear extract. The resultant DSB formation led to DNA dissociating from the histone core and nucleosomal dispersion. Clustered damaged sites containing bistranded AP site/8-oxoG residues showed no retardation of cleavage of the AP site but retardation of 8-oxoG excision, compared to isolated lesions, thus DSB formation was not seen. An increased understanding of processing of clustered DNA damage in a nucleosomal environment may lead to new strategies to enhance the cytotoxic effects of radiotherapeutics. PMID:26439176

  16. 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.

    PubMed Central

    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. Images PMID:6345392

  17. Integral Representation of the Pictorial Proof of Sum of [superscript n][subscript k=1]k[superscript 2] = 1/6n(n+1)(2n+1)

    ERIC Educational Resources Information Center

    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…

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

    SciTech Connect

    Miller, Steven L.; Aroniadou-Anderjaska, Vassiliki; Figueiredo, Taiza H.; Prager, Eric M.; Almeida-Suhett, Camila P.; Apland, James P.; 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

  19. Investigation of diffuse phase transition in ferroelectric Pb2- x K1+ x Li x Nb5O15 (0 ≤ x ≤ 1.5) ceramics

    NASA Astrophysics Data System (ADS)

    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.

  20. Dielectric and structural properties of diffuse ferroelectric phase transition in Pb1.85K1.15Li0.15Nb5O15 ceramic

    NASA Astrophysics Data System (ADS)

    Choukri, E.; Gagou, Y.; Mezzane, D.; Abkhar, Z.; El Moznine, R.; Luk'yanchuk, I.; Saint-Grégoire, P.; Kavokin, A. V.

    2011-02-01

    We studied the structural and dielectric properties of new Tetragonal Tungsten Bronze (TTB) ceramics Pb1.85K1.15Li0.15Nb5O15 that was synthesized by solid-state reaction. We pay a special attention to the diffuse phase transition (DPT) that occurs close to 425 °C. Using dielectric measurements in a frequency range of 10 Hz-1 MHz and in the temperature range 30-560 °C, we have shown that the real permittivity close to DPT is well described by Santos-Eiras phenomenological model. Space-charge polarization, relaxation phenomena and free charges conductivity have been analyzed using dielectric spectroscopy impedance and modulus characterization. Cole-Cole plots show a non-Debye (polydispersive) type relaxation. In paraelectric phase the Arrhenius activation energy was determined as Eτ = 0.72 eV. We demonstrated that frequency dependence of ac conductivity at different temperatures obeys the Jonscher's universal law: σac = σdc + A(ω)n.

  1. KEY COMPARISON: COOMET.RI(I)-K1 comparison of national measurement standards of air kerma for 60Co γ radiation

    NASA Astrophysics Data System (ADS)

    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).

  2. Three predictions: Comet 67P/Churyumov-Gerasimenko, comet C/2012 K1 PANSTARRS, and comet C/2013 V5 Oukaimeden

    NASA Astrophysics Data System (ADS)

    Ferrín, Ignacio

    2014-09-01

    We make the following predictions: (1) The secular light curve (SLC) of comet 67P/Churyumov-Gerasimenko exhibits a photometric anomaly in magnitude that is present in 1982, 1996, 2002 and 2009. Thus it must be real. We interpret this anomaly as a topographic feature on the surface of the nucleus that may be a field of debris, a region made only of dust or an area of solid stones but in any case it is depleted in volatiles. We predict that images taken by spacecraft Rosetta will show a region morphologically different to the rest of the nucleus, at the pole pointing to the Sun near perihelion. (2) Comet C/2012 K1 PANSTARRS exhibits the same Slope Discontinuity Event (SDE)+magnitude dip after the event than other comets listed in Table 1 most of which disintegrated. This group includes comet C/2012 S1 ISON. Thus, it is reasonable to expect that this comet may disintegrate too. The probability of disintegration of this comet is 27%. (3) Comet C/2013 V5 Oukaimeden exhibits the same SDE+standstill signature exhibited by other comets in Table 1. We predict that there is a 93% probability that this comet will disintegrate. (4) Another purpose of this work is to present evidence to conclude that the SLCs have predictive power.

  3. Origin of the crossover between a freezing and a structural transition at low concentration in the relaxor ferroelectric K1 -xLixTaO3

    NASA Astrophysics Data System (ADS)

    Cai, Ling; Toulouse, Jean; Harriger, Leland; Downing, R. Gregory; Boatner, L. A.

    2015-04-01

    The origin of the relaxor behavior in K1 -xLixTaO3(KLT ) and other disordered perovskites is now recognized to be due to the reorientation of the polar nanodomains formed by the correlated dipoles of off-center ions. The collective dynamics of these systems evolve through several temperature stages. On decreasing temperature below the so-called Burns temperature TB, individual dipoles become correlated within nanosized regions. On further cooling, the slow dynamics of these polar regions allows local lattice distortions to take place and the formation of polar nanodomains at T*

  4. Vertical Extraction Process Implemented at the 118-K-1 Burial Ground for Removal of Irradiated Reactor Debris from Silo Structures - 12431

    SciTech Connect

    Teachout, Douglas B.; Adamson, Clinton J.; Zacharias, Ames

    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. UV/Visible Observations of C/2012 K1 PanSTARRS and C/2013 A1 Siding Spring from a Stratospheric Telescope

    NASA Astrophysics Data System (ADS)

    Young, E. F.; Diller, J.; Dinkel, K.; Dischner, Z.; Cheng, A. F.; Hibbitts, C.; Osterman, S. N.

    2014-12-01

    The UV-VIS (Ultra-Violet/Visible) instrument on the BOPPS mission (Balloon Observation Platform for Planetary Science) has two main goals: to demonstrate pointing stability at the 0.1" level, commensurate with the near-space PSF (Point Spread Function) expected from the 80 cm aperture telescope, and to observe targets in wavelengths where the telluric transmission is low. Two potential targets of a BOPPS flight in September 2014 are comets C/2012 K1 PanSTARRS and C/2013 A1 Siding Spring. The UV-VIS science camera has been outfitted with four filters from the NASA's Hale-Bopp filter set: an OH filter at 308 nm, a CN filter at 385 nm and continuum filters at 345 and 445 nm. UV-VIS can potentially measure OH emission from the cometary targets if the BOPPS flight extends past sunset; the sky background due to Rayleigh scattering at 310 nm is too high for daytime observations, even from float altitudes near 35 km. The BOPPS telescope is stabilized at the few arcsecond level by inertial-guidance gyroscopes that control coarse elevation and azimuth reaction wheels. The UV-VIS bench includes a fast guide camera and an FSM (fine steering mirror) - both COTS items - to bridge the pointing gap from a few arcseconds to a tenth of an arcsecond. In two separate hang tests, the guide camera was able to provide a 50 Hz pointing error signal on a star, and the resulting FSM-corrected images were reduced in areal size by 50% (e.g., linear PSF widths reduced from 2.4" to 1.6"). The FSM performance is expected to achieve the 0.1" stability goal in the stratosphere, where the image degradation is caused by motion of the optical system instead of atmospheric turbulence.

  6. The Genome Sequence of Avian Pathogenic Escherichia coli Strain O1:K1:H7 Shares Strong Similarities with Human Extraintestinal Pathogenic E. coli Genomes▿

    PubMed Central

    Johnson, Timothy J.; Kariyawasam, Subhashinie; Wannemuehler, Yvonne; Mangiamele, Paul; Johnson, Sara J.; Doetkott, Curt; Skyberg, Jerod A.; Lynne, Aaron M.; Johnson, James R.; Nolan, Lisa K.

    2007-01-01

    Escherichia coli strains that cause disease outside the intestine are known as extraintestinal pathogenic E. coli (ExPEC) and include human uropathogenic E. coli (UPEC) and avian pathogenic E. coli (APEC). Regardless of host of origin, ExPEC strains share many traits. It has been suggested that these commonalities may enable APEC to cause disease in humans. Here, we begin to test the hypothesis that certain APEC strains possess potential to cause human urinary tract infection through virulence genotyping of 1,000 APEC and UPEC strains, generation of the first complete genomic sequence of an APEC (APEC O1:K1:H7) strain, and comparison of this genome to all available human ExPEC genomic sequences. The genomes of APEC O1 and three human UPEC strains were found to be remarkably similar, with only 4.5% of APEC O1's genome not found in other sequenced ExPEC genomes. Also, use of multilocus sequence typing showed that some of the sequenced human ExPEC strains were more like APEC O1 than other human ExPEC strains. This work provides evidence that at least some human and avian ExPEC strains are highly similar to one another, and it supports the possibility that a food-borne link between some APEC and UPEC strains exists. Future studies are necessary to assess the ability of APEC to overcome the hurdles necessary for such a food-borne transmission, and epidemiological studies are required to confirm that such a phenomenon actually occurs. PMID:17293413

  7. Catalytic properties and crystal structure of thermostable NAD(P)H-dependent carbonyl reductase from the hyperthermophilic archaeon Aeropyrum pernix K1.

    PubMed

    Fukuda, Yudai; Sakuraba, Haruhiko; Araki, Tomohiro; Ohshima, Toshihisa; Yoneda, Kazunari

    2016-09-01

    A gene encoding NAD(P)H-dependent carbonyl reductase (CR) from the hyperthermophilic archaeon Aeropyrum pernix K1 was overexpressed in Escherichia coli. Its product was effectively purified and characterized. The expressed enzyme was the most thermostable CR found to date; the activity remained at approximately 75% of its activity after incubation for 10min up to 90°C. In addition, A. pernix CR exhibited high stability at a wider range of pH values and longer periods of storage compared with CRs previously identified from other sources. A. pernix CR catalyzed the reduction of various carbonyl compounds including ethyl 4-chloro-3-oxobutanoate and 9,10-phenanthrenequinone, similar to the CR from thyroidectomized (Tx) chicken fatty liver. However, A. pernix CR exhibited significantly higher Km values against several substrates than Tx chicken fatty liver CR. The three-dimensional structure of A. pernix CR was determined using the molecular replacement method at a resolution of 2.09Å, in the presence of NADPH. The overall fold of A. pernix CR showed moderate similarity to that of Tx chicken fatty liver CR; however, A. pernix CR had no active-site lid unlike Tx chicken fatty liver CR. Consequently, the active-site cavity in the A. pernix CR was much more solvent-accessible than that in Tx chicken fatty liver CR. This structural feature may be responsible for the enzyme's lower affinity for several substrates and NADPH. The factors contributing to the much higher thermostability of A. pernix CR were analyzed by comparing its structure with that of Tx chicken fatty liver CR. This comparison showed that extensive formation of the intrasubunit ion pair networks, and the presence of the strong intersubunit interaction, is likely responsible for A. pernix CR thermostability. Site-directed mutagenesis showed that Glu99 plays a major role in the intersubunit interaction. This is the first report regarding the characteristics and three-dimensional structure of

  8. Summer (subarctic) versus winter (subtropic) production affects spinach (Spinacia oleracea L.) leaf bionutrients: vitamins (C, E, Folate, K1, provitamin A), lutein, phenolics, and antioxidants.

    PubMed

    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.

  9. RioK1, a New Interactor of Protein Arginine Methyltransferase 5 (PRMT5), Competes with pICln for Binding and Modulates PRMT5 Complex Composition and Substrate Specificity*

    PubMed Central

    Guderian, Gernot; Peter, Christoph; Wiesner, Julia; Sickmann, Albert; Schulze-Osthoff, Klaus; Fischer, Utz; Grimmler, Matthias

    2011-01-01

    Protein arginine methylation plays a critical role in differential gene expression through modulating protein-protein and protein-DNA/RNA interactions. Although numerous proteins undergo arginine methylation, only limited information is available on how protein arginine methyltransferases (PRMTs) identify their substrates. The human PRMT5 complex consists of PRMT5, WD45/MEP50 (WD repeat domain 45/methylosome protein 50), and pICln and catalyzes the symmetrical arginine dimethylation of its substrate proteins. pICln recruits the spliceosomal Sm proteins to the PRMT5 complex for methylation, which allows their subsequent loading onto snRNA to form small nuclear ribonucleoproteins. To understand how the PRMT5 complex is regulated, we investigated its biochemical composition and identified RioK1 as a novel, stoichiometric component of the PRMT5 complex. We show that RioK1 and pICln bind to PRMT5 in a mutually exclusive fashion. This results in a PRMT5-WD45/MEP50 core structure that either associates with pICln or RioK1 in distinct complexes. Furthermore, we show that RioK1 functions in analogy to pICln as an adapter protein by recruiting the RNA-binding protein nucleolin to the PRMT5 complex for its symmetrical methylation. The exclusive interaction of PRMT5 with either pICln or RioK1 thus provides the first mechanistic insight into how a methyltransferase can distinguish between its substrate proteins. PMID:21081503

  10. Variable region sequences and idiotypic expression of a protective human immunoglobulin M antibody to capsular polysaccharides of Neisseria meningitidis group B and Escherichia coli K1.

    PubMed Central

    Azmi, F H; Lucas, A H; Raff, H V; Granoff, D M

    1994-01-01

    We determined the heavy (H)- and light (L)-chain variable (V) region nucleotide and translated amino acid sequences of the human immunoglobulin M(kappa) monoclonal antibody (MAb) 5E1, which is specific for the polysaccharide capsule of Escherichia coli K1 and Neisseria meningitidis group B (poly[alpha(2-->8)-N-acetylneuraminic acid]) and which is protective in animal models of infection. The 5E1 VH gene is a member of the VHIIIb family and is 97% homologous to the 9.1 germ line gene. The 5E1 VL gene is a member of the kappa I subgroup and is 98% homologous to the germ line gene, 15A, also known as KLO12. The VL and/or VH genes used by 5E1 are highly homologous to the V genes encoding antibodies to the Haemophilus influenzae type b polysaccharide and to antibodies reactive with self-antigens such as erythrocyte "i," DNA, and thyroid peroxidase. We also produced three murine anti-idiotype (Id) MAbs against 5E1. All three anti-Ids recognize a minor subset of antimeningococcal B polysaccharide antibodies present in serum from normal adults. Two of the anti-Ids define distinct Ids associated with antibodies having kappa I-15A V regions. These 15A-associated Ids are expressed by some heterologous human antimeningococcal B polysaccharide MAbs, and they also are independently expressed by two human MAbs that are specific for either the H. influenzae b polysaccharide or the i erythrocyte antigen and that utilize the kappa I-15A V region. Taken together, these data indicate that the 5E1 antibody uses V regions that recur in the human antibody repertoires to this polysaccharide and to structurally dissimilar polysaccharides and autoantigens. Thus, the poor immunogenicity of poly[alpha(2-->8)-N-acetylneuraminic acid] cannot be explained by the unavailability of certain critical VH and VL genes required for generation of antibody response. PMID:8168940

  11. Effect of phylloquinone (vitamin K1) supplementation for 12 months on the indices of vitamin K status and bone health in adult patients with Crohn's disease.

