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Sample records for organischen stoffgruppen pak

  1. PAK family kinases

    PubMed Central

    Zhao, Zhuo-shen; Manser, Ed

    2012-01-01

    The p21-activated kinases (PAKs) are a family of Ser/Thr protein kinases that are represented by six genes in humans (PAK 1–6), and are found in all eukaryotes sequenced to date. Genetic and knockdown experiments in frogs, fish and mice indicate group I PAKs are widely expressed, required for multiple tissue development, and particularly important for immune and nervous system function in the adult. The group II PAKs (human PAKs 4–6) are more enigmatic, but their restriction to metazoans and presence at cell-cell junctions suggests these kinases emerged to regulate junctional signaling. Studies of protozoa and fungal PAKs show that they regulate cell shape and polarity through phosphorylation of multiple cytoskeletal proteins, including microtubule binding proteins, myosins and septins. This chapter discusses what we know about the regulation of PAKs and their physiological role in different model organisms, based primarily on gene knockout studies. PMID:23162738

  2. A tale of two Paks.

    PubMed

    Arias-Romero, Luis E; Chernoff, Jonathan

    2008-02-01

    Paks (p21-activated kinases) are effectors for the small GTPases Cdc42 (cell division cycle 42) and Rac that play fundamental roles in a wide range of cellular processes, including cell morphology, motility, survival, gene transcription, apoptosis and hormone signalling. These enzymes are widely expressed in numerous tissues and are activated by extracellular signals through GTPase-dependent and -independent mechanisms. On the basis of structural and biochemical features, the Pak family members have been classified into two groups, comprising three members each. The two Pak groups have generally been considered as two halves of a single entity, but accumulating evidence indicates that this is not so. In this review, we discuss recent advances in our understanding of the structure, regulation and function of these kinases, emphasizing the many differences between these two groups of signalling proteins.

  3. Role of PAK6 in Prostate Cancer

    DTIC Science & Technology

    2007-04-01

    IQGAP1 and PP2C . Increased expression of PAK6 in androgen independent conditions suggests the role of PAK6 in androgen independent prostate cancers. 15...SUBJECT TERMS AR, PAK6, CRIB, PCA, MKK6, Nucleolin, IQGAP1, PP2C . 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF...Nontransfected 293 cells were used as negative control. Another important proteins which were found to interact with PAK6 are IQGAP1 (Fig.2) and PP2C (Fig

  4. Pyrolysis of the tetra pak

    SciTech Connect

    Korkmaz, Ahmet; Yanik, Jale Brebu, Mihai; Vasile, Cornelia

    2009-11-15

    This study deals with pyrolysis of tetra pak which is widely used as an aseptic beverage packaging material. Pyrolysis experiments were carried out under inert atmosphere in a batch reactor at different temperatures and by different pyrolysis modes (one- and two-step). The yields of char, liquid and gas were quantified. Pyrolysis liquids produced were collected as three separate phases; aqueous phase, tar and polyethylene wax. Characterization of wax and the determination of the total amount of phenols in aqueous phase were performed. Chemical compositions of gas and char products relevant to fuel applications were determined. Pure aluminum can be also recovered by pyrolysis.

  5. Structure, biochemistry, and biology of PAK kinases.

    PubMed

    Kumar, Rakesh; Sanawar, Rahul; Li, Xiaodong; Li, Feng

    2017-03-20

    PAKs, p21-activated kinases, play central roles and act as converging junctions for discrete signals elicited on the cell surface and for a number of intracellular signaling cascades. PAKs phosphorylate a vast number of substrates and act by remodeling cytoskeleton, employing scaffolding, and relocating to distinct subcellular compartments. PAKs affect wide range of processes that are crucial to the cell from regulation of cell motility, survival, redox, metabolism, cell cycle, proliferation, transformation, stress, inflammation, to gene expression. Understandably, their dysregulation disrupts cellular homeostasis and severely impacts key cell functions, and many of those are implicated in a number of human diseases including cancers, neurological disorders, and cardiac disorders. Here we provide an overview of the members of the PAK family and their current status. We give special emphasis to PAK1 and PAK4, the prototypes of groups I and II, for their profound roles in cancer, the nervous system, and the heart. We also highlight other family members. We provide our perspective on the current advancements, their growing importance as strategic therapeutic targets, and our vision on the future of PAKs.

  6. Role of PAK6 in Prostate Cancer

    DTIC Science & Technology

    2006-10-01

    IQGAP1 and PP2C . 15. SUBJECT TERMS AR,PAK6,CRIB,PCA,MKK6,Nucleolin,IQGAP1, PP2C . 16. SECURITY CLASSIFICATION OF: 18. NUMBER OF PAGES 19a. NAME...Nontransfected 293 cells were used as negative control Another important proteins which were found to interact with PAK6 are IQGAP1(Fig.2)and PP2C (Fig.3...in MCF7 cells expressing triple FLAG PAK6. Protein phosphatase 1 B isoform 2,a 44-46 kDa protein, a member of the PP2C family of Ser/Thr protein

  7. PAK4–6 in cancer and neuronal development

    PubMed Central

    2012-01-01

    PAKs 4, 5 and 6 are members of the group B family of p21-activated kinases. Among this group, PAK4 has been most extensively studied. While it has essential roles in embryonic development, in adults high levels of PAK4 are frequently associated with cancer. PAK4 is overexpressed in a variety of cancers, and the Pak4 gene is amplified in some cancers. PAK4 overexpression is sufficient to cause oncogenic transformation in cells and in mouse models. The tight connection between PAK4 and cancer make it a promising diagnostic tool as well as a potential drug target. The group B PAKs also have important developmental functions. PAK4 is important for many early developmental processes, while PAK5 and PAK6 play roles in learning and memory in mice. This chapter provides an overview of the roles of the group B PAKs in cancer as well as development, and includes a discussion of PAK mediated signaling pathways and cellular functions. PMID:23125951

  8. Pak2 regulates hematopoietic progenitor cell proliferation, survival and differentiation

    PubMed Central

    Zeng, Yi; Broxmeyer, Hal E.; Staser, Karl; Chitteti, Brahmananda Reddy; Park, Su-Jung; Hahn, Seongmin; Cooper, Scott; Sun, Zejin; Jiang, Li; Yang, XianLin; Yuan, Jin; Kosoff, Rachelle; Sandusky, George; Srour, Edward F.; Chernoff, Jonathan; Clapp, Wade

    2015-01-01

    p21-activated kinase 2 (Pak2), a serine/threonine kinase, has been previously shown to be essential for hematopoietic stem cell (HSC) engraftment. However, Pak2 modulation of long-term hematopoiesis and lineage commitment remain unreported. Utilizing a conditional Pak2 knock out (KO) mouse model, we found that disruption of Pak2 in HSCs induced profound leukopenia and a mild macrocytic anemia. Although loss of Pak2 in HSCs leads to less efficient short- and long-term competitive hematopoiesis than wild type (WT) cells, it does not affect HSC self-renewal per se. Pak2 disruption decreased the survival and proliferation of multi-cytokine stimulated immature progenitors. Loss of Pak2 skewed lineage differentiation toward granulocytopoiesis and monocytopoiesis in mice as evidenced by 1) a three to six-fold increase in the percentage of peripheral blood granulocytes and a significant increase in the percentage of granulocyte-monocyte progenitors (GMPs) in mice transplanted with Pak2-disrupted BM; 2) Pak2-disrupted BM and c-kit+ cells yielded higher numbers of more mature subsets of granulocyte-monocyte colonies and polymophonuclear neutrophils (PMNs), respectively, when cultured in the presence of granulocyte-macrophage colony stimulating factor (GM-CSF). Pak2 disruption resulted respectively in decreased and increased gene expression of transcription factors JunB and c-Myc, which may suggest underlying mechanisms by which Pak2 regulates granulocyte-monocyte lineage commitment. Furthermore, Pak2 disruption led to 1) higher percentage of CD4+CD8+ double positive T cells and lower percentages of CD4+CD8− or CD4−CD8+ single positive T cells in thymus and 2) decreased numbers of mature B cells and increased numbers of Pre-Pro B cells in BM, suggesting defects in lymphopoiesis. PMID:25586960

  9. Herbal therapeutics that block the oncogenic kinase PAK1: a practical approach towards PAK1-dependent diseases and longevity.

    PubMed

    Maruta, Hiroshi

    2014-05-01

    Over 35 years research on PAKs, RAC/CDC42(p21)-activated kinases, comes of age, and in particular PAK1 has been well known to be responsible for a variety of diseases such as cancer (mainly solid tumors), Alzheimer's disease, acquired immune deficiency syndrome and other viral/bacterial infections, inflammatory diseases (asthma and arthritis), diabetes (type 2), neurofibromatosis, tuberous sclerosis, epilepsy, depression, schizophrenia, learning disability, autism, etc. Although several distinct synthetic PAK1-blockers have been recently developed, no FDA-approved PAK1 blockers are available on the market as yet. Thus, patients suffering from these PAK1-dependent diseases have to rely on solely a variety of herbal therapeutics such as propolis and curcumin that block PAK1 without affecting normal cell growth. Furthermore, several recent studies revealed that some of these herbal therapeutics significantly extend the lifespan of nematodes (C. elegans) and fruit flies (Drosophila), and PAK1-deficient worm lives longer than the wild type. Here, I outline mainly pathological phenotypes of hyper-activated PAK1 and a list of herbal therapeutics that block PAK1, but cause no side (harmful) effect on healthy people or animals.

  10. PAKS--Arbeitsbericht Nr. 5. Juli 1970. [PAKS--Working Paper No. 5. July 1970.

    ERIC Educational Resources Information Center

    Stuttgart Univ. (Germany).

    This report, the fifth in a series of working papers issued by the Project on Applied Contrastive Linguistics (PAKS) at the University of Stuttgart, is dedicated to a consideration of error analysis in language learning, here seen as relevant not only for the teacher but for the text book writer and the curriculum planner as well. An introduction…

  11. Signalling to cancer cell invasion through PAK family kinases.

    PubMed

    Whale, Andrew; Hashim, Fariesha Nur; Fram, Sally; Jones, Gareth E; Wells, Claire M

    2011-01-01

    Cancer cell metastasis involves a series of changes in cell behaviour, driven by oncogenic transformation, that leads to local tissue invasion, migration through extracellular matrix, entry into the vascular or lymphatic system and colonisation of distant sites. It is well established that the Rho family GTPases Rho, Rac and Cdc42 orchestrate many of the processes required during metastasis. The Rho family GTPases regulate cellular behaviour through their interaction with downstream effector proteins. The p-21 activated kinases (PAKs), effector proteins for Rac and Cdc42, are known to be important regulators of cell migration and invasion. There are six mammalian PAKs which can be divided into two groups: group I PAKs (PAK1-3) and group II PAKs (PAK4-6). Although the two PAK groups are architecturally similar there are differences in their mode of regulation suggesting their cellular functions are likely to be different. This review will focus on the latest evidence relating to the role of PAK family kinases in the cell signalling pathways that drive cancer cell migration and invasion.

  12. Substrate and Inhibitor Specificity of the Type II p21-Activated Kinase, PAK6

    PubMed Central

    Gao, Jia; Ha, Byung Hak; Lou, Hua Jane; Morse, Elizabeth M.; Zhang, Rong; Calderwood, David A.; Turk, Benjamin E.; Boggon, Titus J.

    2013-01-01

    The p21-activated kinases (PAKs) are important effectors of Rho-family small GTPases. The PAK family consists of two groups, type I and type II, which have different modes of regulation and signaling. PAK6, a type II PAK, influences behavior and locomotor function in mice and has an ascribed role in androgen receptor signaling. Here we show that PAK6 has a peptide substrate specificity very similar to the other type II PAKs, PAK4 and PAK5 (PAK7). We find that PAK6 catalytic activity is inhibited by a peptide corresponding to its N-terminal pseudosubstrate. Introduction of a melanoma-associated mutation, P52L, into this peptide reduces pseudosubstrate autoinhibition of PAK6, and increases phosphorylation of its substrate PACSIN1 (Syndapin I) in cells. Finally we determine two co-crystal structures of PAK6 catalytic domain in complex with ATP-competitive inhibitors. We determined the 1.4 Å co-crystal structure of PAK6 with the type II PAK inhibitor PF-3758309, and the 1.95 Å co-crystal structure of PAK6 with sunitinib. These findings provide new insights into the structure-function relationships of PAK6 and may facilitate development of PAK6 targeted therapies. PMID:24204982

  13. The Retinoblastoma Tumor Suppressor Transcriptionally Represses Pak1 in Osteoblasts

    PubMed Central

    Sosa-García, Bernadette; Vázquez-Rivera, Viviana; González-Flores, Jonathan N.; Engel, Brienne E.; Cress, W. Douglas; Santiago-Cardona, Pedro G.

    2015-01-01

    We previously characterized the retinoblastoma tumor suppressor protein (Rb) as a regulator of adherens junction assembly and cell-to-cell adhesion in osteoblasts. This is a novel function since Rb is predominantly known as a cell cycle repressor. Herein, we characterized the molecular mechanisms by which Rb performs this function, hypothesizing that Rb controls the activity of known regulators of adherens junction assembly. We found that Rb represses the expression of the p21-activated protein kinase (Pak1), an effector of the small Rho GTPase Rac1. Rac1 is a well-known regulator of adherens junction assembly whose increased activity in cancer is linked to perturbations of intercellular adhesion. Using nuclear run-on and luciferase reporter transcription assays, we found that Pak1 repression by Rb is transcriptional, without affecting Pak1 mRNA and protein stability. Pak1 promoter bioinformatics showed multiple E2F1 binding sites within 155 base pairs of the transcriptional start site, and a Pak1-promoter region containing these E2F sites is susceptible to transcriptional inhibition by Rb. Chromatin immunoprecipitations showed that an Rb-E2F complex binds to the region of the Pak1 promoter containing the E2F1 binding sites, suggesting that Pak1 is an E2F target and that the repressive effect of Rb on Pak1 involves blocking the trans-activating capacity of E2F. A bioinformatics analysis showed elevated Pak1 expression in several solid tumors relative to adjacent normal tissue, with both Pak1 and E2F increased relative to normal tissue in breast cancer, supporting a cancer etiology for Pak1 up-regulation. Therefore, we propose that by repressing Pak1 expression, Rb prevents Rac1 hyperactivity usually associated with cancer and related to cytoskeletal derangements that disrupt cell adhesion, consequently enhancing cancer cell migratory capacity. This de-regulation of cell adhesion due to Rb loss could be part of the molecular events associated with cancer progression

  14. Rac1-PAK2 pathway is essential for zebrafish heart regeneration.

    PubMed

    Peng, Xiangwen; He, Quanze; Li, Guobao; Ma, Jinmin; Zhong, Tao P

    2016-04-15

    P-21 activated kinases, or PAKs, are serine-threonine kinases that play important roles in diverse heart functions include heart development, cardiovascular development and function in a range of models; however, the mechanisms by which PAKs mediate heart regeneration are unknown. Here, we demonstrate that PAK2 and PAK4 expression is induced in cardiomyocytes and vessels, respectively, following zebrafish heart injury. Inhibition of PAK2 and PAK4 using a specific small molecule inhibitor impedes cardiomyocyte proliferation/dedifferentiation and cardiovascular regeneration, respectively. Cdc42 is specifically expressed in the ventricle and may function upstream of PAK2 but not PAK4 under normal conditions and that cardiomyocyte proliferentation during heart regeneration relies on Rac1-mediated activation of Pak2. Our results indicate that PAKs play a key role in heart regeneration.

  15. ARROW-PAK Macroencapsulation. Innovative Technology Summary Report

    SciTech Connect

    2002-04-01

    An ARROW-PAK is a high density polyethylene (HDPE) tube, about 21 feet long and 30 inches wide. Each ARROW-PAK can hold the equivalent of 21 55-gallon drums of mixed waste debris. Each tube is fused to HDPE endcaps using localized heating and high pressure contact. The sleeves and encaps form a tube for macroencapsulating mixed waste debris. The ARROW-PAK may achieve a mixed waste debris volume one-fourth that of the conventional macroencapsulation approach. The mixed waste debris is loaded into 55-gallon drums. Once filled a 'supercompactor' crushes the drums into 12-inch thick pucks. Three pucks can be loaded into a standard 85-gallon metal drum known as an 'overpack'. Seven overpacks fit into each ARROW-PAK.

  16. LAT-independent Erk activation via Bam32-PLC-γ1-Pak1 complexes: GTPase-independent Pak1 activation.

    PubMed

    Rouquette-Jazdanian, Alexandre K; Sommers, Connie L; Kortum, Robert L; Morrison, Deborah K; Samelson, Lawrence E

    2012-10-26

    In T cells, the adaptor Bam32 is coupled to Erk activation downstream of the TCR by an unknown mechanism. We characterized in Jurkat cells and primary T lymphocytes a pathway dependent on Bam32-PLC-γ1-Pak1 complexes, in which Pak1 kinase activates Raf-1 and Mek-1, both upstream of Erk. In the Bam32-PLC-γ1-Pak1 complex, catalytically inactive PLC-γ1 is used as a scaffold linking Bam32 to Pak1. PLC-γ1(C-SH2) directly binds S141 of Bam32, preventing LAT-mediated activation of Ras by PLC-γ1. The Bam32-PLC-γ1 interaction enhances the binding of the SH3 domain of the phospholipase with Pak1. The PLC-γ1(SH3)-Pak1 interaction activates Pak1 independently of the small GTPases Rac1/Cdc42, previously described as being the only activators of Pak1 in T cells. Direct binding of the SH3 domain of PLC-γ1 to Pak1 dissociates inactive Pak1 homodimers, a mechanism required for Pak1 activation. We have thus uncovered a LAT/Ras-independent, Bam32-nucleated pathway that activates Erk signaling in T cells.

  17. Development of a Hydronic Rooftop Unit-HyPak-MA

    SciTech Connect

    Lee, Eric; Berman, Mark

    2009-11-14

    The majority of U.S. commercial floor space is cooled by rooftop HVAC units (RTUs). RTU popularity derives chiefly from their low initial cost and relative ease of service access without disturbing building occupants. Unfortunately, current RTUs are inherently inefficient due to a combination of characteristics that unnecessarily increase cooling loads and energy use. 36% percent of annual U.S. energy, and two-thirds of electricity, is consumed in and by buildings. Commercial buildings consume approximately 4.2 quads of energy each year at a cost of $230 billion per year, with HVAC equipment consuming 1.2 quads of electricity. More than half of all U.S. commercial floor space is cooled by packaged HVAC units, most of which are rooftop units (RTUs). Inefficient RTUs create an estimated 3.5% of U.S. CO{sub 2} emissions, thus contributing significantly to global warming5. Also, RTUs often fail to maintain adequate ventilation air and air filtration, reducing indoor air quality. This is the second HyPak project to be supported by DOE through NETL. The prior project, referred to as HyPak-1 in this report, had two rounds of prototype fabrication and testing as well as computer modeling and market research. The HyPak-1 prototypes demonstrated the high performance capabilities of the HyPak concept, but made it clear that further development was required to reduce heat exchanger cost and improve system reliability before HyPak commercialization can commence. The HyPak-1 prototypes were limited to about 25% ventilation air fraction, limiting performance and marketability. The current project is intended to develop a 'mixed-air' product that is capable of full 0-100% modulation in ventilation air fraction, hence it was referred to as HyPak-MA in the proposal. (For simplicity, the -MA has been dropped when referencing the current project.) The objective of the HyPak Project is to design, develop and test a hydronic RTU that provides a quantum improvement over conventional RTU

  18. PAK1 modulates a PPARγ/NF-κB cascade in intestinal inflammation.

    PubMed

    Dammann, Kyle; Khare, Vineeta; Lang, Michaela; Claudel, Thierry; Harpain, Felix; Granofszky, Nicolas; Evstatiev, Rayko; Williams, Jonathan M; Pritchard, D Mark; Watson, Alastair; Gasche, Christoph

    2015-10-01

    P21-activated kinases (PAKs) are multifunctional effectors of Rho GTPases with both kinase and scaffolding activity. Here, we investigated the effects of inflammation on PAK1 signaling and its role in colitis-driven carcinogenesis. PAK1 and p-PAK1 (Thr423) were assessed by immunohistochemistry, immunofluorescence, and Western blot. C57BL6/J wildtype mice were treated with a single intraperitoneal TNFα injection. Small intestinal organoids from these mice and from PAK1-KO mice were cultured with TNFα. NF-κB and PPARγ were analyzed upon PAK1 overexpression and silencing for transcriptional/translational regulation. PAK1 expression and activation was increased on the luminal intestinal epithelial surface in inflammatory bowel disease and colitis-associated cancer. PAK1 was phosphorylated upon treatment with IFNγ, IL-1β, and TNFα. In vivo, mice administered with TNFα showed increased p-PAK1 in intestinal villi, which was associated with nuclear p65 and NF-κB activation. p65 nuclear translocation downstream of TNFα was strongly inhibited in PAK1-KO small intestinal organoids. PAK1 overexpression induced a PAK1-p65 interaction as visualized by co-immunoprecipitation, nuclear translocation, and increased NF-κB transactivation, all of which were impeded by kinase-dead PAK1. Moreover, PAK1 overexpression downregulated PPARγ and mesalamine recovered PPARγ through PAK1 inhibition. On the other hand PAK1 silencing inhibited NF-κB, which was recovered using BADGE, a PPARγ antagonist. Altogether these data demonstrate that PAK1 overexpression and activation in inflammation and colitis-associated cancer promote NF-κB activity via suppression of PPARγ in intestinal epithelial cells.

  19. PAK1 modulates a PPARγ/NF-κB cascade in intestinal inflammation

    PubMed Central

    Dammann, Kyle; Khare, Vineeta; Lang, Michaela; Claudel, Thierry; Harpain, Felix; Granofszky, Nicolas; Evstatiev, Rayko; Williams, Jonathan M.; Pritchard, D. Mark; Watson, Alastair; Gasche, Christoph

    2015-01-01

    P21-activated kinases (PAKs) are multifunctional effectors of Rho GTPases with both kinase and scaffolding activity. Here, we investigated the effects of inflammation on PAK1 signaling and its role in colitis-driven carcinogenesis. PAK1 and p-PAK1 (Thr423) were assessed by immunohistochemistry, immunofluorescence, and Western blot. C57BL6/J wildtype mice were treated with a single intraperitoneal TNFα injection. Small intestinal organoids from these mice and from PAK1-KO mice were cultured with TNFα. NF-κB and PPARγ were analyzed upon PAK1 overexpression and silencing for transcriptional/translational regulation. PAK1 expression and activation was increased on the luminal intestinal epithelial surface in inflammatory bowel disease and colitis-associated cancer. PAK1 was phosphorylated upon treatment with IFNγ, IL-1β, and TNFα. In vivo, mice administered with TNFα showed increased p-PAK1 in intestinal villi, which was associated with nuclear p65 and NF-κB activation. p65 nuclear translocation downstream of TNFα was strongly inhibited in PAK1-KO small intestinal organoids. PAK1 overexpression induced a PAK1–p65 interaction as visualized by co-immunoprecipitation, nuclear translocation, and increased NF-κB transactivation, all of which were impeded by kinase-dead PAK1. Moreover, PAK1 overexpression downregulated PPARγ and mesalamine recovered PPARγ through PAK1 inhibition. On the other hand PAK1 silencing inhibited NF-κB, which was recovered using BADGE, a PPARγ antagonist. Altogether these data demonstrate that PAK1 overexpression and activation in inflammation and colitis-associated cancer promote NF-κB activity via suppression of PPARγ in intestinal epithelial cells. PMID:26036343

  20. Lethality of PAK3 and SGK2 shRNAs to human papillomavirus positive cervical cancer cells is independent of PAK3 and SGK2 knockdown.

    PubMed

    Zhou, Nannan; Ding, Bo; Agler, Michele; Cockett, Mark; McPhee, Fiona

    2015-01-01

    The p21-activated kinase 3 (PAK3) and the serum and glucocorticoid-induced kinase 2 (SGK2) have been previously proposed as essential kinases for human papillomavirus positive (HPV+) cervical cancer cell survival. This was established using a shRNA knockdown approach. To validate PAK3 and SGK2 as potential targets for HPV+ cervical cancer therapy, the relationship between shRNA-induced phenotypes in HPV+ cervical cancer cells and PAK3 or SGK2 knockdown was carefully examined. We observed that the phenotypes of HPV+ cervical cancer cells induced by various PAK3 and SGK2 shRNAs could not be rescued by complement expression of respective cDNA constructs. A knockdown-deficient PAK3 shRNA with a single mismatch was sufficient to inhibit HeLa cell growth to a similar extent as wild-type PAK3 shRNA. The HPV+ cervical cancer cells were also susceptible to several non-human target shRNAs. The discrepancy between PAK3 and SGK2 shRNA-induced apoptosis and gene expression knockdown, as well as cell death stimulation, suggested that these shRNAs killed HeLa cells through different pathways that may not be target-specific. These data demonstrated that HPV+ cervical cancer cell death was not associated with RNAi-induced PAK3 and SGK2 knockdown but likely through off-target effects.

  1. PAK4 interacts with p85 alpha: implications for pancreatic cancer cell migration

    PubMed Central

    King, Helen; Thillai, Kiruthikah; Whale, Andrew; Arumugam, Prabhu; Eldaly, Hesham; Kocher, Hemant M.; Wells, Claire M.

    2017-01-01

    It has been reported that p21-activated kinase 4 (PAK4) is amplified in pancreatic cancer tissue. PAK4 is a member of the PAK family of serine/threonine kinases, which act as effectors for several small GTPases, and has been specifically identified to function downstream of HGF-mediated c-Met activation in a PI3K dependent manner. However, the functionality of PAK4 in pancreatic cancer and the contribution made by HGF signalling to pancreatic cancer cell motility remain to be elucidated. We now find that elevated PAK4 expression is coincident with increased expression levels of c-Met and the p85α subunit of PI3K. Furthermore, we demonstrate that pancreatic cancer cells have a specific motility response to HGF both in 2D and 3D physiomimetic organotypic assays; which can be suppressed by inhibition of PI3K. Significantly, we report a specific interaction between PAK4 and p85α and find that PAK4 deficient cells exhibit a reduction in Akt phosphorylation downstream of HGF signalling. These results implicate a novel role for PAK4 within the PI3K pathway via interaction with p85α. Thus, PAK4 could be an essential player in PDAC progression representing an interesting therapeutic opportunity. PMID:28205613

  2. Development of 2, 4-diaminoquinazoline derivatives as potent PAK4 inhibitors by the core refinement strategy.

    PubMed

    Hao, Chenzhou; Huang, Wanxu; Li, Xiaodong; Guo, Jing; Chen, Meng; Yan, Zizheng; Wang, Kai; Jiang, Xiaolin; Song, Shuai; Wang, Jian; Zhao, Dongmei; Li, Feng; Cheng, Maosheng

    2017-05-05

    Upon analysis of the reported crystal structure of PAK4 inhibitor KY04031 (PAK4 IC50 = 0.790 μM) in the active site of PAK4, we investigated the possibility of changing the triazine core of KY04031 to a quinazoline. Using KY04031 as a starting compound, a library of 2, 4-diaminoquinazoline derivatives were designed and synthesized. These compounds were evaluated for PAK4 inhibition, leading to the identification of compound 9d (PAK4 IC50 = 0.033 μM). Compound 9d significantly induced the cell cycle in the G1/S phase and inhibited migration and invasion of A549 cells that over-express PAK4 via regulation of the PAK4-LIMK1 signalling pathway. A docking study of compound 9d was performed to elucidate its possible binding modes and to provide a structural basis for further structure-guided design of PAK4 inhibitors. Compound 9d may serve as a lead compound for anticancer drug discovery and as a valuable research probe for further biological investigation of PAK4.

  3. Pak1 and PIX regulate contact inhibition during epithelial wound healing.

    PubMed

    Zegers, Mirjam M P; Forget, Marie-Annick; Chernoff, Jonathan; Mostov, Keith E; ter Beest, Martin B A; Hansen, Steen H

    2003-08-15

    Wound healing in epithelia requires coordinated cell migration and proliferation regulated by signaling mechanisms that are poorly understood. Here we show that epithelial cells expressing constitutively active or kinase-dead mutants of the Rac/Cdc42 effector Pak1 fail to undergo growth arrest upon wound closure. Strikingly, this phenotype is only observed when the Pak1 kinase mutants are expressed in cells possessing a free lateral surface, i.e. one that is not engaged in contact with neighboring cells. The Pak1 kinase mutants perturb contact inhibition by a mechanism that depends on the Pak-interacting Rac-GEF PIX. In control cells, endogenous activated Pak and PIX translocate from focal complexes to cell-cell contacts during wound closure. This process is abrogated in cells expressing Pak1 kinase mutants. In contrast, Pak1 mutants rendered defective in PIX binding do not impede translocation of activated Pak and PIX, and exhibit normal wound healing. Thus, recruitment of activated Pak and PIX to cell-cell contacts is pivotal to transduction of growth-inhibitory signals from neighboring cells in epithelial wound healing.

  4. Optimization of a Dibenzodiazepine Hit to a Potent and Selective Allosteric PAK1 Inhibitor

    PubMed Central

    2015-01-01

    The discovery of inhibitors targeting novel allosteric kinase sites is very challenging. Such compounds, however, once identified could offer exquisite levels of selectivity across the kinome. Herein we report our structure-based optimization strategy of a dibenzodiazepine hit 1, discovered in a fragment-based screen, yielding highly potent and selective inhibitors of PAK1 such as 2 and 3. Compound 2 was cocrystallized with PAK1 to confirm binding to an allosteric site and to reveal novel key interactions. Compound 3 modulated PAK1 at the cellular level and due to its selectivity enabled valuable research to interrogate biological functions of the PAK1 kinase. PMID:26191365

  5. PAK4 suppresses PDZ-RhoGEF activity to drive invadopodia maturation in melanoma cells

    PubMed Central

    Nicholas, Nicole S.; Pipili, Aikaterini; Lesjak, Michaela S.; Ameer, Simon M.; Geh, Jenny L. C.; Healy, Ciaran; Ross, Alistair D. MacKenzie; Parsons, Maddy; Nestle, Frank O.; Lacy, Katie E.; Wells, Claire M.

    2016-01-01

    Cancer cells are thought to use actin rich invadopodia to facilitate matrix degradation. Formation and maturation of invadopodia requires the co-ordained activity of Rho-GTPases, however the molecular mechanisms that underlie the invadopodia lifecycle are not fully elucidated. Previous work has suggested a formation and disassembly role for Rho family effector p-21 activated kinase 1 (PAK1) however, related family member PAK4 has not been explored. Systematic analysis of isoform specific depletion using in vitro and in vivo invasion assays revealed there are differential invadopodia-associated functions. We consolidated a role for PAK1 in the invadopodia formation phase and identified PAK4 as a novel invadopodia protein that is required for successful maturation. Furthermore, we find that PAK4 (but not PAK1) mediates invadopodia maturation likely via inhibition of PDZ-RhoGEF. Our work points to an essential role for both PAKs during melanoma invasion but provides a significant advance in our understanding of differential PAK function. PMID:27765920

  6. Outgrowth of Neurites from NIE-115 Neuroblastoma Cells Is Prevented on Repulsive Substrates through the Action of PAK

    PubMed Central

    Marler, Katharine J. M.; Kozma, Robert; Ahmed, Sohail; Dong, Jing-Ming; Hall, Christine; Lim, Louis

    2005-01-01

    In the central nervous system (CNS), damaged axons are inhibited from regeneration by glial scars, where secreted chondroitin sulfate proteoglycan (CSPG) and tenascin repulse outgrowth of neurites, the forerunners of axons and dendrites. During differentiation, these molecules are thought to form boundaries for guiding neurons to their correct targets. In neuroblastoma NIE-115 cells, outgrowth of neurites on laminin could be induced by serum starvation or inhibition of RhoA by Clostridium botulinum C3 toxin. The outgrowing neurites avoided crossing onto the repulsive substrate CSPG or tenascin. This avoidance response was partially overcome on expression of membrane-targeted and kinase-inactive forms of PAK. In these cells, the endogenous PAK isoforms colocalized with actin in distinctive sites, αPAK in the cell center as small clusters and along the neurite shaft and βPAK and γPAK in areas with membrane ruffles and filopodia, respectively. When isoform-specific N-terminal PAK sequences were introduced to interfere with PAK function, substantially more neurites crossed onto CSPG when cells contained a γPAK-derived peptide but not the corresponding αPAK- or βPAK-derived peptide. Thus, while neurite outgrowth can be promoted by RhoA inhibition, overcoming the accompanying repulsive guidance response will require modulation of PAK activity. These results have therapeutic implications for CNS repair processes. PMID:15923637

  7. Pak3 regulates apical-basal polarity in migrating border cells during Drosophila oogenesis.

    PubMed

    Felix, Martina; Chayengia, Mrinal; Ghosh, Ritabrata; Sharma, Aditi; Prasad, Mohit

    2015-11-01

    Group cell migration is a highly coordinated process that is involved in a number of physiological events such as morphogenesis, wound healing and tumor metastasis. Unlike single cells, collectively moving cells are physically attached to each other and retain some degree of apical-basal polarity during the migratory phase. Although much is known about direction sensing, how polarity is regulated in multicellular movement remains unclear. Here we report the role of the protein kinase Pak3 in maintaining apical-basal polarity in migrating border cell clusters during Drosophila oogenesis. Pak3 is enriched in border cells and downregulation of its function impedes border cell movement. Time-lapse imaging suggests that Pak3 affects protrusive behavior of the border cell cluster, specifically regulating the stability and directionality of protrusions. Pak3 functions downstream of guidance receptor signaling to regulate the level and distribution of F-actin in migrating border cells. We also provide evidence that Pak3 genetically interacts with the lateral polarity marker Scribble and that it regulates JNK signaling in the moving border cells. Since Pak3 depletion results in mislocalization of several apical-basal polarity markers and overexpression of Jra rescues the polarity of the Pak3-depleted cluster, we propose that Pak3 functions through JNK signaling to modulate apical-basal polarity of the migrating border cell cluster. We also observe loss of apical-basal polarity in Rac1-depleted border cell clusters, suggesting that guidance receptor signaling functions through Rac GTPase and Pak3 to regulate the overall polarity of the cluster and mediate efficient collective movement of the border cells to the oocyte boundary.

  8. Rho family-associated kinases PAK1 and rock.

    PubMed

    Maruta, Hiroshi; Nheu, Thao V; He, Hong; Hirokawa, Yumiko

    2003-01-01

    Rho family GTPases (Rho, Rac and CDC42) share around 30% sequence identity with RAS family GTPases, and are essential for RAS-induced malignant transformation, i.e., aberrant serum/anchorage-independent growth and actin cytoskeleton-linked morphological changes. Oncogenic RAS mutants such as v-Ha-RAS trigger cell cycle entry (G0-G1 transition) mainly by up-regulating cyclin D1, an activator of cyclin-dependent kinases (CDK), and down-regulating p27, a CDK inhibitor. Although both Rac and CDC42 are clearly activated by RAS, there is so far no evidence that RAS activates Rho. In this chapter, we will discuss the role of these Rho family GTPases and their effectors, in particular the Ser/Thr kinases PAK1 and Rock, in RAS-induced serum/anchorage-independent cell cycling, and discuss several potential therapeutics, peptides or chemical compounds, that could block this oncogenic cell cycle signalling pathway.

  9. Tuning PAK Activity to Rescue Abnormal Myelin Permeability in HNPP.

    PubMed

    Hu, Bo; Arpag, Sezgi; Zhang, Xuebao; Möbius, Wiebke; Werner, Hauke; Sosinsky, Gina; Ellisman, Mark; Zhang, Yang; Hamilton, Audra; Chernoff, Jonathan; Li, Jun

    2016-09-01

    Schwann cells in the peripheral nervous systems extend their membranes to wrap axons concentrically and form the insulating sheath, called myelin. The spaces between layers of myelin are sealed by myelin junctions. This tight insulation enables rapid conduction of electric impulses (action potentials) through axons. Demyelination (stripping off the insulating sheath) has been widely regarded as one of the most important mechanisms altering the action potential propagation in many neurological diseases. However, the effective nerve conduction is also thought to require a proper myelin seal through myelin junctions such as tight junctions and adherens junctions. In the present study, we have demonstrated the disruption of myelin junctions in a mouse model (Pmp22+/-) of hereditary neuropathy with liability to pressure palsies (HNPP) with heterozygous deletion of Pmp22 gene. We observed a robust increase of F-actin in Pmp22+/- nerve regions where myelin junctions were disrupted, leading to increased myelin permeability. These abnormalities were present long before segmental demyelination at the late phase of Pmp22+/- mice. Moreover, the increase of F-actin levels correlated with an enhanced activity of p21-activated kinase (PAK1), a molecule known to regulate actin polymerization. Pharmacological inhibition of PAK normalized levels of F-actin, and completely prevented the progression of the myelin junction disruption and nerve conduction failure in Pmp22+/- mice. Our findings explain how abnormal myelin permeability is caused in HNPP, leading to impaired action potential propagation in the absence of demyelination. We call it "functional demyelination", a novel mechanism upstream to the actual stripping of myelin that is relevant to many demyelinating diseases. This observation also provides a potential therapeutic approach for HNPP.

  10. Tuning PAK Activity to Rescue Abnormal Myelin Permeability in HNPP

    PubMed Central

    Hu, Bo; Zhang, Xuebao; Möbius, Wiebke; Werner, Hauke; Sosinsky, Gina; Ellisman, Mark; Zhang, Yang; Hamilton, Audra; Chernoff, Jonathan; Li, Jun

    2016-01-01

    Schwann cells in the peripheral nervous systems extend their membranes to wrap axons concentrically and form the insulating sheath, called myelin. The spaces between layers of myelin are sealed by myelin junctions. This tight insulation enables rapid conduction of electric impulses (action potentials) through axons. Demyelination (stripping off the insulating sheath) has been widely regarded as one of the most important mechanisms altering the action potential propagation in many neurological diseases. However, the effective nerve conduction is also thought to require a proper myelin seal through myelin junctions such as tight junctions and adherens junctions. In the present study, we have demonstrated the disruption of myelin junctions in a mouse model (Pmp22+/-) of hereditary neuropathy with liability to pressure palsies (HNPP) with heterozygous deletion of Pmp22 gene. We observed a robust increase of F-actin in Pmp22+/- nerve regions where myelin junctions were disrupted, leading to increased myelin permeability. These abnormalities were present long before segmental demyelination at the late phase of Pmp22+/- mice. Moreover, the increase of F-actin levels correlated with an enhanced activity of p21-activated kinase (PAK1), a molecule known to regulate actin polymerization. Pharmacological inhibition of PAK normalized levels of F-actin, and completely prevented the progression of the myelin junction disruption and nerve conduction failure in Pmp22+/- mice. Our findings explain how abnormal myelin permeability is caused in HNPP, leading to impaired action potential propagation in the absence of demyelination. We call it “functional demyelination”, a novel mechanism upstream to the actual stripping of myelin that is relevant to many demyelinating diseases. This observation also provides a potential therapeutic approach for HNPP. PMID:27583434

  11. p21-activated kinase1 (Pak1) is a negative regulator of NADPH-oxidase 2 in ventricular myocytes

    PubMed Central

    DeSantiago, Jaime; Bare, Dan J; Xiao, Lei; Ke, Yunbo; Solaro, R. John; Banach, Kathrin

    2014-01-01

    Ischemic conditions reduce the activity of the p21-activated kinase (Pak1) resulting in increased arrhythmic activity. Triggered arrhythmic activity during ischemia is based on changes in cellular ionic balance and the cells Ca2+ handling properties. In the current study we used isolated mouse ventricular myocytes (VMs) deficient for the expression of Pak1 (Pak1-/-) to determine the mechanism by which Pak1 influences the generation of arrhythmic activity during simulated ischemia. The Ca2+ transient amplitude and kinetics did not significantly change in wild type (WT) and Pak1-/- VMs during 15 min of simulated ischemia. However, Pak1-/- VMs exhibited an exaggerated increase in [Ca2+]i, which resulted in spontaneous Ca2+ release events and waves. The Ca2+ overload in Pak1-/- VMs could be suppressed with a reverse mode blocker (KB-R7943) of the sodium calcium exchanger (NCX), a cytoplasmic scavenger of reactive oxygen species (ROS; TEMPOL) or a RAC1 inhibitor (NSC23766). Measurements of the cytoplasmic ROS levels revealed that decreased Pak1 activity in Pak1-/- VMs or VMs treated with the Pak1 inhibitor (IPA3) enhanced cellular ROS production. The Pak1 dependent increase in ROS was attenuated in VMs deficient for NADPH oxidase 2 (NOX2; p47phox-/-) or in VMs where NOX2 was inhibited (gp91ds-tat). Voltage clamp recordings showed increased NCX activity in Pak1-/- VMs that depended on enhanced NOX2 induced ROS production. The exaggerated Ca2+ overload in Pak1-/- VMs could be mimicked by low concentrations of ouabain. Overall our data show that Pak1 is a critical negative regulator of NOX2 dependent ROS production and that a latent ROS dependent stimulation of NCX activity can predispose VMs to Ca2+ overload under conditions where no significant changes in excitation-contraction coupling are yet evident. PMID:24380729

  12. Early Results from the HexPak and GradPak Variable-Scale Dual-Head IFUs on the WIYN 3.5-meter Telescope

    NASA Astrophysics Data System (ADS)

    Hooper, Eric; Bershady, Matthew A.; Eigenbrot, Arthur; Wood, Corey M.; Buckley, Scott; Smith, Michael; Corson, Charles; Wolf, Marsha J.; Zhu, Guanying Y.; Vang, Andrea; Gallagher, John S.; Sheinis, Andrew; Washburn Astronomical Laboratories

    2015-01-01

    The WIYN Observatory recently installed two new integral field units (IFUs) on its 3.5-meter telescope on Kitt Peak, Arizona. Each IFU is unique in that it contains different sized fibers in the same head to optimize the tradeoff between spatial resolution and surface brightness sensitivity for observations of galaxies. These instruments were designed and constructed (M. Bershady, PI) at the University of Wisconsin's Washburn Astronomical Laboratory. HexPak, with a central core of 1 arcsec fibers surrounded by a halo of 3 arcsec fibers, was designed for early type, face-on disk, and quasar host galaxies. GradPak, with a series of rows of fibers of increasing diameter from 2 arcsec to 6 arcsec (5 different diameters total), was designed for edge-on galaxies, where the small fibers lie along the midplane and larger fibers sample the progressively lower surface brightnesses above the plane. The instruments were installed alongside the existing SparsePak IFU in late 2013 and have been used in several observing runs since. The different fiber sizes present additional data reduction challenges, particularly regarding flux calibration and sky subtraction. Early results on studies of the stellar populations of galaxies are quite promising and demonstrate the advantages of fiber sizes tailored to the objects under study. HexPak and GradPak were built with funds from NSF award ATI-0804576.

  13. PAK6 targets to cell–cell adhesions through its N-terminus in a Cdc42-dependent manner to drive epithelial colony escape

    PubMed Central

    Morse, Elizabeth M.; Sun, Xiaowen; Olberding, Jordan R.; Ha, Byung Hak; Boggon, Titus J.; Calderwood, David A.

    2016-01-01

    ABSTRACT The six serine/threonine kinases in the p21-activated kinase (PAK) family are important regulators of cell adhesion, motility and survival. PAK6, which is overexpressed in prostate cancer, was recently reported to localize to cell–cell adhesions and to drive epithelial cell colony escape. Here we report that PAK6 targeting to cell–cell adhesions occurs through its N-terminus, requiring both its Cdc42/Rac interactive binding (CRIB) domain and an adjacent polybasic region for maximal targeting efficiency. We find PAK6 localization to cell–cell adhesions is Cdc42-dependent, as Cdc42 knockdown inhibits PAK6 targeting to cell–cell adhesions. We further find the ability of PAK6 to drive epithelial cell colony escape requires kinase activity and is disrupted by mutations that perturb PAK6 cell–cell adhesion targeting. Finally, we demonstrate that all type II PAKs (PAK4, PAK5 and PAK6) target to cell–cell adhesions, albeit to differing extents, but PAK1 (a type I PAK) does not. Notably, the ability of a PAK isoform to drive epithelial colony escape correlates with its targeting to cell–cell adhesions. We conclude that PAKs have a broader role in the regulation of cell–cell adhesions than previously appreciated. PMID:26598554

  14. Relationship Between Pak-Mediated Cell Death and Stress-Activated Kinase Signaling in Breast Cancer

    DTIC Science & Technology

    2000-02-01

    part of the cell death execution machinery. Here we show that a correlation exists in breast cancer cells between caspase- dependent cleavage of the...inhibits its activity might allow us to specifically inhibit signaling pathways downstream of Pak and evaluate how the cell death process is affected. In...a biochemical approach screening for substrates and possible mediators of cell death signaling components via Pak kinases we identified a guanine

  15. Reduced PAK1 activity sensitizes FA/BRCA-proficient breast cancer cells to PARP inhibition.

    PubMed

    Villamar Cruz, Olga; Prudnikova, Tatiana Y; Araiza-Olivera, Daniela; Perez-Plasencia, Carlos; Johnson, Neil; Bernhardy, Andrea J; Slifker, Michael; Renner, Catherine; Chernoff, Jonathan; Arias-Romero, Luis E

    2016-11-22

    Cells that are deficient in homologous recombination, such as those that have mutations in any of the Fanconi Anemia (FA)/BRCA genes, are hypersensitive to inhibition of poly(ADP-ribose) polymerase (PARP). However, FA/BRCA-deficient tumors represent a small fraction of breast cancers, which might restrict the therapeutic utility of PARP inhibitor monotherapy. The gene encoding the serine-threonine protein kinase p21-activated kinase 1 (PAK1) is amplified and/or overexpressed in several human cancer types including 25-30% of breast tumors. This enzyme controls many cellular processes by phosphorylating both cytoplasmic and nuclear substrates. Here, we show that depletion or pharmacological inhibition of PAK1 down-regulated the expression of genes involved in the FA/BRCA pathway and compromised the ability of cells to repair DNA by Homologous Recombination (HR), promoting apoptosis and reducing colony formation. Combined inhibition of PAK1 and PARP in PAK1 overexpressing breast cancer cells had a synergistic effect, enhancing apoptosis, suppressing colony formation, and delaying tumor growth in a xenograft setting. Because reduced PAK1 activity impaired FA/BRCA function, inhibition of this kinase in PAK1 amplified and/or overexpressing breast cancer cells represents a plausible strategy for expanding the utility of PARP inhibitors to FA/BRCA-proficient cancers.

  16. PAKs supplement improves immune status and body composition but not muscle strength in resistance trained individuals

    PubMed Central

    2010-01-01

    Mixed formula supplements are very popular among recreational and professional weightlifters. They are usually known as PAKs and they are supposed to have a synergistic effect of their different nutrients. The purpose of this study was to determine the effects of chronic (4 weeks) PAKS supplementation in combination with strength training on body composition, immune status and performance measures in recreationally trained individuals with or without PAKs supplementation. Methods: Twelve male subjects (Placebo n = 6 and PAKs supplement n = 6) were recruited for this study. The body composition, one maximum strength repetition tests and immune status were assessed before and after 4 week supplementation. Our data showed that, 4 week PAK supplementation associated with strength exercise not was effective in change strength than compared with placebo group. However, we observed that, PAK supplement was able to improve immune status and reduced body composition when compared with placebo group. These results indicate that, a mixed formula supplement is able to improve immune status and body composition but not maximum strength in recreational strength trained subjects in a 4 weeks period. PMID:21059194

  17. Reduced PAK1 activity sensitizes FA/BRCA-proficient breast cancer cells to PARP inhibition

    PubMed Central

    Araiza-Olivera, Daniela; Perez-Plasencia, Carlos; Johnson, Neil; Bernhardy, Andrea J.; Slifker, Michael; Renner, Catherine; Chernoff, Jonathan; Arias, Luis E.

    2016-01-01

    Cells that are deficient in homologous recombination, such as those that have mutations in any of the Fanconi Anemia (FA)/BRCA genes, are hypersensitive to inhibition of poly(ADP-ribose) polymerase (PARP). However, FA/BRCA-deficient tumors represent a small fraction of breast cancers, which might restrict the therapeutic utility of PARP inhibitor monotherapy. The gene encoding the serine-threonine protein kinase p21-activated kinase 1 (PAK1) is amplified and/or overexpressed in several human cancer types including 25-30% of breast tumors. This enzyme controls many cellular processes by phosphorylating both cytoplasmic and nuclear substrates. Here, we show that depletion or pharmacological inhibition of PAK1 down-regulated the expression of genes involved in the FA/BRCA pathway and compromised the ability of cells to repair DNA by Homologous Recombination (HR), promoting apoptosis and reducing colony formation. Combined inhibition of PAK1 and PARP in PAK1 overexpressing breast cancer cells had a synergistic effect, enhancing apoptosis, suppressing colony formation, and delaying tumor growth in a xenograft setting. Because reduced PAK1 activity impaired FA/BRCA function, inhibition of this kinase in PAK1 amplified and/or overexpressing breast cancer cells represents a plausible strategy for expanding the utility of PARP inhibitors to FA/BRCA-proficient cancers. PMID:27740936

  18. Group I Paks as therapeutic targets in NF2-deficient meningioma

    PubMed Central

    Duron, Sergio G.; Campbell, David A.; Ong, Christy C.; Hoeflich, Klaus P.; Chang, Long-Sheng; Welling, D. Bradley; Yang, Zeng-jie; Chernoff, Jonathan

    2015-01-01

    Neurofibromatosis type 2 (NF2) is an autosomal dominant disorder characterized by the development of multiple tumors in the central nervous system, most notably schwannomas and meningiomas. Mutational inactivation of NF2 is found in 40–60% of sporadic meningiomas, but the molecular mechanisms underlying malignant changes of meningioma cells remain unclear. Because group I p21-activated kinases (Paks) bind to and are inhibited by the NF2-encoded protein Merlin, we assessed the signaling and anti-tumor effects of three group-I specific Pak inhibitors - Frax597, 716 and 1036 - in NF2−/− meningiomas in vitro and in an orthotopic mouse model. We found that these Pak inhibitors suppressed the proliferation and motility of both benign (Ben-Men1) and malignant (KT21-MG1) meningiomas cells. In addition, we found a strong reduction in phosphorylation of Mek and S6, and decreased cyclin D1 expression in both cell lines after treatment with Pak inhibitors. Using intracranial xenografts of luciferase-expressing KT21-MG1 cells, we found that treated mice showed significant tumor suppression for all three Pak inhibitors. Similar effects were observed in Ben-Men1 cells. Tumors dissected from treated animals exhibited an increase in apoptosis without notable change in proliferation. Collectively, these results suggest that Pak inhibitors might be useful agents in treating NF2-deficient meningiomas. PMID:25596744

  19. Comparison of the SidePak personal monitor with the Aerosol Particle Sizer (APS).

    PubMed

    Sánchez Jiménez, Araceli; van Tongeren, Martie; Galea, Karen S; Steinsvåg, Kjersti; MacCalman, Laura; Cherrie, John W

    2011-06-01

    The aim of this study was to compare the performance of the TSI Aerodynamic Particle Sizer (APS) and the TSI portable photometer SidePak to measure airborne oil mist particulate matter (PM) with aerodynamic diameters below 10 μm, 2.5 μm and 1 μm (PM(10), PM(2.5) and PM(1)). Three SidePaks each fitted with either a PM(10), PM(2.5) or a PM(1) impactor and an APS were run side by side in a controlled chamber. Oil mist from two different mineral oils and two different drilling fluid systems commonly used in offshore drilling technologies were generated using a nebulizer. Compared to the APS, the SidePaks overestimated the concentration of PM(10) and PM(2.5) by one order of magnitude and PM(1) concentrations by two orders of magnitude after exposure to oil mist for 3.3-6.5 min at concentrations ranging from 0.003 to 18.1 mg m(-3) for PM(10), 0.002 to 3.96 mg m(-3) for PM(2.5) and 0.001 to 0.418 mg m(-3) for PM(1) (as measured by the APS). In a second experiment a SidePak monitor previously exposed to oil mist overestimated PM(10) concentrations by 27% compared to measurements from another SidePak never exposed to oil mist. This could be a result of condensation of oil mist droplets in the optical system of the SidePak. The SidePak is a very useful instrument for personal monitoring in occupational hygiene due to its light weight and quiet pump. However, it may not be suitable for the measurement of particle concentrations from oil mist.

  20. New realism in north Korean propaganda: The death of Pak Chong-ch`ol. Final report

    SciTech Connect

    Shaw, W.

    1987-04-01

    North Korea`s Voice of National Salvation (VNS) broadcasts to South Korea in January and February gave considerable attention to the 13 January 1987 death under police torture of Seoul National University student Pak Chong-ch`ol. VNS commentary emphasized the routine nature of police torture in South Korea and sharply criticized the South Korean Government for blocking attendance at memorial services on 7 February and 3 March. Many VNS comments on the Pak case closely paralleled reactions of the South Korean press and parliamentary opposition, demonstrating close North Korean attention to opinion trends in the South. In a new mood of realism, P`yongyang also used the Pak case to urge South Korean radical students to drop their `avant-gardism`, including excessively leftist slogans and violent tactics that alienate the general populace. P` yongyang clearly wants students to take advantage of issues that have wide public appeal and to broaden resistance to the Chon Tu-hwan Government to include social groups beyond the student population. In P`yongyang`s view, such issues include the Pak case and the signature campaign for constitutional revision in early 1986. If South Korea`s radical students take P`yonyyang`s advice, and if the Republic of Korea Government fails to curb and punish abuses like the Pak killing, this shift in tactics could have significant influence during the coming spring demonstration season.

  1. Inhibition of Nischarin Expression Promotes Neurite Outgrowth through Regulation of PAK Activity

    PubMed Central

    Ding, Yuemin; Li, Yuying; Lu, Lingchao; Zhang, Ruyi; Zeng, Linghui; Wang, Linlin; Zhang, Xiong

    2015-01-01

    Nischarin is a cytoplasmic protein expressed in various organs that plays an inhibitory role in cell migration and invasion and the carcinogenesis of breast cancer cells. We previously reported that Nischarin is highly expressed in neuronal cell lines and is differentially expressed in the brain tissue of adult rats. However, the physiological function of Nischarin in neural cells remains unknown. Here, we show that Nischarin is expressed in rat primary cortical neurons but not in astrocytes. Nischarin is localized around the nucleus and dendrites. Using shRNA to knockdown the expression of endogenous Nischarin significantly increases the percentage of neurite-bearing cells, remarkably increases neurite length, and accelerates neurite extension in neuronal cells. Silencing Nischarin expression also promotes dendrite elongation in rat cortical neurons where Nischarin interacts with p21-activated kinase 1/2 (PAK1/2) and negatively regulates phosphorylation of both PAK1 and PAK2. The stimulation of neurite growth observed in cells with decreased levels of Nischarin is partially abolished by IPA3-mediated inhibition of PAK1 activity. Our findings indicate that endogenous Nischarin inhibits neurite outgrowth by blocking PAK1 activation in neurons. PMID:26670864

  2. Remarkable reductions of PAKs in the brain tissues of scrapie-infected rodent possibly linked closely with neuron loss.

    PubMed

    Meng, Ge; Tian, Chan; Wang, Hui; Xu, Yin; Zhang, Bao-Yun; Shi, Qi; Gao, Chen; Chen, Cao; Fan, Xue-Yu; Wang, Jing; Xiao, Kang; Ren, Ke; Xue, Ming-Ming; Dong, Xiao-Ping

    2014-10-01

    Prion diseases are irreversible progressive neurodegenerative diseases characterized in the brain by PrP(Sc) deposits, neuronal degeneration, gliosis and by cognitive, behavioral and physical impairments, leading to severe incapacity and inevitable death. Proteins of the p21-activated kinase (PAK) family are noted for roles in gene transcription, cytoskeletal dynamics, cell cycle progression and survival signaling. In the present study, we aimed to identify the potential roles of PAKs during prion infection, utilizing the brains of scrapie agent-infected hamsters. Western blots and immunohistochemical assays showed that brain levels of PAK3 and PAK1, as well as their upstream activator Rac/cdc42 and downstream substrate Raf1, were remarkably reduced at terminal stage. Double-stained immunofluorescent assay demonstrated that PAK3 was expressed mainly in neurons. Dynamic analyses of the brain samples collected at the different time points during the incubation period illustrated successive decreases of PAK3, PAK1 and Raf1, especially phosphor Raf1, which correlated well with neuron loss. Rac/cdc42 in the brain tissues increased at early stage and reached to the top at mid-late stage, but diminished at final stage. Unlike the alteration of PAKs in vivo, PAK3 and PAK1, as well as Rac/cdc42 and Raf1 in the prion-infected cell line SMB-S15 remained unchanged compared with those of its normal cell line SMB-PS. Our data here indicate that the functions of PAKs and their associated signaling pathways are seriously affected in the brains of prion disease, which appear to associate closely with the extensive neuron loss.

  3. PAK Inactivation Impairs Social Recognition in 3xTg-AD Mice without Increasing Brain Deposition of Tau and Aβ

    PubMed Central

    Arsenault, Dany; Dal-Pan, Alexandre; Tremblay, Cyntia; Bennett, David A.; Guitton, Matthieu J.; De Koninck, Yves; Tonegawa, Susumu

    2013-01-01

    Defects in p21-activated kinase (PAK) are suspected to play a role in cognitive symptoms of Alzheimer's disease (AD). Dysfunction in PAK leads to cofilin activation, drebrin displacement from its actin-binding site, actin depolymerization/severing, and, ultimately, defects in spine dynamics and cognitive impairment in mice. To determine the role of PAK in AD, we first quantified PAK by immunoblotting in homogenates from the parietal neocortex of subjects with a clinical diagnosis of no cognitive impairment (n = 12), mild cognitive impairment (n = 12), or AD (n = 12). A loss of total PAK, detected in the cortex of AD patients (−39% versus controls), was correlated with cognitive impairment (r2 = 0.148, p = 0.027) and deposition of total and phosphorylated tau (r2 = 0.235 and r2 = 0.206, respectively), but not with Aβ42 (r2 = 0.056). Accordingly, we found a decrease of total PAK in the cortex of 12- and 20-month-old 3xTg-AD mice, an animal model of AD-like Aβ and tau neuropathologies. To determine whether PAK dysfunction aggravates AD phenotype, 3xTg-AD mice were crossed with dominant-negative PAK mice. PAK inactivation led to obliteration of social recognition in old 3xTg-AD mice, which was associated with a decrease in cortical drebrin (−25%), but without enhancement of Aβ/tau pathology or any clear electrophysiological signature. Overall, our data suggest that PAK decrease is a consequence of AD neuropathology and that therapeutic activation of PAK may exert symptomatic benefits on high brain function. PMID:23804095

  4. Viekira Pak Induced Fatal Lactic Acidosis: A Case Report of an Unusual Side Effect

    PubMed Central

    2016-01-01

    Viekira Pak is a new direct-acting antiviral agent that has an excellent efficacy in treating patients with chronic HCV. FDA released a safety warning that Viekira Pak can cause serious liver injury mostly in patients with underlying advanced liver disease. We report the first case of fatal lactic acidosis presenting 3 days after initiating therapy with Viekira Pak. Although it is very hard to precisely determine the cause of lactic acidosis, our case highlights an unusual side effect that ensued after starting the medication. Given the complexity of drug-drug interactions that can happen with the new direct-acting antiviral agents and the paucity of data regarding coadministration and methods of monitoring, a thorough review should be pursued prior to initiating these medications. PMID:28044114

  5. Glucosinolates from pak choi and broccoli induce enzymes and inhibit inflammation and colon cancer differently.

    PubMed

    Lippmann, Doris; Lehmann, Carsten; Florian, Simone; Barknowitz, Gitte; Haack, Michael; Mewis, Inga; Wiesner, Melanie; Schreiner, Monika; Glatt, Hansruedi; Brigelius-Flohé, Regina; Kipp, Anna P

    2014-06-01

    High consumption of Brassica vegetables is considered to prevent especially colon carcinogenesis. The content and pattern of glucosinolates (GSLs) can highly vary among different Brassica vegetables and may, thus, affect the outcome of Brassica intervention studies. Therefore, we aimed to feed mice with diets containing plant materials of the Brassica vegetables broccoli and pak choi. Further enrichment of the diets by adding GSL extracts allowed us to analyze the impact of different amounts (GSL-poor versus GSL-rich) and different patterns (broccoli versus pak choi) of GSLs on inflammation and tumor development in a model of inflammation-triggered colon carcinogenesis (AOM/DSS model). Serum albumin adducts were analyzed to confirm the up-take and bioactivation of GSLs after feeding the Brassica diets for four weeks. In agreement with their high glucoraphanin content, broccoli diets induced the formation of sulforaphane-lysine adducts. Levels of 1-methoxyindolyl-3-methyl-histidine adducts derived from neoglucobrassicin were the highest in the GSL-rich pak choi group. In the colon, the GSL-rich broccoli and the GSL-rich pak choi diet up-regulated the expression of different sets of typical Nrf2 target genes like Nqo1, Gstm1, Srxn1, and GPx2. GSL-rich pak choi induced the AhR target gene Cyp1a1 but did not affect Ugt1a1 expression. Both colitis and tumor number were drastically reduced after feeding the GSL-rich pak choi diet while the other three diets had no effect. GSLs can act anti-inflammatory and anti-carcinogenic but both effects depend on the specific amount and pattern of GSLs within a vegetable. Thus, a high Brassica consumption cannot be generally considered to be cancer-preventive.

  6. Fission yeast pak1+ encodes a protein kinase that interacts with Cdc42p and is involved in the control of cell polarity and mating.

    PubMed Central

    Ottilie, S; Miller, P J; Johnson, D I; Creasy, C L; Sells, M A; Bagrodia, S; Forsburg, S L; Chernoff, J

    1995-01-01

    A STE20/p65pak homolog was isolated from fission yeast by PCR. The pak1+ gene encodes a 72 kDa protein containing a putative p21-binding domain near its amino-terminus and a serine/threonine kinase domain near its carboxyl-terminus. The Pak1 protein autophosphorylates on serine residues and preferentially binds to activated Cdc42p both in vitro and in vivo. This binding is mediated through the p21 binding domain on Pak1p and the effector domain on Cdc42p. Overexpression of an inactive mutant form of pak1 gives rise to cells with markedly abnormal shape with mislocalized actin staining. Pak1 overexpression does not, however, suppress lethality associated with cdc42-null cells or the morphologic defeat caused by overexpression of mutant cdc42 alleles. Gene disruption of pak1+ establishes that, like cdc42+, pak1+ function is required for cell viability. In budding yeast, pak1+ expression restores mating function to STE20-null cells and, in fission yeast, overexpression of an inactive form of Pak inhibits mating. These results indicate that the Pak1 protein is likely to be an effector for Cdc42p or a related GTPase, and suggest that Pak1p is involved in the maintenance of cell polarity and in mating. Images PMID:8846783

  7. Cadherins and Pak1 control contact inhibition of proliferation by Pak1-betaPIX-GIT complex-dependent regulation of cell-matrix signaling.

    PubMed

    Liu, Fengming; Jia, Liwei; Thompson-Baine, Ann-Marie; Puglise, Jason M; Ter Beest, Martin B A; Zegers, Mirjam M P

    2010-04-01

    It is crucial for organ homeostasis that epithelia have effective mechanisms to restrict motility and cell proliferation in order to maintain tissue architecture. On the other hand, epithelial cells need to rapidly and transiently acquire a more mesenchymal phenotype, with high levels of cell motility and proliferation, in order to repair epithelia upon injury. Cross talk between cell-cell and cell-matrix signaling is crucial for regulating these transitions. The Pak1-betaPIX-GIT complex is an effector complex downstream of the small GTPase Rac1. We previously showed that translocation of this complex from cell-matrix to cell-cell adhesion sites was required for the establishment of contact inhibition of proliferation. In this study, we provide evidence that this translocation depends on cadherin function. Cadherins do not recruit the complex by direct interaction. Rather, we found that inhibition of the normal function of cadherin or Pak1 leads to defects in focal adhesion turnover and to increased signaling by phosphatidylinositol 3-kinase. We propose that cadherins are involved in regulation of contact inhibition by controlling the function of the Pak1-betaPIX-GIT complex at focal contacts.

  8. Pak1 regulates the orientation of apical polarization and lumen formation by distinct pathways.

    PubMed

    deLeon, Orlando; Puglise, Jason M; Liu, Fengming; Smits, Jos; ter Beest, Martin B; Zegers, Mirjam M

    2012-01-01

    The development of the basic architecture of branching tubules enclosing a central lumen that characterizes most epithelial organs crucially depends on the apico-basolateral polarization of epithelial cells. Signals from the extracellular matrix control the orientation of the apical surface, so that it faces the lumen interior, opposite to cell-matrix adhesion sites. This orientation of the apical surface is thought to be intrinsically linked to the formation of single lumens. We previously demonstrated in three-dimensional cyst cultures of Madin-Darby canine kidney (MDCK) cells that signaling by β1 integrins regulates the orientation of the apical surface, via a mechanism that depends on the activity of the small GTPase Rac1. Here, we investigated whether the Rac1 effector Pak1 is a downstream effector in this pathway. Expression of constitutive active Pak1 phenocopies the effect of β1 integrin inhibition in that it misorients the apical surface and induces a multilumen phenotype. The misorientation of apical surfaces depends on the interaction of active Pak1 with PIX proteins and is linked to defects in basement membrane assembly. In contrast, the multilumen phenotype was independent of PIX and the basement membrane. Therefore, Pak1 likely regulates apical polarization and lumen formation by two distinct pathways.

  9. Pak1 Regulates the Orientation of Apical Polarization and Lumen Formation by Distinct Pathways

    PubMed Central

    Smits, Jos; ter Beest, Martin B.; Zegers, Mirjam M.

    2012-01-01

    The development of the basic architecture of branching tubules enclosing a central lumen that characterizes most epithelial organs crucially depends on the apico-basolateral polarization of epithelial cells. Signals from the extracellular matrix control the orientation of the apical surface, so that it faces the lumen interior, opposite to cell-matrix adhesion sites. This orientation of the apical surface is thought to be intrinsically linked to the formation of single lumens. We previously demonstrated in three-dimensional cyst cultures of Madin-Darby canine kidney (MDCK) cells that signaling by β1 integrins regulates the orientation of the apical surface, via a mechanism that depends on the activity of the small GTPase Rac1. Here, we investigated whether the Rac1 effector Pak1 is a downstream effector in this pathway. Expression of constitutive active Pak1 phenocopies the effect of β1 integrin inhibition in that it misorients the apical surface and induces a multilumen phenotype. The misorientation of apical surfaces depends on the interaction of active Pak1 with PIX proteins and is linked to defects in basement membrane assembly. In contrast, the multilumen phenotype was independent of PIX and the basement membrane. Therefore, Pak1 likely regulates apical polarization and lumen formation by two distinct pathways. PMID:22815903

  10. The p21-activated kinase PAK is negatively regulated by POPX1 and POPX2, a pair of serine/threonine phosphatases of the PP2C family.

    PubMed

    Koh, Cheng-Gee; Tan, E-Jean; Manser, Edward; Lim, Louis

    2002-02-19

    The Rho GTPases are involved in many signaling pathways and cellular functions, including the organization of the actin cytoskeleton, regulation of transcription, cell motility, and cell division. The p21 (Cdc42/Rac)-activated kinase PAK mediates a number of biological effects downstream of these Rho GTPases (reviewed by [1]). The phosphorylation state of mammalian PAK is highly regulated: upon binding of GTPases, PAK is potently activated by autophosphorylation at multiple sites, although the mechanisms of PAK downregulation are not known. We now report two PP2C-like serine/threonine phosphatases (POPX1 and POPX2) that efficiently inactivate PAK. POPX1 was isolated as a binding partner for the PAK interacting guanine nucleotide exchange factor PIX. The dephosphorylating activity of POPX correlates with an ability to block the in vivo effects of active PAK. Consonant with these effects on PAK, POPX can also inhibit actin stress fiber breakdown and morphological changes driven by active Cdc42(V12). The association of the POPX phosphatases with PAK complexes may allow PAK to cycle rapidly between active and inactive states; it represents a unique regulatory component of the signaling pathways of the PAK kinase family.

  11. PAK6 increase chemoresistance and is a prognostic marker for stage II and III colon cancer patients undergoing 5-FU based chemotherapy

    PubMed Central

    Yan, Dongwang; Cui, Feifei; Wang, Xiaoliang; Yu, Fudong; Xue, Yingming; Feng, Xiaodong; Wang, Jingtao; Wang, Xiao; Jiang, Tao; Zhang, Meng; Zhao, Senlin; Yu, Yang; Tang, Huamei; Peng, Zhihai

    2015-01-01

    p21-Activated kinase 6 (PAK6) has been implicated in radiotherapy and docetaxel resistance. We have further evaluated PAK6 as a predictor of 5-fluorouracil (5-FU) treatment response in colon cancer. Here we report that in colon cancer PAK6 promotes tumor progression and chemoresistance both in vitro and in vivo. In the clinical analysis, PAK6 was overexpressed in 104 of 147 (70.75%) stage II and III patients who received 5-FU based chemotherapy after surgery. Multivariate Cox regression analysis indicated that PAK6 was an independent prognostic factor for overall survival (P < 0.001) and disease-free survival (P < 0.001). Colon cancer cell lines showed increased PAK6 expression upon 5-FU treatment. In PAK6-knockdown cells treated with 5-FU, cell viability and phosphorylation of BAD decreased, and the number of apoptotic cells, levels of cleaved caspase 3 and PARP increased compared to control cells. The opposite was observed in PAK6 overexpressing cells. Short hairpin RNA knockdown of PAK6 blocked cells in G2-M phase. Furthermore, Animal experiments results in vivo are consistent with outcomes in vitro. This study demonstrates that PAK6 is an independent prognostic factor for adjuvant 5-FU-based chemotherapy in patients with stage II and stage III colon cancer. PMID:25426562

  12. A PAK1-PIX-PKL complex is activated by the T-cell receptor independent of Nck, Slp-76 and LAT.

    PubMed

    Ku, G M; Yablonski, D; Manser, E; Lim, L; Weiss, A

    2001-02-01

    Given the importance of the Rho GTPase family member Rac1 and the Rac1/Cdc42 effector PAK1 in T-cell activation, we investigated the requirements for their activation by the T-cell receptor (TCR). Rac1 and PAK1 activation required the tyrosine kinases ZAP-70 and Syk, but not the cytoplasmic adaptor Slp-76. Surprisingly, PAK1 was activated in the absence of the transmembrane adaptor LAT while Rac1 was not. However, efficient PAK1 activation required its binding sites for Rho GTPases and for PIX, a guanine nucleotide exchange factor for Rho GTPases. The overexpression of ssPIX that either cannot bind PAK1 or lacks GEF function blocked PAK1 activation. These data suggest that a PAK1-PIX complex is recruited to appropriate sites for activation and that PIX is required for Rho family GTPase activation upstream of PAK1. Furthermore, we detected a stable trimolecular complex of PAK1, PIX and the paxillin kinase linker p95PKL. Taken together, these data show that PAK1 contained in this trimolecular complex is activated by a novel LAT- and Slp-76-independent pathway following TCR stimulation.

  13. miR-129 suppresses tumor cell growth and invasion by targeting PAK5 in hepatocellular carcinoma

    SciTech Connect

    Zhai, Jian; Qu, Shuping; Li, Xiaowei; Zhong, Jiaming; Chen, Xiaoxia; Qu, Zengqiang; Wu, Dong

    2015-08-14

    Emerging evidence suggests that microRNAs (miRNAs) play important roles in regulating HCC development and progression; however, the mechanisms by which their specific functions and mechanisms remained to be further explored. miR-129 has been reported in gastric cancers, lung cancer and colon cancer. In this study, we disclosed a new tumor suppresser function of miR-129 in HCC. We also found the downregulation of miR-129 occurred in nearly 3/4 of the tumors examined (56/76) compared with adjacent nontumorous tissues, which was more importantly, correlated to the advanced stage and vascular invasion. We then demonstrated that miR-129 overexpression attenuated HCC cells proliferation and invasion, inducing apoptosis in vitro. Moreover, we used miR-129 antagonist and found that anti-miR-129 promoted HCC cells malignant phenotypes. Mechanistically, our further investigations revealed that miR-129 suppressed cell proliferation and invasion by targeting the 3’-untranslated region of PAK5, as well as miR-129 silencing up-regulated PAK5 expression. Moreover, miR-129 expression was inversely correlated with PAK5 expression in 76 cases of HCC samples. RNA interference of PAK5 attenuated anti-miR-129 mediated cell proliferation and invasion in HCC cells. Taken together, these results demonstrated that miR-129 suppressed tumorigenesis and progression by directly targeting PAK5, defining miR-129 as a potential treatment target for HCC. - Highlights: • Decreased of miR-129 is found in HCC and associated with advanced stage and metastasis. • miR-129 suppresses proliferation and invasion of HCC cells. • miR-129 directly targets the 3′ UTR of PAK5 and diminishes PAK5 expression. • PAK5 is involved in miR-129 mediated suppression functions.

  14. Effect of Okinawa Propolis on PAK1 Activity, Caenorhabditis elegans Longevity, Melanogenesis, and Growth of Cancer Cells.

    PubMed

    Taira, Nozomi; Nguyen, Binh Cao Quan; Be Tu, Pham Thi; Tawata, Shinkichi

    2016-07-13

    Propolis from different areas has been reported to inhibit oncogenic/aging kinase PAK1, which is responsible for a variety of conditions, including cancer, longevity, and melanogenesis. Here, a crude extract of Okinawa propolis (OP) was tested against PAK1 activity, Caenorhabditis elegans (C. elegans) longevity, melanogenesis, and growth of cancer cells. We found that OP blocks PAK1 and exhibits anticancer activity in the A549 cell (human lung cancer cell) line with IC50 values of 6 μg/mL and 12 μg/mL, respectively. Most interestingly, OP (1 μg/mL) significantly reduces reproduction and prolongs the lifespan of C. elegans by activating the HSP-16.2 gene, as shown in the PAK1-deficient strain. Furthermore, OP inhibits melanogenesis in a melanoma cell line (B16F10) by downregulating intracellular tyrosinase activity with an IC50 of 30 μg/mL. Our results suggest that OP demonstrated a life span extending effect, C. elegans, anticancer, and antimelanogenic effects via PAK1 inactivation; therefore, this can be a potent natural medicinal supplement against PAK1-dependent diseases.

  15. Bistability in the Rac1, PAK, and RhoA Signaling Network Drives Actin Cytoskeleton Dynamics and Cell Motility Switches

    PubMed Central

    Byrne, Kate M.; Monsefi, Naser; Dawson, John C.; Degasperi, Andrea; Bukowski-Wills, Jimi-Carlo; Volinsky, Natalia; Dobrzyński, Maciej; Birtwistle, Marc R.; Tsyganov, Mikhail A.; Kiyatkin, Anatoly; Kida, Katarzyna; Finch, Andrew J.; Carragher, Neil O.; Kolch, Walter; Nguyen, Lan K.; von Kriegsheim, Alex; Kholodenko, Boris N.

    2016-01-01

    Summary Dynamic interactions between RhoA and Rac1, members of the Rho small GTPase family, play a vital role in the control of cell migration. Using predictive mathematical modeling, mass spectrometry-based quantitation of network components, and experimental validation in MDA-MB-231 mesenchymal breast cancer cells, we show that a network containing Rac1, RhoA, and PAK family kinases can produce bistable, switch-like responses to a graded PAK inhibition. Using a small chemical inhibitor of PAK, we demonstrate that cellular RhoA and Rac1 activation levels respond in a history-dependent, bistable manner to PAK inhibition. Consequently, we show that downstream signaling, actin dynamics, and cell migration also behave in a bistable fashion, displaying switches and hysteresis in response to PAK inhibition. Our results demonstrate that PAK is a critical component in the Rac1-RhoA inhibitory crosstalk that governs bistable GTPase activity, cell morphology, and cell migration switches. PMID:27136688

  16. A novel orally bioavailable compound KPT-9274 inhibits PAK4, and blocks triple negative breast cancer tumor growth

    PubMed Central

    Rane, Chetan; Senapedis, William; Baloglu, Erkan; Landesman, Yosef; Crochiere, Marsha; Das-Gupta, Soumyasri; Minden, Audrey

    2017-01-01

    Breast cancer is a heterogeneous disease consisting of several subtypes. Among these subtypes, triple negative breast cancer is particularly difficult to treat. This is due to a lack of understanding of the mechanisms behind the disease, and consequently a lack of druggable targets. PAK4 plays critical roles in cell survival, proliferation, and morphology. PAK4 protein levels are high in breast cancer cells and breast tumors, and the gene is often amplified in basal like breast cancers, which are frequently triple negative. PAK4 is also overexpressed in other types of cancer, making it a promising drug target. However, its inhibition is complicated by the fact that PAK4 has both kinase-dependent and -independent functions. Here we investigate a new clinical compound KPT-9274, which has been shown to inhibit PAK4 and NAMPT. We find that KPT-9274 (and its analog, KPT-8752) can reduce the steady state level of PAK4 protein in triple negative breast cancer cells. These compounds also block the growth of the breast cancer cells in vitro, and stimulate apoptosis. Most importantly, oral administration of KPT-9274 reduces tumorigenesis in mouse models of human triple negative breast cancer. Our results indicate that KPT-9274 is a novel therapeutic option for triple negative breast cancer therapy. PMID:28198380

  17. Dbo/Henji Modulates Synaptic dPAK to Gate Glutamate Receptor Abundance and Postsynaptic Response

    PubMed Central

    Wang, Manyu; Chen, Pei-Yi; Wang, Chien-Hsiang; Lai, Tzu-Ting; Tsai, Pei-I; Cheng, Ying-Ju; Kao, Hsiu-Hua; Chien, Cheng-Ting

    2016-01-01

    In response to environmental and physiological changes, the synapse manifests plasticity while simultaneously maintains homeostasis. Here, we analyzed mutant synapses of henji, also known as dbo, at the Drosophila neuromuscular junction (NMJ). In henji mutants, NMJ growth is defective with appearance of satellite boutons. Transmission electron microscopy analysis indicates that the synaptic membrane region is expanded. The postsynaptic density (PSD) houses glutamate receptors GluRIIA and GluRIIB, which have distinct transmission properties. In henji mutants, GluRIIA abundance is upregulated but that of GluRIIB is not. Electrophysiological results also support a GluR compositional shift towards a higher IIA/IIB ratio at henji NMJs. Strikingly, dPAK, a positive regulator for GluRIIA synaptic localization, accumulates at the henji PSD. Reducing the dpak gene dosage suppresses satellite boutons and GluRIIA accumulation at henji NMJs. In addition, dPAK associated with Henji through the Kelch repeats which is the domain essential for Henji localization and function at postsynapses. We propose that Henji acts at postsynapses to restrict both presynaptic bouton growth and postsynaptic GluRIIA abundance by modulating dPAK. PMID:27736876

  18. Tyrosyl Phosphorylated Serine-Threonine Kinase PAK1 is a Novel Regulator of Prolactin-Dependent Breast Cancer Cell Motility and Invasion

    PubMed Central

    Hammer, Alan

    2015-01-01

    Despite efforts to discover the cellular pathways regulating breast cancer metastasis, little is known as to how prolactin (PRL) cooperates with extracellular environment and cytoskeletal proteins to regulate breast cancer cell motility and invasion. We implicated serine-threonine kinase p21-activated kinase 1 (PAK1) as a novel target for PRL-activated Janus-kinase 2 (JAK2). JAK2-dependent PAK1 tyrosyl phosphorylation plays a critical role in regulation of both PAK1 kinase activity and scaffolding properties of PAK1. Tyrosyl phosphorylated PAK1 facilitates PRL-dependent motility via at least two mechanisms: formation of paxillin/GIT1/βPIX/pTyr-PAK1 complexes resulting in increased adhesion turnover and phosphorylation of actin-binding protein filamin A. Increased adhesion turnover is the basis for cell migration and phosphorylated filamin A stimulates the kinase activity of PAK1 and increases actin-regulating activity to facilitate cell motility. Tyrosyl phosphorylated PAK1 also stimulates invasion of breast cancer cells in response to PRL and three-dimensional (3D) collagen IV via transcription and secretion of MMP-1 and MMP-3 in a MAPK-dependent manner. These data illustrate the complex interaction between PRL and the cell microenvironment in breast cancer cells and suggest a pivotal role for PRL/PAK1 signaling in breast cancer metastasis. PMID:25472536

  19. PAK5 mediates cell: cell adhesion integrity via interaction with E-cadherin in bladder cancer cells.

    PubMed

    Ismail, Ahmad Fahim; Oskay Halacli, Sevil; Babteen, Nouf; De Piano, Mario; Martin, Tracey A; Jiang, Wen G; Khan, Muhammad Shamim; Dasgupta, Prokar; Wells, Claire M

    2017-03-24

    Urothelial bladder cancer is a major cause of morbidity and mortality worldwide, causing an estimated 150 000 deaths per year. Whilst non-muscle-invasive bladder tumours can be effectively treated, with high survival rates, many tumours recur, and some will progress to muscle-invasive disease with a much poorer long-term prognosis. Thus, there is a pressing need to understand the molecular transitions occurring within the progression of bladder cancer to an invasive disease. Tumour invasion is often associated with a down-regulation of E-cadherin expression concomitant with a suppression of cell:cell junctions, and decreased levels of E-cadherin expression have been reported in higher grade urothelial bladder tumours. We find that expression of E-cadherin in a panel of bladder cancer cell lines correlated with the presence of cell:cell junctions and the level of PAK5 expression. Interestingly, exogenous PAK5 has recently been described to be associated with cell:cell junctions and we now find that endogenous PAK5 is localised to cell junctions and interacts with an E-cadherin complex. Moreover, depletion of PAK5 expression significantly reduced junctional integrity. These data suggest a role for PAK5 in maintaining junctional stability and we find that, in both our own patient samples and a commercially available dataset, PAK5mRNA levels are reduced in human bladder cancer compared with normal controls. Taken together, the present study proposes that PAK5 expression levels could be used as a novel prognostic marker for bladder cancer progression.

  20. Smad3 Couples Pak1 With the Antihypertrophic Pathway Through the E3 Ubiquitin Ligase, Fbxo32.

    PubMed

    Tsui, Hoyee; Zi, Min; Wang, Shunyao; Chowdhury, Sanjoy K; Prehar, Sukhpal; Liang, Qiangrong; Cartwright, Elizabeth J; Lei, Ming; Liu, Wei; Wang, Xin

    2015-12-01

    Pathological cardiac hypertrophy is regarded as a critical intermediate step toward the development of heart failure. Many signal transduction cascades are demonstrated to dictate the induction and progression of pathological hypertrophy; however, our understanding in regulatory mechanisms responsible for the suppression of hypertrophy remains limited. In this study, we showed that exacerbated hypertrophy induced by pressure overload in cardiac-deleted Pak1 mice was attributable to a failure to upregulate the antihypertrophic E3 ligase, Fbxo32, responsible for targeting proteins for the ubiquitin-degradation pathway. Under pressure overload, cardiac overexpression of constitutively active Pak1 mice manifested strong resilience against pathological hypertrophic remodeling. Mechanistic studies demonstrated that subsequent to Pak1 activation, the binding of Smad3 on a critical singular AGAC(-286)-binding site on the FBXO32 promoter was crucial for its transcriptional regulation. Pharmacological upregulation of Fbxo32 by Berberine ameliorated hypertrophic remodeling and improved cardiac performance in cardiac-deficient Pak1 mice under pressure overload. Our findings discover Smad3 and Fbxo32 as novel downstream components of the Pak1-dependent signaling pathway for the suppression of hypertrophy. This discovery opens a new venue for opportunities to identify novel targets for the management of cardiac hypertrophy.

  1. PakB binds to the SH3 domain of Dictyostelium Abp1 and regulates its effects on cell polarity and early development.

    PubMed

    Yang, Yidai; de la Roche, Marc; Crawley, Scott W; Li, Zhihao; Furmaniak-Kazmierczak, Emilia; Côté, Graham P

    2013-07-01

    Dictyostelium p21-activated kinase B (PakB) phosphorylates and activates class I myosins. PakB colocalizes with myosin I to actin-rich regions of the cell, including macropinocytic and phagocytic cups and the leading edge of migrating cells. Here we show that residues 1-180 mediate the cellular localization of PakB. Yeast two-hybrid and pull-down experiments identify two proline-rich motifs in PakB-1-180 that directly interact with the SH3 domain of Dictyostelium actin-binding protein 1 (dAbp1). dAbp1 colocalizes with PakB to actin-rich regions in the cell. The loss of dAbp1 does not affect the cellular distribution of PakB, whereas the loss of PakB causes dAbp1 to adopt a diffuse cytosolic distribution. Cosedimentation studies show that the N-terminal region of PakB (residues 1-70) binds directly to actin filaments, whereas dAbp1 exhibits only a low affinity for filamentous actin. PakB-1-180 significantly enhances the binding of dAbp1 to actin filaments. When overexpressed in PakB-null cells, dAbp1 completely blocks early development at the aggregation stage, prevents cell polarization, and significantly reduces chemotaxis rates. The inhibitory effects are abrogated by the introduction of a function-blocking mutation into the dAbp1 SH3 domain. We conclude that PakB plays a critical role in regulating the cellular functions of dAbp1, which are mediated largely by its SH3 domain.

  2. Involvement of alpha-PAK-interacting exchange factor in the PAK1-c-Jun NH(2)-terminal kinase 1 activation and apoptosis induced by benzo[a]pyrene.

    PubMed

    Yoshii, S; Tanaka, M; Otsuki, Y; Fujiyama, T; Kataoka, H; Arai, H; Hanai, H; Sugimura, H

    2001-10-01

    Benzo[a]pyrene [B(a)P], a potent procarcinogen found in combustion products such as diesel exhaust and cigarette smoke, has been recently shown to activate the c-Jun NH(2)-terminal kinase 1 (JNK1) and induce caspase-3-mediated apoptosis in Hepa1c1c7 cells. However, the molecules of the signaling pathway that control the mitogen-activated protein kinase cascades induced by B(a)P and the interaction between those and apoptosis by B(a)P have not been well defined. We report here that B(a)P promoted Cdc42/Rac1, p21-activated kinase 1 (PAK1), and JNK1 activities in 293T and HeLa cells. Moreover, alpha-PAK-interacting exchange factor (alpha PIX) mRNA and its protein expression were upregulated by B(a)P. While overexpression of an active mutant of alpha PIX (DeltaCH) facilitated B(a)P-induced activation of Cdc42/Rac1, PAK1, and JNK1, overexpression of mutated alphaPIX (L383R, L384S), which lacks guanine nucleotide exchange factor activity, SH3 domain-deleted alphaPIX (Delta SH3), which lacks the ability to bind PAK, kinase-negative PAK1 (K299R), and kinase-negative SEK1 (K220A, K224L) inhibited B(a)P-triggered JNK1 activation. Interestingly, overexpression of alphaPIX (Delta CH) and a catalytically active mutant PAK1 (T423E) accelerated B(a)P-induced apoptosis in HeLa cells, whereas alphaPIX (Delta SH3), PAK1 (K299R), and SEK 1 (K220A, K224L) inhibited B(a)P-initiated apoptosis. Finally, a preferential caspase inhibitor, Z-Asp-CH2-DCB, strongly blocked the alphaPIX (Delta CH)-enhanced apoptosis in cells treated with B(a)P but did not block PAK1/JNK1 activation. Taken together, these results indicate that alphaPIX plays a crucial role in B(a)P-induced apoptosis through activation of the JNK1 pathway kinases.

  3. Cadmium accumulation in pak choi (Brassica chinensis L.) and estimated dietary intake in the suburb of Hangzhou city, China.

    PubMed

    Yan, S; Ling, Q; Bao, Z; Chen, Z; Yan, S; Dong, Z; Zhang, B; Deng, B

    2009-01-01

    The total content of cadmium and its distribution between leaves and roots were investigated for pak choi (Brassica chinensis L.), which was grown in a contaminated area of Hangzhou city, China. Results showed that cadmium concentrations in 90% of samples exceeded the Chinese tolerance limit for food. Although roots of pak choi had a higher concentration of cadmium than leaves, leaf tissue (the edible part) accumulated over 80% (on average) of the whole plant cadmium burden due to the greater biomass in leaf tissue. Since pak choi is the staple vegetable in the study area and of great relevance to human health, the dietary intake of cadmium and calculated risk indexes to local residents through the food chain were determined.

  4. MiR-145 regulates PAK4 via the MAPK pathway and exhibits an antitumor effect in human colon cells

    SciTech Connect

    Wang, Zhigang; Zhang, Xiaoping; Yang, Zhili; Du, Hangxiang; Wu, Zhenqian; Gong, Jianfeng; Yan, Jun; Zheng, Qi

    2012-10-26

    Highlights: Black-Right-Pointing-Pointer MiR-145 targets a putative binding site in the 3 Prime UTR of PAK4. Black-Right-Pointing-Pointer MiR-145 played an important role in inhibiting cell growth by directly targeting PAK4. Black-Right-Pointing-Pointer MiR-145 may function as tumor suppressors. -- Abstract: MicroRNAs (miRNAs) are regulators of numerous cellular events; accumulating evidence indicates that miRNAs play a key role in a wide range of biological functions, such as cellular proliferation, differentiation, and apoptosis in cancer. Down-regulated expression of miR-145 has been reported in colon cancer tissues and cell lines. The molecular mechanisms underlying miR-145 and the regulation of colon carcinogenesis remain unclear. In this study, we investigated the levels of miR-145 in human colon cancer cells using qRT-PCR and found markedly decreased levels compared to normal epithelial cells. We identified PAK4 as a novel target of miR-145 using informatics screening. Additionally, we demonstrated that miR-145 targets a putative binding site in the 3 Prime UTR of PAK4 and that its abundance is inversely associated with miR-145 expression in colon cancer cells; we confirmed this relationship using the luciferase reporter assay. Furthermore, restoration of miR-145 by mimics in SW620 cells significantly attenuated cell growth in vitro, in accordance with the inhibitory effects induced by siRNA mediated knockdown of PAK4. Taken together, these findings demonstrate that miR-145 downregulates P-ERK expression by targeting PAK4 and leads to inhibition of tumor growth.

  5. Novel p21-Activated Kinase 4 (PAK4) Allosteric Modulators Overcome Drug Resistance and Stemness in Pancreatic Ductal Adenocarcinoma.

    PubMed

    Aboukameel, Amro; Muqbil, Irfana; Senapedis, William; Baloglu, Erkan; Landesman, Yosef; Shacham, Sharon; Kauffman, Michael; Philip, Philip A; Mohammad, Ramzi M; Azmi, Asfar S

    2017-01-01

    The p21-activated kinase 4 (PAK4) is a key downstream effector of the Rho family GTPases and is found to be overexpressed in pancreatic ductal adenocarcinoma (PDAC) cells but not in normal human pancreatic ductal epithelia (HPDE). Gene copy number amplification studies in PDAC patient cohorts confirmed PAK4 amplification making it an attractive therapeutic target in PDAC. We investigated the antitumor activity of novel PAK4 allosteric modulators (PAM) on a panel of PDAC cell lines and chemotherapy-resistant flow-sorted PDAC cancer stem cells (CSC). The toxicity and efficacy of PAMs were evaluated in multiple subcutaneous mouse models of PDAC. PAMs (KPT-7523, KPT-7189, KPT-8752, KPT-9307, and KPT-9274) show antiproliferative activity in vitro against different PDAC cell lines while sparing normal HPDE. Cell growth inhibition was concurrent with apoptosis induction and suppression of colony formation in PDAC. PAMs inhibited proliferation and antiapoptotic signals downstream of PAK4. Co-immunoprecipitation experiments showed disruption of PAK4 complexes containing vimentin. PAMs disrupted CSC spheroid formation through suppression of PAK4. Moreover, PAMs synergize with gemcitabine and oxaliplatin in vitro KPT-9274, currently in a phase I clinical trial (clinicaltrials.gov; NCT02702492), possesses desirable pharmacokinetic properties and is well tolerated in mice with the absence of any signs of toxicity when 200 mg/kg daily is administered either intravenously or orally. KPT-9274 as a single agent showed remarkable antitumor activity in subcutaneous xenograft models of PDAC cell lines and CSCs. These proof-of-concept studies demonstrated the antiproliferative effects of novel PAMs in PDAC and warrant further clinical investigations. Mol Cancer Ther; 16(1); 76-87. ©2016 AACR.

  6. [Rapid isolation of 4-hydroxyphenylethanol from the bovine brain using Water Sep-Pak Plus cartridges with lipophilic sorbent (Florisil)].

    PubMed

    Panova, N G; Veselovskaia, N V; Medvedev, A E

    1996-01-01

    4-Hydroxypherlylethanol, MAO-A Inhibitory component of endogenous MAO Inhibitory activity, tribulin, was isolated from a small quantity of bovine brain using Maters Sep-Pak Florisil cartridges. Procedure included isolation of crude tribulin fraction, which was passed through the cartridge. Preliminary data revealed that this substance is not absorbed on Florisil. The employment of this type of cartridges essentially accelerates isolation of 4-hydroxyphenyethanol from brain samples for quantitative determination. Pilot experiments also revealed that Sep-Pak Folrisil cartridges could be used for the rapid isolation of isatin, another endogenous MAO (B) inhibitor, from brain tissue.