    PubMed

    O'Connor, Eibhlís M; Grealy, Geraldine; McCarthy, Jane; Desmond, Alan; Craig, Orla; Shanahan, Fergus; Cashman, Kevin D

    2014-10-14

    Although epidemiological findings support a role for vitamin K status in the improvement of bone indices in adult patients with Crohn's disease (CD), this needs to be confirmed in double-blind, randomised controlled trials (RCT) with phylloquinone (vitamin K1). By conducting two RCT, the present study aimed to first establish whether supplementation with 1000 μg of phylloquinone daily near-maximally suppresses the percentage of undercarboxylated osteocalcin in serum (%ucOC; marker of vitamin K status) in adult patients with CD currently in remission as it does in healthy adults and second determine the effect of supplementation with phylloquinone at this dose for 12 months on the indices of bone turnover and bone mass. The initial dose-ranging RCT was conducted in adult patients with CD (n 10 per group) using 0 (placebo), 1000 or 2000 μg of phylloquinone daily for 2 weeks. In the main RCT, the effect of placebo v. 1000 μg vitamin K/d (both co-administered with Ca (500 mg/d) and vitamin D3 (10 μg/d)) for 12 months (n 43 per group) on the biochemical indices of bone turnover (determined by enzyme immunoassay) and bone mass (determined by dual-energy X-ray absorptiometry) were investigated. At baseline, the mean %ucOC was 47 %, and this was suppressed upon supplementation with 1000 μg of phylloquinone daily ( - 81 %; P< 0·01) and not suppressed further by 2000 μg of phylloquinone daily. Compared with the placebo, supplementation with 1000 μg of phylloquinone daily for 12 months had no significant effect (P>0·1) on bone turnover markers or on the bone mass of the lumbar spine or femur, but modestly increased (P< 0·05) the bone mass of the total radius. Despite near maximal suppression of serum %ucOC, supplementation with 1000 μg of phylloquinone daily (with Ca and vitamin D3) had no effect on the indices of bone health in adult CD patients with likely vitamin K insufficiency.

  12. Effects of traditional Chinese medicine Xin-Ji-Er-Kang formula on 2K1C hypertensive rats: role of oxidative stress and endothelial dysfunction

    PubMed Central

    2013-01-01

    circumferential collagen area (PVCA), reduced the content of Hyp in the left ventricular tissue, inhibited the decrease of SOD activity and increase of MDA, Ang II content in serum. Moreover, treatment with XJEK improved endothelial dysfunction (ED) manifested by promoting endothelial-dependent vasodilation of thoracic aortic rings and enhancing the NO activity in serum. Conclusions These findings suggest that administration of XJEK possess protective effects against 2K1C induced hypertension and cardiac remodeling in rats, preserve NO activity and endothelial function. PMID:23849603

  13. The acute effects of strength, endurance and concurrent exercises on the Akt/mTOR/p70(S6K1) and AMPK signaling pathway responses in rat skeletal muscle.

    PubMed

    de Souza, E O; Tricoli, V; Bueno Junior, C; Pereira, M G; Brum, P C; Oliveira, E M; Roschel, H; Aoki, M S; Urginowitsch, C

    2013-04-01

    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/p70(S6K1) 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-Thr(172)AMPK (p-AMPK), Akt, phospho-Ser(473)Akt (p-Akt), p70(S6K1), phospho-Thr(389)-p70(S6K1) (p-p70(S6K1)), mTOR, phospho-Ser(2448)mTOR (p-mTOR), and phospho-Thr(1462)-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 p70(S6K1) 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.

  14. O1, P1, N2 models of the global ocean tide on an elastic earth plus surface potential and spherical harmonic decompositions for M2, S2, and K1

    NASA Technical Reports Server (NTRS)

    Parke, M. E.

    1982-01-01

    The models of M2, S2, and K1 presented in Parke and Hendershott (1980) are supplemented with models of O1, P1, and N2. The models satisfy specified elevation boundary conditions and are generated by fighting a small number of test functions to island data. Maps are presented of the geocentric tide, the induced free space potential, the induced vertical component of the solid earth tide, and the induced vertical component of the gravitational field for each new component. Maps of the tidal potential seen by an observer fixed to the surface of the solid earth are also presented for all six constituents. Spherical harmonic coefficients up to order four and the rms magnitude of the coefficients to order fifteen are presented for each constituent. The rms magnitudes of the P1 and K1 coefficients normalized by their respective equilibrium amplitudes are compared to determine the effect of the diurnal core resonance.

  15. Transitions 2s/2/ 2p/k/ - 2s 2p/k+1/ of the F I, O I, and N I isoelectronic sequences. [in laser plasma spectra

    NASA Technical Reports Server (NTRS)

    Doschek, G. A.; Cowan, R. D.; Cohen, L.; Feldman, U.

    1974-01-01

    Transitions of the type 2s/2/ 2p/k/ - 2s 2p/k+1/ have been identified for the elements from titanium through nickel for ions of the fluorine, oxygen, and nitrogen isoelectronic sequences. Wavelengths, visual intensity estimates, and energies are given. The energy differences of levels of the ground configuration are compared with predictions based on semiempirical equations derived by Edlen. Some of the lines of these isoelectronic sequences should be strong lines in solar-flare spectra.

  16. Zinc stimulates glucose consumption by modulating the insulin signaling pathway in L6 myotubes: essential roles of Akt-GLUT4, GSK3β and mTOR-S6K1.

    PubMed

    Wu, Yuntang; Lu, Huizi; Yang, Huijun; Li, Chunlei; Sang, Qian; Liu, Xinyan; Liu, Yongzhe; Wang, Yongming; Sun, Zhong

    2016-08-01

    The present study was performed to evaluate the insulin-like effects of zinc in normal L6 myotubes as well as its ability to alleviate insulin resistance. Glucose consumption was measured in both normal and insulin-resistant L6 myotubes. Western blotting and immunofluorescence revealed that zinc exhibited insulin-like glucose transporting effects by activating key markers that are involved in the insulin signaling cascade (including Akt, GLUT4 and GSK3β), and downregulating members of the insulin signaling feedback cascade such as mammalian target of rapamycin (mTOR) and ribosomal protein S6 kinase (S6K1). In normal L6 myotubes, zinc enhanced glucose consumption via a mechanism that might involve the activation of Akt phosphorylation, glucose transporter 4 (GLUT4) translocation and GSK3β phosphorylation. In contrast, zinc exerted insulin-mimetic effects in insulin-resistant L6 myotubes by upregulating Akt phosphorylation, GLUT4 translocation and GSK3β phosphorylation, and downregulating the expression of mTOR and S6K1. In conclusion, zinc might enhance glucose consumption by modulating insulin signaling pathways including Akt-GLUT4, GSK3β, mTOR and S6K1.

  17. Update of the BIPM comparison BIPM.RI(II)-K1.F-18 of activity measurements of the radionuclide 18F to include the 2010 result of the LNE-LNHB (France)

    NASA Astrophysics Data System (ADS)

    Michotte, C.; Ratel, G.; Courte, S.; Cassette, P.; Moune, M.

    2016-01-01

    Since 2001, six national metrology institutes (NMI) have submitted seven samples of known activity of 18F 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.F-18. The values of the activity submitted were between about 1 MBq and 18 MBq. The primary standardization result for the LNE-LNHB, France, replaces their earlier result of 2002 and the key comparison reference value (KCRV) has been recalculated. In the frame of the BIPM.RI(II)-K4.F18 comparison, the NPL updated their result in the KCDB. Consequently there are now five results in the BIPM.RI(II)-K1.F-18 comparison. The degrees of equivalence between each equivalent activity measured in the SIR and the updated 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).

  18. Artists Paint ... Fall: Grades K-1

    ERIC Educational Resources Information Center

    Herberholz, Barbara

    2012-01-01

    Artists often paint the different seasonal activities people engage in and the way the world looks as changes take place. The weather for each of the four seasons is different. Farmers plant crops and gardens in the spring and harvest their crops in the fall, just like "The Harvesters" by Pieter Bruegel the Elder. To begin, children will observe…

  19. Hawaiian Studies Curriculum Guide. Grades K-1.

    ERIC Educational Resources Information Center

    Hawaii State Dept. of Education, Honolulu. Office of Instructional Services.

    This curriculum guide suggests activities and educational experiences within a Hawaiian cultural context for kindergarten and Grade 1 students in Hawaiian schools. First, an introduction discusses the contents of the guide, the relationship of the classroom teacher and the kupuna (Hawaiian-speaking elder); the identification and scheduling of…

  20. Hierarchical domain structure of lead-free piezoelectric (Na1/2 Bi1/2)TiO3-(K1/2 Bi1/2)TiO3 single crystals

    NASA Astrophysics Data System (ADS)

    Luo, Chengtao; Wang, Yaojin; Ge, Wenwei; Li, Jiefang; Viehland, Dwight; Delaire, Olivier; Li, Xiaobin; Luo, Haosu

    2016-05-01

    We report a unique hierarchical domain structure in single crystals of (Na1/2Bi1/2)TiO3-xat. %(K1/2Bi1/2)TiO3 for x = 5 and 8 by transmission electron microscopy (TEM). A high density of polar nano-domains with a lamellar morphology was found, which were self-assembled into a quadrant-like configuration, which then assembled into conventional ferroelectric macro-domains. Studies by high resolution TEM revealed that the polar lamellar regions contained a coexistence of in-phase and anti-phase oxygen octahedral tilt regions of a few nanometers in size. Domain frustration over multiple length scales may play an important role in the stabilization of the hierarchy, and in reducing the piezoelectric response of this Pb-free piezoelectric solid solution.

  1. Can PC-9 Zhong chong replace K-1 Yong quan for the acupunctural resuscitation of a bilateral double-amputee? Stating the “random criterion problem” in its statistical analysis

    PubMed Central

    Inchauspe, Adrián Angel

    2016-01-01

    AIM: To present an inclusion criterion for patients who have suffered bilateral amputation in order to be treated with the supplementary resuscitation treatment which is hereby proposed by the author. METHODS: This work is based on a Retrospective Cohort model so that a certainly lethal risk to the control group is avoided. RESULTS: This paper presents a hypothesis on acupunctural PC-9 Zhong chong point, further supported by previous statistical work recorded for the K-1 Yong quan resuscitation point. CONCLUSION: Thanks to the application of the resuscitation maneuver herein proposed on the previously mentioned point, patients with bilateral amputation would have another alternative treatment available in case basic and advanced CPR should fail. PMID:27152257

  2. Transposon-encoded sucrose metabolism in Lactococcus lactis. Purification of sucrose-6-phosphate hydrolase and genetic linkage to N5-(L-1-carboxyethyl)-L-ornithine synthase in strain K1.

    PubMed

    Thompson, J; Nguyen, N Y; Sackett, D L; Donkersloot, J A

    1991-08-01

    Sucrose-6-phosphate hydrolase from Lactococcus lactis subsp. lactis K1-23 (formerly Streptococcus lactis K1-23) has been purified 600-fold to electrophoretic homogeneity. Purification of the enzyme was achieved by DEAE-Sephacel, phosphocellulose P-11, and gel exclusion (Ultrogel AcA 54) chromatography. The purified enzyme (specific activity 31 units/mg) catalyzed the hydrolysis of both 6-O-phosphoryl-alpha-D-glucopyranosyl-1,2-beta-D-fructofuranoside (sucrose 6-phosphate) and sucrose (Km = 0.1 and 100 mM, respectively). Ultracentrifugal analysis of sucrose-6-phosphate hydrolase indicated an Mr = 52,200. The purified enzyme migrated as a single protein during sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr = 52,000). However, four distinct polypeptides were detected by analytical electrofocusing, and all four species hydrolyzed sucrose and sucrose 6-phosphate. The amino acid composition of sucrose-6-phosphate hydrolase, and the sequence of the first 12 amino acids from the NH2 terminus, have been determined. Hybridization studies with oligonucleotide probes show that the genes for sucrose-6-phosphate hydrolase (scrB), Enzyme IIScr of the phosphoenolypyruvate-dependent sucrose:phosphotransferase system (scrA), and N5-(carboxyethyl)ornithine synthase (ceo) are encoded by the same approximately 20-kilobase EcoRI fragment. This fragment is part of a large transposon Tn5306 that also encodes the nisin precursor gene, spaN, and IS904. In L. lactis ATCC 11454, spaN, IS904, scrA, and scrB (but not ceo) are encoded on a related transposon, Tn5307.

  3. Glycosaminoglycans affect heparanase location in CHO cell lines.

    PubMed

    Piva, Maria B R; Suarez, Eloah R; Melo, Carina M; Cavalheiro, Renan P; Nader, Helena B; Pinhal, Maria A S

    2015-09-01

    Glycosaminoglycans (GAG) play a ubiquitous role in tissues and cells. In eukaryotic cells, heparan sulfate (HS) is initially degraded by an endo-β-glucuronidase called heparanase-1 (HPSE). HS oligosaccharides generated by the action of HPSE intensify the activity of signaling molecules, activating inflammatory response, tumor metastasis, and angiogenesis. The aim of the present study was to understand if sulfated GAG could modulate HPSE, since the mechanisms that regulate HPSE have not been completely defined. CHO-K1 cells were treated with 4-methylumbelliferone (4-MU) and sodium chlorate, to promote total inhibition of GAG synthesis, and reduce the sulfation pattern, respectively. The GAG profile of the wild CHO-K1 cells and CHO-745, deficient in xylosyltransferase, was determined after [(35)S]-sulfate labeling. HPSE expression was determined via real-time quantitative polymerase chain reaction. Total ablation of GAG with 4-MU in CHO-K1 inhibited HPSE expression, while the lack of sulfation had no effect. Interestingly, 4-MU had no effect in CHO-745 cells for these assays. In addition, a different enzyme location was observed in CHO-K1 wild-type cells, which presents HPSE mainly in the extracellular matrix, in comparison with the CHO-745 mutant cells, which is found in the cytoplasm. In view of our results, we can conclude that GAG are essential modulators of HPSE expression and location. Therefore, GAG profile could impact cell behavior mediated by the regulation of HPSE.

  4. Structure and ferroelectric properties of Bi(Zn(1/2)Ti(1/2))O3-(Bi(1/2)K(1/2))TiO3 perovskite solid solutions.

    PubMed

    Huang, Chien-Chih; Vittayakorn, Naratip; Cann, David P

    2009-07-01

    Lead-free piezoelectric ceramics based on chiBi(Zn(1/2)Ti(1/2))O3-(1-chi)(Bi(1/2)K(1/2))TiO3 were obtained via solid state processing techniques. A single perovskite phase with tetragonal symmetry was obtained for Bi(Zn(1/2)Ti(1/2))O3 (BZT) substitutions up to 20 mol%. The maximum density was 97.1% at the composition of chi = 0.1. The dielectric measurement indicated that the transition temperature decreased linearly with increasing BZT content. The P-E loops revealed an increase in remanent polarization (P(r)) with the addition of BZT. The maximum planar coupling coefficient, kappa(p), for the chi = 0.1 composition was 21.6 and the piezoelectric coefficient, d33, for chi = 0, chi = 0.05, and chi = 0.1 was 108, 185, and 235 pm/V, respectively. Overall, the dielectric and piezoelectric properties showed significant improvement when BZT was added.

  5. Influence of N-H-O hydrogen bonds on the structure and properties of (K1-x(NH4)xH2PO4) proton glasses: a single crystal neutron diffraction study

    NASA Astrophysics Data System (ADS)

    Ranjan Choudhury, Rajul; Chitra, R.

    2013-02-01

    It has been known for quite some time now that proton dynamics plays a key role in the structural ferroelectric (FE)/antiferroelectric (AFE) phase transition in the crystals belonging to the potassium dihydrogen phosphate crystal family. Mixed crystals belonging to this family having the composition M1-x(NW4)xW2AO4, where M = K, Rb, Cs, W = H, D, and A = P, As, exhibit proton glass behavior due to frustration between FE and AFE ordering; these proton glasses do not undergo any structural phase change but retain their room temperature structure down to very low temperatures. Single crystal neutron diffraction investigations of four mixed crystals with composition (K1-x(NH4)xH2PO4), where x = 0.0, 0.29, 0.67 1.0, were undertaken with the intention to investigate the effect of the local structural deviations on the overall average structure of the crystals and correlate these structural changes to the presence or absence of a structural phase transition in these crystals. Hydrogen bonding is shown to play a key role in the changing nature of the mixed crystals as the composition varies from the potassium rich ferroelectric region to the proton glass region to the ammonium rich antiferroelectric region.

  6. A vibrational spectroscopic study of the silicate mineral harmotome - (Ba,Na,K)1-2(Si,Al)8O16ṡ6H2O - A natural zeolite

    NASA Astrophysics Data System (ADS)

    Frost, Ray L.; López, Andrés; Wang, Lina; Romano, Antônio Wilson; Scholz, Ricardo

    2015-02-01

    The mineral harmotome (Ba,Na,K)1-2(Si,Al)8O16ṡ6H2O is a crystalline sodium calcium silicate which has the potential to be used in plaster boards and other industrial applications. It is a natural zeolite with catalytic potential. Raman bands at 1020 and 1102 cm-1 are assigned to the SiO stretching vibrations of three dimensional siloxane units. Raman bands at 428, 470 and 491 cm-1 are assigned to OSiO bending modes. The broad Raman bands at around 699, 728, 768 cm-1 are attributed to water librational modes. Intense Raman bands in the 3100 to 3800 cm-1 spectral range are assigned to OH stretching vibrations of water in harmotome. Infrared spectra are in harmony with the Raman spectra. A sharp infrared band at 3731 cm-1 is assigned to the OH stretching vibration of SiOH units. Raman spectroscopy with complimentary infrared spectroscopy enables the characterization of the silicate mineral harmotome.

  7. 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.

    PubMed

    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.

  8. Top Seeded Solution Growth, Structural and Vibrational Analyses of K1- x Na x Gd(WO4)2 (0.0 ≤ x ≤ 0.2) Single Crystals

    NASA Astrophysics Data System (ADS)

    Durairajan, A.; Thangaraju, D.; Valente, M. A.; Moorthy Babu, S.

    2016-08-01

    Laser host single crystals of K1- x Na x Gd(WO4)2 (0.0 ≤ x ≤ 0.2) have been grown by high temperature top seeded solution with low viscus K2WO4 as flux. The powder x-ray diffraction pattern of the synthesised charge confirmed the crystalline quality and possible incorporation of sodium in the KGd(WO4)2 (KGW) matrix. Estimated lattice parameters of the grown crystals revealed that there were no anomalous structural changes due to the incorporation of sodium in the KGW matrix. The change in structural phase transition and its thermal stability due to addition of sodium ions have been studied through differential thermal analysis. The homogeneous distribution of sodium and other metal ions in the matrix were qualitatively studied from energy-dispersive x-ray spectroscopy measurements. The Raman and Fourier transform infrared spectroscopy analyses have shown the influence of vibrational changes due to the incorporation of sodium ions in KGW. The optical absorption studies have shown the broad absorption band in the near-ultra violet region corresponding to the W6+ to O2- charge transfer band. These results suggest that incorporation of sodium in a KGW matrix appear to have modulated the physical properties of the grown crystals.

  9. Long-chain omega-3 fatty acids regulate bovine whole-body protein metabolism by promoting muscle insulin signalling to the Akt–mTOR–S6K1 pathway and insulin sensitivity

    PubMed Central

    Gingras, Andrée-Anne; White, Phillip James; Chouinard, P Yvan; Julien, Pierre; Davis, Teresa A; Dombrowski, Luce; Couture, Yvon; Dubreuil, Pascal; Myre, Alexandre; Bergeron, Karen; Marette, André; Thivierge, M Carole

    2007-01-01

    The ability of the skeletal musculature to use amino acids to build or renew constitutive proteins is gradually lost with age and this is partly due to a decline in skeletal muscle insulin sensitivity. Since long-chain omega-3 polyunsaturated fatty acids (LCn–3PUFA) from fish oil are known to improve insulin-mediated glucose metabolism in insulin-resistant states, their potential role in regulating insulin-mediated protein metabolism was investigated in this study. Experimental data are based on a switchback design composed of three 5 week experimental periods using six growing steers to compare the effect of a continuous abomasal infusion of LCn–3PUFA-rich menhaden oil with an iso-energetic control oil mixture. Clamp and insulin signalling observations were combined with additional data from a second cohort of six steers. We found that enteral LCn–3PUFA potentiate insulin action by increasing the insulin-stimulated whole-body disposal of amino acids from 152 to 308 μmol kg−1 h−1 (P = 0.006). The study further showed that in the fed steady-state, chronic adaptation to LCn–3PUFA induces greater activation (P < 0.05) of the Akt–mTOR–S6K1 signalling pathway. Simultaneously, whole-body total flux of phenylalanine was reduced from 87 to 67 μmol kg−1 h−1 (P = 0.04) and oxidative metabolism was decreased (P = 0.05). We conclude that chronic feeding of menhaden oil provides a novel nutritional mean to enhance insulin-sensitive aspects of protein metabolism. PMID:17158167

  10. Long-chain omega-3 fatty acids regulate bovine whole-body protein metabolism by promoting muscle insulin signalling to the Akt-mTOR-S6K1 pathway and insulin sensitivity.

    PubMed

    Gingras, Andrée-Anne; White, Phillip James; Chouinard, P Yvan; Julien, Pierre; Davis, Teresa A; Dombrowski, Luce; Couture, Yvon; Dubreuil, Pascal; Myre, Alexandre; Bergeron, Karen; Marette, André; Thivierge, M Carole

    2007-02-15

    The ability of the skeletal musculature to use amino acids to build or renew constitutive proteins is gradually lost with age and this is partly due to a decline in skeletal muscle insulin sensitivity. Since long-chain omega-3 polyunsaturated fatty acids (LCn-3PUFA) from fish oil are known to improve insulin-mediated glucose metabolism in insulin-resistant states, their potential role in regulating insulin-mediated protein metabolism was investigated in this study. Experimental data are based on a switchback design composed of three 5 week experimental periods using six growing steers to compare the effect of a continuous abomasal infusion of LCn-3PUFA-rich menhaden oil with an iso-energetic control oil mixture. Clamp and insulin signalling observations were combined with additional data from a second cohort of six steers. We found that enteral LCn-3PUFA potentiate insulin action by increasing the insulin-stimulated whole-body disposal of amino acids from 152 to 308 micromol kg(-1) h(-1) (P=0.006). The study further showed that in the fed steady-state, chronic adaptation to LCn-3PUFA induces greater activation (P<0.05) of the Akt-mTOR-S6K1 signalling pathway. Simultaneously, whole-body total flux of phenylalanine was reduced from 87 to 67 micromol kg(-1) h(-1) (P=0.04) and oxidative metabolism was decreased (P=0.05). We conclude that chronic feeding of menhaden oil provides a novel nutritional mean to enhance insulin-sensitive aspects of protein metabolism. PMID:17158167

  11. Influence of varying hydrogen bond strength resulting from compositional variation on the vibration spectra of proton glasses: K1-x(NH4)xH2PO4

    NASA Astrophysics Data System (ADS)

    Choudhury, Rajul Ranjan; Chitra, R.; Abraham, Geogy J.

    2014-07-01

    Single crystal neutron diffraction investigation [Choudhury and Chitra, J. Phys. Condense Matter, 25 (2013) 075902] on four mixed crystals with composition (K1-x(NH4)xH2PO4) where x=0.0, 0.29, 0.67, and 1.0 belonging to the potassium dihydrogen phosphate family of hydrogen bonded ferroelectric crystals had revealed that the compositional variation results in subtle structural differences primarily in the hydrogen bonds of these crystals. The study indicated that there is a change in hydrogen bond strengths with the change in crystal composition. Spectral investigation of the same set of four mixed crystals is undertaken with an intention to study the influence of the varying hydrogen bond strength on the vibrational properties of the crystals. Room temperature Raman spectra for all the four crystals are recorded in the range 100-4000 cm-1. This Raman investigation correlates the structural changes observed from neutron diffraction investigations to the changes in the vibration spectra of the crystals. The varying N-H-O hydrogen bond strength in the mixed crystals is found to have an observable effect on the librational frequencies of the molecular components of these crystals. The strong OHO hydrogen bonds in these crystals give rise to four spectral bands in the 1500-3000 cm-1 spectral region; this is in accordance with the theoretical prediction from the tunneling model for the very strong OHO hydrogen bonds. These OHO bonds can be described by a low barrier double well potential; the vibrational energy levels of the potential are split due to quantum tunneling effects. It is observed that the varying OHO hydrogen bond strength of these crystals results in a variation in the splitting of the vibrational energy levels of the hydrogen bond potential. It is attempted to correlate the varying OHO hydrogen bond strength with the expected variation in the freezing temperature with composition of these proton glasses.

  12. Solid-state conversion of (Na1/2Bi1/2)TiO3-BaTiO3-(K1/2Na1/2)NbO3 single crystals and their piezoelectric properties

    NASA Astrophysics Data System (ADS)

    Park, Ji-Hoon; Lee, Ho-Yong; Kang, Suk-Joong L.

    2014-06-01

    Piezoelectric ceramic with a composition of (94 - x)(Na1/2Bi1/2)TiO3-6BaTiO3-x(K1/2Na1/2)NbO3 (NBT-BT-xKNN) is a promising lead-free piezoelectric material for actuator applications because of its giant electric-field-induced strains, which are comparable with that of soft Pb(ZrxTi1-x)O3 (PZT) ceramics. Using the solid-state single crystal growth method, we succeeded in fabricating usable single crystals of NBT-BT-3KNN (6 mm × 6 mm × 8 mm size) with a uniform chemical composition. The room temperature piezoelectric properties of ⟨001⟩, ⟨110⟩, and ⟨111⟩ oriented single crystals were measured. Single crystals showed strong anisotropic strain characteristics. In particular, ⟨001⟩ oriented single crystals had excellent piezoelectric properties with small hysteresis and a high strain of 0.57% at 7 kV/mm. In addition, the fabricated single crystals exhibited a high converse piezoelectric constant, Smax/Emax, of over 1000 pm/V at 4 kV/mm. These values are greater than those reported for any lead-containing and lead-free ceramics and comparable with those of lead-based single crystals. Our investigation demonstrates the solid-state conversion of lead-free single crystals and their practical usability in replacement of lead-based materials.

  13. Mitochondrial alternative oxidase pathway protects plants against photoinhibition by alleviating inhibition of the repair of photodamaged PSII through preventing formation of reactive oxygen species in Rumex K-1 leaves.

    PubMed

    Zhang, Li-Tao; Zhang, Zi-Shan; Gao, Hui-Yuan; Xue, Zhong-Cai; Yang, Cheng; Meng, Xiang-Long; Meng, Qing-Wei

    2011-12-01

    The purpose of this study was to explore how the mitochondrial AOX (alternative oxidase) pathway alleviates photoinhibition in Rumex K-1 leaves. Inhibition of the AOX pathway decreased the initial activity of NADP-malate dehydrogenase (EC 1.1.1.82, NADP-MDH) and the pool size of photosynthetic end electron acceptors, resulting in an over-reduction of the photosystem I (PSI) acceptor side. The over-reduction of the PSI acceptor side further inhibited electron transport from the photosystem II (PSII) reaction centers to the PSII acceptor side as indicated by an increase in V(J) (the relative variable fluorescence at J-step), causing an imbalance between photosynthetic light absorption and energy utilization per active reaction center (RC) under high light, which led to the over-excitation of the PSII reaction centers. The over-reduction of the PSI acceptor side and the over-excitation of the PSII reaction centers enhanced the accumulation of reactive oxygen species (ROS), which inhibited the repair of the photodamaged PSII. However, the inhibition of the AOX pathway did not change the level of photoinhibition under high light in the presence of the chloroplast D1 protein synthesis inhibitor chloramphenicol, indicating that the inhibition of the AOX pathway did not accelerate the photodamage to PSII directly. All these results suggest that the AOX pathway plays an important role in the protection of plants against photoinhibition by minimizing the inhibition of the repair of the photodamaged PSII through preventing the over-production of ROS.

  14. Inositol metabolism and cell growth in a Chinese hamster ovary cell myo-inositol auxotroph.

    PubMed

    Jackowski, S; Voelker, D R; Rock, C O

    1988-11-15

    The intracellular concentrations of polyphosphoinositides and inositol phosphates were determined, and their role in growth factor-initiated cell division was investigated in a Chinese hamster ovary cell inositol auxotroph (CHO-K1-Ins). Metabolic labeling experiments during inositol starvation of CHO-K1-Ins cells showed that 1) the lipid-linked inositol component was maintained at the expense of the soluble inositol pool, 2) the decreasing cellular content of phosphatidylinositol was replaced by phosphatidylglycerol, and 3) the concentrations of inositol polyphosphates and polyphosphoinositides were conserved at the expense of inositol and phosphatidylinositol. These data show that homeostatic mechanisms exist for the maintenance of the polyphosphoinositide and inositol phosphate pools at the expense of inositol and phosphatidylinositol. The addition of alpha-thrombin to growth-arrested (serum-starved) CHO-K1-Ins cells stimulated the incorporation of [3H]thymidine into DNA to the same extent as that observed following serum readdition. gamma-Thrombin was also an effective mitogen, but active site-inhibited alpha-thrombin was not. Both alpha- and gamma-thrombin, but not catalytic site-inhibited alpha-thrombin, initiated phosphatidylinositol turnover in vivo and increased phosphatidylinositol 4,5-bisphosphate phospholipase C activity in vitro. Serum and insulin were potent CHO-K1-Ins cell mitogens, but neither triggered phosphatidylinositol turnover in vivo nor activated phospholipase C in vitro. The activation of phospholipase C plays a determinant role in thrombin-initiated cell cycle progression in Chinese hamster ovary cells, although other growth factor-signaling pathways exist that are independent of polyphosphoinositide catabolism.