  7. PAK proteins and YAP-1 signalling downstream of integrin beta-1 in myofibroblasts promote liver fibrosis

    PubMed Central

    Martin, Katherine; Pritchett, James; Llewellyn, Jessica; Mullan, Aoibheann F.; Athwal, Varinder S.; Dobie, Ross; Harvey, Emma; Zeef, Leo; Farrow, Stuart; Streuli, Charles; Henderson, Neil C.; Friedman, Scott L.; Hanley, Neil A.; Piper Hanley, Karen

    2016-01-01

    Fibrosis due to extracellular matrix (ECM) secretion from myofibroblasts complicates many chronic liver diseases causing scarring and organ failure. Integrin-dependent interaction with scar ECM promotes pro-fibrotic features. However, the pathological intracellular mechanism in liver myofibroblasts is not completely understood, and further insight could enable therapeutic efforts to reverse fibrosis. Here, we show that integrin beta-1, capable of binding integrin alpha-11, regulates the pro-fibrotic phenotype of myofibroblasts. Integrin beta-1 expression is upregulated in pro-fibrotic myofibroblasts in vivo and is required in vitro for production of fibrotic ECM components, myofibroblast proliferation, migration and contraction. Serine/threonine-protein kinase proteins, also known as P21-activated kinase (PAK), and the mechanosensitive factor, Yes-associated protein 1 (YAP-1) are core mediators of pro-fibrotic integrin beta-1 signalling, with YAP-1 capable of perpetuating integrin beta-1 expression. Pharmacological inhibition of either pathway in vivo attenuates liver fibrosis. PAK protein inhibition, in particular, markedly inactivates the pro-fibrotic myofibroblast phenotype, limits scarring from different hepatic insults and represents a new tractable therapeutic target for treating liver fibrosis. PMID:27535340

  8. IPA-3 Inhibits the Growth of Liver Cancer Cells By Suppressing PAK1 and NF-κB Activation

    PubMed Central

    Wong, Leo Lap-Yan; Lam, Ian Pak-Yan; Wong, Tracy Yuk-Nar; Lai, Wai-Lung; Liu, Heong-Fai; Yeung, Lam-Lung; Ching, Yick-Pang

    2013-01-01

    Hepatocellular carcinoma (HCC) is one of the major malignancies worldwide and is associated with poor prognosis due to the high incidences of metastasis and tumor recurrence. Our previous study showed that overexpression of p21-activated protein kinase 1 (PAK1) is frequently observed in HCC and is associated with a more aggressive tumor behavior, suggesting that PAK1 is a potential therapeutic target in HCC. In the current study, an allosteric small molecule PAK1 inhibitor, IPA-3, was evaluated for the potential in suppressing hepatocarcinogenesis. Consistent with other reports, inhibition of PAK1 activity was observed in several human HCC cell lines treated with various dosages of IPA-3. Using cell proliferation, colony formation and BrdU incorporation assays, we demonstrated that IPA-3 treatment significantly inhibited the growth of HCC cells. The mechanisms through which IPA-3 treatment suppresses HCC cell growth are enhancement of apoptosis and blockage of activation of NF-κB. Furthermore, our data suggested that IPA-3 not only inhibits the HCC cell growth, but also suppresses the metastatic potential of HCC cells. Nude mouse xenograft assay demonstrated that IPA-3 treatment significantly reduced the tumor growth rate and decreased tumor volume, indicating that IPA-3 can suppress the in vivo tumor growth of HCC cells. Taken together, our demonstration of the potential preclinical efficacy of IPA-3 in HCC provides the rationale for cancer therapy. PMID:23894351

  9. Downregulation of microRNA-23a suppresses prostate cancer metastasis by targeting the PAK6-LIMK1 signaling pathway

    PubMed Central

    Li, Xiaojuan; Cai, Yi; Ye, Zhiqiang; Li, Shigeng; Li, Jun; Huang, Huaiqiu; Peng, Shubin; Wang, Jun; Tao, Yiran; Huang, Hongxing; Wen, Xinglai; Mo, Jianfeng; Deng, Zhupeng; Wang, Jian; Zhang, Yangfan; Gao, Xin; Wen, Xingqiao

    2015-01-01

    Here we found that levels of miR-23a were decreased in prostate cancer cell lines and tumor tissues. These low levels were associated with poor patients' prognosis. MiR-23a inhibited migration and invasion of prostate cancer in vivo and in orthotopic prostate cancer mice model. MiR-23a decreased levels of p21-activated kinase 6 (PAK6). Expression of miR-23a inhibited phosphorylation of LIM kinase 1 (LIMK1) and cofilin, in turn suppressing formation of stress fibers and actin filaments, which was required for cell motility and invasion. PAK6 bound to LIMK1 and activated it via phosphorylation at Thr-508. Also, PAK6 and LIMK1 were colocalized in the cytoplasma. Thus, miR-23a regulated cytoskeleton by affecting LIMK1 and cofilin. In summary, we have identified the miR-23a-PAK6-LIMK1 pathway of prostate cancer metastasis. Potential therapeutic approach by targeting miR-23 is suggested. PMID:25714010

  10. Abnormalities of the Duo/Rac-1/PAK1 Pathway Drive Myosin Light Chain Phosphorylation in Frontal Cortex in Schizophrenia

    PubMed Central

    Rubio, María D.; Haroutunian, Vahram; Meador-Woodruff, James H.

    2012-01-01

    BACKGROUND Recent studies on GTPases have suggested that reduced Duo and Cdc42 transcript expression is involved in dendritic spine loss in schizophrenia. In murine models, Duo and Cdc42 phosphorylate PAK1, which modifies the activity of regulatory myosin light chain (MLC) and cofilin by altering their phosphorylation. Therefore, we hypothesized that in schizophrenia abnormal Duo and Cdc42 expression result in changes in MLC and/or cofilin phosphorylation, which may alter actin cytoskeleton dynamics underlying dendritic spine maintenance. METHODS We performed Western blot protein expression analysis in postmortem brains from patients diagnosed with schizophrenia and a comparison group. We focused our studies in the anterior cingulate cortex (ACC) (n=33 comparison group; n=36 schizophrenia) and dorsolateral prefrontal cortex (DLPFC) (n=29 comparison group; n=35 schizophrenia). RESULTS In both ACC and DLPFC, we found a reduction of Duo expression and PAK1 phosphorylation in schizophrenia. Cdc42 protein expression was decreased in ACC, but not in DLPFC. In ACC, we observed decreased PAK1 phosphorylation and increased MLC (pMLC) phosphorylation, while in DLPFC pMLC remained unchanged. DISCUSSION These data suggest a novel mechanism that may underlie dendritic spine loss in schizophrenia. The increase in pMLC seen in ACC may be associated with dendritic spine shrinkage. The lack of an effect on pMLC in DLPFC suggests that in schizophrenia PAK1 downstream pathways are differentially affected in these cortical areas. PMID:22458949

  11. p-21 activated kinase 4 (PAK4) maintains stem cell-like phenotypes in pancreatic cancer cells through activation of STAT3 signaling

    PubMed Central

    Tyagi, Nikhil; Marimuthu, Saravanakumar; Bhardwaj, Arun; Deshmukh, Sachin K.; Srivastava, Sanjeev K.; Singh, Ajay P.; McClellan, Steven; Carter, James E.; Singh, Seema

    2015-01-01

    Pancreatic cancer (PC) remains a highly lethal malignancy due to its unusual chemoresistance and high aggressiveness. A subpopulation of pancreatic tumor cells, known as cancer stem cells (CSCs), is considered responsible not only for tumor-maintenance, but also for its widespread metastasis and therapeutic failure. Here we investigated the role of p-21 activated kinase 4 (PAK4) in driving PC stemness properties. Our data demonstrate that triple-positive (CD24+/CD44+/EpCAM+) subpopulation of pancreatic CSCs exhibits greater level of PAK4 as compared to triple-negative (CD24−/CD44−/EpCAM−) cells. Moreover, PAK4 silencing in PC cells leads to diminished fraction of CD24, CD44, and EpCAM positive cells. Furthermore, we show that PAK4-silenced PC cells exhibit decreased sphere-forming ability and increased chemo-sensitivity to gemcitabine toxicity. PAK4 expression is also associated with enhanced levels of stemness-associated transcription factors (Oct4/Nanog/Sox2 and KLF4). Furthermore, our data show decreased nuclear accumulation and transcriptional activity of STAT3 in PAK4-silenced PC cells and restitution of its activity leads to restoration of stem cell phenotypes. Together, our findings deliver first experimental evidence for the involvement of PAK4 in PC stemness and support its clinical utility as a novel therapeutic target in PC. PMID:26546043

  12. PREX1 Protein Function Is Negatively Regulated Downstream of Receptor Tyrosine Kinase Activation by p21-activated Kinases (PAKs).

    PubMed

    Barrows, Douglas; He, John Z; Parsons, Ramon

    2016-09-16

    Downstream of receptor tyrosine kinase and G protein-coupled receptor (GPCR) stimulation, the phosphatidylinositol 3,4,5-trisphosphate (PIP3)-dependent Rac exchange factor (PREX) family of guanine nucleotide exchange factors (GEFs) activates Rho GTPases, leading to important roles for PREX proteins in numerous cellular processes and diseases, including cancer. PREX1 and PREX2 GEF activity is activated by the second messengers PIP3 and Gβγ, and further regulation of PREX GEF activity occurs by phosphorylation. Stimulation of receptor tyrosine kinases by neuregulin and insulin-like growth factor 1 (IGF1) leads to the phosphorylation of PREX1; however, the kinases that phosphorylate PREX1 downstream of these ligands are not known. We recently reported that the p21-activated kinases (PAKs), which are activated by GTP-bound Ras-related C3 botulinum toxin substrate 1 (Rac1), mediate the phosphorylation of PREX2 after insulin receptor activation. Here we show that certain phosphorylation events on PREX1 after insulin, neuregulin, and IGF1 treatment are PAK-dependent and lead to a reduction in PREX1 binding to PIP3 Like PREX2, PAK-mediated phosphorylation also negatively regulates PREX1 GEF activity. Furthermore, the onset of PREX1 phosphorylation was delayed compared with the phosphorylation of AKT, supporting a model of negative feedback downstream of PREX1 activation. We also found that the phosphorylation of PREX1 after isoproterenol and prostaglandin E2-mediated GPCR activation is partially PAK-dependent and likely also involves protein kinase A, which is known to reduce PREX1 function. Our data point to multiple mechanisms of PREX1 negative regulation by PAKs within receptor tyrosine kinase and GPCR-stimulated signaling pathways that have important roles in diseases such as diabetes and cancer.

  13. The Pakistan National Emergency Department Surveillance Study (Pak-NEDS): Introducing a pilot surveillance

    PubMed Central

    2015-01-01

    Background Evidence-based decision making is essential for appropriate prioritization and service provision by healthcare systems. Despite higher demands, data needs for this practice are not met in many cases in low- and middle-income countries because of underdeveloped sources, among other reasons. Emergency departments (EDs) provide an important channel for such information because of their strategic position within healthcare systems. This paper describes the design and pilot test of a national ED based surveillance system suitable for the Pakistani context. Methods The Pakistan National Emergency Department Surveillance Study (Pak-NEDS) was pilot tested in the emergency departments of seven major tertiary healthcare centres across the country. The Aga Khan University, Karachi, served as the coordinating centre. Key stakeholders and experts from all study institutes were involved in outlining data needs, development of the study questionnaire, and identification of appropriate surveillance mechanisms such as methods for data collection, monitoring, and quality assurance procedures. The surveillance system was operational between November 2010 and March 2011. Active surveillance was done 24 hours a day by data collectors hired and trained specifically for the study. All patients presenting to the study EDs were eligible participants. Over 270,000 cases were registered in the surveillance system over a period of four months. Coverage levels in the final month ranged from 91-100% and were highest in centres with the least volume of patients. Overall the coverage for the four months was 79% and crude operational costs were less than $0.20 per patient. Conclusions Pak-NEDS is the first multi-centre ED based surveillance system successfully piloted in a sample of major EDs having some of the highest patient volumes in Pakistan. Despite the challenges identified, our pilot shows that the system is flexible and scalable, and could potentially be adapted for many other

  14. Clinical evaluation of the HeartPak. A new pneumatic portable driver for use with the HeartMate Implantable Pneumatic Left Ventricular Assist System.

    PubMed

    Tamez, D; Myers, T J; Inman, R W; Miller, K A; Frazier, O H

    1997-01-01

    The HeartPak Portable Pneumatic Driver was designed for use with the HeartMate Implantable Pneumatic Left Ventricular Assist Device (IP-LVAS) (Thermo Cardiosystems, Inc., Woburn, MA). The HeartPak measures 48 x 23 x 15 cm, weighs 9.3 kg with batteries, and can be carried by a handle, by a shoulder strap, or on a trolley. Four 12 V batteries provide power for as long as 8 hr. To test the HeartPak in the hospital environment, seven men were studied who were bridge-to-transplant patients (mean age, 59.8 +/- 8.87 years) undergoing HeartMate IP-LVAS therapy. They were supported by the HeartPak for 429 days with a cycle count of 57,826,560. To normalize the mean pump flow rate, we used the body surface area to obtain a pump flow index in each case. The mean flow rate was 2.65 +/- 0.57 L/min/m2 for the HeartPak vs. 2.64 +/- 0.45 L/min/m2 for the HeartMate 1000, the conventional driver previously used in these patients. The only potentially serious problem with the HeartPak was console failure in one case. The patient took appropriate backup measures, and the HeartPak was replaced. In no case did the device cause any adverse effects or interruption of LVAS support. Compared with HeartMate 1000, the HeartPak was more convenient, easier to operate, and allowed better patient mobility.

  15. PAK1 interacts with beta-catenin and is required for the regulation of the beta-catenin signalling pathway by gastrins.

    PubMed

    He, Hong; Shulkes, Arthur; Baldwin, Graham S

    2008-10-01

    Beta-catenin regulates cell-cell adhesion by binding to E-cadherin at the cell membrane and, when translocated into the nucleus, mediates signalling by activation of transcription factors such as TCF4. Mutations of the components of the Wnt/beta-catenin pathway are found in many gastrointestinal cancers. Gastrins, including amidated (Gamide) and glycine-extended (Ggly) gastrin(17), stimulate the proliferation of gastrointestinal cancer cells. Gastrins also regulate beta-catenin signalling through multiple pathways which seem to converge on p21-activated kinase 1 (PAK1). In this study, we have investigated the role of PAK1 in the regulation of beta-catenin signalling by gastrins. Here we report for the first time that PAK1 associated with beta-catenin. Both Gamide and Ggly stimulated the phosphorylation and activation of beta-catenin in a PAK1-dependent manner. A kinase-inactive mutant PAK1(K299A) blocked the gastrin-stimulated dissociation of beta-catenin from E-cadherin, translocation of beta-catenin from the cell membrane to the nucleus, and association of beta-catenin with the transcription factor TCF4. The PAK1(K299A) mutant also inhibited the stimulation of the expression of c-myc and cyclin D1, and of cell proliferation and migration, by gastrins. The results indicate that gastrins regulate beta-catenin signalling through a PAK1-dependent pathway. PAK1 seems to be the point of convergence of multiple signalling pathways activated by gastrins.

  16. Pak4 Is Required during Epithelial Polarity Remodeling through Regulating AJ Stability and Bazooka Retention at the ZA

    PubMed Central

    Walther, Rhian F.; Nunes de Almeida, Francisca; Vlassaks, Evi; Burden, Jemima J.; Pichaud, Franck

    2016-01-01

    Summary The ability of epithelial cells to assemble into sheets relies on their zonula adherens (ZA), a circumferential belt of adherens junction (AJ) material, which can be remodeled during development to shape organs. Here, we show that during ZA remodeling in a model neuroepithelial cell, the Cdc42 effector P21-activated kinase 4 (Pak4/Mbt) regulates AJ morphogenesis and stability through β-catenin (β-cat/Arm) phosphorylation. We find that β-catenin phosphorylation by Mbt, and associated AJ morphogenesis, is needed for the retention of the apical determinant Par3/Bazooka at the remodeling ZA. Importantly, this retention mechanism functions together with Par1-dependent lateral exclusion of Par3/Bazooka to regulate apical membrane differentiation. Our results reveal an important functional link between Pak4, AJ material morphogenesis, and polarity remodeling during organogenesis downstream of Par3. PMID:27052178

  17. Activity release from damaged fuel during the Paks-2 cleaning tank incident in the spent fuel storage pool

    NASA Astrophysics Data System (ADS)

    Hózer, Zoltán; Szabó, Emese; Pintér, Tamás; Varjú, Ilona Baracska; Bujtás, Tibor; Farkas, Gábor; Vajda, Nóra

    2009-07-01

    During crud removal operations the integrity of 30 fuel assemblies was lost at high temperature at the unit No. 2 of the Paks NPP. Part of the fission products was released from the damaged fuel into the coolant of the spent fuel storage pool. The gaseous fission products escaped through the chimney from the reactor hall. The volatile and non-volatile materials remained mainly in the coolant and were collected on the filters of water purification system. The activity release from damaged fuel rods during the Paks-2 cleaning tank incident was estimated on the basis of coolant activity concentration measurements and chimney activity data. The typical release rate of noble gases, iodine and caesium was 1-3%. The release of non-volatile fission products and actinides was also detected.

  18. Towards the Seismic Hazard Reassessment of Paks NPP (Hungary) Site: Seismicity and Sensitivity Studies

    NASA Astrophysics Data System (ADS)

    Toth, Laszlo; Monus, Peter; Gyori, Erzsebet; Grenerczy, Gyula; Janos Katona, Tamas; Kiszely, Marta

    2015-04-01

    In context of extension of Paks Nuclear Power Plant by new units, a comprehensive site seismic hazard evaluation program has been developed that is already approved by the Hungarian Authorities. This includes a 3D seismic survey, drilling of several deep boreholes, extensive geological mapping, and geophysical investigations at the site and its vicinity, as well as on near regional, and regional scale. Furthermore, all relevant techniques of modern space geodesy (GPS, PSInSAR) will be also utilized to construct a new seismotectonic model. The implementation of the project is still in progress. In the presentation, some important elements of the new seismic hazard assessment are highlighted, and some results obtained in the preliminary phase of the program are presented and discussed. The first and most important component of the program is the compilation of the seismological database that is developed on different time scale zooming on different event recurrence rates such as paleo-earthquakes (10-1/a). In 1995, Paks NPP installed and started to operate a sensitive microseismic monitoring network capable for locating earthquakes as small as magnitude 2.0 within about 100 km of the NPP site. During the two decades of operation, the microseismic monitoring network located some 2,000 earthquakes within the region of latitude 45.5 - 49 N and longitude 16 - 23 E. Out of the total number of events, 130 earthquakes were reported as 'felt events'. The largest earthquake was an event of ML 4.8, causing significant damage in the epicenter area. The results of microseismic monitoring provide valuable data for seismotectonic modelling and results in more accurate earthquake recurrence equations. The first modern PSHA of Paks NPP site was carried out in 1995. Complex site characterization project was implemented and hazard curves had been evaluated for 10-3 - 10-5 annual frequency. As a follow-up, PSHA results have been reviewed and updated in the frame of periodic safety

  19. Overexpression of a novel activator of PAK4, the CDK5 kinase-associated protein CDK5RAP3, promotes hepatocellular carcinoma metastasis.

    PubMed

    Mak, Grace Wing-Yan; Chan, Mandy Man-Lok; Leong, Veronica Yee-Law; Lee, Joyce Man-Fong; Yau, Tai-On; Ng, Irene Oi-Lin; Ching, Yick-Pang

    2011-04-15

    The CDK5 kinase regulatory subunit-associated protein 3 (CDK5RAP3 or C53/LZAP) regulates apoptosis induced by genotoxic stress. Although CDK5RAP3 has been implicated in cancer progression, its exact role in carcinogenesis is not well established. In this article, we report that CDK5RAP3 has an important prometastatic function in hepatocarcinogenesis. An examination of human hepatocellular carcinoma (HCC) samples revealed at least twofold overexpression of CDK5RAP3 transcripts in 58% (39/67) of HCC specimens when compared with corresponding nontumorous livers. CDK5RAP3 overexpression was associated with more aggressive biological behavior. In HCC cell lines, stable overexpression of CDK5RAP3 promoted, and small interfering RNA-mediated knockdown inhibited, tumorigenic activity and metastatic potential. We found that overexpression of CDK5RAP3 and p21-activated protein kinase 4 (PAK4) correlated in human HCCs, and that CDK5RAP3 was a novel binding partner of PAK4, and this binding enhanced PAK4 activity. siRNA-mediated knockdown of PAK4 in CDK5RAP3-expressing HCC cells reversed the enhanced cell invasiveness mediated by CDK5RAP3 overexpression, implying that PAK4 is essential for CDK5RAP3 function. Taken together, our findings reveal that CDK5RAP3 is widely overexpressed in HCC and that overexpression of CDK5RAP3 promotes HCC metastasis through PAK4 activation.

  20. p21-activated Kinases (PAKs) Mediate the Phosphorylation of PREX2 Protein to Initiate Feedback Inhibition of Rac1 GTPase.

    PubMed

    Barrows, Douglas; Schoenfeld, Sarah M; Hodakoski, Cindy; Silkov, Antonina; Honig, Barry; Couvillon, Anthony; Shymanets, Aliaksei; Nürnberg, Bernd; Asara, John M; Parsons, Ramon

    2015-11-27

    Phosphatidylinositol 3,4,5-trisphosphate (PIP3)-dependent Rac exchanger 2 (PREX2) is a guanine nucleotide exchange factor (GEF) for the Ras-related C3 botulinum toxin substrate 1 (Rac1) GTPase, facilitating the exchange of GDP for GTP on Rac1. GTP-bound Rac1 then activates its downstream effectors, including p21-activated kinases (PAKs). PREX2 and Rac1 are frequently mutated in cancer and have key roles within the insulin-signaling pathway. Rac1 can be inactivated by multiple mechanisms; however, negative regulation by insulin is not well understood. Here, we show that in response to being activated after insulin stimulation, Rac1 initiates its own inactivation by decreasing PREX2 GEF activity. Following PREX2-mediated activation of Rac1 by the second messengers PIP3 or Gβγ, we found that PREX2 was phosphorylated through a PAK-dependent mechanism. PAK-mediated phosphorylation of PREX2 reduced GEF activity toward Rac1 by inhibiting PREX2 binding to PIP3 and Gβγ. Cell fractionation experiments also revealed that phosphorylation prevented PREX2 from localizing to the cellular membrane. Furthermore, the onset of insulin-induced phosphorylation of PREX2 was delayed compared with AKT. Altogether, we propose that second messengers activate the Rac1 signal, which sets in motion a cascade whereby PAKs phosphorylate and negatively regulate PREX2 to decrease Rac1 activation. This type of regulation would allow for transient activation of the PREX2-Rac1 signal and may be relevant in multiple physiological processes, including diseases such as diabetes and cancer when insulin signaling is chronically activated.

  1. p21-activated Kinases (PAKs) Mediate the Phosphorylation of PREX2 Protein to Initiate Feedback Inhibition of Rac1 GTPase*

    PubMed Central

    Barrows, Douglas; Schoenfeld, Sarah M.; Hodakoski, Cindy; Silkov, Antonina; Honig, Barry; Couvillon, Anthony; Shymanets, Aliaksei; Nürnberg, Bernd; Asara, John M.; Parsons, Ramon

    2015-01-01

    Phosphatidylinositol 3,4,5-trisphosphate (PIP3)-dependent Rac exchanger 2 (PREX2) is a guanine nucleotide exchange factor (GEF) for the Ras-related C3 botulinum toxin substrate 1 (Rac1) GTPase, facilitating the exchange of GDP for GTP on Rac1. GTP-bound Rac1 then activates its downstream effectors, including p21-activated kinases (PAKs). PREX2 and Rac1 are frequently mutated in cancer and have key roles within the insulin-signaling pathway. Rac1 can be inactivated by multiple mechanisms; however, negative regulation by insulin is not well understood. Here, we show that in response to being activated after insulin stimulation, Rac1 initiates its own inactivation by decreasing PREX2 GEF activity. Following PREX2-mediated activation of Rac1 by the second messengers PIP3 or Gβγ, we found that PREX2 was phosphorylated through a PAK-dependent mechanism. PAK-mediated phosphorylation of PREX2 reduced GEF activity toward Rac1 by inhibiting PREX2 binding to PIP3 and Gβγ. Cell fractionation experiments also revealed that phosphorylation prevented PREX2 from localizing to the cellular membrane. Furthermore, the onset of insulin-induced phosphorylation of PREX2 was delayed compared with AKT. Altogether, we propose that second messengers activate the Rac1 signal, which sets in motion a cascade whereby PAKs phosphorylate and negatively regulate PREX2 to decrease Rac1 activation. This type of regulation would allow for transient activation of the PREX2-Rac1 signal and may be relevant in multiple physiological processes, including diseases such as diabetes and cancer when insulin signaling is chronically activated. PMID:26438819

  2. SparsePak: A Formatted Fiber Field Unit for The WIYN Telescope Bench Spectrograph. II. On-Sky Performance

    NASA Astrophysics Data System (ADS)

    Bershady, Matthew A.; Andersen, David R.; Verheijen, Marc A. W.; Westfall, Kyle B.; Crawford, Steven M.; Swaters, Rob A.

    2005-02-01

    We present a performance analysis of SparsePak and the WIYN Bench Spectrograph for precision studies of stellar and ionized gas kinematics of external galaxies. We focus on spectrograph configurations with echelle and low-order gratings yielding spectral resolutions of ~10,000 between 500 and 900 nm. These configurations are of general relevance to the spectrograph performance. Benchmarks include spectral resolution, sampling, vignetting, scattered light, and an estimate of the system absolute throughput. Comparisons are made to other, existing, fiber feeds on the WIYN Bench Spectrograph. Vignetting and relative throughput are found to agree with a geometric model of the optical system. An aperture-correction protocol for spectrophotometric standard-star calibrations has been established using independent WIYN imaging data and the unique capabilities of the SparsePak fiber array. The WIYN point-spread function is well fitted by a Moffat profile with a constant power-law outer slope of index -4.4. We use SparsePak commissioning data to debunk a long-standing myth concerning sky-subtraction with fibers: by properly treating the multifiber data as a ``long-slit'' it is possible to achieve precision sky-subtraction with a signal-to-noise performance as good or better than conventional long-slit spectroscopy. No beam-switching is required, and hence the method is efficient. Finally, we give several examples of science measurements that SparsePak now makes routine. These include Hα velocity fields of low surface brightness disks, gas and stellar velocity-fields of nearly face-on disks, and stellar absorption-line profiles of galaxy disks at spectral resolutions of ~24,000.

  3. Mangrove Colonization: Mangrove Progression Over the Growing Pak Phanang (SE Thailand) Mud Flat

    NASA Astrophysics Data System (ADS)

    Panapitukkul, N.; Duarte, C. M.; Thampanya, U.; Kheowvongsri, P.; Srichai, N.; Geertz-Hansen, O.; Terrados, J.; Boromthanarath, S.

    1998-07-01

    A combination of remote sensing techniques and in situmeasurements along a chronosequence was used to elucidate the rate of progression of the mangrove forest in the Pak Phanang Bay (SE Thailand), a large bay with an extended and rapidly accreting mud flat. The examination of black and white aerial photographs of the forest in 1966, 1974, 1989 and 1995, and satellite images in 1985, 1990 and 1994 revealed that the mangrove forest located in the eastern bank of the bay was progressing over the mud flat. The rate of progression was estimated, from examination of changes in the position of the forest edge with time in the series of images, to average 38·6 m year -1over the 28-year interval encompassed by the images. Mangrove progression rates were fastest between 1966 and 1974 and slowest between 1974 and 1985, remaining uniform at about 30 m year -1thereafter. The in situexamination of vegetation along transects in the area of fastest mangrove progression showed an average progression rate of 53·12±5·86 m year -1, quite similar to the estimate (48·4 m year -1) derived from remote sensing techniques for the area where the transects were surveyed. Avicennia albawas found to dominate the vegetation at the progressing edge of the mangrove, followed by Sonneratia caseolaris, with Rhizophora apiculatabeing present only occasionally. The fast colonization of A. albaover the mud flat was supported by a large export flux of mangrove propagules from the channels draining the mangrove forest, which averaged 3715±920 and 1900±808 fruits day -1in each of the channels examined. Extrapolation of the long-term mean mangrove progression rate observed along the eastern bank of the Pak Phanang Bay suggested that this mangrove forest will increase by 33 ha year -1. These results provide evidence that natural mangrove colonization can be a rapid process if sufficient propagules of the pioneer species ( A. albaand S. caseolaris) are available, and point, therefore, to alternative

  4. PAK1 and CtBP1 Regulate the Coupling of Neuronal Activity to Muscle Chromatin and Gene Expression

    PubMed Central

    Thomas, Jean-Luc; Ravel-Chapuis, Aymeric; Valente, Carmen; Corda, Daniela; Méjat, Alexandre

    2015-01-01

    Acetylcholine receptor (AChR) expression in innervated muscle is limited to the synaptic region. Neuron-induced electrical activity participates in this compartmentalization by promoting the repression of AChR expression in the extrasynaptic regions. Here, we show that the corepressor CtBP1 (C-terminal binding protein 1) is present on the myogenin promoter together with repressive histone marks. shRNA-mediated downregulation of CtBP1 expression is sufficient to derepress myogenin and AChR expression in innervated muscle. Upon denervation, CtBP1 is displaced from the myogenin promoter and relocates to the cytoplasm, while repressive histone marks are replaced by activating ones concomitantly to the activation of myogenin expression. We also observed that upon denervation the p21-activated kinase 1 (PAK1) expression is upregulated, suggesting that phosphorylation by PAK1 may be involved in the relocation of CtBP1. Indeed, preventing CtBP1 Ser158 phosphorylation induces CtBP1 accumulation in the nuclei and abrogates the activation of myogenin and AChR expression. Altogether, these findings reveal a molecular mechanism to account for the coordinated control of chromatin modifications and muscle gene expression by presynaptic neurons via a PAK1/CtBP1 pathway. PMID:26416879

  5. Reactor Dosimetry Aspects of the Service Life Extension of the Hungarian Paks NPP

    NASA Astrophysics Data System (ADS)

    Zsolnay, Eva M.; Czifrus, Szabolcs; Fehér, Sándor; Hordósy, Gábor; Keresztúri, András; Kresz, Norbert; Oszvald, Ferenc

    2016-02-01

    The service life of the Hungarian Paks Nuclear Power Plant (NPP) will be extended from the originally planned 30 years to 50 years. To improve the reliability of the results obtained in frame of the old reactor pressure vessel (RPV) surveillance programme, new methods have been developed, and based on them, the old exposition data have been re-evaluated for all the four reactor units. At the same time, a new RPV surveillance programme has been developed and introduced, and long term irradiations have been performed to determine the radiation damage of the surveillance specimens due to the high fast neutron exposition. Neutron transport calculations have been performed with a validated neutron transport code system to determine the fast neutron exposition of the RPVs during the extended service life. The cavity dosimetry is in the introductory phase. This paper presents the new developments in the field of the RPV surveillance dosimetry and summarises the results obtained. According to the results the service life of the NPP can safely be extended for the planned 50 years.

  6. Geochemistry and tectonic setting of the Central Loei volcanic rocks, Pak Chom area, Loei, northeastern Thailand

    NASA Astrophysics Data System (ADS)

    Panjasawatwong, Y.; Zaw, Khin; Chantaramee, S.; Limtrakun, P.; Pirarai, K.

    2006-01-01

    The Central Loei volcanic rocks, as evidenced by those in the Pak Chom area, were formed in the Late Devonian-Early Carboniferous and can be separated into three magmatic groups: transitional tholeiitic basalt, tholeiitic microgabbro and calc-alkalic basalt/andesite on the basis of immobile-element contents and ratios of least altered samples. All the tholeiitic microgabbro possibly occurred as dikes. Chemically, the transitional tholeittic basalt and tholeiitic microgabbro have higher abundances of TiO 2, Ni and Cr relative to the calc-alkalic basalt/andesite at similar values for FeO*/MgO; they also contain higher Ti/Zr but lower Zr/Nb. The transitional tholeiitic basalt has higher concentrations of P 2O 5 and Nb relative to the tholeiitic microgabbro at similar levels of FeO*/MgO, and also has higher ratios of Nb/Y and Ti/V, but lower values for Ti/Zr and Zr/Nb. In terms of chondrite normalized REE and N-MORB normalized patterns, the transitional tholeiitic basalt, tholeiitic basalt and calc-alkalic basalt/andesite are analogous to those from North Atlantic, Southwest Indian Ridge and New Britain Arc. On this basis, the Central Loei volcanic rocks are comprised of MORBs and oceanic island-arc lavas. These arc lavas may have erupted on an oceanic basement in the same ocean basin as those in the Chiang Rai-Chiang Mai volcanic belt.

  7. Chronic exposure to cigarette smoke leads to activation of p21 (RAC1)-activated kinase 6 (PAK6) in non-small cell lung cancer cells

    PubMed Central

    Syed, Nazia; Solanki, Hitendra S.; Puttamallesh, Vinuth N.; Balaji, Sai A.; Nanjappa, Vishalakshi; Datta, Keshava K.; Babu, Niraj; Renuse, Santosh; Patil, Arun H.; Izumchenko, Evgeny; Prasad, T.S. Keshava; Chang, Xiaofei; Rangarajan, Annapoorni; Sidransky, David; Pandey, Akhilesh; Gowda, Harsha; Chatterjee, Aditi

    2016-01-01

    Epidemiological data clearly establishes cigarette smoking as one of the major cause for lung cancer worldwide. Recently, targeted therapy has become one of the most preferred modes of treatment for cancer. Though certain targeted therapies such as anti-EGFR are in clinical practice, they have shown limited success in lung cancer patients who are smokers. This demands discovery of alternative drug targets through systematic investigation of cigarette smoke-induced signaling mechanisms. To study the signaling events activated in response to cigarette smoke, we carried out SILAC-based phosphoproteomic analysis of H358 lung cancer cells chronically exposed to cigarette smoke. We identified 1,812 phosphosites, of which 278 phosphosites were hyperphosphorylated (≥ 3-fold) in H358 cells chronically exposed to cigarette smoke. Our data revealed hyperphosphorylation of S560 within the conserved kinase domain of PAK6. Activation of PAK6 is associated with various processes in cancer including metastasis. Mechanistic studies revealed that inhibition of PAK6 led to reduction in cell proliferation, migration and invasion of the cigarette smoke treated cells. Further, siRNA mediated silencing of PAK6 resulted in decreased invasive abilities in a panel of non-small cell lung cancer (NSCLC) cells. Consistently, mice bearing tumor xenograft showed reduced tumor growth upon treatment with PF-3758309 (group II PAK inhibitor). Immunohistochemical analysis revealed overexpression of PAK6 in 66.6% (52/78) of NSCLC cases in tissue microarrays. Taken together, our study indicates that PAK6 is a promising novel therapeutic target for NSCLC, especially in smokers. PMID:27542207

  8. Rainfall Trends over the Indo-Pak Summer Monsoon and Related Large-Scale Dynamics

    NASA Astrophysics Data System (ADS)

    Latif, Muhammad; Syed, Faisal; Hannachi, Abdel

    2016-04-01

    The study of regional rainfall trends over South Asia is critically important for food security and infrastructure. This study investigates the presence of trends in seasonal and sub-seasonal (June through September-JJAS) rainfall obtained from multiple observed datasets. The obtained results identified a dipole-type structure in rainfall trends over the region north of the Indo-Pak subcontinent, where significant increasing trends are seen over the core monsoon region of Pakistan and significant decreasing trends are observed over the central-north India and adjacent areas. The study strongly suggests that strengthening of Vertically Integrated Meridional Moisture Transport (VIMMT) over the Arabian Sea is likely reason for the trend of rainfall in the core monsoon region of Pakistan. In contrast, over the central-north India region, the rainfall trends are significantly decreasing due to the weakening of IMT over the Bay of Bengal. The leading EOF clearly shows the strengthening (weakening) patterns of VIMMT over the Arabian Sea (Bay of Bengal) in seasonal and sub-seasonal interannual time-scales. The regression analysis between the principal components and rainfall confirms the dipole pattern over the region. Our results also suggest that the Circumglobal Teleconnection in upper troposphere influence in maintaining the mean rainfall over Pakistan via cross-equatorial flow of moisture into the Arabian Sea. We also investigate seasonal JJAS rainfall trends using historical and climate change (RCP4.5 and RCP8.5) simulations from a set of regional climate models from Coupled Model Intercomparison Project (CMIP5). Trends and asymmetry of seasonal rainfall show great variability across models. Meridional moisture transport and associated large-scale dynamics will also be discussed.

  9. PAK2 is an effector of TSC1/2 signaling independent of mTOR and a potential therapeutic target for Tuberous Sclerosis Complex

    PubMed Central

    Alves, Maria M.; Fuhler, Gwenny M.; Queiroz, Karla C.S.; Scholma, Jetse; Goorden, Susan; Anink, Jasper; Arnold Spek, C.; Hoogeveen-Westerveld, Marianne; Bruno, Marco J.; Nellist, Mark; Elgersma, Ype; Aronica, Eleonora; Peppelenbosch, Maikel P.

    2015-01-01

    Tuberous sclerosis complex (TSC) is caused by inactivating mutations in either TSC1 or TSC2 and is characterized by uncontrolled mTORC1 activation. Drugs that reduce mTOR activity are only partially successful in the treatment of TSC, suggesting that mTOR-independent pathways play a role in disease development. Here, kinome profiles of wild-type and Tsc2−/− mouse embryonic fibroblasts (MEFs) were generated, revealing a prominent role for PAK2 in signal transduction downstream of TSC1/2. Further investigation showed that the effect of the TSC1/2 complex on PAK2 is mediated through RHEB, but is independent of mTOR and p21RAC. We also demonstrated that PAK2 over-activation is likely responsible for the migratory and cell cycle abnormalities observed in Tsc2−/− MEFs. Finally, we detected high levels of PAK2 activation in giant cells in the brains of TSC patients. These results show that PAK2 is a direct effector of TSC1-TSC2-RHEB signaling and a new target for rational drug therapy in TSC. PMID:26412398

  10. Increased expression of microRNA-221 inhibits PAK1 in endothelial progenitor cells and impairs its function via c-Raf/MEK/ERK pathway

    SciTech Connect

    Zhang, Xiaoping; Mao, Haian; Chen, Jin-yuan; Wen, Shengjun; Li, Dan; Ye, Meng; Lv, Zhongwei

    2013-02-15

    Highlights: ► MicroRNA-221 is upregulated in the endothelial progenitor cells of atherosclerosis patients. ► PAK1 is a direct target of microRNA-221. ► MicroRNA-221 inhibits EPCs proliferation through c-Raf/MEK/ERK pathway. -- Abstract: Coronary artery disease (CAD) is associated with high mortality and occurs via endothelial injury. Endothelial progenitor cells (EPCs) restore the integrity of the endothelium and protect it from atherosclerosis. In this study, we compared the expression of microRNAs (miRNAs) in EPCs in atherosclerosis patients and normal controls. We found that miR-221 expression was significantly up-regulated in patients compared with controls. We predicted and identified p21/Cdc42/Rac1-activated kinase 1 (PAK1) as a novel target of miR-221 in EPCs. We also demonstrated that miR-221 targeted a putative binding site in the 3′UTR of PAK1, and absence of this site was inversely associated with miR-221 expression in EPCs. We confirmed this relationship using a luciferase reporter assay. Furthermore, overexpression of miR-221 in EPCs significantly decreased EPC proliferation, in accordance with the inhibitory effects induced by decreased PAK1. Overall, these findings demonstrate that miR-221 affects the MEK/ERK pathway by targeting PAK1 to inhibit the proliferation of EPCs.

  11. Threonine 209 phosphorylation on RUNX3 by Pak1 is a molecular switch for its dualistic functions.

    PubMed

    Kumar, A; Singhal, M; Chopra, C; Srinivasan, S; Surabhi, R P; Kanumuri, R; Tentu, S; Jagadeeshan, S; Sundaram, S; Ramanathan, K; Shankar Pitani, R; Muthuswamy, B; Abhijit, S; Nair, A S; Venkatraman, G; Rayala, S K

    2016-09-15

    P21 Activated Kinase 1 (Pak1), an oncogenic serine/threonine kinase, is known to have a significant role in the regulation of cytoskeleton and cellular morphology. Runx3 was initially known for its role in tumor suppressor function, but recent studies have reported the oncogenic role of Runx3 in various cancers. However, the mechanism that controls the paradoxical functions of Runx3 still remains unclear. In this study, we show that Runx3 is a physiologically interacting substrate of Pak1. We identified the site of phosphorylation in Runx3 as Threonine 209 by mass spectrometry analysis and site-directed mutagenesis, and further confirmed the same with a site-specific antibody. Results from our functional studies showed that Threonine 209 phosphorylation in Runx3 alters its subcellular localization by protein mislocalization from the nucleus to the cytoplasm and subsequently converses its biological functions. This was further supported by in vivo tumor xenograft studies in nude mouse models which clearly demonstrated that PANC-28 cells transfected with the Runx3-T209E clone showed high tumorigenic potential as compared with other clones. Our results from clinical samples also suggest that Threonine 209 phosphorylation by Pak1 could be a potential therapeutic target and of great clinical relevance with implications for Runx3 inactivation in cancer cells where Runx3 is known to be oncogenic. The findings presented in this study provide evidence of Runx3-Threonine 209 phosphorylation as a molecular switch in dictating the tissue-specific dualistic functions of Runx3 for the first time.

  12. Slit stimulation recruits Dock and Pak to the roundabout receptor and increases Rac activity to regulate axon repulsion at the CNS midline.

    PubMed

    Fan, Xueping; Labrador, Juan Pablo; Hing, Huey; Bashaw, Greg J

    2003-09-25

    Drosophila Roundabout (Robo) is the founding member of a conserved family of repulsive axon guidance receptors that respond to secreted Slit proteins. Here we present evidence that the SH3-SH2 adaptor protein Dreadlocks (Dock), the p21-activated serine-threonine kinase (Pak), and the Rac1/Rac2/Mtl small GTPases can function during Robo repulsion. Loss-of-function and genetic interaction experiments suggest that limiting the function of Dock, Pak, or Rac partially disrupts Robo repulsion. In addition, Dock can directly bind to Robo's cytoplasmic domain, and the association of Dock and Robo is enhanced by stimulation with Slit. Furthermore, Slit stimulation can recruit a complex of Dock and Pak to the Robo receptor and trigger an increase in Rac1 activity. These results provide a direct physical link between the Robo receptor and an important cytoskeletal regulatory protein complex and suggest that Rac can function in both attractive and repulsive axon guidance.

  13. Molluscan associations from the Pak Phanang Bay (SW Gulf of Thailand) as a record of natural and anthropogenic changes

    NASA Astrophysics Data System (ADS)

    Negri, Mauro Pietro; Sanfilippo, Rossana; Basso, Daniela; Rosso, Antonietta; Di Geronimo, Sebastiano Italo

    2014-08-01

    Recent environmental changes in the Pak Phanang Bay (SW Gulf of Thailand) are investigated by means of mollusc assemblages. The present-day water depth within the bay slightly exceeds 2 m at low tide and the seafloor is almost entirely muddy, except for the outermost part of the embayment which is directly influenced by the longshore drift that is building the Laem Talumpuk sand spit. A multivariate analysis of the molluscan fauna recovered at 16 sampling stations within the bay delineates three thanatofacies and two biofacies. The Bay Mouth thanatofacies, including several infralittoral species, is distributed around the bay entrance; the Tidal Flat thanatofacies, characterized by few brackish and freshwater taxa, occurs in the inner part of the bay; the Channel thanatofacies includes a mixed fauna and is found along the long axis of the bay. All thanatofacies are not older than a few decades. The two biofacies are significantly less diverse than their dead counterparts, and are simply identified as Bay Mouth biofacies and Inner Bay biofacies. The faunal evolution, combined with bathymetric and sedimentological data, confirms that the embayment is undergoing a confinement process. The inner bay has evolved into an undifferentiated tidal flat hosting an oligospecific fauna. The confinement trend and the consequent siltation of the bay, mainly due to natural geomorphologic processes acting since centuries, are likely to have sped up in recent years by interaction with some human activities; among these, the deforestation in the upper Pak Phanang basin and the construction of Uthokaviphatprasit Watergate.

  14. Reversed-phase liquid chromatography of radiolabeled peptides using a C18 guard-PAK precolumn system

    SciTech Connect

    Carriere, P.D.; Bennett, H.P. )

    1989-03-01

    In order to avoid radioactive contamination of high-performance liquid chromatography columns and injectors, we have investigated the use of a Guard-PAK precolumn system for the chromatography of ({sup 125}I) labeled peptides. Two gonadotropin-releasing hormone analogs: (1) (D-Ala6-des-Gly10)-GnRH (GnRH-(Ala6)) and (2) (D-Ser(TBu)6-des-Gly10)-GnRH (GnRH-(Ser6)) and rat prolactin (r-PRL) were radiolabeled with {sup 125}I and subjected to reversed-phase liquid chromatography using a C18 Guard-PAK precolumn system. Major peak fractions of purified ({sup 125}I)GnRH-(Ala6), ({sup 125}I)GnRH-(Ser6), and ({sup 125}I)r-PRL eluted at 24%, 28%, and 55% acetonitrile, respectively. Purified ({sup 125}I)GnRH analogs showed specific high affinity binding to rat anterior pituitary gland membranes (specific activity: 1500-1700 Ci/mmol). Purified ({sup 125}I)r-PRL showed high affinity binding to r-PRL antibody by RIA (specific activity: 70-75 microCi/micrograms). This rapid and efficient chromatographic method should be useful in the separation of a wide range of radiolabeled protein and peptide molecules.

  15. Increased Circulating Endothelial Microparticles Associated with PAK4 Play a Key Role in Ventilation-Induced Lung Injury Process

    PubMed Central

    Pan, Shuming; Fei, Aihua; Jing, Lihong; Zhang, Xiangyu

    2017-01-01

    Inappropriate mechanical ventilation (MV) can result in ventilator-induced lung injury (VILI). Probing mechanisms of VILI and searching for effective methods are current areas of research focus on VILI. The present study aimed to probe into mechanisms of endothelial microparticles (EMPs) in VILI and the protective effects of Tetramethylpyrazine (TMP) against VILI. In this study, C57BL/6 and TLR4KO mouse MV models were used to explore the function of EMPs associated with p21 activated kinases-4 (PAK-4) in VILI. Both the C57BL/6 and TLR4 KO groups were subdivided into a mechanical ventilation (MV) group, a TMP + MV group, and a control group. After four hours of high tidal volume (20 ml/kg) MV, the degree of lung injury and the protective effects of TMP were assessed. VILI inhibited the cytoskeleton-regulating protein of PAK4 and was accompanied by an increased circulating EMP level. The intercellular junction protein of β-catenin was also decreased accompanied by a thickening alveolar wall, increased lung W/D values, and neutrophil infiltration. TMP alleviated VILI via decreasing circulating EMPs, stabilizing intercellular junctions, and alleviating neutrophil infiltration. PMID:28261612

  16. Glucosinolate Accumulation and Related Gene Expression in Pak Choi (Brassica rapa L. ssp. chinensis var. communis [N. Tsen & S.H. Lee] Hanelt) in Response to Insecticide Application.