  15. Oxidative stress increases the risk of pancreatic β cell damage in chronic renal hypertensive rats.

    PubMed

    Gao, Shan; Park, Byung M; Cha, Seung A; Bae, Ui J; Park, Byung H; Park, Woo H; Kim, Suhn H

    2016-08-01

    Hypertension often occurs in conjunction with insulin resistance. The purpose of this study was to evaluate whether sustained renal hypertension increases the risk of diabetes mellitus in rats, and to define the underlying mechanisms. Two-kidney, one-clip hypertensive (2K1C) rats received captopril (50 mg/kg/day), α-lipoic acid (100 mg/kg/day), or vehicle treatment for 3 months after surgery. Blood pressure was measured by tail cuff plethysmography. Oral glucose tolerance test (OGTT), immunohistochemistry, and western blotting were performed. In addition, insulin secretion from islet cells was measured. OGTT yielded abnormal results, and the number of islet cells and the size of pancreatic β/α cells were decreased in 2K1C rats. Basal insulin levels were also reduced in the plasma. Insulin secretion from pancreatic islet cells in response to high glucose was also attenuated in 2K1C rats compared with sham rats. The levels of oxidative stress markers, including 8-hydroxydeoxyguanosine and NADPH oxidase-4, were increased in pancreatic tissue and pancreatic islets in 2K1C rats. The abnormalities observed in 2K1C rats were improved by captopril or α-lipoic acid treatment. These findings indicate that sustained renal hypertension may lead to pancreatic dysfunction, increasing oxidative stress in pancreatic islets. PMID:27535482

  16. Colonization process of the Brazilian common vesper mouse, Calomys expulsus (Cricetidae, Sigmodontinae): a biogeographic hypothesis.

    PubMed

    Nascimento, Fabrícia Ferreira do; Pereira, Luciana G; Geise, Lena; Bezerra, Alexandra M R; D'Andrea, Paulo S; Bonvicino, Cibele R

    2011-01-01

    Riverine barriers have been associated to genetic diversification and speciation of several taxa. The Rio São Francisco is one of the largest rivers in South America, representing the third largest river basin in Brazil and operating as a geographic barrier to gene flow of different taxa. To evaluate the influence of the Rio São Francisco in the speciation of small rodents, we investigated the genetic structure of Calomys expulsus with phylogenetic and network analyses of cytochrome b DNA. Our results suggested that C. expulsus can be divided into 3 subpopulations, 2 on the left and another one on the right bank of this river. The time of divergence of these subpopulations, using a Bayesian framework, suggested colonization from the south to the north/northeast. Spatial analysis using a clustering method and the Monmonier's algorithm suggested that the Rio São Francisco is a biogeographic barrier to gene flow and indicated that this river may play a role in the incipient speciation process of these subpopulations. PMID:21441460

  17. [Revision of the Taxonomic Position of the Olkhon Mountain Vole (Rodentia, Cricetidae)].

    PubMed

    Bodrov, S Yu; Kostygov, A Yu; Rudneva, L V; Abramson, N I

    2016-01-01

    An analysis of the phylogenetic position of the Olkhon mountain vole (Alticolaolchonensis Litvinov 1960) using the sequences of four nuclear (BRCA, GHR, LCAT, and IRBP) and one mitochondrial (cyt. b) genes was undertaken. It was noted that, until recently, multiple studies of the systematic position of this vole had been based exclusively on morphological data, while the major taxonomic traits contained contradictory information regarding both the subgeneric status of this species and its genus. It was established that the molecular data and morphology data allow us to attribute the Lake Baikal vole unambiguously to the nominative subgenus Alticola instead of Aschizomys.

  18. DNA barcoding of Murinae (Rodentia: Muridae) and Arvicolinae (Rodentia: Cricetidae) distributed in China.

    PubMed

    Li, Jing; Zheng, Xin; Cai, Yansen; Zhang, Xiuyue; Yang, Min; Yue, Bisong; Li, Jing

    2015-01-01

    Identification of rodents is very difficult mainly due to high similarities in morphology and controversial taxonomy. In this study, mitochondrial cytochrome oxidase subunit I (COI) was used as DNA barcode to identify the Murinae and Arvicolinae species distributed in China and to facilitate the systematics studies of Rodentia. In total, 242 sequences (31 species, 11 genera) from Murinae and 130 sequences (23 species, 6 genera) from Arvicolinae were investigated, of which 90 individuals were novel. Genetic distance, threshold method, tree-based method, online BLAST and BLOG were employed to analyse the data sets. There was no obvious barcode gap. The average K2P distance within species and genera was 2.10% and 12.61% in Murinae, and 2.86% and 11.80% in Arvicolinae, respectively. The optimal threshold was 5.62% for Murinae and 3.34% for Arvicolinae. All phylogenetic trees exhibited similar topology and could distinguish 90.32% of surveyed species in Murinae and 82.60% in Arvicolinae with high support values. BLAST analyses yielded similar results with identification success rates of 92.15% and 93.85% for Murinae and Arvicolinae, respectively. BLOG successfully authenticated 100% of detected species except Leopoldamys edwardsi based on the latest taxonomic revision. Our results support the species status of recently recognized Micromys erythrotis, Eothenomys tarquinius and E. hintoni and confirm the important roles of comprehensive taxonomy and accurate morphological identification in DNA barcoding studies. We believe that, when proper analytic methods are applied or combined, DNA barcoding could serve as an accurate and effective species identification approach for Murinae and Arvicolinae based on a proper taxonomic framework.

  19. Seasonal energy requirements and thermoregulation of growing pouched mice, Saccostomus campestris (Cricetidae)

    NASA Astrophysics Data System (ADS)

    Ellison, G. T. H.; Skinner, J. D.

    1991-06-01

    Pouched mice ( Saccostomus campestris) were born in captivity during January and March and subsequently maintained under long photoperiod (14 h light: 10 h dark) at 25°C. During their first winter (July) and the following summer (January) the pouched mice were exposed to natural photoperiod in an unheated laboratory for 3 weeks prior to measurement. The pouched mice continued to grow during the study, and were significantly heavier after summer exposure than after winter exposure 6 months earlier. Although this increase in body mass would result in a decline in their surface area to volume ratio there was no significant decline in minimal thermal conductance ( C m) and winter-exposed pouched mice had a relatively lower C m than expected. Meanwhile the smaller, winter-exposed animals displayed a significantly higher capacity for non-shivering thermogenesis, together with higher levels of basal metabolism than summer individuals. These differences were not solely attributable to the contrasting body mass of each group and it is therefore clear that S. campestris can increase thermoregulatory heat production, and modify heat loss following exposure to short photoperiod and cold during their first winter. Despite the significant increase in metabolism, the overall energy requirements of small, winter-exposed animals were significantly lower than those for heavier pouched mice following exposure to summer conditions. These results suggest that growing pouched mice can effectively adapt to lower temperature conditions during their first winter, yet accrue considerable overall savings in total energy requirements as a result of their smaller body mass.

  20. Molecular diversification of Trichuris spp. from Sigmodontinae (Cricetidae) rodents from Argentina based on mitochondrial DNA sequences.

    PubMed

    Callejón, Rocío; Robles, María Del Rosario; Panei, Carlos Javier; Cutillas, Cristina

    2016-08-01

    A molecular phylogenetic hypothesis is presented for the genus Trichuris based on sequence data from mitochondrial cytochrome c oxidase 1 (cox1) and cytochrome b (cob). The taxa consisted of nine populations of whipworm from five species of Sigmodontinae rodents from Argentina. Bayesian Inference, Maximum Parsimony, and Maximum Likelihood methods were used to infer phylogenies for each gene separately but also for the combined mitochondrial data and the combined mitochondrial and nuclear dataset. Phylogenetic results based on cox1 and cob mitochondrial DNA (mtDNA) revealed three clades strongly resolved corresponding to three different species (Trichuris navonae, Trichuris bainae, and Trichuris pardinasi) showing phylogeographic variation, but relationships among Trichuris species were poorly resolved. Phylogenetic reconstruction based on concatenated sequences had greater phylogenetic resolution for delimiting species and populations intra-specific of Trichuris than those based on partitioned genes. Thus, populations of T. bainae and T. pardinasi could be affected by geographical factors and co-divergence parasite-host. PMID:27083190

  1. Cytokine mRNA expression in Peromyscus yucatanicus (Rodentia: Cricetidae) infected by Leishmania (Leishmania) mexicana.

    PubMed

    Loria-Cervera, Elsy Nalleli; Sosa-Bibiano, Erika Ivett; Van Wynsberghe, Nicole Raymonde; Saldarriaga, Omar Abdul; Melby, Peter C; Andrade-Narvaez, Fernando Jose

    2016-07-01

    Peromyscus yucatanicus, the main reservoir of Leishmania (Leishmania) mexicana in the Yucatan peninsula of Mexico, reproduces clinical and histological pictures of LCL in human as well as subclinical infection. Thus, we used this rodent as a novel experimental model. In this work, we analyzed cytokine mRNA expression in P. yucatanicus infected with L. (L.) mexicana. Animals were inoculated with either 2.5×10(6) or 1×10(2) promastigotes and cytokine expressions were analyzed by real-time RT-PCR in skin at 4 and 12weeks post-infection (wpi). Independently of the parasite inoculum none of the infected rodents had clinical signs of LCL at 4wpi and all expressed high IFN-γ mRNA. All P. yucatanicus inoculated with 2.5×10(6) promastigotes developed signs of LCL at 12wpi while the mice inoculated with 1×10(2) remained subclinical. At that time, both IFN-γ and IL-10 were expressed in P. yucatanicus with clinical and subclinical infections. Expressions of TNF-α and IL-4 were significantly higher in clinical animals (2.5×10(6)) compared with subclinical ones (1×10(2)). High TGF-β expression was observed in P. yucatanicus with clinical signs when compared with healthy animals. Results suggested that the clinical course of L. (L.) mexicana infection in P. yucatanicus was associated with a specific local pattern of cytokine production at 12wpi. PMID:27155064

  2. Karyotypic variation in the Andean rodent Phyllotis xanthopygus (Waterhouse, 1837) (Rodentia, Cricetidae, Sigmodontinae)

    PubMed Central

    Labaroni, Carolina Alicia; Malleret, Matías Maximiliano; Novillo, Agustina; Ojeda, Agustina; Rodriguez, Daniela; Cuello, Pablo; Ojeda, Ricardo; Dardo Martí; Lanzone, Cecilia

    2014-01-01

    Abstract Phyllotis xanthopygus (Waterhouse, 1837) is an Andean rodent endemic to South America. Despite its wide geographical distribution in Argentina, few individuals have been studied on the cytogenetic level and only through conventional staining. In this work, chromosome characterization of Argentine samples of this species was performed using solid staining, C-banding and base-specific fluorochromes. Twenty two specimens were analyzed, collected in the provinces of Jujuy, Catamarca, and the north and south of Mendoza. All studied specimens showed 2n=38, having mostly the bi-armed autosomes, metacentric or submetacentric. Fundamental Number varied between 70 and 72. These changes were due to the presence of chromosome heteromorphisms in individuals from southern Mendoza and Jujuy. C-banding revealed pericentromeric blocks of constitutive heterochromatin in most chromosomes. Acrocentric chromosomes involved in heteromorphisms showed high variation in the amount of heterochromatin within and among populations. Additionally, banding with fluorochromes (DAPI and chromomycin A3) revealed homologous localization of AT and GC rich regions among chromosomes of the different populations analyzed. Comparisons among heteromorphic pairs suggested, however, that the variation might be the result of complex chromosome rearrangements, involving possibly amplifications and/or deletions of heterochromatic segments. These results are in accordance with molecular studies that indicate genetic variability within and among the populations of this taxon. PMID:25610549

  3. Colonization process of the Brazilian common vesper mouse, Calomys expulsus (Cricetidae, Sigmodontinae): a biogeographic hypothesis.

    PubMed

    Nascimento, Fabrícia Ferreira do; Pereira, Luciana G; Geise, Lena; Bezerra, Alexandra M R; D'Andrea, Paulo S; Bonvicino, Cibele R

    2011-01-01

    Riverine barriers have been associated to genetic diversification and speciation of several taxa. The Rio São Francisco is one of the largest rivers in South America, representing the third largest river basin in Brazil and operating as a geographic barrier to gene flow of different taxa. To evaluate the influence of the Rio São Francisco in the speciation of small rodents, we investigated the genetic structure of Calomys expulsus with phylogenetic and network analyses of cytochrome b DNA. Our results suggested that C. expulsus can be divided into 3 subpopulations, 2 on the left and another one on the right bank of this river. The time of divergence of these subpopulations, using a Bayesian framework, suggested colonization from the south to the north/northeast. Spatial analysis using a clustering method and the Monmonier's algorithm suggested that the Rio São Francisco is a biogeographic barrier to gene flow and indicated that this river may play a role in the incipient speciation process of these subpopulations.

  4. Chromosomal variation in Argentine populations of Akodon montensis Thomas, 1913 (Rodentia, Cricetidae, Sigmodontinae)

    PubMed Central

    Malleret, Matías Maximiliano; Labaroni, Carolina Alicia; García, Gabriela Verónica; Ferro, Juan Martín; Martí, Dardo Andrea; Lanzone, Cecilia

    2016-01-01

    Abstract The genus Akodon Meyen, 1833 is one of the most species-rich among sigmodontine rodents and has great chromosome variability. Akodon montensis has a relatively broad distribution in South America, and Argentine populations are located in the southernmost region of its range. Brazilian populations have important chromosomal variability, but cytogenetic data from Argentina are scarce. We performed a chromosome characterization of natural populations of Akodon montensis using conventional staining, C-banding, Ag-NORs and base-specific fluorochromes. A total of 31 specimens from five localities of Misiones Province, in Argentina, were analyzed. The 2n=24 chromosomes was the most frequently observed karyotype. However, five individuals presented 25 chromosomes due to a supernumerary B-chromosome; and one individual had 2n=26 due to one B plus a trisomy for chromosome 11. Additionally, two XY females and two variants of the X chromosomes were found. C-positive centromeric bands occurred in all chromosomes; additional C-bands were observed in some autosomes, the X, Y and B chromosomes. Ag-NORs were observed in five autosomes, and the B chromosome was frequently marked. Fluorochrome banding was similar among karyotypes of the analyzed populations. Comparisons of cytogenetic data among populations of Argentina and Brazil showed the presence of high intraspecific variability in Akodon montensis and some differences among regions. PMID:27186343

  5. Reservoir competence of Microtus pennsylvanicus (Rodentia: Cricetidae) for the Lyme disease spirochete, Borrelia burgdorferi

    USGS Publications Warehouse

    Markowski, D.; Ginsberg, H.S.; Hyland, K.E.; Hu, R.

    1998-01-01

    The reservoir competence of the meadow vole, Microtus pennsylvanicus Ord, for the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner was established on Patience Island, RI. Meadow voles were collected from 5 locations throughout Rhode Island. At 4 of the field sites, M. pennsylvanicus represented only 4.0% (n = 141) of the animals captured. However, on Patience Island, M. pennsylvanicus was the sole small mammal collected (n = 48). Of the larval Ixodes scapularis Say obtained from the meadow voles on Patience Island, 62% (n = 78) was infected with B. burgdorferi. Meadow voles from all 5 locations were successfully infected with B. burgdorferi in the laboratory and were capable of passing the infection to xenodiagnostic I. scapularis larvae for 9 wk. We concluded that M. pennsylvanicus was physiologically capable of maintaining B. burgdorferi infection. However, in locations where Peromyscus leucopus (Rafinesque) is abundant, the role of M. pennsylvanicus as a primary reservoir for B. burgdorferi was reduced.

  6. [The rodent. Akodon arviculoides, Wagner, 1842 (Cricetidae)--importance in plague foci in Brazil].

    PubMed

    de Almeida, C R; de Almeida, A M; Brasil, D P; Dantas Sobrinho, J; Leal, M A

    1986-01-01

    The occurrence of the rodent Akodon arviculoides Wagner, 1842 in the plague focus of the "Agreste" region of the State of Pernambuco and a report on its ability for survival, reproduction and development in captivity, its susceptibility to Yersinia pestis infection and the role of this rodent species in Brazilian plague foci are reported. PMID:3302594

  7. [Revision of the Taxonomic Position of the Olkhon Mountain Vole (Rodentia, Cricetidae)].

    PubMed

    Bodrov, S Yu; Kostygov, A Yu; Rudneva, L V; Abramson, N I

    2016-01-01

    An analysis of the phylogenetic position of the Olkhon mountain vole (Alticolaolchonensis Litvinov 1960) using the sequences of four nuclear (BRCA, GHR, LCAT, and IRBP) and one mitochondrial (cyt. b) genes was undertaken. It was noted that, until recently, multiple studies of the systematic position of this vole had been based exclusively on morphological data, while the major taxonomic traits contained contradictory information regarding both the subgeneric status of this species and its genus. It was established that the molecular data and morphology data allow us to attribute the Lake Baikal vole unambiguously to the nominative subgenus Alticola instead of Aschizomys. PMID:27396178

  8. The mechanisms of cytotoxicity of urethane dimethacrylate to Chinese hamster ovary cells.

    PubMed

    Chang, Hsiao-Hua; Chang, Mei-Chi; Lin, Li-Deh; Lee, Jang-Jaer; Wang, Tong-Mei; Huang, Chun-Hsun; Yang, Ting-Ting; Lin, Hsueh-Jen; Jeng, Jiiang-Huei

    2010-09-01

    Monomers released from resin-containing products may cause various adverse effects. Urethane dimethacrylate (UDMA) is a principal resin monomer and also a major component released from various dental resin materials. Thus the toxic effects and mechanisms should be elucidated for improving of its safety use. Here we investigated the effects of UDMA on the growth, cell cycle progression, reactive oxygen species (ROS) production and glutathione (GSH) alteration in CHO-K1 cells, and the preventive effects by antioxidants (NAC and catalase) were also evaluated. UDMA elicited growth inhibition (>0.025 mm) of CHO-K1 cells in a clearly dose-dependent manner. Cell cycle perturbation and ROS overproduction were also observed. A 0.1 mm UDMA-induced S-phase cell cycle arrest and ROS accumulation. Cell apoptosis and necrosis became significant when UDMA concentration was 0.25 mm. GSH depletion occurred at cells treated with 0.25 mm UDMA, a highly cytotoxic concentration at which point myriad cells were under apoptosis or necrosis. Thus GSH depletion can be crucial for the death of CHO-K1 cells. Furthermore NAC (0.5-10 mm) and catalase (250-1000 U/ml) obviously attenuated the UDMA-induced toxicity by reducing ROS generation and cell cycle disturbance, and the effects were dose-related. These results suggest that UDMA toxicity is associated with ROS production, GSH depletion, cell cycle disturbance and cell apoptosis/necrosis. PMID:20579731

  9. Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system.

    PubMed

    Nakamura, Tsuyoshi; Omasa, Takeshi

    2015-09-01

    Therapeutic antibodies are commonly produced by high-expressing, clonal and recombinant Chinese hamster ovary (CHO) cell lines. Currently, CHO cells dominate as a commercial production host because of their ease of use, established regulatory track record, and safety profile. CHO-K1SV is a suspension, protein-free-adapted CHO-K1-derived cell line employing the glutamine synthetase (GS) gene expression system (GS-CHO expression system). The selection of high-producing mammalian cell lines is a crucial step in process development for the production of therapeutic antibodies. In general, cloning by the limiting dilution method is used to isolate high-producing monoclonal CHO cells. However, the limiting dilution method is time consuming and has a low probability of monoclonality. To minimize the duration and increase the probability of obtaining high-producing clones with high monoclonality, an automated single cell-based clone selector, the ClonePix FL system, is available. In this study, we applied the high-throughput ClonePix FL system for cell line development using CHO-K1SV cells and investigated efficient conditions for single cell-based clone selection. CHO-K1SV cell growth at the pre-picking stage was improved by optimizing the formulation of semi-solid medium. The efficiency of picking and cell growth at the post-picking stage was improved by optimization of the plating time without decreasing the diversity of clones. The conditions for selection, including the medium formulation, were the most important factors for the single cell-based clone selection system to construct a high-producing CHO cell line.

  10. Differential cytotoxic effects of arsenic on human and animal cells.

    PubMed

    Lee, T C; Ho, I C

    1994-09-01

    Human fibroblasts (HFW) were 10-fold more susceptible than Chinese hamster ovary (CHO-K1) cells to sodium arsenite. Comparison of cellular antioxidant enzyme activities showed that CHO-K1 cells contained 3- and 8-fold more glutathione-peroxidase and catalase activities, respectively, than HFW cells. Since vitamin E, methylamine, and benzyl alcohol could prevent, in part, the arsenite-induced killing of HFW cells, we suggest that arsenite can induce oxidative damage in HFW cells. We have also established arsenic-resistant cells, SA7 and CL3R, from CHO cells and from a human lung adenocarcinoma cell line (CL3), respectively. The arsenic resistance of SA7 cells was attributed mainly to elevation of glutathione S-transferase pi levels, and that of CL3R cells was possibly due to an increase in heme oxygenase activity. Since induction of heme oxygenase is a general response to oxidative stress, we suspect that the differential toxicity of arsenic to human and animal cells could be due to arsenic's more efficient induction of oxidative damage in human cells.

  11. Analysis of the DNA replication competence of the xrs-5 mutant cells defective in Ku86.

    PubMed

    Matheos, Diamanto; Novac, Olivia; Price, Gerald B; Zannis-Hadjopoulos, Maria

    2003-01-01

    The radiosensitive mutant xrs-5, a derivative of the Chinese hamster ovary (CHO) K1 cell line, is defective in DNA double-strand break repair and V(D)J recombination. The defective phenotypes of xrs-5 cells are complemented by the 86 kDa subunit of Ku antigen. OBA is a protein, previously purified from HeLa cells, that binds in a sequence-specific manner to mammalian origins of DNA replication. The DNA-binding subunit of OBA has been identified as Ku86. We tested the xrs-5 cell line for its ability to replicate a mammalian origin-containing plasmid, p186, in vivo and in vitro. In vivo, the p186 episomal DNA replication in transfected xrs-5 cells was reduced by 45% when compared with the CHO K1 cells transfected with p186. In vitro, although total and cytoplasmic cell extracts from xrs-5 cells replicated the p186 with the same efficiency as the parental CHO K1 cell extracts, xrs-5 nuclear extracts did not possess any detectable replication activity. Addition of affinity-purified OBA/Ku restored replication in the xrs-5 nuclear extract reaction. Western blot analyses showed that the levels of other replication proteins (Orc2, PCNA, DNA polymerase epsilon and delta, Primase and Topoisomerase IIalpha) were comparable in both the xrs-5 mutant and CHO K1 wild-type cell lines. In addition, the in vivo association of Ku with the DHFR origin-containing sequence (oribeta) was examined in both the CHO K1 and xrs-5 cell lines by a chromatin immunoprecipitation (ChIP) assay. Anti-Ku antibodies did not immunoprecipitate a detectable amount of Ku from the xrs-5 cells in the origin-containing sequence, in contrast to the CHO K1 cells, wherein Ku was found to be associated with the oribeta origin. The data implicate Ku antigen in in vivo and in vitro DNA replication and suggest the existence of another protein with Ku-like functions in the xrs-5 cells.

  12. Effects of saponins against clinical E. coli strains and eukaryotic cell line.

    PubMed

    Arabski, Michał; Węgierek-Ciuk, Aneta; Czerwonka, Grzegorz; Lankoff, Anna; Kaca, Wiesław

    2012-01-01

    Saponins are detergent-like substances showing antibacterial as well as anticancer potential. In this study, the effects of saponins from Quillaja saponaria were analyzed against prokaryotic and eukaryotic cells. Multidrug-resistant clinical E. coli strains were isolated from human urine. As eukaryotic cells, the CHO-K1 cell lines were applied. Antibacterial effect of ampicillin, streptomycin, and ciprofloxacin in the presence of saponins was measured by cultivation methods. Properties of saponins against CHO-K1 cells were measured by the MTT test, hemolysis assay and flow cytometry. Saponin from Quillaja saponaria has a cytotoxic effect at concentrations higher than 25 μg/mL and in the range of 12-50 μg/mL significantly increases the level of early apoptotic cells. Saponin at dose of 12 μg/mL enhances the six E. coli strains growth. We postulate that saponins increase the influx of nutrients from the medium into E. coli cells. Saponins do not have synergetic effects on antibacterial action of tested antibiotics. In contrary, in the presence of saponins and antibiotics, more CFU/mL E. coli cells were observed. This effect was similar to saponins action alone towards E. coli cells. In conclusion, saponins was cytotoxic against CHO-K1 cells, whereas against E. coli cells this effect was not observed. PMID:22500084

  13. Essential Roles of RNA-binding Protein HuR in Activation of Hepatic Stellate Cells Induced by Transforming Growth Factor-β1

    PubMed Central

    Ge, Jingjing; Chang, Na; Zhao, Zhongxin; Tian, Lei; Duan, Xianghui; Yang, Lin; Li, Liying

    2016-01-01

    RNA-binding protein HuR mediates transforming growth factor (TGF)-β1-induced profibrogenic actions. Up-regulation of Sphingosine kinase 1 (SphK1) is involved in TGF-β1-induced activation of hepatic stellate cells (HSCs) in liver fibrogenesis. However, the molecular mechanism of TGF-β1 regulates SphK1 remains unclear. This study was designed to investigate the role of HuR in TGF-β1-induced SphK1 expression and identify a new molecular mechanism in liver fibrogenensis. In vivo, HuR expression was increased, translocated to cytoplasm, and bound to SphK1 mRNA in carbon tetrachloride- and bile duct ligation-induced mouse fibrotic liver. HuR mRNA expression had a positive correlation with mRNA expressions of SphK1 and fibrotic markers, α-smooth muscle actin (α-SMA) and Collagen α1(I), respectively. In vitro, up-regulation of SphK1 and activation of HSCs stimulated by TGF-β1 depended on HuR cytoplasmic accumulation. The effects of TGF-β1 were diminished when HuR was silenced or HuR cytoplasmic translocation was blocked. Meanwhile, overexpression of HuR mimicked the effects of TGF-β1. Furthermore, TGF-β1 prolonged half-life of SphK1 mRNA by promoting its binding to HuR. Pharmacological or siRNA-induced SphK1 inhibition abrogated HuR-mediated HSC activation. In conclusion, our data suggested that HuR bound to SphK1 mRNA and played a crucial role in TGF-β1-induced HSC activation. PMID:26912347

  14. Essential Roles of RNA-binding Protein HuR in Activation of Hepatic Stellate Cells Induced by Transforming Growth Factor-β1.

    PubMed

    Ge, Jingjing; Chang, Na; Zhao, Zhongxin; Tian, Lei; Duan, Xianghui; Yang, Lin; Li, Liying

    2016-02-25

    RNA-binding protein HuR mediates transforming growth factor (TGF)-β1-induced profibrogenic actions. Up-regulation of Sphingosine kinase 1 (SphK1) is involved in TGF-β1-induced activation of hepatic stellate cells (HSCs) in liver fibrogenesis. However, the molecular mechanism of TGF-β1 regulates SphK1 remains unclear. This study was designed to investigate the role of HuR in TGF-β1-induced SphK1 expression and identify a new molecular mechanism in liver fibrogenensis. In vivo, HuR expression was increased, translocated to cytoplasm, and bound to SphK1 mRNA in carbon tetrachloride- and bile duct ligation-induced mouse fibrotic liver. HuR mRNA expression had a positive correlation with mRNA expressions of SphK1 and fibrotic markers, α-smooth muscle actin (α-SMA) and Collagen α1(I), respectively. In vitro, up-regulation of SphK1 and activation of HSCs stimulated by TGF-β1 depended on HuR cytoplasmic accumulation. The effects of TGF-β1 were diminished when HuR was silenced or HuR cytoplasmic translocation was blocked. Meanwhile, overexpression of HuR mimicked the effects of TGF-β1. Furthermore, TGF-β1 prolonged half-life of SphK1 mRNA by promoting its binding to HuR. Pharmacological or siRNA-induced SphK1 inhibition abrogated HuR-mediated HSC activation. In conclusion, our data suggested that HuR bound to SphK1 mRNA and played a crucial role in TGF-β1-induced HSC activation.

  15. The function and molecular identity of inward rectifier channels in vestibular hair cells of the mouse inner ear.

    PubMed

    Levin, Michaela E; Holt, Jeffrey R

    2012-07-01

    Inner ear hair cells respond to mechanical stimuli with graded receptor potentials. These graded responses are modulated by a host of voltage-dependent currents that flow across the basolateral membrane. Here, we examine the molecular identity and the function of a class of voltage-dependent ion channels that carries the potassium-selective inward rectifier current known as I(K1). I(K1) has been identified in vestibular hair cells of various species, but its molecular composition and functional contributions remain obscure. We used quantitative RT-PCR to show that the inward rectifier gene, Kir2.1, is highly expressed in mouse utricle between embryonic day 15 and adulthood. We confirmed Kir2.1 protein expression in hair cells by immunolocalization. To examine the molecular composition of I(K1), we recorded voltage-dependent currents from type II hair cells in response to 50-ms steps from -124 to -54 in 10-mV increments. Wild-type cells had rapidly activating inward currents with reversal potentials close to the K(+) equilibrium potential and a whole-cell conductance of 4.8 ± 1.5 nS (n = 46). In utricle hair cells from Kir2.1-deficient (Kir2.1(-/-)) mice, I(K1) was absent at all stages examined. To identify the functional contribution of Kir2.1, we recorded membrane responses in current-clamp mode. Hair cells from Kir2.1(-/-) mice had significantly (P < 0.001) more depolarized resting potentials and larger, slower membrane responses than those of wild-type cells. These data suggest that Kir2.1 is required for I(K1) in type II utricle hair cells and contributes to hyperpolarized resting potentials and fast, small amplitude receptor potentials in response to current inputs, such as those evoked by hair bundle deflections.