    PubMed

    Zhu, Biao; Yang, Jing; He, Yong; Zang, Yunxiang; Zhu, Zhujun

    2015-11-11

    Glucosinolates and their breakdown products are well-known for their cancer-chemoprotective functions and biocidal activities against pathogens and generalist herbivores. Insecticides are commonly used in the production of pak choi (Brassica rapa L. ssp. chinensis var. communis [N. Tsen & S.H. Lee] Hanelt). We studied the effects of four commonly used insecticides, namely, β-cypermethrin, acephate, pymetrozine, and imidacloprid, on glucosinolate metabolism in pak choi. All insecticides significantly increased both the transcription of glucosinolate biosynthetic genes and the aliphatic and total glucosinolate accumulations in pak choi. β-Cypermethrin and acephate caused gradual and continuous up-regulation of gene expression from 0.5 to 24 h after treatment, whereas pymetrozine and imidacloprid did so more rapidly, reaching a peak at 1 h and returning to normal at 3 h. Our findings indicate that the four insecticides affect glucosinolate metabolism in pak choi plants to various degrees and suggest that glucosinolates may be involved in plant insecticide metabolism.

  17. Test Plan for Lockheed Idaho Technologies Company (LITCO), ARROW-PAK Packaging, Docket 95-40-7A, Type A Container

    SciTech Connect

    Kelly, D.L.

    1995-10-23

    This report documents the U.S. Department of Transportation Specification 7A Type A (DOT-7A) compliance testing to be followed for qualification of the Lockheed Idaho Technologies Company, ARROW-PAK, for use as a Type A Packaging. The packaging configuration being tested is intended for transportation of radioactive solids, Form No. 1, Form No. 2, and Form No. 3.

  18. ETV TEST REPORT OF CONTROL OF BIOAEROSOLS IN HVAC SYSTEMS GLASFLOSS INDUSTRIES Z-PAK SERIES S, MODEL ZPS24241295BO

    EPA Science Inventory

    The Environmental Technology Verification report discusses the technology and performance of the Z-Pak Series S, Model ZPS24241295B0 air filter for dust and bioaerosol filtration manufactured by Glasfloss Industries, Inc. The pressure drop across the filter was 91 Pa clean and 34...

  19. 1,2,3-Triazolyl ester of Ketorolac: A "Click Chemistry"-based highly potent PAK1-blocking cancer-killer.

    PubMed

    Nguyen, Binh Cao Quan; Takahashi, Hideaki; Uto, Yoshihiro; Shahinozzaman, M D; Tawata, Shinkichi; Maruta, Hiroshi

    2017-01-27

    An old anti-inflammatory/analgesic drug called Toradol is a racemic form of Ketorolac (50% R-form and 50% S-form) that blocks the oncogenic RAC-PAK1-COX-2 (cyclooxygenase-2) signaling, through the direct inhibition of RAC by the R-form and of COX-2 by the S-form, eventually down-regulating the production of prostaglandins. However, due to its COOH moiety which is clearly repulsive to negatively-charged phospholipid-based plasma membrane, its cell-permeability is rather poor (the IC50 against the growth of human cancer cells such as A549 is around 13 μM). In an attempt to boost its anti-cancer activity, hopefully by increasing its cell-permeability through abolishing the negative charge, yet keeping its water-solubility, here we synthesized a 1,2,3-triazolyl ester of Toradol through "Click Chemistry". The resultant water-soluble "azo" derivative called "15K" was found to be over 500 times more potent than Toradol with the IC50 around 24 nM against the PAK1-dependent growth of A549 cancer cells, inactivating PAK1 in cell culture with the apparent IC50 around 65 nM, and inhibiting COX-2 in vitro with the IC50 around 6 nM. Furthermore, the Click Chemistry boosts the anti-cancer activity of Ketorolac by 5000 times against the PAK1-independent growth of B16F10 melanoma cells. Using a multi-drug-resistant (MDR) cancer cell line (EMT6), we found that the esterization of Ketorolac boosts its cell-permeability by at least 10 folds. Thus, the Click Chemistry dramatically boosts the anti-cancer activity of Ketorolac, at least in three ways: increasing its cell-permeability, the anti-PAK1 activity of R-form and anti-COX-2 activity of S-form. The resultant "15K" is so far among the most potent PAK1-blockers, and therefore would be potentially useful for the therapy of many different PAK1-dependent diseases/disorders such as cancers.

  20. A Population Growth Trend Analysis for Neotricula aperta, the Snail Intermediate Host of Schistosoma mekongi, after Construction of the Pak-Mun Dam

    PubMed Central

    Attwood, Stephen W.; Upatham, E. Suchart

    2013-01-01

    Background The Pak-Mun dam is a controversial hydro-power project on the Mun River in Northeast Thailand. The dam is sited in a habitat of the freshwater snail Neotricula aperta, which is the intermediate host for the parasitic blood-fluke Schistosoma mekongi causing Mekong schistosomiasis in humans in Cambodia and Laos. Few data are available which can be used to assess the effects of water resource development on N. aperta. The aim of this study was to obtain data and to analyze the possible impact of the dam on N. aperta population growth. Methodology/Principal Findings Estimated population densities were recorded for an N. aperta population in the Mun River 27 km upstream of Pak-Mun, from 1990 to 2011. The Pak-Mul dam began to operate in 1994. Population growth was modeled using a linear mixed model expression of a modified Gompertz stochastic state-space exponential growth model. The N. aperta population was found to be quite stable, with the estimated growth parameter not significantly different from zero. Nevertheless, some marked changes in snail population density were observed which were coincident with changes in dam operation policy. Conclusions/Significance The study found that there has been no marked increase in N. aperta population growth following operation of the Pak-Mun dam. The analysis did indicate a large and statistically significant increase in population density immediately after the dam came into operation; however, this increase was not persistent. The study has provided the first vital baseline data on N. aperta population behavior near to the Pak-Mun dam and suggests that the operation policy of the dam may have an impact on snail population density. Nevertheless, additional studies are required for other N. aperta populations in the Mun River and for an extended time series, to confirm or refine the findings of this work. PMID:24244775

  1. Final evaluation report for Lockheed Idaho Technologies Company, ARROW-PAK packaging, Docket 95-40-7A, Type A container

    SciTech Connect

    Kelly, D.L.

    1995-11-01

    The report documents the U.S. Department of Transportation Specification 7A Type A (DOT-7A) compliance test results of the ARROW-PAK packaging. The ARROW-PAK packaging system consists of Marlex M-8000 Driscopipe (Series 8000 [gas] or Series 8600 [industrial]) resin pipe, manufactured by Phillips-Driscopipe, Inc., and is sealed with two dome-shaped end caps manufactured from the same materials. The patented sealing process involves the use of electrical energy to heat opposing faces of the pipe and end caps, and hydraulic rams to press the heated surfaces together. This fusion process produces a homogeneous bonding of the end cap to the pipe. The packaging may be used with or without the two internal plywood spacers. This packaging was evaluated and tested in October 1995. The packaging configuration described in this report is designed to ship Type A quantities of solid radioactive materials, Form No. 1, Form No. 2, and Form No. 3.

  2. Molecular evolution, characterization, and expression analysis of SnRK2 gene family in Pak-choi (Brassica rapa ssp. chinensis)

    PubMed Central

    Huang, Zhinan; Tang, Jun; Duan, Weike; Wang, Zhen; Song, Xiaoming; Hou, Xilin

    2015-01-01

    The sucrose non-fermenting 1-related protein kinase 2 (SnRK2) family members are plant-specific serine/threonine kinases that are involved in the plant response to abiotic stress and abscisic acid (ABA)-dependent plant development. Further understanding of the evolutionary history and expression characteristics of these genes will help to elucidate the mechanisms of the stress tolerance in Pak-choi, an important green leafy vegetable in China. Thus, we investigated the evolutionary patterns, footprints and conservation of SnRK2 genes in selected plants and later cloned and analyzed SnRK2 genes in Pak-choi. We found that this gene family was preferentially retained in Brassicas after the Brassica-Arabidopsis thaliana split. Next, we cloned and sequenced 13 SnRK2 from both cDNA and DNA libraries of stress-induced Pak-choi, which were under conditions of ABA, salinity, cold, heat, and osmotic treatments. Most of the BcSnRK2s have eight exons and could be divided into three groups. The subcellular localization predictions suggested that the putative BcSnRK2 proteins were enriched in the nucleus. The results of an analysis of the expression patterns of the BcSnRK2 genes showed that BcSnRK2 group III genes were robustly induced by ABA treatments. Most of the BcSnRK2 genes were activated by low temperature, and the BcSnRK2.6 genes responded to both ABA and low temperature. In fact, most of the BcSnRK2 genes showed positive or negative regulation under ABA and low temperature treatments, suggesting that they may be global regulators that function at the intersection of multiple signaling pathways to play important roles in Pak-choi stress responses. PMID:26557127

  3. FTY720 prevents ischemia/reperfusion injury-associated arrhythmias in an ex vivo rat heart model via activation of Pak1/Akt signaling.

    PubMed

    Egom, E Eroume A; Ke, Yunbo; Musa, Hanny; Mohamed, Tamer M A; Wang, Tao; Cartwright, Elizabeth; Solaro, R John; Lei, Ming

    2010-02-01

    Recent studies demonstrated a role of sphingosine-1-phosphate (S1P) in the protection against the stress of ischemia/reperfusion (I/R) injury. In experiments reported here, we have investigated the signaling through the S1P cascade by FTY720, a sphingolipid drug candidate displaying structural similarity to S1P, underlying the S1P cardioprotective effect. In ex vivo rat heart and isolated sinoatrial node models, FTY720 significantly prevented arrhythmic events associated with I/R injury including premature ventricular beats, VT, and sinus bradycardia as well as A-V conduction block. Real-time PCR and Western blot analysis demonstrated the expression of the S1P receptor transcript pools and corresponding proteins including S1P1, S1P2, and S1P3 in tissues dissected from sinoatrial node, atrium and ventricle. FTY720 (25 nM) significantly blunted the depression of the levels of phospho-Pak1 and phospho-Akt with ischemia and with reperfusion. There was a significant increase in phospho-Pak1 levels by 35%, 199%, and 205% after 5, 10, and 15 min of treatment with 25 nM FTY720 compared with control nontreated myocytes. However, there was no significant difference in the levels of total Pak1 expression between nontreated and FTY720 treated. Phospho-Akt levels were increased by 44%, 63%, and 61% after 5, 10, and 15 min of treatment with 25 nM FTY720, respectively. Our data provide the first evidence that FTY720 prevents I/R injury-associated arrhythmias and indicate its potential significance as an important and new agent protecting against I/R injury. Our data also indicate, for the first time, that the cardioprotective effect of FTY720 is likely to involve activation of signaling through the Pak1.

  4. Phytoavailability of Cadmium (Cd) to Pak Choi (Brassica chinensis L.) Grown in Chinese Soils: A Model to Evaluate the Impact of Soil Cd Pollution on Potential Dietary Toxicity

    PubMed Central

    Yang, Xiaoe; Xiao, Wendan; Stoffella, Peter J.; Saghir, Aamir; Azam, Muhammad; Li, Tingqiang

    2014-01-01

    Food chain contamination by soil cadmium (Cd) through vegetable consumption poses a threat to human health. Therefore, an understanding is needed on the relationship between the phytoavailability of Cd in soils and its uptake in edible tissues of vegetables. The purpose of this study was to establish soil Cd thresholds of representative Chinese soils based on dietary toxicity to humans and develop a model to evaluate the phytoavailability of Cd to Pak choi (Brassica chinensis L.) based on soil properties. Mehlich-3 extractable Cd thresholds were more suitable for Stagnic Anthrosols, Calcareous, Ustic Cambosols, Typic Haplustalfs, Udic Ferrisols and Periudic Argosols with values of 0.30, 0.25, 0.18, 0.16, 0.15 and 0.03 mg kg−1, respectively, while total Cd is adequate threshold for Mollisols with a value of 0.86 mg kg−1. A stepwise regression model indicated that Cd phytoavailability to Pak choi was significantly influenced by soil pH, organic matter, total Zinc and Cd concentrations in soil. Therefore, since Cd accumulation in Pak choi varied with soil characteristics, they should be considered while assessing the environmental quality of soils to ensure the hygienically safe food production. PMID:25386790

  5. Phytoavailability of cadmium (Cd) to Pak choi (Brassica chinensis L.) grown in Chinese soils: a model to evaluate the impact of soil Cd pollution on potential dietary toxicity.

    PubMed

    Rafiq, Muhammad Tariq; Aziz, Rukhsanda; Yang, Xiaoe; Xiao, Wendan; Stoffella, Peter J; Saghir, Aamir; Azam, Muhammad; Li, Tingqiang

    2014-01-01

    Food chain contamination by soil cadmium (Cd) through vegetable consumption poses a threat to human health. Therefore, an understanding is needed on the relationship between the phytoavailability of Cd in soils and its uptake in edible tissues of vegetables. The purpose of this study was to establish soil Cd thresholds of representative Chinese soils based on dietary toxicity to humans and develop a model to evaluate the phytoavailability of Cd to Pak choi (Brassica chinensis L.) based on soil properties. Mehlich-3 extractable Cd thresholds were more suitable for Stagnic Anthrosols, Calcareous, Ustic Cambosols, Typic Haplustalfs, Udic Ferrisols and Periudic Argosols with values of 0.30, 0.25, 0.18, 0.16, 0.15 and 0.03 mg kg-1, respectively, while total Cd is adequate threshold for Mollisols with a value of 0.86 mg kg-1. A stepwise regression model indicated that Cd phytoavailability to Pak choi was significantly influenced by soil pH, organic matter, total Zinc and Cd concentrations in soil. Therefore, since Cd accumulation in Pak choi varied with soil characteristics, they should be considered while assessing the environmental quality of soils to ensure the hygienically safe food production.

  6. Experimental investigation and modelling of tritium washout by precipitation in the area of the nuclear power plant of Paks, Hungary.

    PubMed

    Köllo, Z; Palcsu, L; Major, Z; Papp, L; Molnár, M; Ranga, T; Dombóvári, P; Manga, L

    2011-01-01

    Tritium occurs in nature in trace amounts, but its concentration is changing due to natural and artificial sources. Studies focusing on natural tritium have to take into account the effect of artificial sources. Also, the impact of tritium is an important issue in environmental protection, e.g. in connection with the emissions from nuclear power plants. The present work focuses on the rain washout of tritium emitted from the Paks nuclear power plant in Hungary. Rainwater collectors were placed around the plant and after a period of precipitation, rainwater was collected and analysed for tritium content. Samples were analysed using low-level liquid scintillation counting, with some also subject to the more accurate (3)He ingrowth method. The results clearly show the trace of the tritium plume emitted from the plant; however, values are only about one order of magnitude higher than environmental background levels. A washout model was devised to estimate the distribution of tritium around the plant. The model gives slightly higher concentrations than those measured in the field, but in general the agreement is satisfactory. The modelled values demonstrate that the effect of the plant on rainwater tritium levels is negligible over a distance of some kilometres.

  7. Separation properties of aluminium-plastic laminates in post-consumer Tetra Pak with mixed organic solvent.

    PubMed

    Zhang, S F; Zhang, L L; Luo, K; Sun, Z X; Mei, X X

    2014-04-01

    The separation properties of the aluminium-plastic laminates in postconsumer Tetra Pak structure were studied in this present work. The organic solvent blend of benzene-ethyl alcohol-water was used as the separation reagent. Then triangle coordinate figure analysis was taken to optimize the volume proportion of various components in the separating agent and separation process. And the separation temperature of aluminium-plastic laminates was determined by the separation time, efficiency, and total mass loss of products. The results show that cost-efficient separations perform best with low usage of solvents at certain temperatures, for certain times, and within a certain range of volume proportions of the three components in the solvent agent. It is also found that similar solubility parameters of solvents and polyethylene adhesives (range 26.06-34.85) are a key factor for the separation of the aluminium-plastic laminates. Such multisolvent processes based on the combined-system concept will be vital to applications in the recycling industry.

  8. An approach to Moho discontinuity recovery from on-orbit GOCE data with application over Indo-Pak region

    NASA Astrophysics Data System (ADS)

    Eshagh, Mehdi; Hussain, Matloob

    2016-10-01

    In this research, a modified form of Vening Meinesz-Moritz (VMM) theory of isostasy for the second-order radial derivative of gravitational potential, measured from the Gravity field and steady-state Ocean Circulation Explorer (GOCE), is developed for local Moho depth recovery. An integral equation is organised for inverting the GOCE data to compute a Moho model in combination with topographic/bathymetric heights of SRTM30, sediment and consolidated crystalline basement and the laterally-varying density contrast model of CRUST1.0. A Moho model from EGM2008 to degree and order 180 is also computed based on the same principle for the purpose of comparison. In addition, we compare both of them with the 3 available seismic Moho models; two global and one regional over the Indo-Pak region. Numerical results show that our GOCE-based Moho model is closer to the all seismic models than that of EGM2008. The model is closest to the regional one with a standard deviation of 5.5 km and a root mean squares error of 7.8 km, which is 2.3 km smaller than the corresponding one based on EGM2008.

  9. The role of remote and regional boundary forcing in the evolution of 2010 summer monsoon heavy rainfall over northwest Indo-Pak region

    NASA Astrophysics Data System (ADS)

    Priya, Pattancheri; Mujumdar, Milind; P, Sabin T.; Pascal, Terray; Krishnan, Raghavan

    2015-04-01

    The evolution of sub-tropical south Asian heavy rainfall during 2010 summer and its relationship with tropical boundary forcing, is diagnosed in this study. Interestingly, the summer time Indo-Pacific SST during 2010 evolved as a combination of three dominant modes of variability. In addition to the strongest La Niña event on long-term record, warming over tropical Indian Ocean and a negative phase of the Indian Ocean Dipole (IOD) phenomenon were also prominent during 2010 boreal summer. A high resolution AGCM is used in this study to distinguish the role of remote and regional tropical SST boundary forcing on the heavy rainfall over northwest Indo-Pak region. The remote boundary forcing over Pacific seems to induce a westward shift of the large-scale monsoon circulation and significantly weakens the convection over Bay of Bengal, but is not sufficient for a realistic simulation of the flood event during 2010. The intensification of northward moisture transport from Arabian Sea into the subtropical Indo-Pak region leading to positive rainfall anomalies as observed in 2010 could be attributed to regional boundary forcing over Indian Ocean. The warmer SST anomalies over southeast Indian Ocean significantly strengthen the cyclonic convergence (divergence) over the region (Bay of Bengal and central and western Indian Ocean). Furthermore, divergence over equatorial Indian Ocean together with positive SST anomalies over north Arabian Sea enhances regional convection and promotes northward moisture transport, which is important for explaining the rainfall anomalies over northwest Indo-Pak region during 2010.

  10. Will the Need-Based Planning of Health Human Resources Currently Undertaken in Several Countries Lead to Excess Supply and Inefficiency? A Comment on Basu and Pak.

    PubMed

    Birch, Stephen; Tomblin Murphy, Gail; MacKenzie, Adrian; Whittaker, William; Mason, Thomas

    2016-06-13

    Basu and Pak (2014) argue that need-based workforce planning models would not maximize social welfare, and use of need-based models would result in inefficiency. They propose that planning be based on service utilization to incorporate preferences or other socioeconomic factors. We show that the analysis is based on inappropriate considerations of the nature of healthcare demand, a misrepresentation of need-based approaches and misunderstanding publicly funded healthcare system objectives. We explain how current levels of utilization emerge from workload and income interests of providers that underlie utilization-based models and are incompatible with public goals of maximizing health gains. Copyright © 2016 John Wiley & Sons, Ltd.

  11. Induced production of 1-methoxy-indol-3-ylmethyl glucosinolate by jasmonic acid and methyl jasmonate in sprouts and leaves of pak choi (Brassica rapa ssp. chinensis).

    PubMed

    Wiesner, Melanie; Hanschen, Franziska S; Schreiner, Monika; Glatt, Hansruedi; Zrenner, Rita

    2013-07-18

    Pak choi plants (Brassica rapa ssp. chinensis) were treated with different signaling molecules methyl jasmonate, jasmonic acid, linolenic acid, and methyl salicylate and were analyzed for specific changes in their glucosinolate profile. Glucosinolate levels were quantified using HPLC-DAD-UV, with focus on induction of indole glucosinolates and special emphasis on 1-methoxy-indol-3-ylmethyl glucosinolate. Furthermore, the effects of the different signaling molecules on indole glucosinolate accumulation were analyzed on the level of gene expression using semi-quantitative realtime RT-PCR of selected genes. The treatments with signaling molecules were performed on sprouts and mature leaves to determine ontogenetic differences in glucosinolate accumulation and related gene expression. The highest increase of indole glucosinolate levels, with considerable enhancement of the 1-methoxy-indol-3-ylmethyl glucosinolate content, was achieved with treatments of sprouts and mature leaves with methyl jasmonate and jasmonic acid. This increase was accompanied by increased expression of genes putatively involved in the indole glucosinolate biosynthetic pathway. The high levels of indole glucosinolates enabled the plant to preferentially produce the respective breakdown products after tissue damage. Thus, pak choi plants treated with methyl jasmonate or jasmonic acid, are a valuable tool to analyze the specific protection functions of 1-methoxy-indole-3-carbinole in the plants defense strategy in the future.

  12. BET Bromodomain Suppression Inhibits VEGF-induced Angiogenesis and Vascular Permeability by Blocking VEGFR2-mediated Activation of PAK1 and eNOS

    PubMed Central

    Huang, Mingcheng; Qiu, Qian; Xiao, Youjun; Zeng, Shan; Zhan, Mingying; Shi, Maohua; Zou, Yaoyao; Ye, Yujin; Liang, Liuqin; Yang, Xiuyan; Xu, Hanshi

    2016-01-01

    The tyrosine kinase receptor vascular endothelial growth factor receptor 2 (VEGFR2) is a critical modulator of angiogenesis. Increasing evidence indicate the important role of bromodomain and extra-terminal domain (BET) of chromatin adaptors in regulating tumor growth and inflammatory response. However, whether BET proteins have a role in angiogenesis and endothelial permeability is unclear. In this study, we observed that treatment with JQ1, a specific BET inhibitor, suppressed in vitro tube formation of human umbilical vein endothelial cells (HUVECs) and in vivo angiogenesis in a Matrigel plug and oxygen-induced retinopathy neovascularization. JQ1 attenuated the VEGF-induced decrease in TEER in HUVECs and prevented Evans blue dye leakage in the VEGF-induced Miles assay in athymic Balb/c nude mice. BET inhibition with JQ1 or shRNA for Brd2 or Brd4 suppressed VEGF-induced migration, proliferation, and stress fiber formation of HUVECs. Furthermore, BET inhibition suppressed phosphorylation of VEGFR2 and PAK1, as well as eNOS activation in VEGF-stimulated HUVECs. Inhibition with VEGFR2 and PAK1 also reduced migration and proliferation, and attenuated the VEGF-induced decrease in TEER. Thus, our observations suggest the important role of BET bromodomain in regulating VEGF-induced angiogenesis. Strategies that target the BET bromodomain may provide a new therapeutic approach for angiogenesis-related diseases. PMID:27044328

  13. Comparison of CampyPak II with standard 5% oxygen and candle jars for growth of Campylobacter jejuni from human feces.

    PubMed

    Wang, W L; Luechtefeld, N W; Blaser, M J; Reller, L B

    1982-08-01

    To determine optimal temperature and atmospheric conditions for isolating Campylobacter jejuni from fecal specimens of humans, we studied six laboratory isolates and 19 fecal specimens that were known to contain C. jejuni. We compared incubations in 5% oxygen, the CampyPak II (BBL Microbiology Systems, Cockeysville, Md.) with 6 plates per jar (CP-6) and 12 plates per jar (CP-12), and candle jars at 37 and 42 degrees C. At both temperatures, the colony sizes for the laboratory strains were larger in the 5% O2 and the CP-6 than under the other two conditions. For the primary isolations, CP-12 failed to detect one and two campylobacters at 42 and 37 degrees C, respectively, whereas the candle jar failed to detect one at 42 degrees C and four at 37 degrees C. Colony size was again larger in the 5% O2 and the CP-6. For all four atmospheric conditions tested, colonies were significantly larger at 42 degrees C than at 37 degrees C. These studies showed that incubation at 42 degrees C in either 5% O2 or the CampyPak II with six plates per jar was optimal for primary isolation of C. jejuni from fecal specimens of humans. The candle jars incubated at 42 degrees C appeared to be satisfactory for primary isolation of C. jejuni from human feces, but incubation at 37 degrees C was not acceptable.

  14. Mini-Pak Fencing.

    ERIC Educational Resources Information Center

    Stucky, Franklin; And Others

    The document presents unit plans which offer lists of experiences and competencies to be learned in the construction, planning, and building of fences. The lessons are comprised of seven areas: (1) planning the location and arrangement of fences, (2) fencing and fence building equipment, (3) types of fences, (4) constructing fences, (5) end and…

  15. A protein related to p21-activated kinase (PAK) that is involved in neurogenesis in the Drosophila adult central nervous system.

    PubMed

    Melzig, J; Rein, K H; Schäfer, U; Pfister, H; Jäckle, H; Heisenberg, M; Raabe, T

    1998-11-05

    Brains are organized by the developmental processes generating them. The embryonic neurogenic phase of Drosophila melanogaster has been studied in detail at the genetic, cellular and molecular level. In contrast, much of what is known of postembryonic brain development has been gathered by neuroanatomical and gene expression studies. The molecular mechanisms underlying cellular diversity and structural organisation in the adult brain, such as the establishment of the correct neuroblast number, the spatial and temporal control of neuroblast proliferation, cell fate determination, and the generation of the precise pattern of neuronal connectivity, are largely unknown. In a screen for viable mutations affecting adult central brain structures, we isolated the mushroom bodies tiny (mbt) gene of Drosophila, which encodes a protein related to p21-activated kinase (PAK). We show that mutations in mbt primarily interfere with the generation or survival of the intrinsic cells (Kenyon cells) of the mushroom body, a paired neuropil structure in the adult brain involved in learning and memory.

  16. Paxillin LD4 motif binds PAK and PIX through a novel 95-kD ankyrin repeat, ARF-GAP protein: A role in cytoskeletal remodeling.

    PubMed

    Turner, C E; Brown, M C; Perrotta, J A; Riedy, M C; Nikolopoulos, S N; McDonald, A R; Bagrodia, S; Thomas, S; Leventhal, P S

    1999-05-17

    Paxillin is a focal adhesion adaptor protein involved in the integration of growth factor- and adhesion-mediated signal transduction pathways. Repeats of a leucine-rich sequence named paxillin LD motifs (Brown M.C., M.S. Curtis, and C.E. Turner. 1998. Nature Struct. Biol. 5:677-678) have been implicated in paxillin binding to focal adhesion kinase (FAK) and vinculin. Here we demonstrate that the individual paxillin LD motifs function as discrete and selective protein binding interfaces. A novel scaffolding function is described for paxillin LD4 in the binding of a complex of proteins containing active p21 GTPase-activated kinase (PAK), Nck, and the guanine nucleotide exchange factor, PIX. The association of this complex with paxillin is mediated by a new 95-kD protein, p95PKL (paxillin-kinase linker), which binds directly to paxillin LD4 and PIX. This protein complex also binds to Hic-5, suggesting a conservation of LD function across the paxillin superfamily. Cloning of p95PKL revealed a multidomain protein containing an NH2-terminal ARF-GAP domain, three ankyrin-like repeats, a potential calcium-binding EF hand, calmodulin-binding IQ motifs, a myosin homology domain, and two paxillin-binding subdomains (PBS). Green fluorescent protein- (GFP-) tagged p95PKL localized to focal adhesions/complexes in CHO.K1 cells. Overexpression in neuroblastoma cells of a paxillin LD4 deletion mutant inhibited lamellipodia formation in response to insulin-like growth fac- tor-1. Microinjection of GST-LD4 into NIH3T3 cells significantly decreased cell migration into a wound. These data implicate paxillin as a mediator of p21 GTPase-regulated actin cytoskeletal reorganization through the recruitment to nascent focal adhesion structures of an active PAK/PIX complex potentially via interactions with p95PKL.

  17. Characterization and Comparative Genomic Analyses of Pseudomonas aeruginosa Phage PaoP5: New Members Assigned to PAK_P1-like Viruses

    PubMed Central

    Shen, Mengyu; Le, Shuai; Jin, Xiaolin; Li, Gang; Tan, Yinling; Li, Ming; Zhao, Xia; Shen, Wei; Yang, Yuhui; Wang, Jing; Zhu, Hongbin; Li, Shu; Rao, Xiancai; Hu, Fuquan; Lu, Shuguang

    2016-01-01

    As a potential alternative to antibiotics, phages can be used to treat multi-drug resistant bacteria. As such, the biological characteristics of phages should be investigated to utilize them as effective antimicrobial agents. In this study, phage PaoP5, a lytic virus that infects Pseudomonas aeruginosa PAO1, was isolated and genomically characterized. PaoP5 comprises an icosahedral head with an apex diameter of 69 nm and a contractile tail with a length of 120 nm. The PaoP5 genome is a linear dsDNA molecule containing 93,464 base pairs (bp) with 49.51% G + C content of 11 tRNA genes and a 1,200 bp terminal redundancy. A total of 176 protein-coding genes were predicted in the PaoP5 genome. Nine PaoP5 structural proteins were identified. Three hypothetical proteins were determined as structural. Comparative genomic analyses revealed that seven new Pseudomonas phages, namely, PaoP5, K8, C11, vB_PaeM_C2-10_Ab02, vB_PaeM_C2-10_Ab08, vB_PaeM_C2-10_Ab10, and vB_PaeM_C2-10_Ab15, were similar to PAK_P1-like viruses. Phylogenetic and pan-genome analyses suggested that the new phages should be assigned to PAK_P1-like viruses, which possess approximately 100 core genes and 150 accessory genes. This work presents a detailed and comparative analysis of PaoP5 to enhance our understanding of phage biology. PMID:27659070

  18. Monitoring of tritium, 60Co and 137Cs in the vicinity of the warm water outlet of the Paks Nuclear Power Plant, Hungary.

    PubMed

    Janovics, R; Bihari, Á; Papp, L; Dezső, Z; Major, Z; Sárkány, K E; Bujtás, T; Veres, M; Palcsu, L

    2014-02-01

    Danube water, sediment and various aquatic organisms (snail, mussel, predatory and omnivorous fish) were collected upstream (at a background site) and downstream of the outlet of the warm water channel of Paks Nuclear Power Plant. Gamma emitters, tissue free-water tritium (TFWT) and total organically-bound tritium (T-OBT) measurements were performed. A slight contribution of the power plant to the natural tritium background concentration was measured in water samples from the Danube section downstream of the warm water channel. Sediment samples also contained elevated tritium concentrations, along with a detectable amount of (60)Co. In the case of biota samples, TFWT exhibited only a very slight difference compared to the tritium concentration of the Danube water, however, the OBT was higher than the tritium concentration in the Danube, independent of the origin of the samples. The elevated OBT concentration in the mollusc samples downstream of the warm water channel may be attributed to the excess emission from the nuclear power plant. The whole data set obtained was used for dose rate calculations and will be contributed to the development of the ERICA database.

  19. The metastasis suppressor NDRG1 modulates the phosphorylation and nuclear translocation of β-catenin through mechanisms involving FRAT1 and PAK4.

    PubMed

    Jin, Runsen; Liu, Wensheng; Menezes, Sharleen; Yue, Fei; Zheng, Minhua; Kovacevic, Zaklina; Richardson, Des R

    2014-07-15

    N-myc downstream-regulated gene 1 (NDRG1) is a potent metastasis suppressor that has been demonstrated to inhibit the transforming growth factor β (TGF-β)-induced epithelial-to-mesenchymal transition (EMT) by maintaining the cell-membrane localization of E-cadherin and β-catenin in prostate and colon cancer cells. However, the precise molecular mechanism remains unclear. In this investigation, we demonstrate that NDRG1 inhibits the phosphorylation of β-catenin at Ser33/37 and Thr41 and increases the levels of non-phosphorylated β-catenin at the plasma membrane in DU145 prostate cancer cells and HT29 colon cancer cells. The mechanism of inhibiting β-catenin phosphorylation involves the NDRG1-mediated upregulation of the GSK3β-binding protein FRAT1, which prevents the association of GSK3β with the Axin1-APC-CK1 destruction complex and the subsequent phosphorylation of β-catenin. Additionally, NDRG1 is shown to modulate the WNT-β-catenin pathway by inhibiting the nuclear translocation of β-catenin. This is mediated through an NDRG1-dependent reduction in the nuclear localization of p21-activated kinase 4 (PAK4), which is known to act as a transporter for β-catenin nuclear translocation. The current study is the first to elucidate a unique molecular mechanism involved in the NDRG1-dependent regulation of β-catenin phosphorylation and distribution.

  20. Targeted genetic dependency screen facilitates identification of actionable mutations in FGFR4, MAP3K9, and PAK5 in lung cancer.

    PubMed

    Fawdar, Shameem; Trotter, Eleanor W; Li, Yaoyong; Stephenson, Natalie L; Hanke, Franziska; Marusiak, Anna A; Edwards, Zoe C; Ientile, Sara; Waszkowycz, Bohdan; Miller, Crispin J; Brognard, John

    2013-07-23

    Approximately 70% of patients with non-small-cell lung cancer present with late-stage disease and have limited treatment options, so there is a pressing need to develop efficacious targeted therapies for these patients. This remains a major challenge as the underlying genetic causes of ~50% of non-small-cell lung cancers remain unknown. Here we demonstrate that a targeted genetic dependency screen is an efficient approach to identify somatic cancer alterations that are functionally important. By using this approach, we have identified three kinases with gain-of-function mutations in lung cancer, namely FGFR4, MAP3K9, and PAK5. Mutations in these kinases are activating toward the ERK pathway, and targeted depletion of the mutated kinases inhibits proliferation, suppresses constitutive activation of downstream signaling pathways, and results in specific killing of the lung cancer cells. Genomic profiling of patients with lung cancer is ushering in an era of personalized medicine; however, lack of actionable mutations presents a significant hurdle. Our study indicates that targeted genetic dependency screens will be an effective strategy to elucidate somatic variants that are essential for lung cancer cell viability.

  1. An experimentally based computer search identifies unstructured membrane-binding sites in proteins: application to class I myosins, PAKS, and CARMIL.

    PubMed

    Brzeska, Hanna; Guag, Jake; Remmert, Kirsten; Chacko, Susan; Korn, Edward D

    2010-02-19

    Programs exist for searching protein sequences for potential membrane-penetrating segments (hydrophobic regions) and for lipid-binding sites with highly defined tertiary structures, such as PH, FERM, C2, ENTH, and other domains. However, a rapidly growing number of membrane-associated proteins (including cytoskeletal proteins, kinases, GTP-binding proteins, and their effectors) bind lipids through less structured regions. Here, we describe the development and testing of a simple computer search program that identifies unstructured potential membrane-binding sites. Initially, we found that both basic and hydrophobic amino acids, irrespective of sequence, contribute to the binding to acidic phospholipid vesicles of synthetic peptides that correspond to the putative membrane-binding domains of Acanthamoeba class I myosins. Based on these results, we modified a hydrophobicity scale giving Arg- and Lys-positive, rather than negative, values. Using this basic and hydrophobic scale with a standard search algorithm, we successfully identified previously determined unstructured membrane-binding sites in all 16 proteins tested. Importantly, basic and hydrophobic searches identified previously unknown potential membrane-binding sites in class I myosins, PAKs and CARMIL (capping protein, Arp2/3, myosin I linker; a membrane-associated cytoskeletal scaffold protein), and synthetic peptides and protein domains containing these newly identified sites bound to acidic phospholipids in vitro.

  2. Growth hormone-releasing hormone receptor antagonists inhibit human gastric cancer through downregulation of PAK1–STAT3/NF-κB signaling

    PubMed Central

    Gan, Jinfeng; Ke, Xiurong; Jiang, Jiali; Dong, Hongmei; Yao, Zhimeng; Lin, Yusheng; Lin, Wan; Wu, Xiao; Yan, Shumei; Zhuang, Yixuan; Chu, Wai Kit; Cai, Renzhi; Zhang, Xianyang; Cheung, Herman S.; Block, Norman L.; Pang, Chi Pui; Schally, Andrew V.; Zhang, Hao

    2016-01-01

    Gastric cancer (GC) ranks as the fourth most frequent in incidence and second in mortality among all cancers worldwide. The development of effective treatment approaches is an urgent requirement. Growth hormone-releasing hormone (GHRH) and GHRH receptor (GHRH-R) have been found to be present in a variety of tumoral tissues and cell lines. Therefore the inhibition of GHRH-R was proposed as a promising approach for the treatment of these cancers. However, little is known about GHRH-R and the relevant therapy in human GC. By survival analyses of multiple cohorts of GC patients, we identified that increased GHRH-R in tumor specimens correlates with poor survival and is an independent predictor of patient prognosis. We next showed that MIA-602, a highly potent GHRH-R antagonist, effectively inhibited GC growth in cultured cells. Further, this inhibitory effect was verified in multiple models of human GC cell lines xenografted into nude mice. Mechanistically, GHRH-R antagonists target GHRH-R and down-regulate the p21-activated kinase 1 (PAK1)-mediated signal transducer and activator of transcription 3 (STAT3)/nuclear factor-κB (NF-κB) inflammatory pathway. Overall, our studies establish GHRH-R as a potential molecular target in human GC and suggest treatment with GHRH-R antagonist as a promising therapeutic intervention for this cancer. PMID:27930339

  3. Update of the tectonic model for the Pannonian basin: a contribution to the seismic hazard reassessment of the Paks NPP (Hungary)

    NASA Astrophysics Data System (ADS)

    Horváth, Ferenc; Tóth, Tamás; Wórum, Géza; Koroknai, Balázs; Kádi, Zoltán; Kovács, Gábor; Balázs, Attila; Visnovitz, Ferenc

    2015-04-01

    The planned construction of two new units at the site of the Paks NPP requires a comprehensive site investigation including complete reassessment of the seismic hazard according to the Hungarian as well as international standards. Following the regulations of the Specific Safety Guide no. 9 (IAEA 2010), the approved Hungarian Geological Investigation Program (HGIP) includes integrated geological-geophysical studies at different scales. The regional study aims at to elaborate a new synthesis of all published data for the whole Pannonian basin. This task is nearly completed and the main outcomes have already been published (Horváth et al. 2015). The near regional study is in progress and addresses the construction of a new tectonic model for the circular area with 50 km radius around the NPP using a wealth of unpublished oil company seismic and borehole data. The site vicinity study has also been started with a core activity of 300 km² 3D seismic data acquisition, processing and interpretation assisted by a series of additional geophysical surveys, new drillings and geological mapping. This lecture will present a few important results of the near regional study, which sheds new light on the intricate tectonic evolution of the Mid-Hungarian Fault Zone (MHFZ), which is a strongly deformed belt between the Alcapa and Tisza-Dacia megatectonic units. The nuclear power plant is located at the margin of the Tisza unit near to the southern edge of the MHFZ. Reassessment of seismic hazard at the site of the NPP requires better understanding of the Miocene to Recent tectonic evolution of this region in the central part of the Pannonian basin. Early to Middle Miocene was a period of rifting with formation of 1 to 3 km deep half-grabens filled with terrestrial to marine deposits and large amount of rift-related volcanic material. Graben fill became strongly deformed as a consequence of juxtaposition of the two megatectonic units leading to strong compression and development of

  4. Documentation and Control of Flow Separation on a Low Pressure Turbine Linear Cascade of Pak-B Blades Using Plasma Actuators

    NASA Technical Reports Server (NTRS)

    Corke, Thomas c.; Thomas, FLint, O.; Huang, Junhui

    2007-01-01

    This work involved the documentation and control of flow separation that occurs over low pressure turbine (LPT) blades at low Reynolds numbers. A specially constructed linear cascade was utilized to study the flow field over a generic LPT cascade consisting of Pratt & Whitney "Pak-B" shaped blades. Flow visualization, surface pressure measurements, LDV measurements, and hot-wire anemometry were conducted to examine the flow fields with and without separation control. Experimental conditions were chosen to give a range of chord Reynolds numbers (based on axial chord and inlet velocity) from 10,000 to 100,000, and a range of freestream turbulence intensities from u'/U(infinity) = 0.08 to 2.85 percent. The blade pressure distributions were measured and used to identify the region of separation that depends on Reynolds number and the turbulence intensity. Separation control was performed using dielectric barrier discharge (DBD) plasma actuators. Both steady and unsteady actuation were implemented and found to work well. The comparison between the steady and unsteady actuators showed that the unsteady actuators worked better than the steady ones. For the steady actuators, it was found that the separated region is significantly reduced. For the unsteady actuators, where the signal was pulsed, the separation was eliminated. The total pressure losses (a low Reynolds number) was reduced by approximately a factor of two. It was also found that lowest plasma duty cycle (10 percent in this work) was as effective as the highest plasma duty cycle (50 percent in this work). The mechanisms of the steady and unsteady plasma actuators were studied. It was suggested by the experimental results that the mechanism for the steady actuators is turbulence tripping, while the mechanism for the unsteady actuators is to generate a train of spanwise structures that promote mixing.

  5. From Rocks to Pots. [Project ECOLogy ELE Pak, Grim Pak].

    ERIC Educational Resources Information Center

    Grim, Dale

    This is one of a series of units for environmental education developed by the Highline Public Schools. The unit is designed for use by art classes at the secondary school level; it illustrates the availability of natural clay and provides the student with experiences such as digging the clay, locating desirable clays, preparing it for production,…

  6. Natural or Organic Foods? [Project ECOLogy ELE Pak, Schmidt Pak].

    ERIC Educational Resources Information Center

    Schmidt, Linda

    This is one of a series of units for environmental education developed by the Highline Public Schools. The unit is designed for secondary students in home economics classes. The content of the units focuses on natural and organic foods, characteristics of the foods, and uses of the foods. The seven lessons in this unit are designed to last over a…

  7. Prolactin-Stimulated Activation of ERK1/2 Mitogen-Activated Protein Kinases is Controlled by PI3-Kinase/Rac/PAK Signaling Pathway in Breast Cancer Cells

    PubMed Central

    Aksamitiene, Edita; Achanta, Sirisha; Kolch, Walter; Kholodenko, Boris N.; Hoek, Jan B.; Kiyatkin, Anatoly

    2011-01-01

    There is strong evidence that deregulation of prolactin (PRL) signaling contributes to pathogenesis and chemoresistance of breast cancer. Therefore, understanding cross-talk between distinct signal transduction pathways triggered by activation of the prolactin receptor (PRL-R), is essential for elucidating the pathogenesis of metastatic breast cancer. In this study, we applied a sequential inhibitory analysis of various signaling intermediates to examine the hierarchy of protein interactions within the PRL signaling network and to evaluate the relative contributions of multiple signaling branches downstream of PRL-R to the activation of the extracellular signal-regulated kinases ERK1 and ERK2 in T47D and MCF-7 human breast cancer cells. Quantitative measurements of the phosphorylation/activation patterns of proteins showed that PRL simultaneously activated Src family kinases (SFKs) and the JAK/STAT, phosphoinositide-3 (PI3)-kinase/Akt and MAPK signaling pathways. The specific blockade or siRNA-mediated suppression of SFK/FAK, JAK2/STAT5, PI3-kinase/PDK1/Akt, Rac/PAK or Ras regulatory circuits revealed that (1) the PI3-kinase/Akt pathway is required for activation of the MAPK/ERK signaling cascade upon PRL stimulation; (2) PI3-kinase-mediated activation of the c-Raf-MEK1/2-ERK1/2 cascade occurs independent of signaling dowstream of STATs, Akt and PKC, but requires JAK2, SFKs and FAK activities; (3) activated PRL-R mainly utilizes the PI3-kinase-dependent Rac/PAK pathway rather than the canonical Shc/Grb2/SOS/Ras route to initiate and sustain ERK1/2 signaling. By interconnecting diverse signaling pathways PLR may enhance proliferation, survival, migration and invasiveness of breast cancer cells. PMID:21726627

  8. Hemopoietic cell kinase (Hck) and p21-activated kinase 2 (PAK2) are involved in the down-regulation of CD1a lipid antigen presentation by HIV-1 Nef in dendritic cells.

    PubMed

    Shinya, Eiji; Shimizu, Masumi; Owaki, Atsuko; Paoletti, Samantha; Mori, Lucia; De Libero, Gennaro; Takahashi, Hidemi

    2016-01-01

    Dendritic cells (DCs) play a major role in in vivo pathogenesis of HIV-1 infection. Therefore, DCs may provide a promising strategy to control and eventually overcome the fatal infection. Especially, immature DCs express all CD1s, the non-MHC lipid antigen -presenting molecules, and HIV-1 Nef down-regulates CD1 expression besides MHC. Moreover, CD1d-restricted CD4(+) NKT cells are infected by HIV-1, reducing the number of these cells in HIV-1-infected individuals. To understand the exact role of DCs and CD1-mediated immune response during HIV-1 infection, Nef down-regulation of CD1a-restricted lipid/glycolipid Ag presentation in iDCs was analyzed. We demonstrated the involvement of the association of Nef with hemopoietic cell kinase (Hck) and p21-activated kinase 2 (PAK2), and that Hck, which is expressed strongly in iDCs, augmented this mutual interaction. Hck might be another therapeutic target to preserve the function of HIV-1 infected DCs, which are potential reservoirs of HIV-1 even after antiretroviral therapy.

  9. Untersuchungen zur Sorptionsreversibilität von organischen Schadstoffen in Aktivkohle, Holzkohle und Zeolith Y-200

    NASA Astrophysics Data System (ADS)

    ElHaddad, Engy; Ensinger, Wolfgang; Schüth, Christoph

    2013-09-01

    Numerous studies have shown that sorption of organic contaminants in soils is dominated by the natural organic carbon content ( C org) of the soil. However, it is still under discussion whether sorption processes are fully reversible or whether an irreversibly sorbed contaminant fraction remains in the soil. This is especially important when considering soil remediation measures and its targets. In multi-stage sorption-desorption batch experiments with TCE, PCE, ortho-xylene and para-xylene and with the sorbents activated carbon, charcoal and a hydrophobic zeolite Y-200, the reversibility of sorption was studied. It could be shown that the structural features of the sorbents are of ample importance for the occurrence of a desorption-resistant fraction. While sorption was mainly reversible for the micro-porous zeolite Y-200 with a rigid pore network, charcoal and the activated carbon showed significant desorption hysteresis. However, following a subsequent sorption step, this fraction eventually desorbs and is re-mobilized.

  10. ...About This Problem of Air Pollution... . [Project ECOLogy ELE Pak, Wright Pak].