  16. N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, a calmodulin antagonist, inhibits cell proliferation.

    PubMed Central

    Hidaka, H; Sasaki, Y; Tanaka, T; Endo, T; Ohno, S; Fujii, Y; Nagata, T

    1981-01-01

    N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and its derivatives are putative calmodulin antagonists that bind to calmodulin and inhibit Ca2+/calmodulin-regulated enzyme activities. Autoradiographic studies using tritiated W-7 showed that this compound penetrates the cell membrane, is distributed mainly in the cytoplasm, and inhibits proliferation of Chinese hamster ovary K1 (CHO-K1) cells. Cytoplasmic [3H]W-7 was excluded completely within 6 hr after removal of [3H]W-7 from the culture medium. N-(6-aminohexyl)-1-naphthalenesulfonamide, an analogue of W-7 that interacts only weakly with calmodulin, proved to be a much weaker inhibitor of cell proliferation. CHO-K1 cells were synchronized by shaking during mitosis and then released into the cell cycle in the presence of 25 microM W-7 or 2.5 mM thymidine for 12 hr. Cell division was observed approximately 6 hr later. The results suggest that the effect of W-7 on cell proliferation might be through selective inhibition of the G1/S boundary phase, which is similar to the effect of excess thymidine. This pharmacological demonstration that cytoplasmic calmodulin is involved in cell proliferation is significant; W-7 and its derivatives may be useful tools for research on calmodulin and cell biology-related studies. Images PMID:6945588

  17. Sphingosine kinase-1 mediates TNF-alpha-induced MCP-1 gene expression in endothelial cells: upregulation by oscillatory flow.

    PubMed

    Chen, Xi-Lin; Grey, Janice Y; Thomas, Suzanne; Qiu, Fei-Hua; Medford, Russell M; Wasserman, Martin A; Kunsch, Charles

    2004-10-01

    Atherosclerosis is a focal inflammatory disease and preferentially occurs in areas of low fluid shear stress and oscillatory flow, whereas the risk of atherosclerosis is decreased in regions of high fluid shear stress and steady laminar flow. Sphingosine kinase-1 (SphK1) catalyzes the conversion of sphingosine to sphingosine-1 phosphate (S1P), a sphingolipid metabolite that plays important roles in angiogenesis, inflammation, and cell growth. In the present study, we demonstrated that exposure of human aortic endothelial cells to oscillatory flow (shear stress, +/-5 dyn/cm(2) for 48 h) resulted in a marked increase in SphK1 mRNA levels compared with endothelial cells kept in static culture. In contrast, laminar flow (shear stress, 20 dyn/cm(2) for 48 h) decreased SphK1 mRNA levels. We further investigated the role of SphK1 in TNF-alpha-induced expression of inflammatory genes, such as monocyte chemoattractant protein-1 (MCP-1) and VCAM-1 by using small interfering RNA (siRNA) specifically for SphK1. Treatment of endothelial cells with SphK1 siRNA suppressed TNF-alpha-induced increase in MCP-1 mRNA levels, MCP-1 protein secretion, and activation of p38 MAPK. SphK1 siRNA also inhibited TNF-alpha-induced cell surface expression of VCAM-1, but not ICAM-1, protein. Exposure of endothelial cells to S1P led to an increase in MCP-1 protein secretion and MCP-1 mRNA levels and activation of NF-kappaB-mediated transcriptional activity. Treatment of endothelial cells with the p38 MAPK inhibitor SB-203580 suppressed S1P-induced MCP-1 protein secretion. These data suggest that SphK1 mediates TNF-alpha-induced MCP-1 gene expression through a p38 MAPK-dependent pathway and may participate in oscillatory flow-mediated proinflammatory signaling pathway in the vasculature. PMID:15191888

  18. Construction of photoenergetic mitochondria in cultured mammalian cells

    PubMed Central

    Hara, Kiyotaka Y.; Wada, Takeyoshi; Kino, Kuniki; Asahi, Toru; Sawamura, Naoya

    2013-01-01

    The proton motive force (PMF) is bio-energetically important for various cellular reactions to occur. We developed PMF-photogenerating mitochondria in cultured mammalian cells. An archaebacterial rhodopsin, delta-rhodopsin, which is a light-driven proton pump derived from Haloterrigena turkmenica, was expressed in the mitochondria of CHO-K1 cells. The constructed stable CHO-K1 cell lines showed suppression of cell death induced by rotenone, a pesticide that inhibits mitochondrial complex I activity involved in PMF generation through the electron transport chain. Delta-rhodopsin was also introduced into the mitochondria of human neuroblastoma SH-SY5Y cells. The constructed stable SH-SY5Y cell lines showed suppression of dopaminergic neuronal cell death induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), an inducer of Parkinson's disease models, which acts through inhibition of complex I activity. These results suggest that the light-activated proton pump functioned as a PMF generator in the mitochondria of mammalian cells, and suppressed cell death induced by inhibition of respiratory PMF generation. PMID:23567447

  19. Improved transduction efficiencies of adeno-associated virus vectors by synthetic cell-permeable peptides.

    PubMed

    Tabata, Kitako; Sugano, Eriko; Murakami, Fumika; Yamashita, Tetsuro; Ozaki, Taku; Tomita, Hiroshi

    2016-09-30

    Various serotypes of adeno-associated virus (AAV) vectors have been used for gene therapy and as research tools. Among these serotypes, the AAV type 2 vector has been used successfully in human gene therapies. However, the transduction efficiency of AAV2 depends on the cell type, and this poses a problem in the efficacy of gene therapy. To improve the transduction efficiency of AAV2, we designed a small peptide consisting of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor peptide and the HIV-Tat sequence Tat-Y1068. Pre- or co-treatment of CYNOM-K1 cells from cynomolgus monkey embryo skin with Tat-Y1068 increased the transduction efficiencies in a dose-dependent manner and caused p38 phosphorylation. The transduction efficiency of AAV2 into the rat fibroblast cell line RAT-1 highly expressing EGFR was less than the transduction efficiency of AAV2 into CYNOM-K1 cells. Tat-Y1068 increased the transduction efficiency in RAT-1 cells in the same manner as in CYNOM-K1 cells. In conclusion, cell-permeable peptides possessing the EGFR tyrosine kinase inhibitor function might serve as a useful ingredient of AAV2 vector solution for increasing the transduction efficiency of gene therapies.

  20. Extracellular Matrix Rigidity-dependent Sphingosine-1-phosphate Secretion Regulates Metastatic Cancer Cell Invasion and Adhesion

    PubMed Central

    Ko, Panseon; Kim, Daehwan; You, Eunae; Jung, Jangho; Oh, Somi; Kim, Jaehyun; Lee, Kwang-Ho; Rhee, Sangmyung

    2016-01-01

    Dynamic interaction between cancer cells and the surrounding microenvironment is critical for cancer progression via changes in cellular behavior including alteration of secreted molecules. However, the molecular mechanisms underlying the influence exerted by the cancer microenvironment on secretion of molecules during cancer progression remain largely unknown. In this study, we report that secretion of spingsine-1-phosphate (S1P) and its regulator, SphK1 expression is dependent of the substrate rigidity, which is critical for the balance between cancer cell invasion and adhesion. Conditioned media (CM) of MDA-MB-231, an aggressive breast cancer cell obtained from soft substrate (~0.5 kPa) induced chemo-attractive invasion, while CM obtained from stiff substrate (~2.5 kPa) increased cell adhesion instead. We found that the expression of SphK1 is upregulated in the stiff substrate, resulting in an increase in S1P levels in the CM. We also found that upregulation of SphK1 expression in the stiff substrate is dominant in metastatic cancer cells but not in primary cancer cells. These results suggest that alterations in the mechanical environment of the ECM surrounding the tumor cells actively regulate cellular properties such as secretion, which in turn, may contribute to cancer progression. PMID:26877098

  1. Improved transduction efficiencies of adeno-associated virus vectors by synthetic cell-permeable peptides.

    PubMed

    Tabata, Kitako; Sugano, Eriko; Murakami, Fumika; Yamashita, Tetsuro; Ozaki, Taku; Tomita, Hiroshi

    2016-09-30

    Various serotypes of adeno-associated virus (AAV) vectors have been used for gene therapy and as research tools. Among these serotypes, the AAV type 2 vector has been used successfully in human gene therapies. However, the transduction efficiency of AAV2 depends on the cell type, and this poses a problem in the efficacy of gene therapy. To improve the transduction efficiency of AAV2, we designed a small peptide consisting of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor peptide and the HIV-Tat sequence Tat-Y1068. Pre- or co-treatment of CYNOM-K1 cells from cynomolgus monkey embryo skin with Tat-Y1068 increased the transduction efficiencies in a dose-dependent manner and caused p38 phosphorylation. The transduction efficiency of AAV2 into the rat fibroblast cell line RAT-1 highly expressing EGFR was less than the transduction efficiency of AAV2 into CYNOM-K1 cells. Tat-Y1068 increased the transduction efficiency in RAT-1 cells in the same manner as in CYNOM-K1 cells. In conclusion, cell-permeable peptides possessing the EGFR tyrosine kinase inhibitor function might serve as a useful ingredient of AAV2 vector solution for increasing the transduction efficiency of gene therapies. PMID:27614311

  2. Activity measurements of the radionuclide 99mTc for the VNIIM, Russian Federation and ENEA-INMRI, Italy, in the ongoing comparison BIPM.RI(II)-K4.Tc-99m and KCRV update in the BIPM.RI(II)-K1.Tc-99m comparison

    NASA Astrophysics Data System (ADS)

    Michottea, C.; Nonisa, M.; Alekseevb, I. V.; Kharitonovb, I. A.; Tereshchenkob, E. E.; Zanevskiyb, A. V.; Capognic, M.; De Felicec, P.; Fazioc, A.; Carconic, P.

    2016-01-01

    In 2014, comparisons of activity measurements of 99mTc using the Transfer Instrument of the International Reference System (SIRTI) took place at the VNIIM (Russian Federation) and at the ENEA-INMRI (Italy), with comparison identifier BIPM.RI(II)-K4.Tc-99m. Ampoules containing about 32 kBq of 99mTc solutions were measured in the SIRTI for three half-lives. There are now nine results in the BIPM.RI(II)-K4.Tc-99m comparison. This comparison is linked to the BIPM.RI(II)-K1.Tc-99m comparison and the key comparison reference value (KCRV) of the BIPM.RI(II)-K1.Tc-99m has been updated to include eligible results from BIPM.RI(II)-K4.Tc-99m. The degrees of equivalence between each equivalent activity measured in the SIR or the SIRTI 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).

  3. Update of the BIPM comparison BIPM.RI(II)-K1.Sn-113 of activity measurements of the radionuclide 113Sn to include the 2010 results of the PTB (Germany) and the LNE-LNHB (France), and the 2011 result of the CIEMAT (Spain)

    NASA Astrophysics Data System (ADS)

    Michotte, C.; Ratel, G.; Courte, S.; Kossert, K.; Nähle, O.; Bobin, C.; Lépy, M.-C.; Moune, M.; Garcia-Toraño, E.; Peyres, V.; Roteta, M.

    2016-01-01

    Since 1975, seven national metrology institutes (NMI) have submitted eleven samples of known activity of 113Sn 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.Sn-113. The values of the activity submitted were between about 0.7 MBq and 22 MBq. The present primary standardization results for the PTB, Germany and the LNE-LNHB, France, replace their earlier results of 1989 and 1992, respectively. A key comparison reference value (KCRV) has been evaluated for the first time for 113Sn. There are only three results remaining in the BIPM.RI(II)-K1.Sn-113 comparison, all the previously published ones being outdated. The degrees of equivalence between each of these three equivalent activities 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. Update of the BIPM comparison BIPM.RI(II)-K1.Cu-64 of activity measurements of the radionuclide 64Cu to include the 2009 results of the CMI-IIR (Czech Rep.) and the NPL (UK), the 2010 result of the LNE-LNHB (France) and the 2011 result of the ENEA-INMRI (Italy)

    NASA Astrophysics Data System (ADS)

    Michotte, C.; Ratel, G.; Courte, S.; Sochorovà, J.; Auerbach, P.; Keightley, J.; Johansson, L.; Bakhshandeiar, E.; Cassette, P.; Moune, M.; Capogni, M.; De Felice, P.

    2013-01-01

    Since 2009, four national metrology institutes (NMI) have submitted four samples of known activity of 64Cu 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.Cu-64. The values of the activity submitted were between about 3 MBq and 260 MBq. There are now five results in the BIPM.RI(II)-K1.Cu-64 comparison. A key comparison reference value (KCRV) has been calculated for the first time for this nuclide. 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).

  5. Rhyming Words. Fun with Phonics! Book 5. Grades K-1.

    ERIC Educational Resources Information Center

    Eaton, Deborah

    Intended for teachers of kindergarten and first grade, this book offers hands-on activities that make phonics instruction easy and fun for teachers and children in the classroom. The book provides methods for practice, reinforcement, and assessment of phonics skills. A poem is used to introduce the phonics element of this book, rhyming words. The…

  6. Korean Language Maintenance in Los Angeles. Professional Papers K-1.

    ERIC Educational Resources Information Center

    Kim, Kenneth Kong-On; And Others

    Characteristics of the Korean population in Los Angeles, intergenerational cultural problems, and efforts to promote language maintenance are described. The majority of Koreans in Los Angeles have been in the United States less than 10 years. A high percentage are from middle class and professional backgrounds. The traditional hierarchical family…

  7. Phylloquinone (vitamin K1): occurrence, biosynthesis and functions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phylloquinone is a prenylated naphthoquinone that is synthesized exclusively by plants, green algae, and some species of cyanobacteria, where it serves as a vital electron carrier in photosystem I and as an electron acceptor for the formation of protein disulfide bonds. In humans and other vertebrat...

  8. K 1- xLa xCa 2- xNb 3O 10, a Layered Perovskite Series with Variable Interlayer Cation Density, and LaCaNb 3O 10, a Novel Layered Perovskite Oxide with No Interlayer Cations

    NASA Astrophysics Data System (ADS)

    Uma, S.; Gopalakrishnan, J.