    ERIC Educational Resources Information Center

    Wright, Jan

    This is one of a series of units for environmental education developed by the Highline Public Schools. The lessons in this unit are designed to help students discover causes, effects, and results of air pollution through involvement in various activities; it is recommended for intermediate grade elementary school pupils. The unit can be used…

  11. Eco-Kids: Experiment with Air on Spaceship Earth. [Project ECOLogy ELE Pak, Bell Pak].

    ERIC Educational Resources Information Center

    Bell, Loretta

    This is one of a series of units for environmental education developed by the Highline Public Schools. The unit, designed for intermediate grades in the elementary schools, is concerned with the study of air, air pollution, effects of air pollution, and ways of improving the quality of the air. Six lessons are included in the unit; most of the…

  12. Eco-Kids Fly Off to the Forests. [Project ECOLogy ELE Pak, Bell Pak].

    ERIC Educational Resources Information Center

    Bell, Loretta

    This is one of a series of units for environmental education developed by the Highline Public Schools. This unit was designed for use with fourth-grade students; it focuses on three forest biomes. Each of the biomes has characteristics of its own. The unit includes eight lessons, as well as additional activities. The unit, which incorporates…

  13. Exploration with Garbage. [Project ECOLogy ELE Pak, Lund and Wolff Pak].

    ERIC Educational Resources Information Center

    Lund, Cherie; Wolff, Chanelle

    This is one of a series of units for environmental education developed by the Highline Public Schools. This unit is concerned with the topic of garbage. The eleven lessons explore what garbage is, problems of littering, ways to reduce garbage, and ways to use garbage. The materials were designed to be used with kindergarten pupils, but could be…

  14. Food: The Challenge to Manage. [Project ECOLogy ELE Pak, Roush Pak].

    ERIC Educational Resources Information Center

    Roush, Judy

    This is one of a series of units for environmental education developed by the Highline Public Schools. The unit is designed for students at the senior high level who have a basic knowledge of nutrition, some experience in menu planning, and who are ready to put this knowledge of nutrition to work in selecting foods to attain maximum nutrition with…

  15. This Land Is Your Land. [Project ECOLogy ELE Pak, Weber Pak].

    ERIC Educational Resources Information Center

    Weber, Lee

    This is one of a series of units for environmental education developed by the Highline Public Schools. The unit has been constructed for use by intermediate grade elementary school pupils. The seven lessons are designed to inform students about the land to develop a land ethic. The unit should be able to be completed in two to three weeks. The…

  16. The Teacher's Planning Pak and Guide to Individualized Instruction.

    ERIC Educational Resources Information Center

    Forte, Imogene; MacKenzie, Joy

    This book is for both elementary school teachers and students. Suggestions are provided for the teacher on ways to organize time, space, and materials with the emphasis on individualized instruction for the pupils. Materials for the students include activities and small questionnaires structured to reveal to the teacher each child's attitudes,…

  17. OPTIMIZING THE PAKS METHOD FOR MEASURING AIRBORNE ACROLEIN

    EPA Science Inventory

    Airborne acrolein is produced from the combustion of fuel and tobacco and is of concern due to its potential for respiratory tract irritation and other adverse health effects. DNPH active-sampling is a method widely used for sampling airborne aldehydes and ketones (carbonyls); ...

  18. Ever Stop to Think Man's Survival Is Dependent on His Use of Food Resources? [Project ECOLogy ELE Pak, Nelson Pak].

    ERIC Educational Resources Information Center

    Nelson, Judy

    This is one of a series of units for environmental education developed by the Highline Public Schools. This unit is designed for senior high school students who have a basic knowledge of nutrition and some experience in menu planning. The five lessons provide experiences in selecting, preparing, and storing foods to attain maximum nutrition with a…

  19. The Pak-U.S. Alliance in the Fight Against Terrorism: A Cost-Benefit Analysis

    DTIC Science & Technology

    2011-12-01

    97 3. Cultural and Environmental Blocks ................................................97 F. CHANGING PERCEPTIONS THROUGH MICRO...bringing substantial changes . Each sector and sub-sector should have centralized accountability at the USAID level. Besides development, it is evident...are able to promote commonalities, understand each other‘s limitations, change their policies and approach the issues with converging interests

  20. Interaction Between a Novel p21 Activated Kinase (PAK6) and Androgen Receptor in Prostate Cancer

    DTIC Science & Technology

    2005-02-01

    not altered by . 2o treatment with DHT (data not shown). Overexpressed 3-cate- nin protein with AR vector, in the absence of DHT, showed the same...Taken together, we due primarily to loss or decreased expression E-cadherin, is conclude that overexpression of E-cadherin in TSU.pr-1 in- frequently...such as glycogen synthase of AR activity by LY294002 is mediated through phos- kinase (GSK3), Bad, and caspase9 and the forkhead transcrip

  1. UTILIZING THE PAKS METHOD FOR MEASURING ACROLEIN AND OTHER ALDEHYDES IN DEARS

    EPA Science Inventory

    Acrolein is a hazardous air pollutant of high priority due to its high irritation potency and other potential adverse health effects. However, a reliable method is currently unavailable for measuring airborne acrolein at typical environmental levels. In the Detroit Exposure and A...

  2. PAKS: Parents-and-Kids Science. 24 Activities for Kids and Adults To Share.

    ERIC Educational Resources Information Center

    McKenzie, Danny L.

    This activity book designed for grades 1-3 provides teachers with ready-to-use materials designed to get parents and children excited about science, help establish a home-school connection, and provide interesting learning activities for children to share with adults. This program gets parents involved in developing their children's science…

  3. Relationship Between Pak-Mediated Cell Death and Stress-Activated Kinase Signaling Breast Cancer

    DTIC Science & Technology

    2001-02-01

    Jurkat: T-lymphoblast cell line; HeLa: human cervical carcinoma; CHO : chinese hamster ovary ; ZR75, MDA23 1, SKBR-3: human breast cancer cell lines 16 RhoA...activation in Jurkat cells . J Immunol 1998 Jan 1; 160(1):7-1 1 12 Genomic Locus of GEF/H1/KIAA0651 GI 11427616: 870353 864048 861008 858631 858224...1: Schematic representation of the genomic locus of GEF-H1i/KIAA0651 as deduced from the working draft sequence of the GI 11427616 contig derived from

  4. Satellite based classification (haze, fog) and affected area estimation over Indo - Pak Sub-Continent

    NASA Astrophysics Data System (ADS)

    Ghauri, Badar; Zafar, Sumaira

    2016-07-01

    Northern Pakistan and bordering Indian Punjab experience intense smog and fog during fall and winters. Environmentalists have been raising their voices over the situation and demanded control over regional emissions to save the livelihood of millions of dwellers whose trade, commerce and agriculture is at stake because of long smog/ fog spells.. This paper estimates the area affected by haze, smog and fog during 2006- 2010. MODIS (geo-referenced MODIS subsets India1, 2 &3) of the area in Pakistan and India from 2006 to 2010 for the period October to February) were analyzed using state of the art software ENVI 4.2 and ArcGIS 10.2. This process resulted in area belonging to each class that is; haze, smog and fog. On the basis of density, haze and fog cover was determined. Variations in fog cover, its density and identification of location of fog initiation process were also determined using near real time (30 minutes) METEOSAT-7 IODC data where actually fog formation started and then extended to the area of favorable conditions. Haze has been noticed to intensify due to massive burning of agricultural waste (rice husk) in India and Pakistan towards the end of October each year. MODIS thermal anomalies/fire data (MYD 14) were also used to verify this activity on the ground, which results in hazy conditions at regional level during fall months. Haze-affected area during 2006 to 2010 in Pakistan ranged from 155,000 Km2 to 354,000 Km2 and in India it ranged from 333,000 Km2 to 846,000 Km2. Similarly winter fog cover during this period in Pakistan varied from 136,000 Km2 to 381,000 Km2 and in India it was estimated at 327,000 Km2 to 566,000 Km2. This phenomenon was more prominent in India than in Pakistan where and fog cover was at least twice than that was observed in Pakistan. It has been noted that area covered by fog, smog and haze doubled during the study period in the region. Atmospheric dimming during autumn/ fall also reduces the mixing height leading to greater pollutants accumulation. So far no mitigation steps have been taken to combat this regional issue. Reduction in local emissions is highly recommended to save at least the lives of vulnerable (children, elderly, patients etc).

  5. Prevalence of Leishmania tropica in school boys of khyber agency, FATA near Pak-Afghan border.

    PubMed

    Qureshi, Naveeda Akhtar; Ali, Abid; Rashid, Umer; Tayyab-Ur-Rehman; Ali, Naeem

    2016-12-01

    In Pakistan leishmaniasis occurs periodically throughout the year and various out breaks are reported frequently. In continuation of our research on this neglected disease, the aim of present study is to explore: (1) the prevalence of cutaneous leishmaniasis in school boys; (2) Leishmania species identification in order to epidemiology and dynamics of the disease; (3) Identification of risk factors for Leishmaniasis especially for CL. The data was collected in August 2014 The experimental strategy involved a questionnaire for data collection and along with clinical diagnosis of 134 out of 9368 students for incidence of Leishmania spp. in 7 square kilometres area in the schoolboys at Tehsil Landi Kotal, District Khyber Agency, FATA Pakistan. The parasitological and molecular diagnosis of clinically suspected cutaneous leishmaniasis cases were performed using microscopical examination of Giemsa-stained smears of lesion exudates and minicircle kDNA semi nested PCR, respectively. Microscopy (x=1000) positive cases were 84/134 (62.6%) and 50/134 (37.4%) slides did not showed any presence of amastigotes of Leishmania spp. The samples were amplified using kDNA semi nested PCR and confirmed the presence of L. Tropica (Ac.no KT 985473). PCR positive cases were 97/134 (72.4%) and 37(27.6%) were negative. The prevalence of L. tropica in school boys was 1.4% (134/9368) in the total population studied (n=9368). The parasite prevalence might be greater as only male students were considered in the study due to ethical and social issues and limitations.

  6. The closure of Pak1-dependent macropinosomes requires the phosphorylation of CtBP1/BARS.

    PubMed

    Liberali, Prisca; Kakkonen, Elina; Turacchio, Gabriele; Valente, Carmen; Spaar, Alexander; Perinetti, Giuseppe; Böckmann, Rainer A; Corda, Daniela; Colanzi, Antonino; Marjomaki, Varpu; Luini, Alberto

    2008-04-09

    Membrane fission is an essential process in membrane trafficking and other cellular functions. While many fissioning and trafficking steps are mediated by the large GTPase dynamin, some fission events are dynamin independent and involve C-terminal-binding protein-1/brefeldinA-ADP ribosylated substrate (CtBP1/BARS). To gain an insight into the molecular mechanisms of CtBP1/BARS in fission, we have studied the role of this protein in macropinocytosis, a dynamin-independent endocytic pathway that can be synchronously activated by growth factors. Here, we show that upon activation of the epidermal growth factor receptor, CtBP1/BARS is (a) translocated to the macropinocytic cup and its surrounding membrane, (b) required for the fission of the macropinocytic cup and (c) phosphorylated on a specific serine that is a substrate for p21-activated kinase, with this phosphorylation being essential for the fission of the macropinocytic cup. Importantly, we also show that CtBP1/BARS is required for macropinocytic internalization and infection of echovirus 1. These results provide an insight into the molecular mechanisms of CtBP1/BARS activation in membrane fissioning, and extend the relevance of CtBP1/BARS-induced fission to human viral infection.

  7. Pattern of fall injuries in Pakistan: the Pakistan National Emergency Department Surveillance (Pak-NEDS) study

    PubMed Central

    2015-01-01

    Background We aimed to analyse the frequency and patterns of fall-related injuries presenting to the emergency departments (EDs) across Pakistan. Methods Pakistan National Emergency Departments surveillance system collected data from November 2010 to March 2011 on a 24/7 basis using a standardized tool in seven major EDs (five public and two private hospitals) in six major cities of Pakistan. For all patients presenting with fall-related injuries, we analysed data by intent with focus on unintentional falls. Simple frequencies were run for basic patient demographics, mechanism of falls, outcomes of fall injuries, mode of arrival to ED, investigations, and procedures with outcomes. Results There were 3335 fall-related injuries. In cases where intent was available, two-thirds (n = 1186, 65.3%) of fall injuries were unintentional. Among unintentional fall patients presenting to EDs, the majority (76.9%) were males and between 15-44 years of age (69%). The majority of the unintentional falls (n = 671, 56.6%) were due to slipping, followed by fall from height (n = 338, 28.5%). About two-thirds (n = 675, 66.6%) of fall injuries involved extremities, followed by head/neck (n = 257, 25.4%) and face (n = 99, 9.8%). Most of the patients were discharged from the hospital (n = 1059, 89.3%). There were 17 (1.3%) deaths among unintentional fall cases. Conclusion Falls are an important cause of injury-related visits to EDs in Pakistan. Most of the fall injury patients were men and in a productive age group. Fall injuries pose a burden on the healthcare system, especially emergency services, and future studies should therefore focus on safety measures at home and in workplaces to reduce this burden. PMID:26691821

  8. Changes in the distributions of juvenile horseshoe crabs (Arthropoda: Chelicerata) (2002-2014) related to environmental perturbations at Pak Nai and Ha Pak Nai, Deep Bay, Hong Kong SAR, China.

    PubMed

    Lee, Christine Nga-Wing; Morton, Brian

    2016-07-15

    A survey of juvenile Asian horseshoe crabs in 2002 on the mudflats along Hong Kong's north-western shoreline abutting Deep Bay identified two species, Tachypleus tridentatus and Carcinoscorpius rotundicauda, and assessed their population characteristics. Since the 2002 survey, there have been significant habitat changes to this natal site for the two species. By employing the same, but expanded, sampling protocol, a further survey was, therefore, conducted twelve years later in 2014. A general population decline was recorded for T. tridentatus whereas for C. rotundicauda there was an increase and its distribution had become more widespread. The distribution patterns of the two species were also shown to have changed. The potential factors that might be responsible for recorded changes in the species' population characteristics between 2002 and 2014 are related to anthropogenic perturbations, including environmental habitat alterations notably the building of a bridge linking Hong Kong to Shenzhen in China.

  9. High performance thin layer chromatographic screening for aflatoxins in poultry feed by using silica Sep-Paks

    SciTech Connect

    Kozloski, R.P.

    1986-06-01

    The need for a convenient, sensitive and highly selective method to test for aflatoxins in poultry feed samples led to the development of a method based on the CB procedure. The CB procedure requires large volumes of solvents, the preparation of silica gel of a specified moisture content and the use of large TLC plates, which have long development times. The desirability of running fortified control samples to monitor aflatoxin recovery and to serve as reference standards doubles the already high volume of solvents required. To overcome these difficulties, a number of changes in the CB method were explored. Sample size reduction was used in an attempt to decrease the volume of solvents required. The practicality of using commercially available silica Sep-Paps was investigated. The use of high performance thin layer chromatographic (HPTLC) plates with preadsorbent zones was tested to decrease spotting and development time. The capability of readily detecting aflatoxins B/sub 1/, G/sub 1/, B/sub 2/ and G/sub 2/ at 10, 10, 5 and 5 ppb (10/sup 9/), respectively, was set as the goal.

  10. Cooperative measures to support the Indo-Pak Agreement Reducing Risk from Accidents Relating to Nuclear Weapons.

    SciTech Connect

    Mishra, Sitakanta; Ahmed, Mansoor

    2014-04-01

    In 2012, India and Pakistan reaffirmed the Agreement on Reducing the Risk from Accidents Relating to Nuclear Weapons. Despite a history of mutual animosity and persistent conflict between the two countries, this agreement derives strength from a few successful nuclear confidence building measures that have stood the test of time. It also rests on the hope that the region would be spared a nuclear holocaust from an accidental nuclear weapon detonation that might be misconstrued as a deliberate use of a weapon by the other side. This study brings together two emerging strategic analysts from South Asia to explore measures to support the Agreement and further develop cooperation around this critical issue. This study briefly dwells upon the strategic landscape of nuclear South Asia with the respective nuclear force management structures, doctrines, and postures of India and Pakistan. It outlines the measures in place for the physical protection and safety of nuclear warheads, nuclear materials, and command and control mechanisms in the two countries, and it goes on to identify the prominent, emerging challenges posed by the introduction of new weapon technologies and modernization of the respective strategic forces. This is followed by an analysis of the agreement itself leading up to a proposed framework for cooperative measures that might enhance the spirit and implementation of the agreement.

  11. A School Improvement-Accountability Process Kit. PAK No. 3.2--Developing Educational Programs: Structures--Objectives--Budgets.

    ERIC Educational Resources Information Center

    Villars, Gerald K.

    The purpose of this Personalized Activity Kit is to provide an introduction to the concept of program budgeting by focusing on several key elements to be considered in the development of a program budgeting system. These elements are an examination of the school's organizational patterns in defining and designing the district's educational…

  12. Group II p21-activated kinases as therapeutic targets in gastrointestinal cancer

    PubMed Central

    Shao, Yang-Guang; Ning, Ke; Li, Feng

    2016-01-01

    P21-activated kinases (PAKs) are central players in various oncogenic signaling pathways. The six PAK family members are classified into group I (PAK1-3) and group II (PAK4-6). Focus is currently shifting from group I PAKs to group II PAKs. Group II PAKs play important roles in many fundamental cellular processes, some of which have particular significance in the development and progression of cancer. Because of their important functions, group II PAKs have become popular potential drug target candidates. However, few group II PAKs inhibitors have been reported, and most do not exhibit satisfactory kinase selectivity and “drug-like” properties. Isoform- and kinase-selective PAK inhibitors remain to be developed. This review describes the biological activities of group II PAKs, the importance of group II PAKs in the development and progression of gastrointestinal cancer, and small-molecule inhibitors of group II PAKs for the treatment of cancer. PMID:26811660

  13. 75 FR 9028 - Quarterly Publication of Individuals, Who Have Chosen To Expatriate, as Required by Section 6039G

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-26

    ... KAREN-HELENA SABRINA HASSAN ABDELMONEM HAYDAY ADRIAN C ] HAYDAY SANDRA ANN HELBIG INGO M. HELD BETTINA... PAHLSSON REX HENRIK PAHR-IVERSEN KAREN-HELENA SABRINA HART PAK CHIN YONG PAK SUN YUL PAK YONG MAE PARK...

  14. Beilstein Without Tears: Education in the Use of the Literature of Organic Chemistry.

    ERIC Educational Resources Information Center

    Callaghan, Patricia M.; And Others

    1986-01-01

    The use of Beilstein ("Handbuch der Organischen Chemie") in the early stages of a second-year, one semester course in organic chemistry is described. Student literature projects, evaluation, use of ancillary literature, and a sample search are included. (JN)

  15. Signaling, Regulation, and Specificity of the Type II p21-activated Kinases*

    PubMed Central

    Ha, Byung Hak; Morse, Elizabeth M.; Turk, Benjamin E.; Boggon, Titus J.

    2015-01-01

    The p21-activated kinases (PAKs) are a family of six serine/threonine kinases that act as key effectors of RHO family GTPases in mammalian cells. PAKs are subdivided into two groups: type I PAKs (PAK1, PAK2, and PAK3) and type II PAKs (PAK4, PAK5, and PAK6). Although these groups are involved in common signaling pathways, recent work indicates that the two groups have distinct modes of regulation and have both unique and common substrates. Here, we review recent insights into the molecular level details that govern regulation of type II PAK signaling. We also consider mechanisms by which signal transduction is regulated at the level of substrate specificity. Finally, we discuss the implications of these studies for clinical targeting of these kinases. PMID:25855792

  16. Multidrug resistant gram-negative bacteria in clinical isolates from Karachi.

    PubMed

    Saeed, Asma; Khatoon, Hajra; Ansari, Fasihuddin Ahmed

    2009-01-01

    A total of 54 gram-negative bacteria obtained from various pathological labs and hospitals of Karachi were screened for their resistance to ampicillin, chloramphenicol, gentamycin, kanamycin, neomycin, streptomycin and tetracycline antibiotics. Of the 54 bacteria, 50 were resistant to one or more antibiotics. Among the resistant bacteria, 13 out of 28 were found to transfer their resistances by conjugation. This indicates that at least 46% of clinical gram-negative bacteria in Karachi possess various types of transferable R plasmids, such as pAK5, pAK9, pAK10, pAK11, pAK12, pAK13, pAK14, pAK15, pAK16, pAK17, pAK18, pAK19, pAK20 and pAK21. The non-conjugative R plasmids included pMT14 and pZ26. Only pAK15 showed 26% segregation even after 20 consecutive transfers in plain broth (spontaneous segregation) whereas only pAK15 and pAK16 showed any significant loss of their markers in curing by acridine orange. The stability of R plasmids is more dangerous from clinical point of view.

  17. Myungshin Im, Won-Kee Park, Changsu Choi, Yiseul Jeon, Ji-Hoon Kim (CEOU/SNU), Giseon Baek, Young-Seok Oh, Soojong Pak (Kyunghee Univ.)

    NASA Astrophysics Data System (ADS)

    Im, Myungshin; Park, Won-Kee; Choi, Changsu; Jeon, Yiseul; Kim, Ji-Hoon; Baek, Giseon; Oh, Young-Seok; Pak, Soojong

    2011-08-01

    On August 30 02:05 UT, we started observations of SN2011fe=PTF11kly (ATel#3581) in grizY and is/iz-bands using CQUEAN on the 2.1m telescope at McDonald observatory, and in zYJHK-bands using UKIRT. SN2011fe is found to have J~11.58 and K~11.55 mag (error~0.05 mag), still brightening in NIR with respect to ATel#3605.

  18. The Botrytis cinerea PAK kinase BcCla4 mediates morphogenesis, growth and cell cycle regulating processes downstream of BcRac.

    PubMed

    Minz-Dub, Anna; Sharon, Amir

    2017-02-06

    Rac proteins are involved in a variety of cellular processes. Effector proteins that interact with active Rac convey the GTPase-generated signal to downstream developmental cascades and processes. Here we report on the analysis of the main effector and signal cascade downstream of BcRac, the Rac homolog of the grey mold fungus Botrytis cinerea. Several lines of evidence highlighted the p21-activated kinase Cla4 as an important effector of Rac in fungi. Analysis of Δbccla4 strains revealed that the BcCla4 protein was sufficient to mediate all of the examined BcRac-driven processes, including hyphal growth and morphogenesis, conidia production and pathogenicity. In addition, the Δbccla4 strains had altered nuclei content, a phenomenon that was previously observed in Δbcrac isolates, thus connecting the BcRac/BcCla4 module with cell cycle control. Further analyses revealed that BcRac/BcCla4 control mitotic entry through changes in phosphorylation status of the cyclin dependent kinase BcCdk1. The complete cascade includes the kinase BcWee1, which is downstream of BcCla4 and upstream of BcCdk1. These results provide a mechanistic insight on the connection of cell cycle, morphogenesis and pathogenicity in fungi, and position BcCla4 as the most essential effector and central regulator of all of these processes downstream of BcRac.

  19. Heat sinking for printed circuitry

    DOEpatents

    Wilson, S.K.; Richardson, G.; Pinkerton, A.L.

    1984-09-11

    A flat pak or other solid-state device mounted on a printed circuit board directly over a hole extends therethrough so that the bottom of the pak or device extends beyond the bottom of the circuit board. A heat sink disposed beneath the circuit board contacts the bottom of the pak or device and provides direct heat sinking thereto. Pressure may be applied to the top of the pak or device to assure good mechanical and thermal contact with the heat sink.

  20. Inhibition of group 1 p21-activated kinases suppresses pancreatic stellate cell activation and increases survival of mice with pancreatic cancer.

    PubMed

    Yeo, Dannel; Phillips, Phoebe; Baldwin, Graham S; He, Hong; Nikfarjam, Mehrdad

    2017-05-01

    Pancreatic cancer remains one of the most lethal of all solid tumors. Pancreatic stellate cells (PSCs) are primarily responsible for the fibrosis that constitutes the stroma and p21-activated kinase 1 (PAK1) may have a role in signalling pathways involving PSCs. This study aimed to examine the role of PAK1 in PSCs and in the interaction of PSCs with pancreatic cancer cells. Human PSCs were isolated using the modified outgrowth method. The effect of inhibiting PAK1 with group 1 PAK inhibitor, FRAX597, on cell proliferation and apoptosis in vitro was measured by thymidine incorporation and annexin V assays, respectively. The effect of depleting host PAK1 on the survival of mice with pancreatic Pan02 cell tumors was evaluated using PAK1 knockout (KO) mice. PAK1 was expressed in isolated PSCs. FRAX597 reduced the activation of PSCs, inhibited PSC proliferation, and increased PSC apoptosis at least in partial by inhibiting PAK1 activity. The decreased expression and activity of PAK1 in PAK1 KO mice tumors was associated with an increased mouse survival. These results implicate PAK1 as a regulator of PSC activation, proliferation and apoptosis. Targeting stromal PAK1 could increase therapeutic response and survival of patients with pancreatic cancer.

  1. Parameters for the Evaluation of the Fate, Transport, and Environmental Impacts of Chemical Agents in Marine Environments

    DTIC Science & Technology

    2007-07-01

    ein Beitrag zur Kenntniss des Schwefels und seiner ringförmigen Verbindungen, Justus Liebigs Ann. Chem., 1908, 362, 133-173. 36 Paterno, E...Bloemer, H., Einige beiträgr zur kenntnis der organischen arsenverbindung. II. Arsentrichlorid und acetylen, Justus Liebigs Ann. Chem., 1923, 431

  2. Interdisciplinary Educational Collaborations: Chemistry and Computer Science 967 Ronald S. Haines, Daniel T. Woo, Benjamin T. Hudson, Joji C. Mori, Evey S. M. Ngan, and Wing-Yee Pak

    NASA Astrophysics Data System (ADS)

    Goedhart, Martin J.

    2007-06-01

    While chemists are usually aware of the possibilities of interdisciplinary collaboration in chemical research they may be less aware of the possibilities of such collaboration in education. This article documents an ongoing collaboration between a chemist and computer scientist to co-supervise computer science students engaged in developing software for chemical education, highlights the benefits to both the chemistry and computer science students, notes some unexpected outcomes, and provides guidance to those planning such collaborations. The experiences described in this work should motivate chemistry educators to approach their colleagues in other disciplines with proposals for joint research projects. The collaboration described here initially resulted in the development of student-friendly software for operating a spectrophotometer. Recent co-supervised students have begun developing other software for chemical education.

  3. Survival Escherichia coli O157:H7 transformed with either the pAK1-lux or pXEN-13 plasmids in in vitro bovine ruminal and fecal microbial fermentations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of luminescent technology may serve as a viable model for the real-time validation of various pre-harvest interventions on the colonization or shedding of Escherichia coli O157:H7 within cattle. The objective of this study was to determine if the growth of E. coli O157:H7 (ATCC 43888) in ru...

  4. The role of p21-activated kinases in hepatocellular carcinoma metastasis

    PubMed Central

    2014-01-01

    The p21-activated kinases (PAKs) are downstream effectors of the Rho family small GTPases as well as a wide variety of mitogenic factors and have been implicated in cancer formation, development and metastasis. PAKs phosphorylate a wide spectrum of substrates to mediate extracellular signals and regulate cytoskeletal remodeling, cell motility and survival. In this review, we aim to summarize the findings regarding the oncogenic role and the underlying mechanisms of PAKs signaling in various cancers, and in particular highlight the prime importance of PAKs in hepatocellular carcinoma (HCC) progression and metastasis. Recent studies exploring the potential therapeutic application of PAK inhibitors will also be discussed. PMID:25093037

  5. New system enables fleets to tailor maintenance programs

    SciTech Connect

    Fiscor, S.

    2007-08-15

    The Lubrizol Corporation has developed FluiPak and FluiPak Lite systems to pull engine oils and sample them continuously, helping to maintain mining equipment between oil change intervals. The FluiPak system has a control box, a sensor box and a reserve system. FluiPak Lite is simply a cut-down version of FluiPak. The article explains how the system works to detect in real time oil quality and contaminants and hence optimize drain intervals and also prolong engine component life and reduce waste oil disposal costs. 100 units are currently installed in coal operations. 5 figs., 5 photos.

  6. P21 activated kinases

    PubMed Central

    Rane, Chetan K; Minden, Audrey

    2014-01-01

    The p21 activated kinases (Paks) are well known effector proteins for the Rho GTPases Cdc42 and Rac. The Paks contain 6 members, which fall into 2 families of proteins. The first family consists of Paks 1, 2, and 3, and the second consists of Paks 4, 5, and 6. While some of the Paks are ubiquitously expressed, others have more restrictive tissue specificity. All of them are found in the nervous system. Studies using cell culture, transgenic mice, and knockout mice, have revealed important roles for the Paks in cytoskeletal organization and in many aspects of cell growth and development. This review discusses the basic structures of the Paks, and their roles in cell growth, development, and in cancer. PMID:24658305

  7. Paxillin-dependent paxillin kinase linker and p21-activated kinase localization to focal adhesions involves a multistep activation pathway.

    PubMed

    Brown, Michael C; West, Kip A; Turner, Christopher E

    2002-05-01

    The precise temporal-spatial regulation of the p21-activated serine-threonine kinase PAK at the plasma membrane is required for proper cytoskeletal reorganization and cell motility. However, the mechanism by which PAK localizes to focal adhesions has not yet been elucidated. Indirect binding of PAK to the focal adhesion protein paxillin via the Arf-GAP protein paxillin kinase linker (PKL) and PIX/Cool suggested a mechanism. In this report, we demonstrate an essential role for a paxillin-PKL interaction in the recruitment of activated PAK to focal adhesions. Similar to PAK, expression of activated Cdc42 and Rac1, but not RhoA, stimulated the translocation of PKL from a generally diffuse localization to focal adhesions. Expression of the PAK regulatory domain (PAK1-329) or the autoinhibitory domain (AID 83-149) induced PKL, PIX, and PAK localization to focal adhesions, indicating a role for PAK scaffold activation. We show PIX, but not NCK, binding to PAK is necessary for efficient focal adhesion localization of PAK and PKL, consistent with a PAK-PIX-PKL linkage. Although PAK activation is required, it is not sufficient for localization. The PKL amino terminus, containing the PIX-binding site, but lacking paxillin-binding subdomain 2 (PBS2), was unable to localize to focal adhesions and also abrogated PAK localization. An identical result was obtained after PKLDeltaPBS2 expression. Finally, neither PAK nor PKL was capable of localizing to focal adhesions in cells overexpressing paxillinDeltaLD4, confirming a requirement for this motif in recruitment of the PAK-PIX-PKL complex to focal adhesions. These results suggest a GTP-Cdc42/GTP-Rac triggered multistep activation cascade leading to the stimulation of the adaptor function of PAK, which through interaction with PIX provokes a functional PKL PBS2-paxillin LD4 association and consequent recruitment to focal adhesions. This mechanism is probably critical for the correct subcellular positioning of PAK, thereby

  8. Cytotaxonomy of Eurypyga helias (Gruiformes, Eurypygidae): First Karyotypic Description and Phylogenetic Proximity with Rynochetidae

    PubMed Central

    dos Santos, Michelly da Silva; Tagliarini, Marcella Mergulhão; O´Brien, Patricia C. M.; Ferguson-Smith, Malcolm A.; de Oliveira, Edivaldo H. C.

    2015-01-01

    The sunbittern (Eurypyga helias) is a South American Gruiformes, the only member of Family Eurypigidae. In most phylogenetic proposals, it is placed in a more distant position than other families of the so-called “core Gruiformes”. Different studies based on molecular, morphological and biogeographical data suggest that the Eurypigidae is closely related to the kagu (Rhynochetos jubatus), the only species in Rynochetidae, another family not included in the core Gruiformes. Here, the karyotype of the sunbittern is described for the first time, by classical and molecular cytogenetics, using whole chromosome probes derived from Gallus gallus and Leucopternis albicollis. We found a diploid number of 80, with only one pair of biarmed autosomal macrochromosomes, similar to that observed in the kagu. Chromosome painting revealed that most syntenies found in the avian putative ancestral karyotype (PAK) were conserved in the sunbittern. However, PAK1, PAK2, and PAK5 corresponded to two chromosome pairs each. Probes derived from L. albicollis confirm that fissions in PAK1 and PAK2 were centric, whereas in PAK5 the fission is interstitial. In addition, there is fusion of segments homologous to PAK2q and PAK5. From a phylogenetic point of view, comparisons of our results with two other Gruiformes belonging to family Rallidae suggest that the PAK5q fission might be a synapomorphy for Gruiformes. Fissions in PAK1 and PAK2 are found only in Eurypigidae, and might also occur in Rynochetidae, in view of the similar chromosomal morphology between the sunbittern and the kagu. This suggests a close phylogenetic relationship between Eurypigidae and Rynochetidae, whose common ancestor was separated by the Gondwana vicariancy in South America and New Caledonia, respectively. PMID:26624624

  9. Biphasic regulation of myosin light chain phosphorylation by p21-activated kinase modulates intestinal smooth muscle contractility.

    PubMed

    Chu, Ji; Pham, Ngoc T; Olate, Nicole; Kislitsyna, Karina; Day, Mary-Clare; LeTourneau, Phillip A; Kots, Alexander; Stewart, Randolph H; Laine, Glen A; Cox, Charles S; Uray, Karen

    2013-01-11

    Supraphysiological mechanical stretching in smooth muscle results in decreased contractile activity. However, the mechanism is unclear. Previous studies indicated that intestinal motility dysfunction after edema development is associated with increased smooth muscle stress and decreased myosin light chain (MLC) phosphorylation in vivo, providing an ideal model for studying mechanical stress-mediated decrease in smooth muscle contraction. Primary human intestinal smooth muscle cells (hISMCs) were subjected to either control cyclical stretch (CCS) or edema (increasing) cyclical stretch (ECS), mimicking the biophysical forces in non-edematous and edematous intestinal smooth muscle in vivo. ECS induced significant decreases in phosphorylation of MLC and MLC phosphatase targeting subunit (MYPT1) and a significant increase in p21-activated kinase (PAK) activity compared with CCS. PAK regulated MLC phosphorylation in an activity-dependent biphasic manner. PAK activation increased MLC and MYPT1 phosphorylation in CCS but decreased MLC and MYPT1 phosphorylation in hISMCs subjected to ECS. PAK inhibition had the opposite results. siRNA studies showed that PAK1 plays a critical role in regulating MLC phosphorylation in hISMCs. PAK1 enhanced MLC phosphorylation via phosphorylating MYPT1 on Thr-696, whereas PAK1 inhibited MLC phosphorylation via decreasing MYPT1 on both Thr-696 and Thr-853. Importantly, in vivo data indicated that PAK activity increased in edematous tissue, and inhibition of PAK in edematous intestine improved intestinal motility. We conclude that PAK1 positively regulates MLC phosphorylation in intestinal smooth muscle through increasing inhibitory phosphorylation of MYPT1 under physiologic conditions, whereas PAK1 negatively regulates MLC phosphorylation via inhibiting MYPT1 phosphorylation when PAK activity is increased under pathologic conditions.

  10. Chinese Policy Toward South Asia: Implications and Prospects for Nepal

    DTIC Science & Technology

    2013-03-01

    India, Pakistan, Nepal and Bhutan), while the other three ( Bangladesh , Sri Lanka and Maldives) do not share borders with China (Map 1). Nepal and...India; contingent cooperation with Pakistan, Bangladesh and Sri Lanka; and secretive cooptation with Nepal and Maldives.16 Pursuing this strategy at...Pak War of 1965, China sided with Pakistan. Again during the Indo-Pak War in 1971, which led to the birth of Bangladesh , Sino-Pak cooperation was

  11. Identifying Breast Cancer Oncogenes

    DTIC Science & Technology

    2011-10-01

    which is a study of 3131 human tumor samples and cancer cell lines including 243 breast samples. Tumorscape showed that PAK1 is located in an...chromosome 11q of human tumor samples and cancer cell lines that exhibit highest level of PAK1 amplification divided according to cancer type...breast, non-small cell (NSC) lung, ovarian (Ov), small cell lung (SCL), melanoma (Mel) and esophageal squamous (Esq). PAK1 and CCND1 1oci are marked . B

  12. P21-activated kinase 1 regulates resistance to BRAF inhibition in human cancer cells.

    PubMed

    Babagana, Mahamat; Johnson, Sydney; Slabodkin, Hannah; Bshara, Wiam; Morrison, Carl; Kandel, Eugene S

    2017-01-04

    BRAF is a commonly mutated oncogene in various human malignancies and a target of a new class of anti-cancer agents, BRAF-inhibitors (BRAFi). The initial enthusiasm for these agents, based on the early successes in the management of metastatic melanoma, is now challenged by the mounting evidence of intrinsic BRAFi-insensitivity in many BRAF-mutated tumors, by the scarcity of complete responses, and by the inevitable emergence of drug resistance in initially responsive cases. These setbacks put an emphasis on discovering the means to increase the efficacy of BRAFi and to prevent or overcome BRAFi-resistance. We explored the role of p21-activated kinases (PAKs), in particular PAK1, in BRAFi response. BRAFi lowered the levels of active PAK1 in treated cells. An activated form of PAK1 conferred BRAFi-resistance on otherwise sensitive cells, while genetic or pharmacologic suppression of PAK1 had a sensitizing effect. While activation of AKT1 and RAC1 proto-oncogenes increased BRAFi-tolerance, the protective effect was negated in the presence of PAK inhibitors. Furthermore, combining otherwise ineffective doses of PAK- and BRAF-inhibitors synergistically affected intrinsically BRAFi-resistant cells. Considering the high incidence of PAK1 activation in cancers, our findings suggests PAK inhibition as a strategy to augment BRAFi therapy and overcome some of the well-known resistance mechanisms.

  13. IPA3 non-specifically enhances phosphorylation of several proteins in human platelets.

    PubMed

    Badolia, Rachit; Manne, Bhanu Kanth; Dangelmaier, Carol; Kunapuli, Satya P

    2015-01-01

    PAK (p21-Activation kinase), a serine-threonine protein kinase contains an autoinhibitory domain that suppresses the catalytic activity of its kinase domain. This autoregulatory domain found within PAK kinase provides a unique target for chemical inhibitors. IPA3, a small molecule allosteric inhibitor of PAK activation, binds covalently to the PAK regulatory domain and prevents binding to its upstream activators. IPA3 has been used in various cells including platelets to evaluate the role of PAK in signaling. In a recent study, PAK functions in platelet aggregation and lamellipodia formation were evaluated using IPA3 as the PAK inhibitor. Herein, we investigated the specificity and selectivity of IPA3 as a PAK inhibitor in the human platelets. Stimulation of platelets pretreated with IPA3 using a PAR-4 or GPV1 agonist resulted in a concentration-dependent inhibition of aggregation, as was suggested by earlier studies. Interestingly, we found that incubation of washed human platelets with IPA3 lead to a non-specific increase in phosphorylation of several proteins in absence of any agonist. However, this phosphorylation is not sufficient for aggregation of platelets by IPA3. In summary, we demonstrate that IPA3 by itself can phosphorylate several proteins in human platelets and thus its use is not an appropriate strategy for investigating PAK function in platelets.

  14. p-21-Activated kinase 1 mediates gastrin-stimulated proliferation in the colorectal mucosa via multiple signaling pathways.

    PubMed

    Huynh, Nhi; Yim, Mildred; Chernoff, Jonathan; Shulkes, Arthur; Baldwin, Graham S; He, Hong

    2013-03-15

    Gastrins, including amidated (Gamide) and glycine-extended (Ggly) forms, function as growth factors for the gastrointestinal mucosa. The p-21-activated kinase 1 (PAK1) plays important roles in growth factor signaling networks that control cell motility, proliferation, differentiation, and transformation. PAK1, activated by both Gamide and Ggly, mediates gastrin-stimulated proliferation and migration, and activation of β-catenin, in gastric epithelial cells. The aim of this study was to investigate the role of PAK1 in the regulation by gastrin of proliferation in the normal colorectal mucosa in vivo. Mucosal proliferation was measured in PAK1 knockout (PAK1 KO) mice by immunohistochemistry. The expression of phosphorylated and unphosphorylated forms of the signaling molecules PAK1, extracellular signal-regulated kinase (ERK), and protein kinase B (AKT), and the expression of β-catenin and its downstream targets c-Myc and cyclin D1, were measured in gastrin knockout (Gas KO) and PAK1 KO mice by Western blotting. The expression and activation of PAK1 are decreased in Gas KO mice, and these decreases are associated with reduced activation of ERK, AKT, and β-catenin. Proliferation in the colorectal mucosa of PAK1 KO mice is reduced, and the reduction is associated with reduced activation of ERK, AKT, and β-catenin. In compensation, antral gastrin mRNA and serum gastrin concentrations are increased in PAK1 KO mice. These results indicate that PAK1 mediates the stimulation of colorectal proliferation by gastrins via multiple signaling pathways involving activation of ERK, AKT, and β-catenin.

  15. Infrared emission features: probing the interstellar PAH population and circumstellar environment of Herbig Ae/Be stars

    NASA Astrophysics Data System (ADS)

    Boersma, Christiaan

    2009-12-01

    AKs zijn alom vertegenwoordigd en bieden een uitstekend middel om de veelheid aan objecten verspreid over het heelal te bestuderen. Met name in gebieden waar zich sterren en planeten vormen, helpen ze bij het ontwarren van de grootschalige structuur. PAKs staat voor polycyclische aromatische koolwaterstoffen en ze vormen een familie van grote kippengaasvormige moleculen opgebouwd uit koolstof en waterstof. Op aarde worden ze onderander aangetroffen in de verbrandingsproducten van fossiele brandstoffen. PAKs vormen het overgangsgebied van stofdeeltjes ter grote van een micron naar moleculair "gas". PAKs zijn uniek op twee manieren. Allereerst, PAKs fluoresceren na de absorptie van een enkel ultraviolet foton, waardoor ze te zien zijn in zeer koude gebieden, ver weg van de aanstralende bron. In de tweede plaats, gegeven de complexiteit van deze moleculen, kunnen ze een belangrijke rol hebben gespeeld in het ontstaan van leven. Met behulp van topklasse ruimte- en grond gestationeerde observatoria, zoals bijvoorbeeld Spitzer en de 10-meter klasse telescopen in Chili, is de PAK-emissie afkomstig van middelzware, zich vormende, sterren onderzocht. Door gebruik van zowel beeldvorming als spectroscopie, zijn morfologische en evolutionaire aspecten van de PAK-emissie vastgesteld. De NASA Ames PAK IR Spectroscopische Database is een verzameling van meer dan 600 berekend en ongeveer 200 experimenteel bepaalde spectra. Deze unieke database gaat eind 2009 publiek. Gebruikmakend van deze database is een systematische zoektocht gedaan naar kandidaten die verantwoordelijk kunnen worden gehouden voor de emissie in twee, tot op heden, niet goed bestudeerde regio's van het PAKs-spectra.

  16. Group I p21-activated kinases facilitate Tax-mediated transcriptional activation of the human T-cell leukemia virus type 1 long terminal repeats

    PubMed Central

    2013-01-01

    Background Human T-cell leukemia virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia and tropical spastic paraparesis. HTLV-1 encodes transactivator protein Tax that interacts with various cellular factors to modulate transcription and other biological functions. Additional cellular mediators of Tax-mediated transcriptional activation of HTLV-1 long terminal repeats (LTR) remain to be identified and characterized. Results In this study, we investigated the regulatory role of group I p21-activated kinases (Paks) in Tax-induced LTR activation. Both wild-type and kinase-dead mutants of Pak3 were capable of potentiating the activity of Tax to activate LTR transcription. The effect of Paks on the LTR was attributed to the N-terminal regulatory domain and required the action of CREB, CREB-regulating transcriptional coactivators (CRTCs) and p300/CREB-binding protein. Paks physically associated with Tax and CRTCs. Paks were recruited to the LTR in the presence of Tax. siRNAs against either Pak1 or Pak3 prevented the interaction of Tax with CRTC1 and the recruitment of Tax to the LTR. These siRNAs also inhibited LTR-dependent transcription in HTLV-1-transformed MT4 cells and in cells transfected with an infectious clone of HTLV-1. Conclusion Group I Paks augment Tax-mediated transcriptional activation of HTLV-1 LTR in a kinase-independent manner. PMID:23622267

  17. Leucine-rich repeat kinase 2 interacts with p21-activated kinase 6 to control neurite complexity in mammalian brain.

    PubMed

    Civiero, Laura; Cirnaru, Maria Daniela; Beilina, Alexandra; Rodella, Umberto; Russo, Isabella; Belluzzi, Elisa; Lobbestael, Evy; Reyniers, Lauran; Hondhamuni, Geshanthi; Lewis, Patrick A; Van den Haute, Chris; Baekelandt, Veerle; Bandopadhyay, Rina; Bubacco, Luigi; Piccoli, Giovanni; Cookson, Mark R; Taymans, Jean-Marc; Greggio, Elisa

    2015-12-01

    Leucine-rich repeat kinase 2 (LRRK2) is a causative gene for Parkinson's disease, but the physiological function and the mechanism(s) by which the cellular activity of LRRK2 is regulated are poorly understood. Here, we identified p21-activated kinase 6 (PAK6) as a novel interactor of the GTPase/ROC domain of LRRK2. p21-activated kinases are serine-threonine kinases that serve as targets for the small GTP binding proteins Cdc42 and Rac1 and have been implicated in different morphogenetic processes through remodeling of the actin cytoskeleton such as synapse formation and neuritogenesis. Using an in vivo neuromorphology assay, we show that PAK6 is a positive regulator of neurite outgrowth and that LRRK2 is required for this function. Analyses of post-mortem brain tissue from idiopathic and LRRK2 G2019S carriers reveal an increase in PAK6 activation state, whereas knock-out LRRK2 mice display reduced PAK6 activation and phosphorylation of PAK6 substrates. Taken together, these results support a critical role of LRRK2 GTPase domain in cytoskeletal dynamics in vivo through the novel interactor PAK6, and provide a valuable platform to unravel the mechanism underlying LRRK2-mediated pathophysiology. We propose p21-activated kinase 6 (PAK6) as a novel interactor of leucine-rich repeat kinase 2 (LRRK2), a kinase involved in Parkinson's disease (PD). In health, PAK6 regulates neurite complexity in the brain and LRRK2 is required for its function, (a) whereas PAK6 is aberrantly activated in LRRK2-linked PD brain (b) suggesting that LRRK2 toxicity is mediated by PAK6.

  18. A new role for cofilin in retinal neovascularization

    PubMed Central

    Kumar, Raj; Janjanam, Jagadeesh; Singh, Nikhlesh K.; Rao, Gadiparthi N.

    2016-01-01

    ABSTRACT Pak1 plays an important role in several cellular processes, including cell migration, but its role in pathological angiogenesis is not known. Here, we have determined its role in pathological retinal angiogenesis using an oxygen-induced retinopathy (OIR) model. VEGFA induced phosphorylation of Pak1 and its effector cofilin in a manner that was dependent on time as well as p38MAPKβ (also known as MAPK11) in human retinal microvascular endothelial cells (HRMVECs). Depletion of the levels of any of these molecules inhibited VEGFA-induced HRMVEC F-actin stress fiber formation, migration, proliferation, sprouting and tube formation. In accordance with these observations, hypoxia induced Pak1 and cofilin phosphorylation with p38MAPKβ being downstream to Pak1 and upstream to cofilin in mouse retina. Furthermore, Pak1 deficiency abolished hypoxia-induced p38MAPKβ and cofilin phosphorylation and abrogated retinal endothelial cell proliferation, tip cell formation and neovascularization. In addition, small interfering RNA (siRNA)-mediated downregulation of p38MAPKβ or cofilin levels in the wild-type mouse retina also diminished endothelial cell proliferation, tip cell formation and neovascularization. Taken together, these observations suggest that, although the p38MAPKβ–Pak1–cofilin axis is required for HRMVEC migration, proliferation, sprouting and tubulogenesis, Pak1–p38MAPKβ–cofilin signaling is also essential for hypoxia-induced mouse retinal endothelial cell proliferation, tip cell formation and neovascularization. PMID:26857814

  19. LeftyA decreases Actin Polymerization and Stiffness in Human Endometrial Cancer Cells

    PubMed Central

    Salker, Madhuri S.; Schierbaum, Nicolas; Alowayed, Nour; Singh, Yogesh; Mack, Andreas F.; Stournaras, Christos; Schäffer, Tilman E.; Lang, Florian

    2016-01-01

    LeftyA, a cytokine regulating stemness and embryonic differentiation, down-regulates cell proliferation and migration. Cell proliferation and motility require actin reorganization, which is under control of ras-related C3 botulinum toxin substrate 1 (Rac1) and p21 protein-activated kinase 1 (PAK1). The present study explored whether LeftyA modifies actin cytoskeleton, shape and stiffness of Ishikawa cells, a well differentiated endometrial carcinoma cell line. The effect of LeftyA on globular over filamentous actin ratio was determined utilizing Western blotting and flow cytometry. Rac1 and PAK1 transcript levels were measured by qRT-PCR as well as active Rac1 and PAK1 by immunoblotting. Cell stiffness (quantified by the elastic modulus), cell surface area and cell volume were studied by atomic force microscopy (AFM). As a result, 2 hours treatment with LeftyA (25 ng/ml) significantly decreased Rac1 and PAK1 transcript levels and activity, depolymerized actin, and decreased cell stiffness, surface area and volume. The effect of LeftyA on actin polymerization was mimicked by pharmacological inhibition of Rac1 and PAK1. In the presence of the Rac1 or PAK1 inhibitor LeftyA did not lead to significant further actin depolymerization. In conclusion, LeftyA leads to disruption of Rac1 and Pak1 activity with subsequent actin depolymerization, cell softening and cell shrinkage. PMID:27404958

  20. The case for pancreas after kidney transplantation.

    PubMed

    Fridell, Jonathan A; Mangus, Richard S; Hollinger, Edward F; Taber, Tim E; Goble, Michelle L; Mohler, Elaine; Milgrom, Martin L; Powelson, John A

    2009-01-01

    Pancreas after kidney (PAK) transplantation has historically demonstrated inferior pancreas allograft survival compared to simultaneous pancreas and kidney (SPK) transplantation. Under our current immunosuppression protocol, we have noted excellent outcomes and rare immunological graft loss. The goal of this study was to compare pancreas allograft survival in PAK and SPK recipients using this regimen. This was a single center retrospective review of all SPK and PAK transplants performed between January 2003 and November 2007. All transplants were performed with systemic venous drainage and enteric exocrine drainage. Immunosuppression included induction with rabbit anti-thymocyte globulin (thymoglobulin), early steroid withdrawal, and maintenance with tacrolimus and sirolimus or mycophenolate mofetil. Study end points included graft and patient survival and immunosuppression related complications. Transplants included PAK 61 (30%) and SPK 142 (70%). One-yr patient survival was PAK 98% and SPK 95% (p = 0.44) and pancreas graft survival was PAK 95% and SPK 90% (p = 0.28). Acute cellular rejection was uncommon with 2% requiring treatment in each group. Survival for PAK using thymoglobulin induction, early steroid withdrawal and tacrolimus-based immunosuppression is at least comparable to SPK and should be pursued in the recipient with a potential living donor.

  1. p21-activated kinase 1 restricts tonic endocannabinoid signaling in the hippocampus

    PubMed Central

    Xia, Shuting; Zhou, Zikai; Leung, Celeste; Zhu, Yuehua; Pan, Xingxiu; Qi, Junxia; Morena, Maria; Hill, Matthew N; Xie, Wei; Jia, Zhengping

    2016-01-01

    PAK1 inhibitors are known to markedly improve social and cognitive function in several animal models of brain disorders, including autism, but the underlying mechanisms remain elusive. We show here that disruption of PAK1 in mice suppresses inhibitory neurotransmission through an increase in tonic, but not phasic, secretion of endocannabinoids (eCB). Consistently, we found elevated levels of anandamide (AEA), but not 2-arachidonoylglycerol (2-AG) following PAK1 disruption. This increased tonic AEA signaling is mediated by reduced cyclooxygenase-2 (COX-2), and COX-2 inhibitors recapitulate the effect of PAK1 deletion on GABAergic transmission in a CB1 receptor-dependent manner. These results establish a novel signaling process whereby PAK1 upregulates COX-2, reduces AEA and restricts tonic eCB-mediated processes. Because PAK1 and eCB are both critically involved in many other organ systems in addition to the brain, our findings may provide a unified mechanism by which PAK1 regulates these systems and their dysfunctions including cancers, inflammations and allergies. DOI: http://dx.doi.org/10.7554/eLife.14653.001 PMID:27296803

  2. The Cossidae (Lepidoptera) of Afghanistan with description of three new species and special notes on the fauna of Bande-Amir National Park.

    PubMed

    Yakovlev, Roman V; Pljustch, Igor G; Skrylnik, Yuriy; Pak, Oleg; Witt, Thomas J

    2015-07-23

    The annotated list of Cossidae of Afghanistan consists of 44 species in 17 genera from the four subfamilies Catoptinae, Cossinae, Zeuzerinae, and Mehariinae. Three new species are described: Cossulus habibae Yakovlev, Pljustch, Skrylnik & Pak, sp. nov., Semagystia bamiani Yakovlev, Pljustch, Skrylnik & Pak, sp. nov., Phragmacossia bandeamiri Yakovlev, Pljustch, Skrylnik & Pak, sp. nov.; all from Band-e-Amir National Park in Bamian Province. Three species (Dervishiya cadambae (Moore, 1865), Semagystia cossoides (Graeser, 1892), Phragmacossia territa (Staudinger, 1879)) are reported for the first time from Afghanistan. A brief biogeographical analysis of the Cossidae of Afghanistan is given.

  3. Precision Nanoparticles

    ScienceCinema

    John Hemminger

    2016-07-12

    A revolutionary technology that efficiently produces nanoparticles in uniform and prescribed sizes (1-100 nanometers) using supercritical fluids. INL researcher Robert Fox was joined by Idaho State University researchers Rene Rodriquez and Joshua Pak in d

  4. Precision Nanoparticles

    SciTech Connect

    John Hemminger

    2009-07-21

    A revolutionary technology that efficiently produces nanoparticles in uniform and prescribed sizes (1-100 nanometers) using supercritical fluids. INL researcher Robert Fox was joined by Idaho State University researchers Rene Rodriquez and Joshua Pak in d

  5. Role of pili in adhesion of Pseudomonas aeruginosa to human respiratory epithelial cells.

    PubMed Central

    Doig, P; Todd, T; Sastry, P A; Lee, K K; Hodges, R S; Paranchych, W; Irvin, R T

    1988-01-01

    The ability of pili from Pseudomonas aeruginosa K (PAK) to act as an adhesin to human respiratory epithelial cells was examined using an in vitro adhesion assay. Equilibrium analysis of PAK binding to human buccal epithelial cells (BECs) and tracheal epithelial cells (TECs) by means of a Langmuir adsorption isotherm revealed that the maximum numbers of binding sites per epithelial cell (N) were 255 for BECs and 236 for TECs, with apparent association constants (Ka) of 2.8 x 10(-9) and 5.8 x 10(-9) ml/CFU, respectively. Trypsinization of the BECs before the binding assay increased N to 605 and decreased the Ka to 1.7 x 10(-9) ml/CFU. Addition of homologous pili to the binding assay with BECs or TECs or the addition of anti-pilus Fab fragments inhibited PAK adherence. Binding of purified pili to BECs was shown to reach saturation. Purified pili and PAK competed for the same receptor on the BEC surface. Further, by using peptide fragments of PAK pilin (derived from the native pili or produced synthetically) in the binding assay for PAK to BECs, we have presumptively identified the pilus binding domain in the C-terminal region of the pilin and shown that the C-terminal disulfide bridge is important in maintaining the functionality of the binding domain. PMID:2897336

  6. Nuclear factor of activated T cells c1 mediates p21-activated kinase 1 activation in the modulation of chemokine-induced human aortic smooth muscle cell F-actin stress fiber formation, migration, and proliferation and injury-induced vascular wall remodeling.

    PubMed

    Kundumani-Sridharan, Venkatesh; Singh, Nikhlesh K; Kumar, Sanjay; Gadepalli, Ravisekhar; Rao, Gadiparthi N

    2013-07-26

    Recent literature suggests that cyclin-dependent kinases (CDKs) mediate cell migration. However, the mechanisms were not known. Therefore, the objective of this study is to test whether cyclin/CDKs activate Pak1, an effector of Rac1, whose involvement in the modulation of cell migration and proliferation is well established. Monocyte chemotactic protein 1 (MCP1) induced Pak1 phosphorylation/activation in human aortic smooth muscle cells (HASMCs) in a delayed time-dependent manner. MCP1 also stimulated F-actin stress fiber formation in a delayed manner in HASMCs, as well as the migration and proliferation of these cells. Inhibition of Pak1 suppressed MCP1-induced HASMC F-actin stress fiber formation, migration, and proliferation. MCP1 induced cyclin D1 expression as well as CDK6 and CDK4 activities, and these effects were dependent on activation of NFATc1. Depletion of NFATc1, cyclin D1, CDK6, or CDK4 levels attenuated MCP1-induced Pak1 phosphorylation/activation and resulted in decreased HASMC F-actin stress fiber formation, migration, and proliferation. CDK4, which appeared to be activated downstream of CDK6, formed a complex with Pak1 in response to MCP1. MCP1 also activated Rac1 in a time-dependent manner, and depletion/inhibition of its levels/activation abrogated MCP1-induced NFATc1-cyclin D1-CDK6-CDK4-Pak1 signaling and, thereby, decreased HASMC F-actin stress fiber formation, migration, and proliferation. In addition, smooth muscle-specific deletion of NFATc1 led to decreased cyclin D1 expression and CDK6, CDK4, and Pak1 activities, resulting in reduced neointima formation in response to injury. Thus, these observations reveal that Pak1 is a downstream effector of CDK4 and Rac1-dependent, NFATc1-mediated cyclin D1 expression and CDK6 activity mediate this effect. In addition, smooth muscle-specific deletion of NFATc1 prevented the capacity of vascular smooth muscle cells for MCP-1-induced activation of the cyclin D1-CDK6-CDK4-Pak1 signaling axis, affecting

  7. p21-activated kinase regulates mast cell degranulation via effects on calcium mobilization and cytoskeletal dynamics

    PubMed Central

    Allen, Jayme D.; Jaffer, Zahara M.; Park, Su-Jung; Burgin, Sarah; Hofmann, Clemens; Sells, Mary Ann; Chen, Shi; Derr-Yellin, Ethel; Michels, Elizabeth G.; McDaniel, Andrew; Bessler, Waylan K.; Ingram, David A.; Atkinson, Simon J.; Travers, Jeffrey B.

    2009-01-01

    Mast cells are key participants in allergic diseases via activation of high-affinity IgE receptors (FcϵRI) resulting in release of proinflammatory mediators. The biochemical pathways linking IgE activation to calcium influx and cytoskeletal changes required for intracellular granule release are incompletely understood. We demonstrate, genetically, that Pak1 is required for this process. In a passive cutaneous anaphylaxis experiment, Wsh/Wsh mast cell–deficient mice locally reconstituted with Pak1−/− bone marrow–derived mast cells (BMMCs) experienced strikingly decreased allergen-induced vascular permeability compared with controls. Consistent with the in vivo phenotype, Pak1−/− BMMCs exhibited a reduction in FcϵRI-induced degranulation. Further, Pak1−/− BMMCs demonstrated diminished calcium mobilization and altered depolymerization of cortical filamentous actin (F-actin) in response to FcϵRI stimulation. These data implicate Pak1 as an essential molecular target for modulating acute mast cell responses that contribute to allergic diseases. PMID:19124833

  8. The impact of method on kidney graft and patient survival in kidney-pancreas transplantations for type I diabetes mellitus.

    PubMed

    Dinckan, Ayhan; Aliosmanoglu, Ibrahim; Kocak, Huseyin; Mesci, Ayhan; Altunbas, Hasan; Gurkan, Alihan

    2015-01-01

    Patients who develop end-stage renal disease (ESRD) associated with Type I Diabetes Mellitus may receive kidney alone (KA) transplantation, simultaneous pancreas-kidney (SPK) transplantation, or a pancreas after kidney (PAK) transplantation. The goal of this study is to examine the long-term impact of pancreas transplantation on kidney graft and patient survival rates. A total of 85 transplantation cases, consisting of 30 that received living donor KA, 21 that received SPK, and 34 that received PAK, from 2003-2010 at Akdeniz University Organ Transplantation Institute were retrospectively screened. There was a graft loss in 4 cases from the KA group, and in 1 case from each of the SPK and PAK groups. The five-year kidney graft survival rates were 86.7% in KA, 95.2% in SPK, and 97.1% in PAK. There was a single patient loss in both KA and SPK. The kidney survival percentages were higher in SPK and PAK groups compared to the KA group. Therefore, SPK should be the primary preference in these patients; however, for the cases that have a living donor, pancreas transplantation should be considered after kidney transplantation, or the patients can be followed-up on with close blood sugar control.

  9. Complete nucleotide sequence and taxonomy of Sugarcane streak mosaic virus, member of a novel genus in the family Potyviridae.

    PubMed

    Xu, D-L; Zhou, G-H; Xie, Y-J; Mock, R; Li, R

    2010-06-01

    The complete genomic sequence of a Pakistani isolate of Sugarcane streak mosaic virus (SCSMV-PAK) is determined to be 9782 nucleotides in length, excluding the 3' poly(A) tail, and it comprises a large open reading frame encoding a polyprotein of 3130 amino acid residues. The deduced polyprotein is likely to be cleaved at nine putative protease sites by three viral proteases to ten mature proteins. Conserved motifs of orthologous proteins of other potyviruses are identified in corresponding positions of SCSMV-PAK. The genomic organization is virtually identical to the genera Ipomovirus, Potyvirus, Rymovirus, and Tritimovirus in the family Potyviridae. Sequence analyses indicate that the SCSMV-PAK genomic sequence is different from those of Sugarcane mosaic virus and Sorghum mosaic virus, two viruses with very similar symptoms and host range to SCSMV-PAK. SCSMV-PAK shares 52.7% identity with Triticum mosaic virus (TriMV) and 26.4-31.5% identities with species of the existing genera and unassigned viruses in the Potyviridae at the polyprotein sequence level. Phylogenetic analyses of the polyprotein and deduced mature protein amino acid sequences reveal that SCSMV, together with TriMV, forms a distinct group in the family at the genus level. Therefore, SCSMV should represent a new genus, Susmovirus, in the Potyviridae.

  10. Effect of plant species on nitrogen recovery in aquaponics.

    PubMed

    Hu, Zhen; Lee, Jae Woo; Chandran, Kartik; Kim, Sungpyo; Brotto, Ariane Coelho; Khanal, Samir Kumar

    2015-01-01

    Nitrogen transformations in aquaponics with different edible plant species, i.e., tomato (Lycopersicon esculentum) and pak choi (Brassica campestris L. subsp. chinensis) were systematically examined and compared. Results showed that nitrogen utilization efficiencies (NUE) of tomato- and pak choi-based aquaponic systems were 41.3% and 34.4%, respectively. The abundance of nitrifying bacteria in tomato-based aquaponics was 4.2-folds higher than that in pak choi-based aquaponics, primarily due to its higher root surface area. In addition, tomato-based aquaponics had better water quality than that of pak choi-based aquaponics. About 1.5-1.9% of nitrogen input were emitted to atmosphere as nitrous oxide (N2O) in tomato- and pak choi-based aquaponic systems, respectively, suggesting that aquaponics is a potential anthropogenic source of N2O emission. Overall, this is the first intensive study that examined the role plant species played in aquaponics, which could provide new strategy in designing and operating an aquaponic system.

  11. Activity-dependent regulation of genes implicated in X-linked non-specific mental retardation.

    PubMed

    Boda, B; Mas, C; Muller, D

    2002-01-01

    X-linked forms of non-specific mental retardation are complex disorders, for which mutations in several genes have recently been identified. These include OPHN1, GDI1, PAK3, IL1RAPL, TM4SF2, FMR2 and RSK2. To investigate the mechanisms through which alterations of these gene products could result in cognitive impairment, we analyzed their expression using quantitative PCR technique in two in vitro models of activity-dependent gene regulation: kainate-induced seizures and long-term synaptic potentiation (LTP). We found that the level of expression of four genes, PAK3, IL1RAPL, RSK2 and TM4SF2, was significantly up-regulated following kainate treatment. Furthermore we observed a significant increase in mRNA levels of PAK3 and IL1RAPL following LTP induction. These results suggest a possible role for these four genes in activity-dependent brain plasticity.

  12. Glucose-regulated protein 78 (GRP78) binds directly to PIP3 phosphatase SKIP and determines its localization.

    PubMed

    Ijuin, Takeshi; Hatano, Naoya; Takenawa, Tadaomi

    2016-05-01

    Skeletal muscle and kidney-enriched inositol polyphosphate phosphatase (SKIP), a PIP3 phosphatase, has been implicated in the regulation of insulin signaling in skeletal muscle. SKIP interacts with Pak1 and glucose-regulated protein 78 (GRP78), both of which are necessary for the regulation of insulin signaling. In this study, we showed that GRP78 directly binds to the SKIP C-terminal homology (SKICH) domain of SKIP and that this binding is necessary for the localization of SKIP at the ER. In addition, in vitro binding analysis showed that GRP78 and Pak1 competitively bind to SKIP. Taken together, these findings suggest a model by which GRP78 regulates intracellular localization of SKIP and how SKIP binds to Pak1 on insulin stimulation.

  13. Glioblastoma following treatment with fingolimod for relapsing-remitting multiple sclerosis.

    PubMed

    Sharim, Justin; Tashjian, Randy; Golzy, Nima; Pouratian, Nader

    2016-08-01

    Glioblastoma is an uncommon and aggressive primary brain tumor with incidence of 3 per 100,000 annually. We report a 50-year-old woman diagnosed with glioblastoma within threeyears of induction of fingolimod therapy for relapsing-remitting multiple sclerosis. Fingolimod, an immunomodulating agent used in the treatment of relapsing-remitting multiple sclerosis, has also been suggested to impart a cardioprotective role in heart failure and arrhythmia via activation of P21-activated kinase-1 (Pak1). In the brain, Pak1 activation has been shown to correlate with decreased survival time amongst patients with glioblastoma. A molecular mechanism underlying a link between fingolimod use and glioblastoma development may involve activation of Pak1. To our knowledge, this is the first report of a potential association between fingolimod use and glioblastoma development.

  14. P21-Activated Kinase Inhibitors FRAX486 and IPA3: Inhibition of Prostate Stromal Cell Growth and Effects on Smooth Muscle Contraction in the Human Prostate

    PubMed Central

    Wang, Yiming; Gratzke, Christian; Tamalunas, Alexander; Wiemer, Nicolas; Ciotkowska, Anna; Rutz, Beata; Waidelich, Raphaela; Strittmatter, Frank; Liu, Chunxiao; Stief, Christian G.; Hennenberg, Martin

    2016-01-01

    Prostate smooth muscle tone and hyperplastic growth are involved in the pathophysiology and treatment of male lower urinary tract symptoms (LUTS). Available drugs are characterized by limited efficacy. Patients’ adherence is particularly low to combination therapies of 5α-reductase inhibitors and α1-adrenoceptor antagonists, which are supposed to target contraction and growth simultaneously. Consequently, molecular etiology of benign prostatic hyperplasia (BPH) and new compounds interfering with smooth muscle contraction or growth in the prostate are of high interest. Here, we studied effects of p21-activated kinase (PAK) inhibitors (FRAX486, IPA3) in hyperplastic human prostate tissues, and in stromal cells (WPMY-1). In hyperplastic prostate tissues, PAK1, -2, -4, and -6 may be constitutively expressed in catecholaminergic neurons, while PAK1 was detected in smooth muscle and WPMY-1 cells. Neurogenic contractions of prostate strips by electric field stimulation were significantly inhibited by high concentrations of FRAX486 (30 μM) or IPA3 (300 μM), while noradrenaline- and phenylephrine-induced contractions were not affected. FRAX486 (30 μM) inhibited endothelin-1- and -2-induced contractions. In WPMY-1 cells, FRAX486 or IPA3 (24 h) induced concentration-dependent (1–10 μM) degeneration of actin filaments. This was paralleled by attenuation of proliferation rate, being observed from 1 to 10 μM FRAX486 or IPA3. Cytotoxicity of FRAX486 and IPA3 in WPMY-1 cells was time- and concentration-dependent. Stimulation of WPMY-1 cells with endothelin-1 or dihydrotestosterone, but not noradrenaline induced PAK phosphorylation, indicating PAK activation by endothelin-1. Thus, PAK inhibitors may inhibit neurogenic and endothelin-induced smooth muscle contractions in the hyperplastic human prostate, and growth of stromal cells. Targeting prostate smooth muscle contraction and stromal growth at once by a single compound is principally possible, at least under

  15. Macroencapsulation of mixed waste debris at the Hanford Nuclear Reservation -- Final project report by AST Environmental Services, LLC

    SciTech Connect

    Baker, T.L.

    1998-02-25

    This report summarizes the results of a full-scale demonstration of a high density polyethylene (HDPE) package, manufactured by Arrow Construction, Inc. of Montgomery, Alabama. The HDPE package, called ARROW-PAK, was designed and patented by Arrow as both a method to macroencapsulation of radioactively contaminated lead and as an improved form of waste package for treatment and interim and final storage and/or disposal of drums of mixed waste. Mixed waste is waste that is radioactive, and meets the criteria established by the United States Environmental Protection Agency (US EPA) for a hazardous material. Results from previous testing conducted for the Department of Energy (DOE) at the Idaho National Engineering Laboratory in 1994 found that the ARROW-PAK fabrication process produces an HDPE package that passes all helium leak tests and drop tests, and is fabricated with materials impervious to the types of environmental factors encountered during the lifetime of the ARROW-PAK, estimated to be from 100 to 300 years. Arrow Construction, Inc. has successfully completed full-scale demonstration of its ARROW-PAK mixed waste macroencapsulation treatment unit at the DOE Hanford Site. This testing was conducted in accordance with Radiological Work Permit No. T-860, applicable project plans and procedures, and in close consultation with Waste Management Federal Services of Hanford, Inc.`s project management, health and safety, and quality assurance representatives. The ARROW-PAK field demonstration successfully treated 880 drums of mixed waste debris feedstock which were compacted and placed in 149 70-gallon overpack drums prior to macroencapsulation in accordance with the US EPA Alternate Debris Treatment Standards, 40 CFR 268.45. Based on all of the results, the ARROW-PAK process provides an effective treatment, storage and/or disposal option that compares favorably with current mixed waste management practices.

  16. Field Evaluation of a Rapid Immunochromatographic Assay for Detection of Trypanosoma cruzi Infection by Use of Whole Blood▿

    PubMed Central

    Roddy, Paul; Goiri, Javier; Flevaud, Laurence; Palma, Pedro Pablo; Morote, Silvia; Lima, Nines; Villa, Luis; Torrico, Faustino; Albajar-Viñas, Pedro

    2008-01-01

    Laboratory and clinical diagnostic classification of seropositive individuals, followed by treatment and supportive therapy, is an established component of Chagas' disease control in areas where this disease is endemic. However, most Chagas' disease patients live in remote areas where neither equipped laboratories nor skilled human resources are widely available. Employing a rapid diagnostic test (RDT), when using whole blood samples, is the best option for Chagas' disease control. A high sensitivity and specificity for the Chagas Stat-Pak RDT (Chembio Diagnostic Systems, Inc., Medford, NY) has been reported for assays using serum and plasma, but its validity for the detection of antibodies to Trypanosoma cruzi infection in whole blood is unknown. This cross-sectional study measured the sensitivity and specificity of the Chagas Stat-Pak with whole blood, using conventional serological assays for comparison. The interobserver reliability in the interpretation of the Chagas Stat-Pak results and “ease-of-use” criterion needed to perform the Chagas Stat-Pak and conventional assays were also measured. The Chagas Stat-Pak yielded a high specificity (99.0%, 95% confidence interval [CI] = 98.4 to 99.4%) but a relatively low sensitivity (93.4%, 95% CI = 87.4 to 97.1%). The interobserver reliability was excellent (kappa [n = 1,913] = 0.999, P < 0.0001), and the quantified ease-of-use criterion suggested that the RDT is simple to perform. Despite the attributes of the Chagas Stat-Pak, it is not an ideal diagnostic test for the population investigated in the present study due to its relatively low sensitivity and high cost. The RDT manufacturer is called upon to improve the test if the international community hopes to make progress in controlling Chagas infections in areas where this disease is endemic. PMID:18400910

  17. Prevailing PA Mutation K356R in Avian Influenza H9N2 Virus Increases Mammalian Replication and Pathogenicity

    PubMed Central

    Xu, Guanlong; Zhang, Xuxiao; Gao, Weihua; Wang, Chenxi; Wang, Jinliang; Sun, Honglei; Sun, Yipeng; Guo, Lu; Zhang, Rui; Chang, Kin-Chow; Liu, Jinhua

    2016-01-01

    ABSTRACT Adaptation of the viral polymerase complex comprising PB1, PB2, and PA is necessary for efficient influenza A virus replication in new host species. We found that PA mutation K356R (PA-K356R) has become predominant since 2014 in avian H9N2 viruses in China as with seasonal human H1N1 viruses. The same mutation is also found in most human isolates of emergent avian H7N9 and H10N8 viruses whose six internal gene segments are derived from the H9N2 virus. We further demonstrated the mammalian adaptive functionality of the PA-K356R mutation. Avian H9N2 virus with the PA-K356R mutation in human A549 cells showed increased nuclear accumulation of PA and increased viral polymerase activity that resulted in elevated levels of viral transcription and virus output. The same mutant virus in mice also enhanced virus replication and caused lethal infection. In addition, combined mutation of PA-K356R and PB2-E627K, a well-known mammalian adaptive marker, in the H9N2 virus showed further cooperative increases in virus production and severity of infection in vitro and in vivo. In summary, PA-K356R behaves as a novel mammalian tropism mutation, which, along with other mutations such as PB2-E627K, might render avian H9N2 viruses adapted for human infection. IMPORTANCE Mutations of the polymerase complex (PB1, PB2, and PA) of influenza A virus are necessary for viral adaptation to new hosts. This study reports a novel and predominant mammalian adaptive mutation, PA-K356R, in avian H9N2 viruses and human isolates of emergent H7N9 and H10N8 viruses. We found that PA-356R in H9N2 viruses causes significant increases in virus replication and severity of infection in human cells and mice and that PA-K356R cooperates with the PB2-E627K mutation, a well-characterized human adaptive marker, to exacerbate mammalian infection in vitro and in vivo. Therefore, the PA-K356R mutation is a significant adaptation in H9N2 viruses and related H7N9 and H10N8 reassortants toward human

  18. The summertime "heat" low over Pakistan/northwestern India: evolution and origin

    NASA Astrophysics Data System (ADS)

    Bollasina, Massimo; Nigam, Sumant

    2011-09-01

    A deep low in sea-level pressure is present from May to September over Pakistan and northwestern India (hereafter, the Pak-India low). It is often referred as the "heat" low to convey the significance of surface thermal effects reckoned to be important for its origin. The present analysis, rooted in observations and diagnostic modeling, suggests that the Pak-India low is influenced both by regional and remote forcing. Regionally, the influence of Hindu Kush mountains is found to be stronger than the impact of land-surface heating and attendant sensible heating of the planetary boundary layer, questioning the suitability of the "heat" label in canonical references to this circulation feature. Observational analysis indicates that the notable May-to-June deepening of the Pak-India low and its further deepening in July, however, arises from remote forcing—the development of monsoon deep-convection over the Bay of Bengal and eastern India in June and July. It is hypothesized that the associated upstream descent over Iran-Turkmenistan-Afghanistan (i.e. east of the Caspian Sea) and related low-level northerlies over the Elburz-Zagros-Hindu Kush mountains contribute to the strengthening of the Pak-India low in June (and July) from interaction with regional orography.

  19. Regulation of microtubule destabilizing activity of Op18/stathmin downstream of Rac1.

    PubMed

    Wittmann, Torsten; Bokoch, Gary M; Waterman-Storer, Clare M

    2004-02-13

    In the leading edge of migrating cells, a subset of microtubules exhibits net growth in a Rac1- and p21-activated kinase-dependent manner. Here, we explore the possibility of whether phosphorylation and inactivation of the microtubule-destabilizing protein Op18/stathmin could be a mechanism regulating microtubule dynamics downstream of Rac1 and p21-activated kinases. We find that, in vitro, Pak1 phosphorylates Op18/stathmin specifically at serine 16 and inactivates its catastrophe promoting activity in biochemical and time lapse microscopy microtubule assembly assays. Furthermore, phosphorylation of either serine 16 or 63 is sufficient to inhibit Op18/stathmin in vitro. In cells, the microtubule-destabilizing effect of an excess of Op18/stathmin can be partially overcome by expression of constitutively active Rac1(Q61L), which is dependent on Pak activity, suggesting that the microtubule cytoskeleton can be regulated through inactivation of Op18/stathmin downstream of Rac1 and Pak in vivo. However, in vivo, Pak1 activity alone is not sufficient to phosphorylate Op18, indicating that additional pathways downstream of Rac1 are required for Op18 regulation.

  20. [Enantioseparation behavior of chiral stationary phases AD, AS and OD].

    PubMed

    Li, Liqun; Fan, Jun; Zhang, Jing; Chen, Xiaodong; Wang, Tai; He, Jianfeng; Zhang, Weiguang

    2016-01-01

    Over the past decades, HPLC enantioseparation with chiral stationary phases (CSPs) has been widely applied in chiral analysis and preparation of new pharmaceuticals, pesticides, food, etc. Herein, enantioseparation of 20 chiral compounds have been carried out on three polysaccharide-based CSPs (EnantioPak AD, AS and OD) with normal phases by HPLC, separately. The influences of skeletal structure and the kinds of derivative groups on separation behaviors of these CSPs have been studied in detail. As results indicated, except for compound 13, the other compounds were baseline separated on EnantioPak AD, with most of resolution over 2. 0; in addition, better separation for acidic or basic compounds was achieved through adding acidic/basic additives into the mobile phase of hexane-alcohol. For four aromatic alcohols (compounds 13-16), their retention in the EnantioPak AD column showed a weakening tendency with increase of carbon number in side chain group, and the reverse trend of their resolution was observed. Furthermore, EnantioPak AD showed much better separation performance for eight compounds (13-20) than the others. In short, these results have provided some references for further investigation of separation behavior and applications of polysaccharide-based CSPs.

  1. Contributions to the Mosquito Fauna of Southeast Asia. V. Genus Aedes, Subgenus Diceromyia Theobald in Southeast Asia

    DTIC Science & Technology

    1970-01-01

    Diceromyia) in Southeast Asia 19 paratype, NB681, from an island in the Chao Phraya River , Pak Kret, No&ha- buri, THAILAND, 16 June 1964, light trap... River (3 miles inland), Palawan and 1 from Cape Melville, Balabac to this species or Heimnannia scintillans Ludlow. Mattingly (1957: 3 2) states the

  2. The effects of control release fertilizer (CRF) on palm growth

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nutri-Pak is a slow release fertilizer in a micro-pore polyethylene packet where moisture enters the packet through micro-pores located on both sides of the packet. Water dissolves the fertilizer and it slowly seeps through the same micro-pores as a vapor into the soil gradually providing nutrients ...

  3. Tamoxifen Dependent Interaction Between in Estrogen Receptor and a Novel p21 Activated Kinase

    DTIC Science & Technology

    2004-06-01

    phosphorylation of Tyr 566 by MKK6, a dual-specificity phosphatase, MKP-1 (Lmitogen kinase phosphotase -1), which can dephosphorylate both threonine and tyrosine... phosphotase -1; WT, wild type; HA, hemagglutinin; MAP, mitogen-activated protein 23 Kaur et al. - PAK6 activation via P38 MAP kinase/MKK6 pathway Figure

  4. Postmortem Photonic Imaging of Lux-Modified Salmonella Typhimuium Within the Gastrointestinal Tract of Swine Following Oral Inoculation In Vivo

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The study objective was to monitor Salmonella progression by photonic detection through segments of the gastrointestinal tract after oral inoculation. Pigs (~80 kg) were inoculated orally with 3.1 or 4.1 x 1010 cfu of Salmonella Typhimurium transformed with plasmid pAK1-lux for a 6-h (n = 6) or 12-h...

  5. Postmortem photonic imaging of lux-modified Salmonella typhimurium within the gastrointestinal tract of swine following oral inoculation in vivo

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The study objective was to monitor Salmonella progression by photonic detection through segments of the gastrointestinal tract following oral inoculation. Pigs (~ 80 kg) were inoculated orally with 3.1 or 4.1×10*10 colony forming units (cfu) of Salmonella typhimurium transformed with plasmid pAK1-lu...

  6. The Effect of an Instructional Unit Incorporating Live Animals on Knowledge of Nutrition for Different Age Levels.

    ERIC Educational Resources Information Center

    Roth, Anne I.; Wunderlich, Kenneth W.

    A nutrition education unit, Rat Pak, developed by Dairy Council, Inc., is an attempt to influence students to make wise food choices. It consists of eleven lessons in an instructional sequence which incorporates the use of white rats as a means of illustrating the effect of improper diet while teaching proper diet. The purpose of this…

  7. Actions of Rho family small G proteins and p21-activated protein kinases on mitogen-activated protein kinase family members.

    PubMed Central

    Frost, J A; Xu, S; Hutchison, M R; Marcus, S; Cobb, M H

    1996-01-01

    The mitogen-activated protein (MAP) kinases are a family of serine/threonine kinases that are regulated by distinct extracellular stimuli. The currently known members include extracellular signal-regulated protein kinase 1 (ERK1), ERK2, the c-Jun N-terminal kinase/stress-activated protein kinases (JNK/SAPKs), and p38 MAP kinases. We find that overexpression of the Ste20-related enzymes p21-activated kinase 1 (PAK1) and PAK2 in 293 cells is sufficient to activate JNK/SAPK and to a lesser extent p38 MAP kinase but not ERK2. Rat MAP/ERK kinase kinase 1 can stimulate the activity of each of these MAP kinases. Although neither activated Rac nor the PAKs stimulate ERK2 activity, overexpression of either dominant negative Rac2 or the N-terminal regulatory domain of PAK1 inhibits Ras-mediated activation of ERK2, suggesting a permissive role for Rac in the control of the ERK pathway. Furthermore, constitutively active Rac2, Cdc42hs, and RhoA synergize with an activated form of Raf to increase ERK2 activity. These findings reveal a previously unrecognized connection between Rho family small G proteins and the ERK pathway. PMID:8668187

  8. A Robot to Help Make the Rounds

    NASA Technical Reports Server (NTRS)

    2003-01-01

    This paper presents a discussion on the Pyxis HelpMate SecurePak (SP) trackless robotic courier designed by Transitions Research Corporation, to navigate autonomously throughout medical facilities, transporting pharmaceuticals, laboratory specimens, equipment, supplies, meals, medical records, and radiology films between support departments and nursing floors.

  9. Surface Water Investigations in Afghanistan: A Summary of Activities from 1952 to 1969

    DTIC Science & Technology

    1969-03-01

    Chakhansur Registan Spin Baldak Chaman Zahedran Gizab Qala-Hazar Qadam Qala Ahangaran Deshu Sistan Basin Adraskan Shindand Kajaki (32°l6-65...Chakhansur Rigestan(region) Spin Buidak Chaman (Pak) Zahedan (Iran) Gizab Hazar Qadam Ahangaran Deh Shu Seistan(region

  10. Complete genome sequence of a recent panzootic virulent Newcastle disease virus from Pakistan

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Complete genome sequence of a new strain of Newcastle disease virus (NDV) (chicken/Pak/Lahore-611/2013) is reported. The strain was isolated from a vaccinated chicken flock in Pakistan in 2013 and has panzootic features. The genome is 15192 nucleotides in length and is classified as sub-genotype V...

  11. Narrow-Band Imaging and Spatially Resolved Spectra of Nova Shells

    NASA Astrophysics Data System (ADS)

    Hillwig, T. C.; Honeycutt, R. K.; Shore, S. N.

    2000-12-01

    Observations of nova shells were made at the WIYN Observatory using the WIYN Imager, the ``naked'' DensePak fiber array, and a Barlow 4x magnifying assembly used with DensePak. DensePak was used to obtain spatially resolved spectra of several nova shells at wavelengths including the Hα , Hβ , [OIII], and [NII] emission lines. The purpose is to derive true shapes and sizes of the nova shells, velocity structure, and abundance structure. The ability to spatially resolve the shell with spectroscopy, with the accuracy and resolution available to DensePak is a useful and unique tool. The velocity structure of the shell provides data which can be compared to models of expected shell structure. Measuring abundances in different, spatially resolved portions of the shell can give indications of the cause of the structure. For example, in shaping by a fast wind, we may expect to see different abundances in the slowly moving ejected material than in the material comprising the fast wind (which becomes apparent in planetary nebulae with wind-blown bubbles). Imaging also provides, along with comparison to velocity structure, an additional constraint on the determination of parallax distances, and the narrow-band imaging can supply estimates of excitation levels in various regions of the shells. All of these are important contributors to the determination of the physical mechanism responsible for the nova shell structure. The first phase of this research is presented here.

  12. Field application of serodiagnostics to identify elephants with Tuberculosis prior to case confirmation by culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three serologic methods for antibody detection in elephant tuberculosis (TB), multiantigen print immunoassay (MAPIA), ElephantTB STAT-PAK kit, and DPP VetTB test, were validated prospectively using serial serum samples from 14 captive elephants in 5 countries which were diagnosed with TB by positive...

  13. ENVIRONMENTAL TECHNOLOGY VERIFICATON: TEST REPORT OF CONTROL OF BIOAEROSOLS IN HVAC SYSTEMS AAF INTERNATIONAL DRIPAK 90/95%

    EPA Science Inventory

    The Environmental Technology Verification report discusses the technology and performance of the DriPak 90/95% air filter for dust and bioaerosol filtration manufactured by AAF International. The pressure drop across the filter was 104 Pa clean and 348 Pa dust loaded, and the fil...

  14. 9 CFR 77.34 - Official tuberculosis tests.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... used in individual captive elk, red deer, white-tailed deer, fallow deer, and reindeer, and in herds of... CervidTB Stat-Pak® test is a primary test that may be used in affected herds of captive elk, red deer, white-tailed deer, fallow deer, and reindeer, and in herds of these species that have received...

  15. How Does Culture Shape Students' Perceptions of Scientists? Cross-National Comparative Study of American and Chinese Elementary Students

    ERIC Educational Resources Information Center

    Farland-Smith, Donna

    2009-01-01

    For decades, researchers have been convinced that one stereotypic image of scientists existed among children worldwide (Chambers, 1983; Chiang & Guo, 1996; Fung, 2002; Maoldomhnaigh & Hunt, 1988; Newton & Newton, 1992, 1998; She, 1998; Song, Pak, & Jang, 1992). This study, however, moves beyond that stereotypic image and examines…

  16. Working with Children to Protect Our Future

    ERIC Educational Resources Information Center

    Williams, Nick

    2011-01-01

    The author has been teaching primary school children for 20 years, and has always been passionate about teaching young people about the environment. In this article, he describes his work with Tetra Pak and WWF-UK to develop a national, school-based competition and teaching programme to help children understand the importance of using renewable…

  17. Area Handbook Series: Thailand. A Country Study

    DTIC Science & Technology

    1987-09-01

    Paleolithic culture in the region and continuous human habitation for at least 20,000 years. The pace of economic and social development was uneven and...Pakistanis, 107 National Security Council, 192, 255 Pak Mai (New Party), 222 National Socialist Party, 37 Paleolithic culture, 4 National Union (Sahachat

  18. The Impact of Chinese Development of Nuclear Weapons on the Pakistan-Indian Dispute

    DTIC Science & Technology

    1966-04-08

    Conference of Dec. 10, 1962." Dept of State Bulletin, Vol. 47, 31 Dec. 1962, p. 998. (Statement of recent US efforts on Kashmir.) 65. Salam , Abdus ...34 RoyaJ Central Asaln Journal, Viii. 51, Oct. 1964, p. 223. 39 Abdus Sal am, !Pak i sLa I- -The Case for Tecict I Cal leve I plmielt , BHlleLtln of

  19. Indole-3-Acetic Acid Produced by Burkholderia heleia Acts as a Phenylacetic Acid Antagonist to Disrupt Tropolone Biosynthesis in Burkholderia plantarii

    PubMed Central

    Wang, Mengcen; Tachibana, Seiji; Murai, Yuta; Li, Li; Lau, Sharon Yu Ling; Cao, Mengchao; Zhu, Guonian; Hashimoto, Makoto; Hashidoko, Yasuyuki

    2016-01-01

    Burkholderia heleia PAK1-2 is a potent biocontrol agent isolated from rice rhizosphere, as it prevents bacterial rice seedling blight disease caused by Burkholderia plantarii. Here, we isolated a non-antibacterial metabolite from the culture fluid of B. heleia PAK1-2 that was able to suppress B. plantarii virulence and subsequently identified as indole-3-acetic acid (IAA). IAA suppressed the production of tropolone in B. plantarii in a dose-dependent manner without any antibacterial and quorum quenching activity, suggesting that IAA inhibited steps of tropolone biosynthesis. Consistent with this, supplementing cultures of B. plantarii with either L-[ring-2H5]phenylalanine or [ring-2H2~5]phenylacetic acid revealed that phenylacetic acid (PAA), which is the dominant metabolite during the early growth stage, is a direct precursor of tropolone. Exposure of B. plantarii to IAA suppressed production of both PAA and tropolone. These data particularly showed that IAA produced by B. heleia PAK1-2 disrupts tropolone production during bioconversion of PAA to tropolone via the ring-rearrangement on the phenyl group of the precursor to attenuate the virulence of B. plantarii. B. heleia PAK1-2 is thus likely a microbial community coordinating bacterium in rhizosphere ecosystems, which never eliminates phytopathogens but only represses production of phytotoxins or bacteriocidal substances. PMID:26935539

  20. Out of Area or Out of Reach? European Military Support for Operations in Southwest Asia.

    DTIC Science & Technology

    1995-01-01

    Watson, Bruce W. Watson, Jr., David Dunphy, Richard Dejong, Brian Gagne, Michael Kirsch , and Yong Pak, "The Effects of the War on Other Nations," in...1994-95 (London: Butler and Tanner Ltd., 1994), p. 219. 12Washington Times, 13 June 1994, p. 14. 13The narrow margin of victory for Helmut Kohl’s CDU

  1. 79 FR 43923 - Approved Tests for Bovine Tuberculosis in Cervids

    Federal Register 2010, 2011, 2012, 2013, 2014

    2014-07-29

    ... Animal and Plant Health Inspection Service 9 CFR Part 77 Approved Tests for Bovine Tuberculosis in... comments. SUMMARY: We are amending the regulations regarding official tuberculosis tests for captive cervids to remove the CervidTB Stat- Pak as an official bovine tuberculosis test for the following...

  2. 78 FR 1718 - Approved Tests for Bovine Tuberculosis in Cervids

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-09

    ... Animal and Plant Health Inspection Service 9 CFR Part 77 Approved Tests for Bovine Tuberculosis in... comments. SUMMARY: We are adding the CervidTB Stat-Pak and DPP tests as official tuberculosis tests for the... of antibodies to bovine tuberculosis in certain species of captive cervids. This action is...

  3. Drug Induced Pneumonitis Secondary to Treatment with Paritaprevir/Ritonavir/Ombitasvir and Dasabuvir (VIEKIRA PAK®) for Chronic Hepatitis C: Case Report of an Unexpected Life-Threatening Adverse Reaction

    PubMed Central

    Faire, Bridget; Gane, Edward

    2017-01-01

    VIEKIRA PAK (ritonavir-boosted paritaprevir/ombitasvir and dasabuvir) is an approved treatment for compensated patients with genotype 1 (GT1) chronic hepatitis C virus (HCV) infection. This oral regimen has minimal adverse effects and is well tolerated. Cure rates are 97% in patients infected with HCV GT 1a and 99% in those with HCV GT 1b. We report the first case of life-threatening allergic pneumonitis associated with VIEKIRA PAK. This unexpected serious adverse event occurred in a 68-year-old Chinese female with genotype 1b chronic hepatitis C and Child-Pugh A cirrhosis. One week into treatment with VIEKIRA PAK without ribavirin, she was admitted to hospital with respiratory distress and acute kidney injury requiring intensive care input. She was initially diagnosed with community acquired pneumonia and improved promptly with intravenous antibiotics and supported care. No bacterial or viral pathogens were cultured. Following complete recovery, she recommenced VIEKIRA PAK but represented 5 days later with more rapidly progressive respiratory failure, requiring intubation and ventilation, inotropic support, and haemodialysis. The final diagnosis was drug induced pneumonitis.