    1993-02-01

    A series of layered perovskite oxides of the formula K1-xLaxCa2-xNb3O10 for 0 < x ≤ 1.0 have been prepared. All the members are isostructural, possessing the structure of KCa2Nb3O10. The interlayer potassium ions in the new series can be ion-exchanged with protons to give H1-xLaxCa2-xNb3O10. The latter readily forms intercalation compounds of the formula (CnH2n+1NH3)1-x LaxCa2-xNb3O10, just as the parent solid acid HCa2Nb3O10. The end member LaCaNb3O10 containing no interlayer cations is a novel layered perovskite oxide, being a n = 3 member of the series An-1BnX3n+1.

  9. Phospholipase D signaling in serotonin-induced mitogenesis of pulmonary artery smooth muscle cells.

    PubMed

    Liu, Y; Fanburg, B L

    2008-09-01

    We have previously reported the participation of mitogen-activated protein, Rho, and phosphoinositide-3 (PI3) kinases in separate pathways in serotonin (5-HT)-induced proliferation of pulmonary artery smooth muscle cells (SMCs). In this study, we investigated the possible participation of phospholipase D (PLD) and phosphatidic acid (PA) in this growth process. 5-HT stimulated a time-dependent increase in [(3)H]phosphatidylbutanol and PA generation. Exposure of SMCs to 1-butanol or overexpression of an inactive mutant of human PLD1R898R blocked 5-HT-induced proliferation. Furthermore, 1-butanol inhibited 5-HT activation of S6K1 and S6 protein, downstream effectors of mammalian target of rapamycin (mTOR), by 80 and 72%, respectively, and partially blocked activation of extracellular signal-regulated kinase (ERK) by 30% but had no effect on other associated signaling pathways. Exogenous PA caused cellular proliferation and revitalized cyclin D1 expression by 5-HT of the 1-butanol-treated cells. PA also reproduced activations by 5-HT of mTOR, S6K1, and ERK. Transfection with inactive human PLD1 reduced 5-HT-induced activation of S6K1 by approximately 50%. Inhibition of 5-HT receptor 2A (R 2A) with ketaserin blocked PLD activation by 5-HT. Inhibition with PI3-kinase inhibitor failed to block either activation of PLD by 5-HT or PA-dependent S6K1 phosphorylation. Taken together, these results indicate that ligation of the 5-HTR 2A by 5-HT initiates PLD activation in SMCs, and that its product, PA, is an early signaling molecule in 5-HT-induced pulmonary artery SMC proliferation. Signaling by PA produces its downstream effects primarily through the mTOR/S6K1 pathway and to a lesser extent through the ERK pathway. Hydrolysis of cell membrane lipid may be important in vascular effects of 5-HT. PMID:18621911

  10. Phospholipase D signaling in serotonin-induced mitogenesis of pulmonary artery smooth muscle cells.

    PubMed

    Liu, Y; Fanburg, B L

    2008-09-01

    We have previously reported the participation of mitogen-activated protein, Rho, and phosphoinositide-3 (PI3) kinases in separate pathways in serotonin (5-HT)-induced proliferation of pulmonary artery smooth muscle cells (SMCs). In this study, we investigated the possible participation of phospholipase D (PLD) and phosphatidic acid (PA) in this growth process. 5-HT stimulated a time-dependent increase in [(3)H]phosphatidylbutanol and PA generation. Exposure of SMCs to 1-butanol or overexpression of an inactive mutant of human PLD1R898R blocked 5-HT-induced proliferation. Furthermore, 1-butanol inhibited 5-HT activation of S6K1 and S6 protein, downstream effectors of mammalian target of rapamycin (mTOR), by 80 and 72%, respectively, and partially blocked activation of extracellular signal-regulated kinase (ERK) by 30% but had no effect on other associated signaling pathways. Exogenous PA caused cellular proliferation and revitalized cyclin D1 expression by 5-HT of the 1-butanol-treated cells. PA also reproduced activations by 5-HT of mTOR, S6K1, and ERK. Transfection with inactive human PLD1 reduced 5-HT-induced activation of S6K1 by approximately 50%. Inhibition of 5-HT receptor 2A (R 2A) with ketaserin blocked PLD activation by 5-HT. Inhibition with PI3-kinase inhibitor failed to block either activation of PLD by 5-HT or PA-dependent S6K1 phosphorylation. Taken together, these results indicate that ligation of the 5-HTR 2A by 5-HT initiates PLD activation in SMCs, and that its product, PA, is an early signaling molecule in 5-HT-induced pulmonary artery SMC proliferation. Signaling by PA produces its downstream effects primarily through the mTOR/S6K1 pathway and to a lesser extent through the ERK pathway. Hydrolysis of cell membrane lipid may be important in vascular effects of 5-HT.

  11. Stem Cells

    MedlinePlus

    Stem cells are cells with the potential to develop into many different types of cells in the body. They serve as a repair ... body. There are two main types of stem cells: embryonic stem cells and adult stem cells. Stem ...

  12. Cell Structure

    MedlinePlus

    ... Cells, Tissues, & Membranes Cell Structure & Function Cell Structure Cell Function Body Tissues Epithelial Tissue Connective Tissue Muscle Tissue ... apparatus , and lysosomes . « Previous (Cell Structure & Function) Next (Cell Function) » Contact Us | Privacy Policy | Accessibility | FOIA | File Formats ...

  13. Viral susceptibility, transfection and growth of SPB--a fish neural progenitor cell line from the brain of snubnose pompano, Trachinotus blochii (Lacépède).

    PubMed

    Wen, C-M; Ku, C-C; Wang, C-S

    2013-07-01

    This study investigates the susceptibilities of the SPB cell line to fish viruses including giant seaperch iridovirus (GSIV-K1), red sea bream iridovirus (RSIV-Ku), grouper nervous necrosis virus (GNNV-K1), chum salmon reovirus (CSV) and eel herpesvirus (HVA). GSIV-K1, RSIV-Ku and CSV replicated well in SPB cells, with a significant cytopathic effect and virus production. However, the cells were HVA and GNNV refractory. To examine the ability of SPB cells to stably express foreign protein, expression vectors encoding GNNV B1 and B2 fused to enhanced green fluorescent protein (EGFP) and GSIV ORF35L fused to DsRed were constructed and introduced by transfection into SPB cells. Stable transfectants displayed different morphologies compared with SPB and with each other. EGFP-B1 was predominantly localized in the nuclei, EFPF-B2 was distributed throughout the cytoplasm and nucleus, and granular 35L-DsRed was localized with secreted vesicles. The expression of EFPF-B2 in SPB cells produced blebs on the surface, but the cells showing stable expression of EGFP, EGFP-B1 or 35L-DsRed showed normal morphologies. Results show the SPB cells and the transfected cells grow well at temperatures between 20 and 35 °C and with serum-dependent growth. SPB cells are suitable for studies on foreign protein expression and virology.

  14. Enhanced photo-transfection efficiency of mammalian cells on graphene coated substrates

    NASA Astrophysics Data System (ADS)

    Mthunzi, Patience; He, Kuang; Ngcobo, Sandile; Warner, Jamie W.

    2014-03-01

    Literature reports graphene, an atomic-thick sheet of carbon atoms as one of the promising biocompatible scaffolds that promotes cellular proliferation in human mesenchymal stem cells. On the other hand, different mammalian cell lines including the induced pluripotent stem cells exhibited an accelerated proliferation rate when cultured on graphene or graphene oxide coated substrates. These findings provide strong motivation to explore the full capability of graphene in further pluripotent stem cell research activities as there exists an urgent requirement to preserve their therapeutic potential. This therefore calls for non-invasive procedures for handling stem cells in-vitro. For example, resent literature has shown successful laser light driven transfection in both multipotent and pluripotent stem cells. In order to explore the non-invasive nature of optical transfection alongside biocompatible qualities of graphene, in this work we investigated the impact of optically transfecting mouse embryonic stem (mES) cells plated on graphene coated sample chambers. Using Chinese Hamster Ovary cells (CHO-K1), we further studied the influence of graphene on cell viability as well as cell cytotoxicity through assessing changes in levels of mitochondrial adenosine triphosphate (ATP) activity and the release of cytosolic lactate dehydrogenase (LHD) respectively. Our results showed that compared to those treated on plain glass, CHO-K1 cells optically treated while plated on graphene coated substrates exhibited a higher production of ATP and a milder release of LDH. In addition there was enhanced photo-transfection efficiency in both CHO-K1 and mES cells irradiated on graphene sample chambers.

  15. A kinetic study of the oxidation by molecular oxygen of the cytochrome chain of intact yeast cells, Acetobacter suboxydans cells, and of particulate suspensions of heart muscle.

    PubMed

    Ludwig, G D; Kuby, S A; Edelman, G M; Chance, B

    1983-01-01

    The pre-steady state kinetics of the cytochrome c oxidase reaction with oxygen were studied by a variation in the reaction time between approximately 6 and 25 ms at oxygen concentrations less than 6 mumol/l. For baker's yeast, a pseudo-first-order velocity constant of approximately 150 s-1 at 1.3 mumol/l O2 was obtained corresponding to a second-order reaction between O2 and a3 at a forward velocity constant (k+1) of approximately 3 X 10(7) liter equiv.-1s-1. Thus, the membrane-bound oxidase in the intact cell exhibits one of the most rapid enzyme-substrate reactions to be reported. The value is identical with that of Greenwood and Gibson on an isolated, solubilized cytochrome c oxidase. Similar values of k+1 are calculated from the turnover numbers [k+2 (a+2)] divided by the Km values (formula; see text) measured for these yeast preparations, which points to an almost negligible reverse reaction (k-1) compared to k+2(a+2). Similar calculations for the membrane-bound cytochrome c oxidase of heart muscle give a value of k+1 approximately equal to 10(7) liter equiv.-1s-1. The concordance of the different values of k+1 supports the view that the yeast cell wall does not impart a significant diffusion barrier to the transport of molecular oxygen. In contrast, Acetobacter suboxydans exhibits a much larger value for Km, and has a terminal oxidase of different kinetic parameters.

  16. Effects of Plasma Membrane Cholesterol Level and Cytoskeleton F-Actin on Cell Protrusion Mechanics

    PubMed Central

    Khatibzadeh, Nima; Spector, Alexander A.; Brownell, William E.; Anvari, Bahman

    2013-01-01

    Protrusions are deformations that form at the surface of living cells during biological activities such as cell migration. Using combined optical tweezers and fluorescent microscopy, we quantified the mechanical properties of protrusions in adherent human embryonic kidney cells in response to application of an external force at the cell surface. The mechanical properties of protrusions were analyzed by obtaining the associated force-length plots during protrusion formation, and force relaxation at constant length. Protrusion mechanics were interpretable by a standard linear solid (Kelvin) model, consisting of two stiffness parameters, k0 and k1 (with k0>k1), and a viscous coefficient. While both stiffness parameters contribute to the time-dependant mechanical behavior of the protrusions, k0 and k1 in particular dominated the early and late stages of the protrusion formation and elongation process, respectively. Lowering the membrane cholesterol content by 25% increased the k0 stiffness by 74%, and shortened the protrusion length by almost half. Enhancement of membrane cholesterol content by nearly two-fold increased the protrusion length by 30%, and decreased the k0 stiffness by nearly two-and-half-fold as compared with control cells. Cytoskeleton integrity was found to make a major contribution to protrusion mechanics as evidenced by the effects of F-actin disruption on the resulting mechanical parameters. Viscoelastic behavior of protrusions was further characterized by hysteresis and force relaxation after formation. The results of this study elucidate the coordination of plasma membrane composition and cytoskeleton during protrusion formation. PMID:23451167

  17. Separation of CHO cells using hydrocyclones

    PubMed Central

    Pinto, Rodrigo C.V.; Castilho, Leda R.

    2007-01-01

    Hydrocyclones are simple and robust separation devices with no moving parts. In the past few years, their use in animal cell separation has been proposed. In this work, the use of different hydrocyclone configurations for Chinese hamster ovary (CHO) cell separation was investigated following an experimental design. It was shown that cell separation efficiencies for cultures of the wild-type CHO.K1 cell line and of a recombinant CHO cell line producing granulocyte-macrophage colony stimulating factor (GM-CSF) were kept above 97%. Low viability losses were observed, as measured by trypan blue exclusion and by determination of intracellular lactate dehydrogenase (LDH) released to the culture medium. Mathematical models were proposed to predict the flow rate, flow ratio and separation efficiency as a function of hydrocyclone geometry and pressure drop. When cells were monitored for any induction of apoptosis upon passage through the hydrocyclones, no increase in apoptotic cell concentration was observed within 48 h of hydrocycloning. Thus, based on the high separation efficiencies, the robustness of the equipment, and the absence of apoptosis induction, hydrocyclones seem to be specially suited for use as cell retention devices in long-term perfusion runs. PMID:19002842

  18. Separation of CHO cells using hydrocyclones.

    PubMed

    Pinto, Rodrigo C V; Medronho, Ricardo A; Castilho, Leda R

    2008-01-01

    Hydrocyclones are simple and robust separation devices with no moving parts. In the past few years, their use in animal cell separation has been proposed. In this work, the use of different hydrocyclone configurations for Chinese hamster ovary (CHO) cell separation was investigated following an experimental design. It was shown that cell separation efficiencies for cultures of the wild-type CHO.K1 cell line and of a recombinant CHO cell line producing granulocyte-macrophage colony stimulating factor (GM-CSF) were kept above 97%. Low viability losses were observed, as measured by trypan blue exclusion and by determination of intracellular lactate dehydrogenase (LDH) released to the culture medium. Mathematical models were proposed to predict the flow rate, flow ratio and separation efficiency as a function of hydrocyclone geometry and pressure drop. When cells were monitored for any induction of apoptosis upon passage through the hydrocyclones, no increase in apoptotic cell concentration was observed within 48 h of hydrocycloning. Thus, based on the high separation efficiencies, the robustness of the equipment, and the absence of apoptosis induction, hydrocyclones seem to be specially suited for use as cell retention devices in long-term perfusion runs.

  19. Extensive Chromosomal Reorganization in the Evolution of New World Muroid Rodents (Cricetidae, Sigmodontinae): Searching for Ancestral Phylogenetic Traits.