  4. The Genus Culex, Subgenus Eumelanomyia Theobald in Southeast Asia and Adjacent Areas

    DTIC Science & Technology

    1972-01-01

    tergal surface; subapical lobe short or elon or sma 1; distimere slender, sickle shaped and long; B ate; proximal division with 3 stout rods...Nok, 1 P. Tak: Huey Lan Saeng, 6d, 69, 12 p. Nakhon Ratchasima: Pak Chong; Musk Lek; Ban Tha Ma Prang; Khlong Pai; Khao Suan Horn; 7~r, 9?, 1 P, 7 p

  5. JPRS Report, East Europe

    DTIC Science & Technology

    1990-10-22

    Crisis Contingency Plan; Possible Scenarios Described [FIGYELO 20 Sep] ■» Paks Nuclear Plant: Origins of Control System Described ^G...Joint Venture in Methane Gas Production Discussed [PRZEGLAD TYGODNIOWY 2 Sep] • w SOCIAL HUNGARY Interior Minister on Police Reform, Public...undertaking. The principal task of the intelligence operatives will involve political and social contacts. Regarding the activities of foreign

  6. Psychoanalysis and Transformation of Heroes in Mohsin Hamid's Novels "Moth Smoke" and "The Reluctant Fundamentalist"

    ERIC Educational Resources Information Center

    Awan, Abdul Ghafoor; Andleeb, Shaista; Yasin, Farhat

    2016-01-01

    Mohsin Hamid is an exponent of postcolonial characterization and possesses a specific touch of today's hero in local Asian context. His pen is fluent on social fiction portraying Indo-Pak culture. Few writers could rise to the height of fame right with only a couple of preliminary works. Hamid secured it through his first novel "Moth…

  7. An Analysis of the Fulton Industrial Area: A Market for Parkway’s Industrial Medicine Program.

    DTIC Science & Technology

    1983-05-30

    are not waiting for proof of the hypothesis related to the benefits of health promotion programs. Many accept the empirical evidence of employee...offered at Tee-Pak. *Interviewee did not desire to respond to the benefits of health promotion programs, because of such programs must originate from the

  8. From LAMP to Koha: Case Study of the Pakistan Legislative Assembly Libraries

    ERIC Educational Resources Information Center

    Shafi-Ullah, Farasat; Qutab, Saima

    2012-01-01

    Purpose: This paper aims to elaborate the library data migration process from LAMP (Library Automation Management Program) to the open source software Koha's (2.2.8 Windows based) Pakistani flavour PakLAG-Koha in six legislative assembly libraries of Pakistan. Design/methodology/approach: The paper explains different steps of the data migration…

  9. A Neural Network for Estimation of Aortic Pressure from the Radial Artery Pressure Pulse

    DTIC Science & Technology

    2001-10-25

    from periphery to artery: a model based study, American Journal of Physiology, 1998,274:43, pp H1386-92 [9] C. Chen, E. Nevo , B Fetics, P Pak, F, Yin, L...36. [10] B Fetics, E Nevo , C. Chen, D Kass, Parametric model derivation of transfer function for noninvasive estimation of aortic pressure by radial

  10. Symptom Assessment in Knee Osteoarthritis Needs to Account for Physical Activity Level

    PubMed Central

    Lo, Grace H.; McAlindon, Timothy E.; Hawker, Gillian A.; Driban, Jeffrey B.; Price, Lori Lyn; Song, Jing; Eaton, Charles B.; Hochberg, Marc C.; Jackson, Rebecca D.; Kwoh, C. Kent; Nevitt, Michael C.; Dunlop, Dorothy D.

    2015-01-01

    Objective Pain is not always correlated with radiographic osteoarthritis (OA) severity possibly because people modify activities to manage symptoms. Measures of symptoms that consider pain in the context of activity level may therefore provide greater discrimination than pain alone. Our objective was to compare discrimination of a measure of pain alone with combined measures of pain relative to physical activity across radiographic OA levels. Methods This is a cross-sectional study of the Osteoarthritis Initiative accelerometer substudy, including those with and without knee OA. Two composite pain and activity knee symptom (PAKS) scores were calculated as Western Ontario and McMaster (WOMAC) Universities Osteoarthritis Pain Scale plus one divided by physical activity measures (step and activity counts). Symptom score discrimination across Kellgren and Lawrence (KL) grades were evaluated using histograms and quantile regression. Results 1806 participants, mean age 65.1 (9.1) years, mean BMI 28.4 (4.8) kg/m2, and 55.6% female, were included. WOMAC, but not PAKS scores, exhibited a floor effect. Adjusted median WOMAC by KL grades 0 – 4 were 0, 0, 1, 1, and 3 respectively. Median PAKS1 and PAKS2 were 24.9, 26.0, 32.4, 46.1, 97.9, and 7.2, 7.2, 9.2, 12.9, 23.8, respectively. PAKS scores had more statistically significant comparisons between KL grades compared with WOMAC. Conclusions Symptom assessments incorporating pain and physical activity did not exhibit a floor effect and were better able to discriminate radiographic severity than pain alone, particularly in milder disease. Pain in the context of physical activity level should be used to assess knee OA symptoms. PMID:26407008

  11. Integrated analysis of breast cancer cell lines reveals unique signaling pathways

    PubMed Central

    Heiser, Laura M; Wang, Nicholas J; Talcott, Carolyn L; Laderoute, Keith R; Knapp, Merrill; Guan, Yinghui; Hu, Zhi; Ziyad, Safiyyah; Weber, Barbara L; Laquerre, Sylvie; Jackson, Jeffrey R; Wooster, Richard F; Kuo, Wen Lin; Gray, Joe W; Spellman, Paul T

    2009-01-01

    Background Cancer is a heterogeneous disease resulting from the accumulation of genetic defects that negatively impact control of cell division, motility, adhesion and apoptosis. Deregulation in signaling along the EgfR-MAPK pathway is common in breast cancer, though the manner in which deregulation occurs varies between both individuals and cancer subtypes. Results We were interested in identifying subnetworks within the EgfR-MAPK pathway that are similarly deregulated across subsets of breast cancers. To that end, we mapped genomic, transcriptional and proteomic profiles for 30 breast cancer cell lines onto a curated Pathway Logic symbolic systems model of EgfR-MAPK signaling. This model was composed of 539 molecular states and 396 rules governing signaling between active states. We analyzed these models and identified several subtype-specific subnetworks, including one that suggested Pak1 is particularly important in regulating the MAPK cascade when it is over-expressed. We hypothesized that Pak1 over-expressing cell lines would have increased sensitivity to Mek inhibitors. We tested this experimentally by measuring quantitative responses of 20 breast cancer cell lines to three Mek inhibitors. We found that Pak1 over-expressing luminal breast cancer cell lines are significantly more sensitive to Mek inhibition compared to those that express Pak1 at low levels. This indicates that Pak1 over-expression may be a useful clinical marker to identify patient populations that may be sensitive to Mek inhibitors. Conclusions All together, our results support the utility of symbolic system biology models for identification of therapeutic approaches that will be effective against breast cancer subsets. PMID:19317917

  12. Integrated analysis of breast cancer cell lines reveals unique signaling pathways

    SciTech Connect

    Heiser, Laura M.; Wang, Nicholas J.; Talcott, Carolyn L.; Laderoute, Keith R.; Knapp, Merrill; Guan, Yinghui; Hu, Zhi; Ziyad, Safiyyah; Weber, Barbara L.; Laquerre, Sylvie; Jackson, Jeffrey R.; Wooster, Richard F.; Kuo, Wen-Lin; Gray, Joe W.; Spellman, Paul T.

    2009-03-31

    Cancer is a heterogeneous disease resulting from the accumulation of genetic defects that negatively impact control of cell division, motility, adhesion and apoptosis. Deregulation in signaling along the EGFR-MAPK pathway is common in breast cancer, though the manner in which deregulation occurs varies between both individuals and cancer subtypes. We were interested in identifying subnetworks within the EGFR-MAPK pathway that are similarly deregulated across subsets of breast cancers. To that end, we mapped genomic, transcriptional and proteomic profiles for 30 breast cancer cell lines onto a curated Pathway Logic symbolic systems model of EGFR-MEK signaling. This model was comprised of 539 molecular states and 396 rules governing signaling between active states. We analyzed these models and identified several subtype specific subnetworks, including one that suggested PAK1 is particularly important in regulating the MAPK cascade when it is over-expressed. We hypothesized that PAK1 overexpressing cell lines would have increased sensitivity to MEK inhibitors. We tested this experimentally by measuring quantitative responses of 20 breast cancer cell lines to three MEK inhibitors. We found that PAK1 over-expressing luminal breast cancer cell lines are significantly more sensitive to MEK inhibition as compared to those that express PAK1 at low levels. This indicates that PAK1 over-expression may be a useful clinical marker to identify patient populations that may be sensitive to MEK inhibitors. All together, our results support the utility of symbolic system biology models for identification of therapeutic approaches that will be effective against breast cancer subsets.

  13. The Role of Startups

    SciTech Connect

    Babinec, Sue

    2015-02-11

    Many ARPA-E-funded universities and research institutions have created start-up companies to further catalyze their next-generation technologies. Ambri and BlackPak are two examples of ARPA-E projects that were spun out by other institutions—Massachusetts Institute of Technology and SRI International, respectively—in an effort to get their technologies out of the lab and into the market quickly. This video features remarks from ARPA-E Senior Commercialization Advisor Sue Babinec and interviews with technologists at Ambri and BlackPak, who each tell the story of how their new companies spun out of the lab and have become agile startups capable of delivering real products to the marketplace.

  14. Myxoma and vaccinia viruses exploit different mechanisms to enter and infect human cancer cells

    SciTech Connect

    Villa, Nancy Y.; Bartee, Eric; Mohamed, Mohamed R.; Rahman, Masmudur M.; Barrett, John W.; McFadden, Grant

    2010-06-05

    Myxoma (MYXV) and vaccinia (VACV) viruses have recently emerged as potential oncolytic agents that can infect and kill different human cancer cells. Although both are structurally similar, it is unknown whether the pathway(s) used by these poxviruses to enter and cause oncolysis in cancer cells are mechanistically similar. Here, we compared the entry of MYXV and VACV-WR into various human cancer cells and observed significant differences: 1 - low-pH treatment accelerates fusion-mediated entry of VACV but not MYXV, 2 - the tyrosine kinase inhibitor genistein inhibits entry of VACV, but not MYXV, 3 - knockdown of PAK1 revealed that it is required for a late stage event downstream of MYXV entry into cancer cells, whereas PAK1 is required for VACV entry into the same target cells. These results suggest that VACV and MYXV exploit different mechanisms to enter into human cancer cells, thus providing some rationale for their divergent cancer cell tropisms.

  15. Myxoma and vaccinia viruses exploit different mechanisms to enter and infect human cancer cells.

    PubMed

    Villa, Nancy Y; Bartee, Eric; Mohamed, Mohamed R; Rahman, Masmudur M; Barrett, John W; McFadden, Grant

    2010-06-05

    Myxoma (MYXV) and vaccinia (VACV) viruses have recently emerged as potential oncolytic agents that can infect and kill different human cancer cells. Although both are structurally similar, it is unknown whether the pathway(s) used by these poxviruses to enter and cause oncolysis in cancer cells are mechanistically similar. Here, we compared the entry of MYXV and VACV-WR into various human cancer cells and observed significant differences: 1--low-pH treatment accelerates fusion-mediated entry of VACV but not MYXV, 2--the tyrosine kinase inhibitor genistein inhibits entry of VACV, but not MYXV, 3--knockdown of PAK1 revealed that it is required for a late stage event downstream of MYXV entry into cancer cells, whereas PAK1 is required for VACV entry into the same target cells. These results suggest that VACV and MYXV exploit different mechanisms to enter into human cancer cells, thus providing some rationale for their divergent cancer cell tropisms.

  16. Optimization of the treatment of wheat samples for the determination of phytic acid by HPLC with refractive index detection.

    PubMed

    Amaro, Rosa; Murillo, Miguel; González, Zurima; Escalona, Andrés; Hernández, Luís

    2009-01-01

    The treatment of wheat samples was optimized before the determination of phytic acid by high-performance liquid chromatography with refractive index detection. Drying by lyophilization and oven drying were studied; drying by lyophilization gave better results, confirming that this step is critical in preventing significant loss of analyte. In the extraction step, washing of the residue and collection of this water before retention of the phytates in the NH2 Sep-Pak cartridge were important. The retention of phytates in the NH2 Sep-Pak cartridge and elimination of the HCI did not produce significant loss (P = 0.05) in the phytic acid content of the sample. Recoveries of phytic acid averaged 91%, which is a substantial improvement with respect to values reported by others using this methodology.

  17. The Role of Startups

    ScienceCinema

    Babinec, Sue

    2016-07-12

    Many ARPA-E-funded universities and research institutions have created start-up companies to further catalyze their next-generation technologies. Ambri and BlackPak are two examples of ARPA-E projects that were spun out by other institutions—Massachusetts Institute of Technology and SRI International, respectively—in an effort to get their technologies out of the lab and into the market quickly. This video features remarks from ARPA-E Senior Commercialization Advisor Sue Babinec and interviews with technologists at Ambri and BlackPak, who each tell the story of how their new companies spun out of the lab and have become agile startups capable of delivering real products to the marketplace.

  18. Validation of a Rapid and Reliable Test for Diagnosis of Chagas' Disease by Detection of Trypanosoma cruzi-Specific Antibodies in Blood of Donors and Patients in Central America

    PubMed Central

    Ponce, Carlos; Ponce, Elisa; Vinelli, Elizabeth; Montoya, Alberto; de Aguilar, Vilma; Gonzalez, Antonio; Zingales, Bianca; Rangel-Aldao, Rafael; Levin, Mariano J.; Esfandiari, Javan; Umezawa, Eufrosina S.; Luquetti, Alejandro O.; da Silveira, José Franco

    2005-01-01

    In this study we compared the performance of the Chagas Stat-Pak rapid immunochromatographic test with a standard enzyme-linked immunosorbent assay (ELISA) in the serodiagnosis of Chagas' disease in Central America. Out of 3,400 blood donor samples, 156 (4.6%) were positive in both assays. Three sera out of 2,084 samples from reference laboratories were negative with the rapid test but positive with the ELISA (99.8% agreement). Agreement of 100% between the two tests was observed with 339 additional sera from patients with cardiopathies and 175 sera from potential blood donors in emergency surgical cases occurring on weekends or at night. In conclusion, Chagas Stat-Pak showed 99.6% and 99.9% sensitivity and specificity, respectively, when assayed with 5,998 serum samples. It is a sensitive and specific alternative to the ELISA, as required in medical emergencies and blood screenings in Central America. PMID:16207963

  19. Analysis of Kidney Stones in the Submarine Force

    DTIC Science & Technology

    1992-12-28

    the decreased the incidence of Diagnosis of urinary calculus by 86%. Our Urolithiasis . population is extremely Unpublished, 1989. motivated and would...of 13. Frank M, et al. North America Prevention of 1990;19:773-803. Urolithiasis . Archives of Environmental Health 7. Ryall RL, Marshall yR. 1966;13...625-30. The Investigation and Management of Idiopathic 14. Pak CY. Prevention of Urolithiasis . In: Recurrent Wickham JE, Buck AC, Nephrolithiasis. In

  20. Post Admission Cognitive Therapy (PACT) for the Inpatient Treatment of Military Personnel with Suicidal Behaviors: A Multi-Site Randomized Controlled Trial

    DTIC Science & Technology

    2015-02-01

    direct stress of a military career. 15. SUBJECT TERMS Suicide Prevention, Acute Care, Inpatient Treatment, Cognitive Behavior Therapy 16. SECURITY...future suicide behavior, specifically young, psychiatrically hospitalized adults under the direct stress of a military career. The development and...for suicide . In Comprehensive Guide to Post-Traumatic Stress Disorders. London: Springer. 7. Tucker, J., Neely, L. L., Colborn, V., Tylor, S., Pak, K

  1. Simultaneous extraction of. beta. -endorphin and leu- and met-enkephalins from human and rat plasma

    SciTech Connect

    Bhathena, S.J.; Smith, P.M.; Kennedy, B.W. ); Voyles, N.R.; Recant, L. )

    1989-01-01

    A simple, rapid and reliable procedure is described to simultaneously concentrated and purify {beta}-endorphin, leu-and met-enkephalins from small volumes of human and rat plasma before radioimmunoassay is performed. It uses C{sub 18} Sep-Pak reverse phase cartridges. The effectiveness of different protease inhibitors in preventing degradation of opiates by plasma and different solvent systems for eluting opiates is also evaluated.

  2. Complete Genome Sequence of a Velogenic Newcastle Disease Virus Strain Isolated from a Clinically Healthy Exotic Parakeet (Melopsittacus undulatus) in Pakistan

    PubMed Central

    Wajid, Abdul; Basharat, Asma; Khan, Taseer Ahmed; Wasim, Muhammad

    2017-01-01

    ABSTRACT The complete genome sequence of a virulent Newcastle disease virus (vNDV) strain isolated from an exotic parakeet (Melopsittacus undulatus) is described here. The virulent strain parakeet/Pak/R-Pindi/SFR-16/2016 was isolated from a bird reared as a pet in the province of Punjab in the northern region of Pakistan in 2016. Phylogenetic analysis classified the isolate as a member of NDV class II, subgenotype VIIi, in genotype VII. PMID:28183762

  3. Security and the Environment in Pakistan

    DTIC Science & Technology

    2010-08-03

    es/ess/compendium_2006/pdf/PAK_ESS_E.pdf. GDP estimates may vary depending in part on the inclusion of cotton production and fisheries . 15 Muhammad...essential in order to prevent salt water intrusion from destroying coastal mangrove stands and farm land. An estimated 85% of the Indus River Delta... mangroves , which are important spawning grounds for fish and crustaceans, have been lost. Flow diversion, leading to increased salinity levels, as well

  4. Stratus Ocean Reference Station (20 deg S, 85 deg W) Mooring Recovery and Deployment Cruise, R/V Ronald H. Brown Cruise 06-07, October 9-October 27, 2006

    DTIC Science & Technology

    2007-02-01

    47 IV.B.5. Time series of downward solar flux ............................................................ 48 IV.B.6. Time series of downward IR...flux ............................................................... 48 IV.B.7. Time series of non- solar surface heat flux components...and a Systron-Donner motion- pak. (2) A mean T/RH sensor in an aspirator on the jackstaff. (3) Solar and IR radiometers (Eppley pyranometers and

  5. Delayed fluorescence spectra of intact leaves photoexcited by sunlight measured with a multichannel Fourier-transform chemiluminescence spectrometer

    NASA Astrophysics Data System (ADS)

    Akita, Saeka; Yano, Ayako; Ishii, Hiroshi; Satoh, Chikahiro; Akai, Nobuyuki; Nakata, Munetaka

    2013-06-01

    Delayed fluorescence spectra of intact leaves of Green pak choi (Brassica rapa var. chinensis) were measured with a multichannel Fourier-transform chemiluminescence spectrometer, which we developed recently. The intact samples, photoexcited by sunlight without artificial light sources, showed delayed fluorescence around 740 nm with a lifetime of ˜6 s. The observed spectra were deconvoluted into two Gaussian bands: the delayed fluorescence from photosystem II and photosystem I complexes. Their relative intensities depended on the chlorophyll concentration, but their wavelengths were unchanged.

  6. Conference Proceedings of NASA/DoD Controls-Structures Interaction Technology Held in San Diego, California on 29 January-2 February 1989,

    DTIC Science & Technology

    1989-08-01

    Suite H 2015 Neil Avenue San Diego, CA 92024 Columbus. OH1 43210 Young H. Pak Richard S. Pappa Bus. Phor e: 714-896-4682 Bus. Phone: 804-864-4321...of the FMI was inspired by the work of Mike Shao of the Harvard Smithsonian Astrophysical Observatory. Dr. Shao currently has in operation, on Mount...1995 to 2015 (Astronomy and Astrophysics), National Academy Press, Washington, DC, 1988. 2. NASA Office of Space Science and Applications 1988

  7. The Soviet Union in Afghanistan: Benefits and Costs.

    DTIC Science & Technology

    1980-06-12

    COLLEGE Carlisle Barracks, Pennsylvania THE SOVIET NION IN4FGHANISTAN: S BENFITS AND COSTS, by SS-hirin/Fahir-Kheli ,/;/ 12 June480./ -/ . DISTRIBUTION...1973. The Soviet Union committed itself to 20 major projects in agriculture, irrigation, electric power, oil and gas exploration , mineral and metal...regime spread, Pak-Afghan relations deteriorated. Pakistan counted 56 violations of its air and ground space (penetratioins of up to three miles above

  8. Behavior of Materials at Cold Regions Temperatures. Part 1. Program Rationale and Test Plan

    DTIC Science & Technology

    1988-07-01

    10 15 i" 0% I 0 0 0 2 3 4 5a 0 1 2 3 4 5 6 7 lf CSTRAIN (IN/INbST AI OW N V, Figure B47. Static stress vs strain curves for resilient expanded ... polystyrene foam (Resilo-Pak) , at temperature extremes (0.5, 0. 75 and 1.3 lb Ifft3 densities) (Titus 1967). so ,eSF 50 -- 8 * TEMPERATURE. K 0 so 100 is

  9. Evaluating the validity of the serologic test for detecting Helicobacter pylori infection in Mongolian gerbils.

    PubMed

    Kuo, Chao-Hung; Yu, Fang-Jung; Tsai, Pei-Yun; Yang, Sheau-Fang; Chang, Lin-Li; Jan, Chang-Ming; Wang, Wen-Ming; Wu, Deng-Chyang

    2007-11-01

    A strong correlation between Helicobacter pylori infection and gastric cancer has been reported. Mongolian gerbils are regarded as the most suitable animal model in which to study carcinogenesis associated with H. pylori. The aim of our study was to evaluate the accuracy of the serologic test for detecting H. pylori infection in Mongolian gerbils. The model was developed as follows: the H. pylori colony (vacuolating cytotoxin A (+)/cytotoxin-associated gene A (+)) was cultured from the mucosas of previously H. pylori-fed gerbils. These colonies were cultured in broth. Then,we fed the gerbils with 0.5-1 mL of broth (about 10(9) CFU/mL) (intragastric administration) twice within a 3-day period. After inoculation for 6 or 26 weeks, the gerbils were sacrificed and their gastric mucosas were sampled for a series of examinations. Blood samples for serologic testing (STAT-PAK) were collected. H. pylori infection was confirmed. Statistical analysis was performed using the Chi-square test. Differences were regarded as significant when the p value was less than 0.05. A total of 50 gerbils were inoculated with H. pylori and the success rate reached 88%. All 10 gerbils in the control group showed a negative result. Damage to the mucosas was more obvious following increasing periods of inoculation. The rates of sensitivity and specificity, as determined by the STAT-PAK test, were 90.9% and 100%, respectively. The positive and negative predictive values were 100% and 60%, respectively. The STAT-PAK test seemed to be more sensitive and accurate (p < 0.05) in high H. pylori densities. In conclusion, the STAT-PAK test (blood-sampling) showed acceptable results and was suitable for long-term observation of H. pylori infection.

  10. JPRS Report, Soviet Union, International Affairs.

    DTIC Science & Technology

    1987-11-27

    Revolutionaries" by B. Pak; the conclud- ing part of I. Belyaev’s article "Egypt. The Revolution and the President" "Mahatmah Gandhi as Described by...O. Martyshin. Mahatmah Gandhi in the Evaluations of Soviet Researchers (p 22) Problems and Opinions VI. Li, G. Mirskiy. Socialist Orientation in...Scientific Life V. Rozhkov. A Forum for Mongolian Scholars (p 44) Culture, Literature, Art V. Ozhogin. Lev Tolstoy and Japanese Culture (p 46

  11. Contingency Pest Management Guide. 2010 Edition

    DTIC Science & Technology

    2010-08-01

    Medicine (USACHPPM) in accordance with DODI 6490.03, Deployment Health, for archiving in the Defense Occupational and Environmental Health Readiness ...Strip (Starbar) 1% Nithiazine 6840-01-467-0994 BX of 12 Ready to Use as directed on label Primarily For Outdoor use. May be used in stables...glyphosate, ( Roundup Dry Pak) 0.96%, water soluble packets 6840-01-399-0673 BX (25 packets) 1 packet/gal, or As directed on label Provides

  12. Neurotoxin Mitigation

    DTIC Science & Technology

    2007-11-01

    biotinamidopentyldecanamide HPLC high performance liquid chromatography ip intraperitoneal LC/MS/MS liquid chromatography coupled to tandem mass...outdated human plasma by ion exchange chromatography at pH 4.0, affinity chromatography on procainamide- Sepharose, and HPLC on a Protein Pak DEAE Anion...porcine trypsin (V5113C sequencing grade modified trypsin in 50 mM acetic acid, Promega) at 37˚C for 5.5 h. HPLC to purify the active site peptide

  13. 7 CFR 361.5 - Sampling of seeds.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Cress, garden 50 5 10 Cress, upland 35 5 10 Cress, water 25 5 10 Cucumber 500 25 50 Dandelion 35 5 10 Dill 50 5 10 Eggplant 50 5 10 Endive 50 5 10 Gherkin, West India 160 25 50 Kale 50 5 10 Kale, Chinese..., India 50 25 100 Mustard, spinach 50 5 10 Okra 500 25 50 Onion 50 5 10 Onion, Welsh 50 5 10 Pak-choi 50...

  14. Performance of converted pressure cookers and two conventional jars for anaerobic bacterial culture.

    PubMed

    Gargan, R A; Phillips, I

    1978-05-01

    The simple conversion of commercial pressure cookers into inexpensive anaerobic jars is described. These containers were shown to be as good as the small conventional BBL polycarbonate GasPak and large vented 150 gas-replacement jars when assessed by means of three biological indicators: Pseudomonas aeruginosa, Bacteroides melaninogenicus, and Bacteroides fragilis. Ps. aeruginosa seeded on Simmond's citrate agar was shown to be the most sensitive indicator of the three for traces of oxygen.

  15. LPP is Required for TGF-Beta Induced Motility and Invasion of Neu/ErbB-2 Expressing Breast Cancer Cells

    DTIC Science & Technology

    2012-09-01

    Gorska, A. E., Moses, H. L. and Arteaga , C. L. (2003). Increased Malignancy of Neu-Induced Mammary Tumors Overexpressing Active Transforming Growth Factor...National Academy of Sciences 100, 8430-8435. 5) Ueda, Y., Wang, S., Dumont, N., Yi, J. Y., Koh, Y. and Arteaga , C. L. (2004). Overexpression of HER2...B. and Arteaga , C. L. (2006). HER2/Neu (ErbB2) signaling to Rac1-Pak1 is temporally and spatially modulated by transforming growth factor beta

  16. Pressure Vessel Calculations for VVER-440 Reactors

    NASA Astrophysics Data System (ADS)

    Hordósy, G.; Hegyi, Gy.; Keresztúri, A.; Maráczy, Cs.; Temesvári, E.; Vértes, P.; Zsolnay, É.

    2003-06-01

    Monte Carlo calculations were performed for a selected cycle of the Paks NPP Unit II to test a computational model. In the model the source term was calculated by the core design code KARATE and the neutron transport calculations were performed by the MCNP. Different forms of the source specification were examined. The calculated results were compared with measurements and in most cases fairly good agreement was found.

  17. Pakistan: Frontline State Again?

    DTIC Science & Technology

    1995-12-01

    leader Mohammed Ali Jinnah embarked on a new strategy against Indian Congress domination stating that Islam was in danger and the congress was...A. OUTLINE OF THE STUDY ............................. 4 II. A COLD W AR W ITHIN ...................................... 9 A. LEGACY OF PARTITION ...in the region, one absolute of Indo-Pak relations remains after the end of the cold war. Since the partition of India in 1947, Pakistan has been

  18. JPRS Report, Near East & South Asia

    DTIC Science & Technology

    2007-11-02

    Garment Industries Look for Quota-Free Bases in Gulf [GULF NEWS 1 May] 36 SOUTH ASIA BANGLADESH Students Riot Over Expulsions [AFP] 38...4,500 Chakma Tribes People Flee Bangladesh Violence [AFP] 52 Editorial Examines Obstacles in Indo-Pak Relations [ANANDA BAZAR PATRIKA 10 Feb] 52...initiated dealing with the engineer- ing geology requirements for building a road across terrain with many sinkholes and solution cavities, for an

  19. Dynamics of Flagellum- and Pilus-Mediated Association of Pseudomonas aeruginosa with Contact Lens Surfaces▿

    PubMed Central

    Tran, Victoria B.; Fleiszig, Suzanne M. J.; Evans, David J.; Radke, Clayton J.

    2011-01-01

    Flagella and pili are appendages that modulate attachment of Pseudomonas aeruginosa to solid surfaces. However, previous studies have mostly reported absolute attachment. Neither the dynamic roles of these appendages in surface association nor those of attachment phenotypes have been quantified. We used video microscopy to address this issue. Unworn, sterile, soft contact lenses were placed in a laminar-flow optical chamber. Initial lens association kinetics for P. aeruginosa strain PAK were assessed in addition to lens-surface association phenotypes. Comparisons were made to strains with mutations in flagellin (fliC) or pilin (pilA) or those in flagellum (motAB) or pilus (pilU) function. PAK and its mutants associated with the contact lens surface at a constant rate according to first-order kinetics. Nonswimming mutants associated ∼30 to 40 times slower than the wild type. PAK and its pilA mutant associated at similar rates, but each ∼4 times faster than the pilU mutant. Lens attachment by wild-type PAK induced multiple phenotypes (static, lateral, and rotational surface movement), each showing only minor detachment. Flagellin (fliC) and flagellar-motility (motAB) mutants did not exhibit surface rotation. Conversely, strains with mutations in pilin (pilA) and pilus retraction (pilU) lacked lateral-surface movement but displayed enhanced surface rotation. Slower surface association of swimming-incapable P. aeruginosa mutants was ascribed to lower convective-diffusion-arrival rates, not to an inability to adhere. Flagellum function (swimming) enhanced lens association, attachment, and rotation; hyperpiliation hindered lens association. P. aeruginosa bound through three different adhesion sites: flagellum, pili, and body. Reduction of bacterial attachment to contact lenses thus requires blockage of multiple adhesion phenotypes. PMID:21498762

  20. DNA studies are necessary for accurate patient diagnosis in compound heterozygosity for Hb Adana (HBA2:c.179>A) with deletional or nondeletional α-thalassaemia

    PubMed Central

    Tan, Jin Ai Mary Anne; Kho, Siew Leng; Ngim, Chin Fang; Chua, Kek Heng; Goh, Ai Sim; Yeoh, Seoh Leng; George, Elizabeth

    2016-01-01

    Haemoglobin (Hb) Adana (HBA2:c.179>A) interacts with deletional and nondeletional α-thalassaemia mutations to produce HbH disorders with varying clinical manifestations from asymptomatic to severe anaemia with significant hepatosplenomegaly. Hb Adana carriers are generally asymptomatic and haemoglobin subtyping is unable to detect this highly unstable α-haemoglobin variant. This study identified 13 patients with compound heterozygosity for Hb Adana with either the 3.7 kb gene deletion (-α3.7), Hb Constant Spring (HbCS) (HBA2:c.427T>C) or Hb Paksé (HBA2:429A>T). Multiplex Amplification Refractory Mutation System was used for the detection of five deletional and six nondeletional α-thalassaemia mutations. Duplex-PCR was used to confirm Hb Paksé and HbCS. Results showed 84.6% of the Hb Adana patients were Malays. Using DNA studies, compound heterozygosity for Hb Adana and HbCS (αcodon 59α/αCSα) was confirmed in 11 patients. A novel point in this investigation was that DNA studies confirmed Hb Paksé for the first time in a Malaysian patient (αcodon 59α/αPakséα) after nine years of being misdiagnosis with Hb Adana and HbCS (αcodon 59α/αCSα). Thus, the reliance on haematology studies and Hb subtyping to detect Hb variants is inadequate in countries where thalassaemia is prevalent and caused by a wide spectrum of mutations. PMID:27271331

  1. In vivo yeast cell morphogenesis is regulated by a p21-activated kinase in the human pathogen Penicillium marneffei.

    PubMed

    Boyce, Kylie J; Schreider, Lena; Andrianopoulos, Alex

    2009-11-01

    Pathogens have developed diverse strategies to infect their hosts and evade the host defense systems. Many pathogens reside within host phagocytic cells, thus evading much of the host immune system. For dimorphic fungal pathogens which grow in a multicellular hyphal form, a central attribute which facilitates growth inside host cells without rapid killing is the capacity to switch from the hyphal growth form to a unicellular yeast form. Blocking this transition abolishes or severely reduces pathogenicity. Host body temperature (37 degrees C) is the most common inducer of the hyphal to yeast transition in vitro for many dimorphic fungi, and it is often assumed that this is the inducer in vivo. This work describes the identification and analysis of a new pathway involved in sensing the environment inside a host cell by a dimorphic fungal pathogen, Penicillium marneffei. The pakB gene, encoding a p21-activated kinase, defines this pathway and operates independently of known effectors in P. marneffei. Expression of pakB is upregulated in P. marneffei yeast cells isolated from macrophages but absent from in vitro cultured yeast cells produced at 37 degrees C. Deletion of pakB leads to a failure to produce yeast cells inside macrophages but no effect in vitro at 37 degrees C. Loss of pakB also leads to the inappropriate production of yeast cells at 25 degrees C in vitro, and the mechanism underlying this requires the activity of the central regulator of asexual development. The data shows that this new pathway is central to eliciting the appropriate morphogenetic response by the pathogen to the host environment independently of the common temperature signal, thus clearly separating the temperature- and intracellular-dependent signaling systems.

  2. Estrogen and the Dietary Phytoestrogen Tesveratrol as Regulators of the Rho GTPase Rac in Breast Cancer Research

    DTIC Science & Technology

    2008-06-01

    AD_________________ Award Number: W81XWH-07-1-0330 TITLE: Estrogen and the Dietary Phytoestrogen ...COVERED 7 May 2007 – 6 May 2008 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Estrogen and the Dietary Phytoestrogen Tesveratrol as Regulators of the... estrogen (E2), or resveratrol (Res). PAK-PBD-GST beads were used to pull-down active GTP- bound Rac from the cell lysates. Active and total Rac levels

  3. Notification levels related to the control of radioactive discharges.

    PubMed

    Kanyár, B; Volent, G; Tóth, K

    2008-11-01

    In relation to the management of elevated discharges of radioactive materials from the Paks Nuclear Power Plant, two different types of notification level are proposed. They relate to discharges which are above normal but below those associated with an emergency. The first is technically based (reference level) while the second (reporting level) is mainly based on considerations of public concern. The actions expected to be implemented if either of the levels is exceeded are summarized.

  4. Korean Affairs Report No. 321

    DTIC Science & Technology

    2007-11-02

    the incident might be caused by an internal foe as in the case of the assassination of Pak Jung -hi in 1979. Another paper of the country MERDEKA 11...Comrade Henryk Jablonski, president of the Council of State of the Polish People’s Republic; Comrade Gustav Husak, general secretary of the Central...president, and Dominic Mintoff, prime minister, of the Republic of Malta; Carl Gustaf, king of Sweden; Rudolf Kirchschlager, president of the

  5. Positive skin and serologic test results of diagnostic assays for bovine tuberculosis and subsequent isolation of Mycobacterium interjectum in a pygmy hippopotamus (Hexaprotodon liberiensis).

    PubMed

    Bouts, Tim; Vordermeier, Martin; Flach, Edmund; Routh, Andrew

    2009-09-01

    A 20-yr-old male pygmy hippopotamus (Hexaprotodon liberiensis), weighing 250 kg, arrived at Zoological Society London Whipsnade Zoo (United Kingdom) from a captive collection in Portugal. A quarantine health check was performed including a comparative intradermal tuberculosis (IDTB) test. Assessment of the comparative IDTB test at 72 hr revealed a strong positive reaction at the bovine site. Serum was tested with a rapid immunochromatographic assay (TB STAT-PAK) and was positive for tuberculosis antibodies. The tuberculosis tests were repeated 6 wk later with the same positive test outcome. In addition, a broncho-alveolar lavage (BAL) was submitted for mycobacterial culture. The positive IDTB test and TB STAT-PAK results were supported by multiantigen print immunoassay (MAPIA). Based on these results, the animal was suspected to be infected with Mycobacterium tuberculosis complex organisms and was euthanized. No gross or histologic signs of tuberculosis were found at postmortem examination. Mycobacterium interjectum was cultured from the BAL but not from necropsy samples. The antigens used in the TB STAT-PAK and MAPIA tests are reportedly specific for the M. tuberculosis complex, and so it is possible this animal presented with a latent case of tuberculosis or had a previous tuberculosis infection that resolved prior to testing. Cross-reactions with nontuberculous mycobacteria have been described with TB STAT-PAK and MAPIA tests. However, Western blotting analysis using serum from this animal did not recognize M. interjectum proteins of equivalent size to the M. tuberculosis-Mycobacterium bovis proteins recognized in the MAPIA. Thus, antigenic cross-reactivity with M. interjectum can be deemed less likely, but other nontuberculous mycobacterial proteins cannot be ruled out. It is therefore possible that false-positive reactions were obtained. These results highlight the difficulty of diagnosing tuberculosis in the absence of pathology and the presence of

  6. Indicators of NGO Security in Afghanistan

    DTIC Science & Technology

    2004-12-14

    warlord in charge of the province, the aid group is less likely to come under attack. Despite their many faults , the Taliban government curbed the...province share a boundary with Afghanistan border?(Yes/No) Source: Afghanistan Information Management System shapefile 7. BorderPak- Does the...province share a boundary with Pakistan? (Yes/No) Source: Afghanistan Information Management System shapefile 8. Foodpop#- Beneficiary population of

  7. USNA: A Dual-Classifier Approach to Contextual Sentiment Analysis

    DTIC Science & Technology

    2013-06-01

    algorithms use emoticons as seman- tic indicators of polarity. For instance, a tweet that contains a sad face likely contains a negative polar- ity (Read...2005; Go et al., 2009; Bifet and Frank, 2010; Pak and Paroubek, 2010; Davidov et al., 2010; Kouloumpis et al., 2011). In a similar vein, hash- tags ...topic (e.g., “US President”), and retrieve tweets that contain the word (O’Connor et al., 2010; Tumasjan et al., 2010; Tan et al., 2011). These sys

  8. Amino acid sequence of atrial natriuretic peptides in human coronary sinus plasma.

    PubMed

    Yandle, T; Crozier, I; Nicholls, G; Espiner, E; Carne, A; Brennan, S

    1987-07-31

    Two atrial natriuretic peptides were purified from pooled human coronary sinus plasma by Sep-Pak extraction, immunoaffinity chromatography and reverse phase HPLC. The amino acid sequences of the two peptides were homologous with 99-126 human atrial natriuretic peptide (hANP) and 106-126 hANP, the latter being most probably linked to 99-105 ANP by the disulphide bond. The molar ratio of the peptides in plasma, as assessed by radioimmunoassay was 10:3.

  9. Radial Artery Pulse Change During Cold Pressor Test

    DTIC Science & Technology

    2007-11-02

    KRF-2000-015-PP0177) REFERENCES [1] C.H. Chen, C.T. Ting, A. Nussbacher, E. Nevo , D. A. Kass, P. Pak, S. P. Wang, M. S Chang, C.P. Frank, “Validation...E. Nevo , B. Fetics, H. Peter, F. C.P. Yin, W. L. Maughan,s D. A. Kass “Estimation of Central Aortic Pressure Waveform by Mathematical Transformation

  10. Microtubule-dependent transport of vimentin filament precursors is regulated by actin and by the concerted action of Rho- and p21-activated kinases.

    PubMed

    Robert, Amélie; Herrmann, Harald; Davidson, Michael W; Gelfand, Vladimir I

    2014-07-01

    Intermediate filaments (IFs) form a dense and dynamic network that is functionally associated with microtubules and actin filaments. We used the GFP-tagged vimentin mutant Y117L to study vimentin-cytoskeletal interactions and transport of vimentin filament precursors. This mutant preserves vimentin interaction with other components of the cytoskeleton, but its assembly is blocked at the unit-length filament (ULF) stage. ULFs are easy to track, and they allow a reliable and quantifiable analysis of movement. Our results show that in cultured human vimentin-negative SW13 cells, 2% of vimentin-ULFs move along microtubules bidirectionally, while the majority are stationary and tightly associated with actin filaments. Rapid motor-dependent transport of ULFs along microtubules is enhanced ≥ 5-fold by depolymerization of actin cytoskeleton with latrunculin B. The microtubule-dependent transport of vimentin ULFs is further regulated by Rho-kinase (ROCK) and p21-activated kinase (PAK): ROCK inhibits ULF transport, while PAK stimulates it. Both kinases act on microtubule transport independently of their effects on actin cytoskeleton. Our study demonstrates the importance of the actin cytoskeleton to restrict IF transport and reveals a new role for PAK and ROCK in the regulation of IF precursor transport.-Robert, A., Herrmann, H., Davidson, M. W., and Gelfand, V. I. Microtubule-dependent transport of vimentin filament precursors is regulated by actin and by the concerted action of Rho- and p21-activated kinases.

  11. Nutritional performance and activity of some digestive enzymes of the cotton bollworm, Helicoverpa armigera, in response to seven tested bean cultivars.

    PubMed

    Namin, Foroogh Rahimi; Naseri, Bahram; Razmjou, Jabraeil

    2014-01-01

    Nutritional performance and activity of some digestive enzymes (protease and α-amylase) of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in response to feeding on bean (Phaseolus vulgaris L. (Fabales: Fabaceae)) cultivars (Shokufa, Akhtar, Sayyad, Naz, Pak, Daneshkadeh, and Talash) were evaluated under laboratory conditions (25 ± 1°C, 65 ± 5% RH, and a 16:8 L:D photoperiod). The highest and lowest respective values of approximate digestibility were observed when fourth, fifth, and sixth larval instar H. armigera were fed red kidney bean Akhtar and white kidney bean Daneshkadeh. The efficiency of conversion of ingested and digested food was highest when H. armigera was fed red kidney beans Akhtar and Naz and lowest when they were fed white kidney bean Pak. The highest protease activity of fifth instars was observed when they were fed red kidney bean Naz, and the highest amylase activity of fifth instars was observed when they were fed red kidney bean Sayyad. Sixth instar larvae that fed on red kidney bean Sayyad showed the highest protease activity. Larvae reared on common bean Talash and white kidney bean Pak showed the highest amylase activity. Among bean cultivars tested, red kidney bean Sayyad was the most unsuitable host for feeding H. armigera.

  12. Filamin, a synaptic organizer in Drosophila, determines glutamate receptor composition and membrane growth

    PubMed Central

    Lee, GaYoung; Schwarz, Thomas L

    2016-01-01

    Filamin is a scaffolding protein that functions in many cells as an actin-crosslinker. FLN90, an isoform of the Drosophila ortholog Filamin/cheerio that lacks the actin-binding domain, is here shown to govern the growth of postsynaptic membrane folds and the composition of glutamate receptor clusters at the larval neuromuscular junction. Genetic and biochemical analyses revealed that FLN90 is present surrounding synaptic boutons. FLN90 is required in the muscle for localization of the kinase dPak and, downstream of dPak, for localization of the GTPase Ral and the exocyst complex to this region. Consequently, Filamin is needed for growth of the subsynaptic reticulum. In addition, in the absence of filamin, type-A glutamate receptor subunits are lacking at the postsynapse, while type-B subunits cluster correctly. Receptor composition is dependent on dPak, but independent of the Ral pathway. Thus two major aspects of synapse formation, morphological plasticity and subtype-specific receptor clustering, require postsynaptic Filamin. DOI: http://dx.doi.org/10.7554/eLife.19991.001 PMID:27914199

  13. The assembly of a GTPase–kinase signalling complex by a bacterial catalytic scaffold

    PubMed Central

    Selyunin, Andrey S.; Sutton, Sarah E.; Weigele, Bethany A.; Reddick, L. Evan; Orchard, Robert C.; Bresson, Stefan M.; Tomchick, Diana R.; Alto, Neal M.

    2011-01-01

    The fidelity and specificity of information flow within a cell is controlled by scaffolding proteins that assemble and link enzymes into signalling circuits1,2. These circuits can be inhibited by bacterial effector proteins that post-translationally modify individual pathway components3–6. However, there is emerging evidence that pathogens directly organize higher-order signalling networks through enzyme scaffolding7,8, and the identity of the effectors and their mechanisms of action are poorly understood. Here we identify the enterohaemorrhagic Escherichia coli O157:H7 type III effector EspG as a regulator of endomembrane trafficking using a functional screen, and report ADP-ribosylation factor (ARF) GTPases and p21-activated kinases (PAKs) as its relevant host substrates. The 2.5 Å crystal structure of EspG in complex with ARF6 shows how EspG blocks GTPase-activating-protein-assisted GTP hydrolysis, revealing a potent mechanism of GTPase signalling inhibition at organelle membranes. In addition, the 2.8 Å crystal structure of EspG in complex with the autoinhibitory Iα3-helix of PAK2 defines a previously unknown catalytic site in EspG and provides an allosteric mechanism of kinase activation by a bacterial effector. Unexpectedly, ARF and PAKs are organized on adjacent surfaces of EspG, indicating its role as a ‘catalytic scaffold’ that effectively reprograms cellular events through the functional assembly of GTPase-kinase signalling complex. PMID:21170023

  14. DEVELOPMENT OF A HYDRONIC ROOFTOP UNIT -- HYPAK

    SciTech Connect

    Eric Lee; Dick Bourne; Mark Berman

    2004-03-25

    The majority of US commercial floor space is cooled by rooftop HVAC units (RTU's). RTU popularity derives chiefly from their low initial cost and relative ease of service access without disturbing building occupants. Unfortunately, current RTU's are inherently inefficient due to a combination of characteristics that unnecessarily increase cooling loads and energy use. Existing RTU's in the U.S. consume an estimated 2.4 quads annually. Inefficient RTU's create an estimated 3.5% of U.S. CO{sub 2} emissions, thus contributing significantly to global warming. Also, RTU's often fail to maintain adequate ventilation air and air filtration. This project was developed to evaluate the feasibility of a radically new ''HyPak'' RTU design that significantly and cost-effectively increases RTU performance and delivered air quality. The objective of the HyPak Project was to design, develop and test a hydronic RTU that provides a quantum improvement over conventional RTU performance. Our proposal targeted 60% and 50% reduction in electrical energy use by the HyPak RTU for dry and humid climates, respectively, when compared with a conventional unit.

  15. Fission products from the damaged Fukushima reactor observed in Hungary.

    PubMed

    Bihari, Árpád; Dezső, Zoltán; Bujtás, Tibor; Manga, László; Lencsés, András; Dombóvári, Péter; Csige, István; Ranga, Tibor; Mogyorósi, Magdolna; Veres, Mihály

    2014-01-01

    Fission products, especially (131)I, (134)Cs and (137)Cs, from the damaged Fukushima Dai-ichi nuclear power plant (NPP) were detected in many places worldwide shortly after the accident caused by natural disaster. To observe the spatial and temporal variation of these isotopes in Hungary, aerosol samples were collected at five locations from late March to early May 2011: Institute of Nuclear Research, Hungarian Academy of Sciences (ATOMKI, Debrecen, East Hungary), Paks NPP (Paks, South-Central Hungary) as well as at the vicinity of Aggtelek (Northeast Hungary), Tapolca (West Hungary) and Bátaapáti (Southwest Hungary) settlements. In addition to the aerosol samples, dry/wet fallout samples were collected at ATOMKI, and airborne elemental iodine and organic iodide samples were collected at Paks NPP. The peak in the activity concentration of airborne (131)I was observed around 30 March (1-3 mBq m(-3) both in aerosol samples and gaseous iodine traps) with a slow decline afterwards. Aerosol samples of several hundred cubic metres of air showed (134)Cs and (137)Cs in detectable amounts along with (131)I. The decay-corrected inventory of (131)I fallout at ATOMKI was 2.1±0.1 Bq m(-2) at maximum in the observation period. Dose-rate contribution calculations show that the radiological impact of this event at Hungarian locations was of no considerable concern.