    PubMed

    Pereira, Adenilson Leão; Malcher, Stella Miranda; Nagamachi, Cleusa Yoshiko; O'Brien, Patricia Caroline Mary; Ferguson-Smith, Malcolm Andrew; Mendes-Oliveira, Ana Cristina; Pieczarka, Julio Cesar

    2016-01-01

    Sigmodontinae rodents show great diversity and complexity in morphology and ecology. This diversity is accompanied by extensive chromosome variation challenging attempts to reconstruct their ancestral genome. The species Hylaeamys megacephalus--HME (Oryzomyini, 2n = 54), Necromys lasiurus--NLA (Akodontini, 2n = 34) and Akodon sp.--ASP (Akodontini, 2n = 10) have extreme diploid numbers that make it difficult to understand the rearrangements that are responsible for such differences. In this study we analyzed these changes using whole chromosome probes of HME in cross-species painting of NLA and ASP to construct chromosome homology maps that reveal the rearrangements between species. We include data from the literature for other Sigmodontinae previously studied with probes from HME and Mus musculus (MMU) probes. We also use the HME probes on MMU chromosomes for the comparative analysis of NLA with other species already mapped by MMU probes. Our results show that NLA and ASP have highly rearranged karyotypes when compared to HME. Eleven HME syntenic blocks are shared among the species studied here. Four syntenies may be ancestral to Akodontini (HME2/18, 3/25, 18/25 and 4/11/16) and eight to Sigmodontinae (HME26, 1/12, 6/21, 7/9, 5/17, 11/16, 20/13 and 19/14/19). Using MMU data we identified six associations shared among rodents from seven subfamilies, where MMU3/18 and MMU8/13 are phylogenetic signatures of Sigmodontinae. We suggest that the associations MMU2entire, MMU6proximal/12entire, MMU3/18, MMU8/13, MMU1/17, MMU10/17, MMU12/17, MMU5/16, MMU5/6 and MMU7/19 are part of the ancestral Sigmodontinae genome.

  20. Parasite diversity at the Holarctic nexus: species of Arostrilepis (Eucestoda: Hymenolepididae) in voles and lemmings (Cricetidae: Arvicolinae) from greater Beringia.

    PubMed

    Makarikov, Arseny A; Galbreath, Kurt E; Hoberg, Eric P

    2013-01-01

    Previously unrecognized species of hymenolepidid cestodes attributable to Arostrilepis Mas-Coma & Tenora, 1997 in arvicoline rodents from the greater Beringian region and western North America are described. Discovery and characterization of these tapeworms contributes to the recognition of a complex of cryptic species distributed across the Holarctic region. Three species are proposed: Arostrilepis gulyaevi sp. n. is named for cestodes in Myodes rufocanus from the Republic of Buryatia, southeastern Siberia and from the Khabarovskiy Kray, Chukotka Autonomous Okrug, and Magadanskaya Oblast', Russian Far East (western Beringia); A. cooki sp. n. is named for cestodes in Myodes gapperi from British Columbia, Canada and Montana, USA; and A. rauschorum sp. n. is named for cestodes in Microtus oeconomus, M. longicaudus, M. pennsylvanicus and M. xanthognathus from the Brooks Range, Seward Peninsula, north-central interior, and Arctic coastal plains of Alaska (eastern Beringia) and Montana, USA. Consistent with recent studies defining diversity in the genus, the form, size, and spination (pattern, shape and size) of the cirrus are diagnostic; species are further distinguished by the relative position and length of the cirrus sac, and arrangement of the testes. Assessment of genetic data from the cytochrome b gene of mitochondrial DNA complements differentiation of this complex based on morphological attributes and confirms known species diversity within the genus. New data for geographical distribution and host specificity of known Arostrilepis spp. indicate that 3 of 12 recognized species have Holarctic distributions extending across Beringia. These include Arostrilepis beringiensis (Kontrimavichus & Smirnova, 1991) in lemmings (species of Lemmus and Synaptomys), A. cf. janickii Makarikov & Kontrimavichus, 2011 in root voles (M. oeconomus) MAKARIKOV ET AL. 402 · Zootaxa 3608 (6) © 2013 Magnolia Press and A. macrocirrosa Makarikov, Gulyaev & Kontrimavichus, 2011 in red backed voles (species of Myodes) and less often other rodent host species.

  1. Analysis of meiotic chromosome structure and behavior in Robertsonian heterozygotes of Ellobius tancrei (Rodentia, Cricetidae): a case of monobrachial homology

    PubMed Central

    Matveevsky, Sergey; Bakloushinskaya, Irina; Tambovtseva, Valentina; Romanenko, Svetlana; Kolomiets, Oxana

    2015-01-01

    Abstract Synaptonemal complex (SC) chains were revealed in semisterile intraspecific F1 hybrids of Ellobius tancrei Blasius, 1884 (2n = 49, NF=56 and 2n=50, NF=56), heterozygous for Robertsonian (Rb) translocations. Chains were formed by Rb submetacentrics with monobrachial homology. Chromosome synapsis in spermatocytes of these hybrids was disturbed, apparently because of the problematic release of the chromosomes from the SC chains. These hybrids suffer from low fertility, and our data support the opinion that this is because a formation of Rb metacentrics with monobrachial homology within different races of the same species might be an initial event for the divergence of chromosomal forms. PMID:26752380

  2. The valid generic name for red-backed voles (Muroidea: Cricetidae: Arvicolinae): restatement of the case for Myodes Pallas, 1811

    USGS Publications Warehouse

    Carleton, Michael D.; Gardner, Alfred L.; Pavlinov, Igor Ya.; Musser, Guy G.

    2014-01-01

    In view of contradictions in the recent literature, the valid genus-group name to be applied to northern red-backed voles—Myodes Pallas, 1811, or Clethrionomys Tilesius, 1850—is reviewed. To develop the thesis that Myodes (type species, Mus rutilus Pallas, 1779) is the correct name, our discussion explores the 19th-century taxonomic works that bear on the relevant taxa, the transition in zoological codes apropos the identification of type species, and past nomenclatural habits in cases where no type species was originally indicated. We conclude that Myodes is the senior name to use for the genus-group taxon that includes the Holarctic species rutilus and frame this conclusion within a synonymy of the genus.

  3. Extensive Chromosomal Reorganization in the Evolution of New World Muroid Rodents (Cricetidae, Sigmodontinae): Searching for Ancestral Phylogenetic Traits

    PubMed Central

    Pereira, Adenilson Leão; Malcher, Stella Miranda; Nagamachi, Cleusa Yoshiko; O’Brien, Patricia Caroline Mary; Ferguson-Smith, Malcolm Andrew; Mendes-Oliveira, Ana Cristina; Pieczarka, Julio Cesar

    2016-01-01

    Sigmodontinae rodents show great diversity and complexity in morphology and ecology. This diversity is accompanied by extensive chromosome variation challenging attempts to reconstruct their ancestral genome. The species Hylaeamys megacephalus–HME (Oryzomyini, 2n = 54), Necromys lasiurus—NLA (Akodontini, 2n = 34) and Akodon sp.–ASP (Akodontini, 2n = 10) have extreme diploid numbers that make it difficult to understand the rearrangements that are responsible for such differences. In this study we analyzed these changes using whole chromosome probes of HME in cross-species painting of NLA and ASP to construct chromosome homology maps that reveal the rearrangements between species. We include data from the literature for other Sigmodontinae previously studied with probes from HME and Mus musculus (MMU) probes. We also use the HME probes on MMU chromosomes for the comparative analysis of NLA with other species already mapped by MMU probes. Our results show that NLA and ASP have highly rearranged karyotypes when compared to HME. Eleven HME syntenic blocks are shared among the species studied here. Four syntenies may be ancestral to Akodontini (HME2/18, 3/25, 18/25 and 4/11/16) and eight to Sigmodontinae (HME26, 1/12, 6/21, 7/9, 5/17, 11/16, 20/13 and 19/14/19). Using MMU data we identified six associations shared among rodents from seven subfamilies, where MMU3/18 and MMU8/13 are phylogenetic signatures of Sigmodontinae. We suggest that the associations MMU2entire, MMU6proximal/12entire, MMU3/18, MMU8/13, MMU1/17, MMU10/17, MMU12/17, MMU5/16, MMU5/6 and MMU7/19 are part of the ancestral Sigmodontinae genome. PMID:26800516

  4. Shippingport, Kentucky, is the type locality for the white-footed mouse, Peromyscus leucopus (Rafinesque, 1818) (Mammalia: Rodentia: Cricetidae)

    USGS Publications Warehouse

    Woodman, Neal

    2015-01-01

    The white-footed mouse, Musculus leucopus Rafinesque, 1818 (= Peromyscus leucopus), is a common small mammal that is widespread in the eastern and central United States. Its abundance in many habitats renders it ecologically important, and its status as a reservoir for hantavirus and Lyme disease gives the species medical and economic significance. The recognition of two cytotypes and up to 17 morphological subspecies of P. leucopus indicates considerable variation in the species, and to understand this variation, it is important that the nominate subspecies be adequately defined so as to act as a standard for comparison. Relevant to this standard for the white-footed mouse is its type locality, which has generally been accepted to be either the vague "pine barrens of Kentucky" or the mouth of the Ohio River. Newly assembled information regarding the life and travels of Constantine S. Rafinesque, the North American naturalist who described P. leucopus, establishes that Rafinesque observed this species in July 1818 while visiting Shippingport, Kentucky, which is now within the city limits of Louisville, Jefferson Co., Kentucky. Shippingport is therefore the actual type locality for this species.

  5. Bionomics of Cuterebra austeni (Diptera: Cuterebridae) and its association with Neotoma albigula (Rodentia: Cricetidae) in the southwestern United States.

    PubMed

    Baird, C R

    1997-11-01

    Cuterebra austeni Sabrosky causes cutaneous myiasis in white-throated woodrats, Neotoma albigula, in the southwestern United States. In central and southern Arizona and southwestern New Mexico, this species is bivoltine. Adult flies are active at hilltop aggregation sites from early spring through mid-May and again to a lesser extent in the fall months. Eggs produced from laboratory matings of adult flies hatched in response to warm breath (34-36 degrees C) 6-8 d after oviposition. Oviposition takes place around burrow entrances and near the bases of Opuntia cacti. In the wild, myiasis occurs in woodrats primarily during the spring months, with a small second peak during the fall. Larvae develop in cutaneous warbles in the sternal and the ventral cervical area of N. albigula and complete development in 33 d. Woodrats do not appear to be affected seriously by the presence of 1-5 larvae. Morphological changes in larvae and pupae are described through to adult eclosion.

  6. A new genus and species of chigger mite (Trombidiformes: Trombiculidae) from Loxodontomys pikumche (Rodentia: Cricetidae) in Chile.

    PubMed

    Fuente, María Carolina Silva-de La; Casanueva, María Eugenia; Salas, Lucila Moreno; González-Acuña, Daniel

    2016-01-01

    The family Trombiculidae is one of the most diverse and cosmopolitan (Walter et al. 2009). In Chile, the family Trombiculidae is represented by six genera associated with reptiles: Eutrombicula Ewing; Microtrombicula Ewing; Paratrombicula Goff & Whitaker; Whartonacarus (Brennan & Jones); Diaguitacarus Stekolnikov & González-Acuña and Proschoengastia Vercammen-Grandjean and two genera associated with rodents Chilacarus Webb, Bennett & Loomis and Poliremotus Brennan & Goff (Stekolnikov & González-Acuña 2015). PMID:27394465

  7. Diversity at the Holarctic nexus: species of Arostrilepis (Eucestoda: Hymenolepididae) in arvicoline rodents (Cricetidae: Arvicolinae) from greater Beringia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previously unrecognized species of hymenolepidid cestodes attributable to Arostrilepis Mas-Coma & Tenora, 1997 in arvicoline rodents from the greater Beringian region and western North America are described. Discovery and characterization of these tapeworms contributes to the recognition of a compl...

  8. New species of Arostrilepis (Eucestoda: Hymenolepididae) in members of Cricetidae and Geomyidae (Rodentia) from the western Nearctic.

    PubMed

    Makarikov, Arseny A; Gardner, Scott L; Hoberg, Eric P

    2012-06-01

    Abstract : Specimens originally identified as Arostrilepis horrida from the Nearctic are revised, contributing to the recognition of a complex of cryptic species distributed across the Holarctic region. Previously unrecognized species are described based on specimens in cricetid (Neotominae) and geomyid rodents. Arostrilepis mariettavogeae n. sp. in Peromyscus californicus from Monterey County, California and Arostrilepis schilleri n. sp. in Thomomys bulbivorus from Corvallis, Oregon are characterized. Consistent with recent studies defining diversity in the genus, form, size, and spination (pattern, shape, and size) of the cirrus are diagnostic; species are further distinguished by the relative position and length of the cirrus sac and arrangement of the testes. Species of Arostrilepis have not previously been described in rodents outside of the Arvicolinae or from localities in the Nearctic. These studies emphasize the need for routine deposition of archival specimens and information, from survey, ecological, and biogeographic studies, in museum collections to serve as self-correcting records for biodiversity at local, regional, and continental scales. PMID:22097959

  9. Solid-state conversion of (94-x)(Na1/2Bi1/2)TiO3-6BaTiO3-x(K1/2Na1/2)NbO3 single crystals and their enhanced converse piezoelectric properties

    NASA Astrophysics Data System (ADS)

    Park, Ji-Hoon; Kang, Suk-Joong L.

    2016-01-01

    (94-x)(Na1/2Bi1/2)TiO3-6BaTiO3-x(K1/2Na1/2)NbO3 (NBT-6BT-xKNN) piezoelectric ceramics have notable potential for replacing lead containing piezoelectric ceramics in actuator applications due to their exceptionally large strain. However, a high electric field for producing a large strain and a large hysteresis are critical issues that should be resolved for practical actuator applications. In an attempt to address these issues and optimize the piezoelectric performance, we fabricated NBT-6BT-xKNN (x = 0 - 5) piezoelectric single crystals with a size of 8 x 8x 10 mm by the solid-state single crystal growth method and systematically measured their electrical properties. With increased addition of KNN to replace NBT, the ferroelectricity and piezoelectricity of the fabricated [001] NBT-6BT-xKNN single crystals decreased, but their unipolar strain and hysteresis were considerably improved. For NBT-6BT-5KNN single crystals, the largest maximum strain (Smax) was 0.57% at 6 kV/mm, showing a converse piezoelectric constant (Smax/Emax) of 950 pm/V, and their hysteresis in the unipolar S-E curve was 12% at 6kV/mm, which would be appropriate for some actuator applications. Our results demonstrate the applicability of the produced single crystals as lead-free piezoelectric actuator components.

  10. KEY COMPARISON: Comparisons CCRI(II)-K3.F-18 and APMP.RI(II)-K3.F-18 of activity measurements of the radionuclide 18F and links to the key comparison reference value of the BIPM.RI(II)-K1.F-18 comparison

    NASA Astrophysics Data System (ADS)

    Ratel, G.; Michotte, C.; Woods, M. J.

    2005-01-01

    In 2003, the CCRI(II) decided that an indirect comparison of 18F measurements piloted by the National Physical Laboratory (NPL), UK in 2001 was sufficiently well constructed that it could be converted into a CCRI(II) comparison, with comparison identifier CCRI(II)-K3.F-18. At the same time, the pilot laboratory made a bilateral comparison with the institute in Chinese Taipei, comparison identifier APMP.RI(II)-K3.F-18. The results of the comparisons have been reported and the