  16. Inhibition of Pseudomonas aeruginosa biofilm formation by 2,2’-bipyridyl, lipoic, kojic and picolinic acids

    PubMed Central

    Çevik, Kübra; Ulusoy, Seyhan

    2015-01-01

    Objective(s): The inhibitory effects of iron chelators, and FeCl3 chelation on biofilm formation and swarming motility were investigated against an opportunistic human pathogen Pseudomonas aeruginosa. Materials and Methods: The inhibitory activity of 2,2’-bipyridyl, lipoic acid, kojic acid and picolinic acid on biofilm formation of P. aeruginosa strain PAO1 and three clinical isolates (P. aeruginosa PAK01, P. aeruginosa PAK02 and P. aeruginosa PAK03) were investigated, based on crystal violet assay, and swarming motility test. Results: The kojic, lipoic and picolinic acid inhibited biofilm formation by 5-33% in all tested P. aeruginosa isolates. When chelated iron was added, biofilm inhibition rates were determined to be 39-57%. Among the tested chelators against P. aeruginosa, lipoic acid (84%) and kojic acid (68%) presented the highest inhibition of swarming motility. This is the first study to report the inhibitory effect of lipoic acid on biofilm formation and swarming motility of P. aeruginosa. Conclusion: It is considered that lipoic and picolinic acids can serve as alternatives for the treatment of the P. aeruginosa infections by inhibiting biofilm formation. PMID:26557964

  17. Die Kometenmission Rosetta

    NASA Astrophysics Data System (ADS)

    Krüger, Harald

    2016-11-01

    Die Rosetta-Mission ist ein Meilenstein in der Erforschung der Kometen und ihrer Entstehung. Eine der größten üerraschungen war die unregelmäßge hantelförmige Gestalt des Zielkometen 67P/Tschurjumow-Gerassimenko. Er besteht wahrscheinlich aus zwei Einzelkörpern, die durch ihre Schwerkraft aneinander gehalten werden. Seine Oberfläche ist sehr rau und zeigt eine sehr vielf ältige Morphologie, die auf eine Vielzahl von ablaufenden Prozessen hindeutet. Der Kometenkern ist vermutlich auf Gr ößnskalen von mehr als etwa 10 bis 100 Metern homogen, Inhomogenitäten auf kleineren Skalen k nnten f r seine Aktivä t verantwortlich sein. Diese ist auf kleine Gebiete konzentriert, und auch Oberflächenveränderungen, die sich innerhalb von einigen Tagen bis wenigen Wochen abspielen, sind lokal. Im Kometenmaterial wurde eine Vielzahl an organischen Substanzen gemessen, die zum Teil als Schlüsselmoleküle für die Synthese der Grundbausteine des Lebens gelten, wie wir es kennen.

  18. Animal Protection and Structural Studies of a Consensus Sequence Vaccine Targeting the Receptor Binding Domain of the Type IV Pilus of Pseudomonas aeruginosa

    SciTech Connect

    Kao, Daniel J.; Churchill, Mair E.A.; Irvin, Randall T.; Hodges, Robert S.

    2008-09-23

    One of the main obstacles in the development of a vaccine against Pseudomonas aeruginosa is the requirement that it is protective against a wide range of virulent strains. We have developed a synthetic-peptide consensus-sequence vaccine (Cs1) that targets the host receptor-binding domain (RBD) of the type IV pilus of P. aeruginosa. Here, we show that this vaccine provides increased protection against challenge by the four piliated strains that we have examined (PAK, PAO, KB7 and P1) in the A.BY/SnJ mouse model of acute P. aeruginosa infection. To further characterize the consensus sequence, we engineered Cs1 into the PAK monomeric pilin protein and determined the crystal structure of the chimeric Cs1 pilin to 1.35 {angstrom} resolution. The substitutions (T130K and E135P) used to create Cs1 do not disrupt the conserved backbone conformation of the pilin RBD. In fact, based on the Cs1 pilin structure, we hypothesize that the E135P substitution bolsters the conserved backbone conformation and may partially explain the immunological activity of Cs1. Structural analysis of Cs1, PAK and K122-4 pilins reveal substitutions of non-conserved residues in the RBD are compensated for by complementary changes in the rest of the pilin monomer. Thus, the interactions between the RBD and the rest of the pilin can either be mediated by polar interactions of a hydrogen bond network in some strains or by hydrophobic interactions in others. Both configurations maintain a conserved backbone conformation of the RBD. Thus, the backbone conformation is critical in our consensus-sequence vaccine design and that cross-reactivity of the antibody response may be modulated by the composition of exposed side-chains on the surface of the RBD. This structure will guide our future vaccine design by focusing our investigation on the four variable residue positions that are exposed on the RBD surface.

  19. Metabolomics differentiation of canola genotypes: toward an understanding of canola allelochemicals

    PubMed Central

    Asaduzzaman, M.; Pratley, James E.; An, Min; Luckett, David J.; Lemerle, Deirdre

    2015-01-01

    Allelopathy is one crop attribute that could be incorporated in an integrated weed management system as a supplement to synthetic herbicides. However, the underlying principles of crop allelopathy and secondary metabolite production are still poorly understood including in canola. In this study, an allelopathic bioassay and a metabolomic analysis were conducted to compare three non-allelopathic and three allelopathic canola genotypes. Results from the laboratory bioassay showed that there were significant differences among canola genotypes in their ability to inhibit root and shoot growth of the receiver annual ryegrass; impacts ranged from 14% (cv. Atr-409) to 76% (cv. Pak85388-502) and 0% (cv. Atr-409) to 45% (cv. Pak85388-502) inhibition respectively. The root length of canola also differed significantly between genotypes, there being a non-significant negative interaction (r = -0.71; y = 0.303x + 21.33) between the root length of donor canola and of receiver annual ryegrass. Variation in chemical composition was detected between organs (root extracts, shoot extracts) and root exudates and also between canola genotypes. Root extracts contained more secondary metabolites than shoot extracts while fewer compounds were recorded in the root exudates. Individual compound assessments identified a total of 14 secondary metabolites which were identified from the six tested genotypes. However, only Pak85388-502 and Av-opal exuded sinapyl alcohol, p-hydroxybenzoic acid and 3,5,6,7,8-pentahydroxy flavones in agar growth medium, suggesting that the synergistic effect of these compounds playing a role for canola allelopathy against annual ryegrass in vitro. PMID:25620971

  20. Retrospective serological investigation of bovine tuberculosis in two gemsbok (Oryx gazelle gazelle) and an onager (Equus hemionus onager).

    PubMed

    Schaftenaar, Willem; Lécu, Alex; Greenwald, Rena; Lyashchenko, Konstantin P

    2013-12-01

    In 1997 a 26-yr-old gemsbok (Oryx gazelle gazelle) died of bovine tuberculosis in a zoo. Three remaining gemsbok were administered the comparative tuberculin skin test repeatedly over a period of 5 mo. Two animals showed inconclusive results on the second test. All three gemsbok were euthanatized. Mycobacterium bovis was isolated from one of those with an inconclusive skin test result, whereas Mycobacterium fortuitum was detected in the other gemsbok. Eight years later, an onager (Equus hemionus onager) died of bovine tuberculosis. This animal had been kept in the same building as the gemsbok. Three herd mates were culled after administering the comparative tuberculin skin test. They were all nonreactors and produced no evidence of tuberculosis at postmortem examination. Retrospectively, using plasma samples collected from the gemsbok and onagers, three antibody tests, Elephant TB STAT-PAK, multiantigen print immunoassay (MAPIA), and dual-path platform (DPP) VetTB (Chembio Diagnostic Systems Inc., Medford, New York, 11763, USA), were used to assess their diagnostic value for these species. The M. bovis-infected gemsbok tested strongly positive by Elephant TB STAT-PAK at the time of euthanasia and 5 mo earlier when the skin test was negative. This animal was not antibody reactive in MAPIA and DPP VetTB. No M. bovis-specific antibody was detected in the other two gemsboks by any of the immunoassays. Among the onagers, Elephant TB STAT-PAK, MAPIA, and DPP VetTB revealed gradually increasing antibody response in the animal that died of bovine tuberculosis, but not in the three disease-free herd mates euthanatized. Seroconversion in the M. bovis-infected onager was first noticed 5 yr before death when the tuberculin skin test was negative.

  1. LIM Kinase, a Newly Identified Regulator of Presynaptic Remodeling by Rod Photoreceptors After Injury

    PubMed Central

    Wang, Weiwei; Townes-Anderson, Ellen

    2015-01-01

    Purpose Rod photoreceptors retract their axon terminals and develop neuritic sprouts in response to retinal detachment and reattachment, respectively. This study examines the role of LIM kinase (LIMK), a component of RhoA and Rac pathways, in the presynaptic structural remodeling of rod photoreceptors. Methods Phosphorylated LIMK (p-LIMK), the active form of LIMK, was examined in salamander retina with Western blot and confocal microscopy. Axon length within the first 7 hours and process growth after 3 days of culture were assessed in isolated rod photoreceptors treated with inhibitors of upstream regulators ROCK and p21-activated kinase (Pak) (Y27632 and IPA-3) and a direct LIMK inhibitor (BMS-5). Porcine retinal explants were also treated with BMS-5 and analyzed 24 hours after detachment. Because Ca2+ influx contributes to axonal retraction, L-type channels were blocked in some experiments with nicardipine. Results Phosphorylated LIMK is present in rod terminals during retraction and in newly formed processes. Axonal retraction over 7 hours was significantly reduced by inhibition of LIMK or its regulators, ROCK and Pak. Process growth was reduced by LIMK or Pak inhibition especially at the basal (axon-bearing) region of the rod cells. Combining Ca2+ channel and LIMK inhibition had no additional effect on retraction but did further inhibit sprouting after 3 days. In detached porcine retina, LIMK inhibition reduced rod axonal retraction and improved retinal morphology. Conclusions Thus structural remodeling, in the form of either axonal retraction or neuritic growth, requires LIMK activity. LIM kinase inhibition may have therapeutic potential for reducing pathologic rod terminal plasticity after retinal injury. PMID:26658506

  2. Evaluation of a rapid serological test for the determination of Mycobacterium bovis infection in badgers (Meles meles) found dead.

    PubMed

    Chambers, Mark A; Lyashchenko, Konstantin P; Greenwald, Rena; Esfandiari, Javan; James, Eurig; Barker, Leslie; Jones, Jeff; Watkins, Gavin; Rolfe, Simon

    2010-03-01

    Between October 2005 and May 2006, a total of 727 badgers found dead in Wales were reported, and 550 were delivered to the Regional Laboratories of the Veterinary Laboratories Agency (VLA). Of the 459 carcasses suitable for examination, 55 were deemed to be infected with Mycobacterium bovis on the basis of culture, spoligotyping, and variable-number tandem repeat typing. Acid-fast bacteria were observed histologically in a further six badgers, but these bacteria were not confirmed as M. bovis by culture. A rapid serological test (BrockTB Stat-Pak) performed on thoracic blood showed a sensitivity of 35% and a specificity of 99%. Presence of M. bovis infection was 45 times more likely to be confirmed postmortem by culture in BrockTB Stat-Pak-reactive animals than in seronegative ones. Using visible carcass lesions as a marker of bovine tuberculosis (bTB) infection had a similar sensitivity (38%) but was significantly less specific (84%) than serology. The overall accuracy of the antibody detection was 93% (346 correct results from 374 tests), whereas the accuracy of regarding visible lesions as a marker for bTB infection was 78% (354 correct from 453 carcasses examined). Culture remains the gold standard method for detecting M. bovis infection in badgers. However, where resources are limited and/or an instant result is preferred, the BrockTB Stat-Pak could be used in field surveillance efforts to identify animals which should be examined further by only submitting test-negative animals to more detailed postmortem examination and culture.

  3. Spleen tyrosine kinase mediates high glucose-induced transforming growth factor-{beta}1 up-regulation in proximal tubular epithelial cells

    SciTech Connect

    Yang, Won Seok; Chang, Jai Won; Han, Nam Jeong; Lee, Sang Koo; Park, Su-Kil

    2012-09-10

    The role of spleen tyrosine kinase (Syk) in high glucose-induced intracellular signal transduction has yet to be elucidated. We investigated whether Syk is implicated in high glucose-induced transforming growth factor-{beta}1 (TGF-{beta}1) up-regulation in cultured human proximal tubular epithelial cells (HK-2 cell). High glucose increased TGF-{beta}1 gene expression through Syk, extracellular signal-regulated kinase (ERK), AP-1 and NF-{kappa}B. High glucose-induced AP-1 DNA binding activity was decreased by Syk inhibitors and U0126 (an ERK inhibitor). Syk inhibitors suppressed high glucose-induced ERK activation, whereas U0126 had no effect on Syk activation. High glucose-induced NF-{kappa}B DNA binding activity was also decreased by Syk inhibitors. High glucose increased nuclear translocation of p65 without serine phosphorylation of I{kappa}B{alpha} and without degradation of I{kappa}B{alpha}, but with an increase in tyrosine phosphorylation of I{kappa}B{alpha} that may account for the activation of NF-{kappa}B. Both Syk inhibitors and Syk-siRNA attenuated high glucose-induced I{kappa}B{alpha} tyrosine phosphorylation and p65 nuclear translocation. Depletion of p21-activated kinase 2 (Pak2) by transfection of Pak2-siRNA abolished high glucose-induced Syk activation. In summary, high glucose-induced TGF-{beta}1 gene transcription occurred through Pak2, Syk and subsequent ERK/AP-1 and NF-{kappa}B pathways. This suggests that Syk might be implicated in the diabetic kidney disease.

  4. Heat shock increases oxidative stress to modulate growth and physico-chemical attributes in diverse maize cultivars

    NASA Astrophysics Data System (ADS)

    Hussain, Iqbal; Ashraf, Muhammad Arslan; Rasheed, Rizwan; Iqbal, Muhammad; Ibrahim, Muhammad; Ashraf, Shamila

    2016-10-01

    The present investigation was conducted to appraise the physiochemical adjustments in contrasting maize cultivars, namely, PakAfgoi (tolerant) and EV-5098 (sensitive) subjected to heat shock. Seven-day-old seedlings were exposed to heat shock for different time intervals (1, 3, 6, 24, 48 and 72 h) and data for various physiochemical attributes determined to appraise time course changes in maize. After 72 h of heat shock, the plants were grown under normal conditions for 5 d and data for different growth attributes and photosynthetic pigments recorded. Exposure to heat shock reduced growth and photosynthetic pigments in maize cultivars. The plants exposed to heat shock for up to 3 h recovered growth and photosynthetic pigments when stress was relieved. A time course rise in the relative membrane permeability, hydrogen peroxide (H2O2) and malondialdehyde contents was recorded particularly in the EV-5098 indicating that heat shock-induced oxidative stress. Activities of different enzymatic antioxidants greatly altered due to heat shock. For instance, an increase in superoxide dismutase activity was recorded in both maize cultivars. The activity of ascorbate peroxidase was greater in Pak-Afgoi. However, the peroxidase and catalase activities were higher in plants of EV-5098. Heat shock caused a significant rise in the proline and decline in the total free amino acids. Overall, the performance of Pak-Afgoi was better in terms of having lesser oxidative damage and greater cellular levels of proline. The results suggested that oxidative stress indicators (relative membrane permeability, H2O2 and malondialdehyde) and proline can be used as markers for heat shock tolerant plants.

  5. Role of Hsl7 in Morphology and Pathogenicity and Its Interaction with Other Signaling Components in the Plant Pathogen Ustilago maydis ▿ †

    PubMed Central

    Lovely, C. Ben; Aulakh, Kavita Burman; Perlin, Michael H.

    2011-01-01

    The phytopathogenic fungus Ustilago maydis undergoes a dimorphic transition in response to mating pheromone, host, and environmental cues. On a solid medium deficient in ammonium (SLAD [0.17% yeast nitrogen base without ammonium sulfate or amino acids, 2% dextrose, 50 μM ammonium sulfate]), U. maydis produces a filamentous colony morphology, while in liquid SLAD, the cells do not form filaments. The p21-activated protein kinases (PAKs) play a substantial role in regulating the dimorphic transition in fungi. The PAK-like Ste20 homologue Smu1 is required for a normal response to pheromone, via upregulation of pheromone expression, and virulence, and its disruption affects both processes. Our experiments suggest that Smu1 also regulates cell length and the filamentous response on solid SLAD medium. Yeast two-hybrid analysis suggested an Hsl7 homologue as a potential interacting partner of Smu1, and a unique open reading frame for such an arginine methyltransferase was detected in the U. maydis genome sequence. Hsl7 regulates cell length and the filamentous response to solid SLAD in a fashion opposite to that of Smu1, but neither overexpression nor disruption of hsl7 attenuates virulence. Simultaneous disruption of hsl7 and overexpression of smu1 lead to a hyperfilamentous response on solid SLAD. Moreover, only this double mutant strain forms filaments in liquid SLAD. The double mutant strain was also significantly reduced in virulence. A similar filamentous response in both solid and liquid SLAD was observed in strains lacking another PAK-like protein kinase involved in cytokinesis and polar growth, Cla4. Our data suggest that Hsl7 may regulate cell cycle progression, while both Smu1 and Cla4 appear to be involved in the filamentous response in U. maydis. PMID:21622903

  6. p21-activated kinase 4 regulates mitotic spindle positioning and orientation.

    PubMed

    Bompard, Guillaume; Morin, Nathalie

    2012-01-01

    During mitosis, microtubules (MTs) are massively rearranged into three sets of highly dynamic MTs that are nucleated from the centrosomes to form the mitotic spindle. Tight regulation of spindle positioning in the dividing cell and chromosome alignment at the center of the metaphase spindle are required to ensure perfect chromosome segregation and to position the cytokinetic furrow that will specify the two daughter cells. Spindle positioning requires regulation of MT dynamics, involving depolymerase activities together with cortical and kinetochore-mediated pushing and pulling forces acting on astral MTs and kinetochore fibres. These forces rely on MT motor activities. Cortical pulling forces exerted on astral MTs depend upon dynein/dynactin complexes and are essential in both symmetric and asymmetric cell division. A well-established spindle positioning pathway regulating the cortical targeting of dynein/dynactin involves the conserved LGN (Leu-Gly-Asn repeat-enriched-protein) and NuMA (microtubule binding nuclear mitotic apparatus protein) complex. Spindle orientation is also regulated by integrin-mediated cell adhesion and actin retraction fibres that respond to mechanical stress and are influenced by the microenvironment of the dividing cell. Altering the capture of astral MTs or modulating pulling forces affects spindle position, which can impair cell division, differentiation and embryogenesis. In this general scheme, the activity of mitotic kinases such as Auroras and Plk1 (Polo-like kinase 1) is crucial. Recently, the p21-activated kinases (PAKs) emerged as novel important players in mitotic progression. In our recent article, we demonstrated that PAK4 regulates spindle positioning in symmetric cell division. In this commentary, and in light of recent published studies, we discuss how PAK4 could participate in the regulation of mechanisms involved in spindle positioning and orientation.

  7. Influence of thermal maturity on the hydrogen isotope content of extractable hydrocarbons

    NASA Astrophysics Data System (ADS)

    Radke, J.; Bechtel, A.; Püttmann, W.; Gleixner, G.

    2003-04-01

    Based on hydrogen isotope analysis of hydrocarbons from recent sediments it is suggested that compound specific hydrogen isotope ratios are a new proxy to reconstruct the palaeoclimate (Sauer et al., 2001). However, it remains unclear if transformation of carbon bound hydrogen with environmental water during maturation or thermal methanogenesis might influence the observed values. Short-term experiments excluded exchange reactions of deuterium from alkanes (Schimmelmann et al., 1999), however, thermally stressed kerogens are enriched in deuterium (Schoell, 1984). Therefore, we investigated the influence of maturity on the deltaD-values of alkanes and acyclic isoprenoids. In the Kupferschiefer horizon from the Polish Zechstein Basin thermal maturity of organic matter is correlated to burial depth yielding a natural long-term exchange experiment. The deltaD-values of extracted hydrocarbons linearly correlated with thermal maturity. These results enable the correction of deltaD values from biomarkers with known maturity and therefore expanding palaeoclimatic reconstructions using deltaD values to the geological past. References: SAUER, P.E., EGLINTON, T.I., HAYES, J.M., SCHIMMELMANN, A. &SESSIONS, A.L. (2001) Compound specific D/H ratios of lipid biomarkers from sediments as a proxy for environmental and climatic conditions. Geochimica et Cosmochimica Acta, 65(2), 213-222. SCHIMMELMANN, A., LEWAN, M.D. &WINTSCH, R.P. (1999) D/H isotope ratios of kerogen, bitumen, oil, and water in hydrous pyrolysis of source rocks containing kerogen types I, II, IIS, and III. Geochimica et Cosmochimica Acta, 63(22), 3751-3766. SCHOELL, M. (1984) Wasserstoff- und Kohlenstoffisotope in organischen Substanzen, Erdölen und Erdgasen. Schweitzerbart'sche Verlagsbuchhandlung, Stuttgart. Reihe D (67), 161pp.

  8. Design of a Matching Network for Dipole Antennas

    DTIC Science & Technology

    1993-03-01

    ZXSS0660 C VALUE GIVES A NEWTON STEP, WHILE A LARGE ZXSS0b7O C VALUE GIVES A STEEPEST DESCENT STE?. ZXSS0680 C THE DEFAULT VALUE FOR PAkM(1) IS 0.01...ITEfI J ISAC ,IK L,It JS, 1,LJ,1ONZ517 * ~IZERb LECEL MUD6 LOzxss1bo REAL AL CONS2 MEN~EIt F0 4GHL ZXSSI890 *ERL2 ERLM F0 tos h ,6G AF ZXSS1900 *HR

  9. Response of Selected Microorganisms to Experimental Planetary Environments

    NASA Technical Reports Server (NTRS)

    Foster, T. L.; Winans, L., Jr.

    1977-01-01

    Results of studies in anaerobic phosphorus metabolism are presented. Specific topics discussed include: (1) anaerobic utilization of PH3; (2) reduction of phosphate or phosphite; (3) isolation of organisms which utilize phosphite or phosphate anaerobically as a final hydrogen acceptor; and (4) the toxicity of PH3 to the organisms. Techniques of anaerobic microbiology associated with space hardware were also studied. These include: (1) the Brewer anaerobe jar/GasPak system; (2) a new procedure to grow aerobes and anaerobes simultaneously; (3) a culture medium to differentiate oblagate from facultative anaerobes; and (4) a procedure to quantitate O2 sensitivity of anaerobes.

  10. Biochar - A chemical view on formation and properties

    NASA Astrophysics Data System (ADS)

    Kruse, Andrea; Rodriguez Correa, Catalina; Becker, Gero C.; Wüst, Domink

    2016-04-01

    In a lot of studies concerning the use of biochar, the carboneous material is not clearly specified. Often, this leads to not comparable results. In this presentation the different processes to produce carbonizates are introduced, the chemical processes to form the product and the consequences for the properties of the char/coke are discussed. The focus is on coke from gasification, char from fast and slow pyrolysis and hydrochar form hydrothermal carbonization. Aspects like the surface properties, stability and contamination e.g. by PAK are illustrated by the chemical processes occurring during the production processes.

  11. Korean Affairs Report

    DTIC Science & Technology

    2007-11-02

    fierce whirlwind of struggle, and had the military turned the muzzle of their guns against Chongwadae by siding with the masses. [Kim] You are right...Korea 283.40 3. Japan 279.55 4. Hong Kong 189.25 Individual Competition Name Points 1. Yang Yueshan (China) 58.10 2. Pak Chong- hun (Korea) 57.35...for exports amounting to $500 million. In addition, Trade-Industry Minister Kim Chin -ho is scheduled to visit Japan next week as a presidential

  12. Determination of rhoifolin and daidzin in human plasma by high-performance liquid chromatography.

    PubMed

    Ishii, K; Urano, S; Furuta, T; Kasuya, Y

    1994-05-13

    A method for determining flavonoids in human plasma is presented for application to pharmacokinetic studies of two flavonoids, rhoifolin and daidzin. Isocratic reversed-phase high-performance liquid chromatography (HPLC) was used with genistin as an internal standard and solid-phase extraction using a Sep-Pak C18 cartridge. The mobile phases were acetonitrile-0.1 M ammonium acetate solution (20:80, v/v) for rhoifolin and methanol-0.1 M ammonium acetate solution (33:67, v/v) for daidzin. The detection limits on-column were 2 ng for rhoifolin and 0.5 ng for daidzin.

  13. KINEMATIC AND PHOTOMETRIC EVIDENCE FOR A BAR IN NGC 2683

    SciTech Connect

    Kuzio de Naray, Rachel; Zagursky, Matthew J.; McGaugh, Stacy S. E-mail: mzagursk@umd.edu

    2009-10-15

    We present optical long-slit and SparsePak Integral Field Unit emission line spectroscopy along with optical broadband and near-IR images of the edge-on spiral galaxy NGC 2683. We find a multi-valued, figure-of-eight velocity structure in the inner 45'' of the long-slit spectrum and twisted isovelocity contours in the velocity field. We also find, regardless of wavelength, that the galaxy isophotes are boxy. We argue that taken together, these kinematic and photometric features are evidence for the presence of a bar in NGC 2683. We use our data to constrain the orientation and strength of the bar.

  14. A 10-year experience with 290 pancreas transplants at a single institution.

    PubMed Central

    Sutherland, D E; Dunn, D L; Goetz, F C; Kennedy, W; Ramsay, R C; Steffes, M W; Mauer, S M; Gruessner, R; Moudry-Munns, K C; Morel, P

    1989-01-01

    Since our report at the 1984 American Surgical Association meeting of 100 pancreas transplants from 1966 through 1983, another 190 have been performed. The current series, begun in 1978, now numbers 276 cases, and includes 133 nonuremic recipients of pancreas transplants alone (PTA), 46 simultaneous pancreas/kidney transplants (SPK), and 97 pancreas tranplants after a kidney transplant (PAK). Duct management techniques used were free intraperitoneal drainage in 44 cases, duct occlusion in 44, enteric drainage in 89, and bladder drainage in 128. The 1-year patient and graft survival rates in the entire cohort of 276 were 91% and 42%. One-year patient survival rates were 88% in the first 100, 91% in the second 100, and 92% in the last 76 cases; corresponding 1-year graft survival rates were 28%, 47%, and 56% (p less than 0.05). A prospective comparison of bladder drainage (n = 82) versus enteric drainage (n = 46) in PAK/PTA cases since November 1, 1984 favored bladder drainage (1-year graft survival rates of 52% vs. 41%) because of urinary amylase monitoring. The best results were in recipients of primary SPK bladder-drained transplants (n = 39), with a 1-year pancreas graft survival rate of 75%, kidney graft survival rate of 80%, and patient survival rate of 95%. Logistic regression analysis, with 1-year graft function as the independent variable, showed significant (p less than 0.05) predictors of success (odds ratio) to be technique: bladder drainage (5.8) versus enteric drainage (2.5) versus duct injection (1.0); category: SPK (6.0) versus PAK from same donor (3.2) versus PAK from different donor (1.2) versus PTA (1.0); and donor HLA DR mismatch: 0 (5.0) versus 1 (2.5) versus 2 (1.0) antigens. On April 1, 1989, 90 patients had functioning grafts (60 euglycemic and insulin-free for more than 1 year, 10 for 5 to 10 years); these, along with 24 others whose grafts functioned for 1 to 6 years before failing, are part of an expanding cohort in whom the influence of

  15. Development of an automated optical inspection system for determining percent area coverage for spacecraft contamination control

    NASA Astrophysics Data System (ADS)

    Hogue, Patrick; Coopersmith, Jonathan D.

    2004-10-01

    This paper describes the use of an automated optical inspection system to determine the percent area coverage of particle matter on surfaces for determination of surface cleanliness in accordance with IEST-STD-CC1246 (formerly MIL-STD-1246). This system consists of an optical microscope, motor-driven X-Y stage, digital camera, and software and can be used to scan surfaces for particles with selected features; an example will be given of a scan for glass beads on Gel-Pak surfaces used to tape-lift spacecraft structure prior to shipment to a propulsion system supplier who prohibits glass beads from entering their facility.

  16. Inherent controllability in modular ALMRs

    SciTech Connect

    Sackett, J.I.; Sevy, R.H.; Wei, T.Y.C.

    1989-01-01

    As part of recent development efforts on advanced reactor designs ANL has proposed the IFR (Integral Fast Reactor) concept. The IFR concept is currently being applied to modular sized reactors which would be built in multiple power paks together with an integrated fuel cycle facility. It has been amply demonstrated that the concept as applied to the modular designs has significant advantages in regard to ATWS transients. Attention is now being focussed on determining whether or not those advantages deriving from the traits of the IFR can be translated to the operational/DBA (design basis accident) class of transients. 5 refs., 3 figs., 3 tabs.

  17. Glufosinate ammonium clean-up procedure from water samples using SPE

    NASA Astrophysics Data System (ADS)

    Tayeb M., A.; Ismail B., S.; Mardiana-Jansar, K.; Ta, Goh Choo; Agustar, Hani Kartini

    2015-09-01

    For the determination of glufosinate ammonium residue in soil and water samples, different solid phase extraction (SPE) sorbent efficiency was studied. Four different SPE sorbents i.e.: CROMABOND PS-H+, CROMABOND PS-OH-, ISOLUTE ENV+, Water Sep-Pak and OASIS HLB were used. Sample clean-up performance was evaluated using high performance liquid chromatography (Agilent 1220 infinity LC) with fluorescence detector. Detection of FMO-derivatives was done at λ ex = 260 nm and λ em= 310 nm. OASIS HLB column was the most suitable for the clean-up in view of the overall feasibility of the analysis.

  18. Pancreas Transplantation of US and Non-US Cases from 2005 to 2014 as Reported to the United Network for Organ Sharing (UNOS) and the International Pancreas Transplant Registry (IPTR).

    PubMed

    Gruessner, Angelika C; Gruessner, Rainer W G

    2016-01-01

    This report is an update of pancreas and kidney transplant activities in the US and non-US region in two periods, 2005-2009 and 2010-2014. The aim of the report was to analyze transplant progress and success in the US compared to non-US countries, and to compare trends between the two periods. Between 2005-2009 and 2010-2014, the number of US pancreas transplants declined by over 20%, while the overall number of pancreas transplants performed outside the US has increased. The decline in US numbers is predominantly due to the decline in primary and secondary pancreas after kidney transplants (PAK). During the time period studied, the number of PAK transplants dropped by 50%. In contrast, the number of simultaneous pancreas/kidney transplants (SPK) declined by only 10%, and the number of pancreas transplants alone (PTA) by 20%. Over 90% of pancreas transplants worldwide were performed, with a simultaneous kidney transplant and excellent results. Transplant outcomes in SPK improved significantly because of a decrease in the rates of technical and immunologic graft loss. In 2010-2014 vs. 2005-2009, US SPK transplant patient survival at 1 year post-transplant increased from 95.7% to 97.4%, pancreas graft function from 88.3% to 91.3%, and kidney function from 93.6% to 95.5%. A significant improvement was also noted in PAK transplants. One-year patient survival increased from 96.4% to 97.9% and pancreas graft function from 81.0% to 86.0%. PTA 1-year patient survival remained constant at 97%, and pancreas 1-year graft survival improved from 81.0% to 85.7%. With the decline in the number of transplants, a change towards better pancreas donor selection was observed. In solitary transplants, the donors were primarily young trauma victims, and the pancreas preservation time was relatively short. A general tendency towards transplanting older recipients was noted. In 2010-2014 vs. 2005-2009, PTA recipients 50 years of age or older accounted for 32% vs. 22%, PAK for 28% vs. 22

  19. Sirolimus induces apoptosis and reverses multidrug resistance in human osteosarcoma cells in vitro via increasing microRNA-34b expression

    PubMed Central

    Zhou, Yan; Zhao, Rui-hua; Tseng, Kuo-Fu; Li, Kun-peng; Lu, Zhi-gang; Liu, Yuan; Han, Kun; Gan, Zhi-hua; Lin, Shu-chen; Hu, Hai-yan; Min, Da-liu

    2016-01-01

    Aim: Multi-drug resistance poses a critical bottleneck in chemotherapy. Given the up-regulation of mTOR pathway in many chemoresistant cancers, we examined whether sirolimus (rapamycin), a first generation mTOR inhibitor, might induce human osteosarcoma (OS) cell apoptosis and increase the sensitivity of OS cells to anticancer drugs in vitro. Methods: Human OS cell line MG63/ADM was treated with sirolimus alone or in combination with doxorubicin (ADM), gemcitabine (GEM) or methotrexate (MTX). Cell proliferation and apoptosis were detected using CCK-8 assay and flow cytometry, respectively. MiRNAs in the cells were analyzed with miRNA microarray. The targets of miR-34b were determined based on TargetScan analysis and luciferase reporter assays. The expression of relevant mRNA and proteins was measured using qRT-PCR and Western blotting. MiR-34, PAK1 and ABCB1 levels in 40 tissue samples of OS patients were analyzed using qRT-PCR and in situ hybridization assays. Results: Sirolimus (1–100 nmol/L) dose-dependently suppressed the cell proliferation (IC50=23.97 nmol/L) and induced apoptosis. Sirolimus (10 nmol/L) significantly sensitized the cells to anticancer drugs, leading to decreased IC50 values of ADM, GEM and MTX (from 25.48, 621.41 and 21.72 μmol/L to 4.93, 73.92 and 6.77 μmol/L, respectively). Treatment of with sirolimus increased miR-34b levels by a factor of 7.5 in the cells. Upregulation of miR-34b also induced apoptosis and increased the sensitivity of the cells to the anticancer drugs, whereas transfection with miR-34b-AMO, an inhibitor of miR-34b, reversed the anti-proliferation effect of sirolimus. Two key regulators of cell cycle, apoptosis and multiple drug resistance, PAK1 and ABCB1, were demonstrated to be the direct targets of miR-34b. In 40 tissue samples of OS patients, significantly higher miR-34 ISH score and lower PAK5 and ABCB1 scores were detected in the chemo-sensitive group. Conclusion: Sirolimus increases the sensitivity of human OS

  20. Studies in Hawaiian Diptera III: New Distributional Records for Canacidae and a New Endemic Species of Procanace

    PubMed Central

    2016-01-01

    Abstract The distributions of Hawaiian Canacidae, comprising nearly 800 individual collection events, are reviewed and a total of four new island records are reported. These include Canaceoides angulatus from Kahoolawae and Procanace bifurcata from Molokai and Maui, and Procanace constricta from Oahu. A new species from Kauai, Procanace hardyi O'Grady and Pak, is described. This species is closely related to P. constricta from Oahu, Maui, Molokai and Hawaii and shares a similar constriction of the abdomen between tergites four and five but differs in the configuration of the seventh abdominal tergite. Detailed distribution maps for all species are included. PMID:27226743

  1. Preparative isolation by high performance liquid chromatography of human insulin B chain produced in escherichia coli

    SciTech Connect

    Cruz, N.; Antonio, S.; De Anda, R.; Gosset, G.; Bolivar, F. )

    1990-01-01

    This paper reports on a simple method developed for the analytical and preparative purification of human insulin B chain from recombinant origin. Three solvent systems: acetonitrile, isopropanol and methanol, were studied to determine their capacity to resolve the insulin B chain from a mixture of cyanogen bromide generated bacterial peptides. Using a {mu}Bondapak C18 column, it was possible to resolve the insulin B chain in all three systems. On a preparative scale, using a PrePak 500 C18 column with the isopropanol system, it was possible to purify insulin B chain and to obtain a 95% protein recovery.

  2. Bibliography of Soviet Laser Developments, Number 59, May - June 1982.

    DTIC Science & Technology

    1983-06-01

    ZhPS, v. 36 , no. 5, 1982, 811-816. 4. Semiconductor a. GaAs 24. Mineyeva, M.A., Ye.G. Mukhina, V.A. Plotnikov, and G.I. Vidro (0). Obtaining multiiayer...and S.N. Sokolov (4). Degradation processes in optoelectronic instruments. Sb 6, 98-141. e. Theory 36 . Kononenko, V.K., and G.T. Pak (3). Heating of...absorption spectra of organic molecules. ZhPS, v. 36 , no. 5, 1982, 776-780. 55. Kempe, N., H. Orzegowski, C. Peschel, and G. Thiede (NS). Pumping source for

  3. Angiogenesis and Therapeutic Approaches to NF1 Tumors

    DTIC Science & Technology

    2007-04-01

    Ferguson and Muir, 2000). Cultures grown on laminin-coated chamber slides were fixed with 2% paraformal- dehyde in 0.1 M phosphate buffer (pH 7.2; PBS...Erlandson RA, Woodruff JM. 1982. Peripheral nerve sheath tumors: an electron microscopic study of 43 cases. Cancer 49:273–287. Ferguson TA, Muir D...tumor xenograft in mice by the anti-PAK1 drug FK228. Cancer Biol Ther 2005;4:379–381. 14. Hirokawa Y, Nheu T, Grimm K, et al. Sichuan pepper extracts

  4. Microdesigning of Lightweight/High Strength Ceramic Materials

    DTIC Science & Technology

    1989-07-31

    pp. 199-204. Mat. Res. Sx). Symp. Proc., 32 Amsterdam: North-Hlolland. 23. See, e.g., Matijevic , E~. 1976. Progr. Coloid & Polymer Sd. 61:24-35. 24...Johnson, D,. W, 1987. Adv. Ceram. 21:3. 25. Matijevic , E. 1984. Ultrastructure Processing of Ceramics, Classes, and Composites, pp. 334-352. L. L...Ilench, 1). R. Ulrich, ed., New York: Wiley. 26. Matijevic , E. 1987. Personal communication. 27. Pak, S. F)88. M.S. Thesis. University of Washington

  5. Cerebral palsy, epilepsy, and severe intellectual disability in a patient with 3q29 microduplication syndrome.

    PubMed

    Fernández-Jaén, Alberto; Castellanos, María del Carmen; Fernández-Perrone, Ana Laura; Fernández-Mayoralas, Daniel Martín; de la Vega, Alberto González; Calleja-Pérez, Beatriz; Fernández, Ester Corbacho; Albert, Jacobo; Hombre, María Carmen Sánchez

    2014-08-01

    Interstitial microduplication of 3q29 has been recently described. Individuals with this syndrome have widely variable phenotypes. We describe the first clinical case with a 1.607 Mb duplication at 3q29 (chr3: 195,731,956-197,339,329), accompanied by severe intellectual disability, epilepsy, and cerebral palsy. This duplication involves 22 genes; PAK2, DLG1, BDH1, and FBXO45 are implicated in neuronal development and synaptic function and could play an important role in this syndrome. We propose considering genetic studies, particularly array comparative genomic hybridization, in patients with epilepsy and/or cerebral palsy of unknown etiology when dysmorphic features are present.

  6. A rapid means of separating A14-/sup 125/I-insulin from heterogeneously labeled insulin molecules for biologic studies

    SciTech Connect

    Stentz, F.B.; Wright, R.K.; Kitabchi, A.E.

    1982-12-01

    We have used two methods for the preparation of a highly homogeneous insulin with high specific activity. After iodination with chloramine T, the labeled peptides were retained on a disposable Sep Pak cartridge and subsequently eluted. The eluted labeled insulins were further purified by either DEAE cellulose or high performance liquid chromatography (HPLC) to separate A14-/sup 125/I- from A19-/sup 125/I-insulin. Both methods of chromatography were effective, but HPLC offered the advantage of better resolution in less time and higher yields of A14-/sup 125/I-insulin, which is suitable for biologic studies in various target tissues.

  7. Analysis of United States (US) and non-US pancreas transplants reported to the United network for organ sharing (UNOS) and the international pancreas transplant registry (IPTR) as of October 2001.

    PubMed

    Gruessner, A C; Sutherland, D E

    2001-01-01

    As of October 10, 2001, > 17,000 pancreas transplant had been reported to the IPTR, > 11,500 in the US and > 4,700 outside the US. An era analysis of US cases from 1987 to 2001 showed a progressive improvement in outcome (p < 0.04), with pancreas transplant graft survival rates (GSRs) going from 75% to 83% at one year for SPK cases, from 50% to 78% for PAK cases, and from 49% to 78% for PTA cases. The improvements were due both to decreases in technical failure (TF) rates (from 14% to 7% in SPK, 23% to 8% in PAK, and 23% to 10% in PTA) and immunological failure rates (going from 7% to 2% for SPK, from 25% to 2% for PAK, and from 28% to 4% for PTA cases). The proportion of recipients > 45 years old increased from 5% to 25%, and the improved outcomes encompassed the older patients as well. In patients > 45 years old, SPK pancreas GSRs at one year increased from 74% to 80% (p < 0.007). Pancreas GSRs were also similar for recipients reported to have Type 1 or Type 2 diabetes (at one year, 84% and 83%, respectively for 1997-2001 SPK transplants), the latter designated in 5% of the recipients. Contemporary pancreas transplant outcomes were calculated separately for 1997-2001 US and non-US cases. US patient survival rates at one year were > or = 95% in each recipient category, with one year pancreas GSRs of 83% for SPK (n = 3885), 79% for PAK (n = 630), and 78% for PTA (n = 240) (p = 0.0002). The immunological graft failure rates for 1997-2001 US SPK, PAK and PTA cases were 4%, 6%, and 8% at one year (p = 0.0001). There was a progressive increase in the use of ED (as opposed to BD) for duct management, to 67% for 1997-2001 US SPK transplants, 51% for PAK and 42% for PTA. Of US SPK ED transplants, 22% had venous drainage via the portal system. US pancreas GSRs were not significantly different for 1997-2001 ED (n = 2,519) and BD (n = 1,260) US SPK transplants (82% and 85%, respectively, at one year), nor was there a difference in pancreas GSRs for systemic (n = 1

  8. Nonmethane organic-carbon concentrations in air masses advected into urban areas in the United States. Final report

    SciTech Connect

    Westberg, H.; MacGregor, L.

    1987-11-01

    This report describes recent aircraft sampling programs to measure nonmethane organic carbon (NMOC) levels aloft in the upwind vicinity of several U.S. cities during the morning period of 0600 - 0900 A.M. The cities studied included Dallas-Ft. Worth, Tulsa, Atlanta, Birmingham, Philadelphia, and New York City. Individual hydrocarbon compounds were measured using gas-chromatographic procedures. The organic carbonyl compounds were collected on silica-packed SEP-PAK cartridges impregnated with 2,4-dinitro phenyl hydrazine (DNPH) and analyzed by HPLC. The data collected will be used in photochemical ozone models, such as OZIPP, to determine ozone control strategies.

  9. Factors to Consider in Blending Active and Reserve Manpower Within Military Units

    DTIC Science & Technology

    2007-01-01

    Consultant, Wex- ford Group; LtCol Charles Q. Brown, U.S. Air Force Reserve; Col. John Nicholson, U.S. Marine Corps Reserve; and CAPT Lou Farrell...Bradley, Lt. Gen. John A., “Future Total Force Means More Reserve Opportunities,” Citizen Airman. As of May 2, 2007: http://fi ndarticles.com/p...articles/mi_m0PAK/is_2_57/ai_n13803229 Brinkerhoff , John R., and Stanley A. Horowitz, Active-Reserve Integration in the Coast Guard, Alexandria, Va

  10. American Jihadist Terrorism: Combating a Complex Threat

    DTIC Science & Technology

    2010-09-20

    and Abdulhakim Muhammed’s shooting at the U.S. Army-Navy Career Center in Little Rock , Arkansas—and produced 14 deaths. By comparison, in more than...Major Nidal Hasan’s alleged assault at Fort Hood in Texas and Abdulhakim Muhammed’s shooting at the U.S. Army-Navy Career Center in Little Rock ... Art Keller, “Why Was Faisal Shahzad a Bad Bombmaker?” AfPak Channel, a special project of Foreign Policy and the New America Foundation, May 14, 2010

  11. American Jihadist Terrorism: Combating a Complex Threat

    DTIC Science & Technology

    2010-12-07

    Texas and Abdulhakim Muhammed’s shooting at the U.S. Army-Navy Career Center in Little Rock , AR—and produced 14 deaths. By comparison, in more than...Muhammed’s shooting at the U.S. Army-Navy Career Center in Little Rock , AR— produced 14 deaths. By comparison, in more than seven years from the...Mumbai, India, and Implications for U.S. Interests, by K. Alan Kronstadt. 18 Art Keller, “Why Was Faisal Shahzad a Bad Bombmaker?” AfPak Channel, a

  12. A simple and sensitive quantitation of N,N-dimethyltryptamine by gas chromatography with surface ionization detection.

    PubMed

    Ishii, A; Seno, H; Suzuki, O; Hattori, H; Kumazawa, T

    1997-01-01

    A simple and sensitive method for determination of N,N-dimethyltryptamine (DMT) by gas chromatography (GC) with surface ionization detection (SID) is presented. Whole blood or urine, containing DMT and gramine (internal standard), was subjected to solid-phase extraction with a Sep-Pak C18 cartridge before analysis by GC-SID. The calibration curve was linear in the DMT range of 1.25-20 ng/mL blood or urine. The detection limit of DMT was about 0.5 ng/mL (10 pg on-column). The recovery of both DMT and gramine spiked in biological fluids was above 86%.

  13. A fuzzy goal programming model for the simple U-line balancing problem with multiple objectives

    NASA Astrophysics Data System (ADS)

    Toklu, Bilal; Ozcan, Uğur

    2008-03-01

    A fuzzy goal programming model for the simple U-line balancing (SULB) problem with multiple objectives is presented. In real life applications of the SULB problem with multiple objectives, it is difficult for the decision-maker(s) to determine the goal value of each objective precisely as the goal values are imprecise, vague, or uncertain. Therefore a fuzzy goal programming model is developed for this purpose. The proposed model is the first fuzzy multi-objective decision-making approach to the SULB problem with multiple objectives which aims at simultaneously optimizing several conflicting goals. The proposed model is illustrated using an example. A computational study is conducted by solving a large number of test problems to investigate the relationship between the fuzzy goals and to compare them with the goal programming model proposed by Gokcen and Ağpak (Gokcen, H. and Ağpak, K., European Journal of Operational Research, 171, 577-585, 2006). The results of the computational study show that the proposed model is more realistic than the existing models for the SULB problem with multiple objectives and also provides increased flexibility for the decision-maker(s) to determine different alternatives.

  14. Characterization of Novel Molecular Mechanisms Favoring Rac1 Membrane Translocation.

    PubMed

    Castro-Castro, Antonio; Muriel, Olivia; Del Pozo, Miguel A; Bustelo, Xosé R

    2016-01-01

    The Rac1 GTPase plays key roles in cytoskeletal organization, cell motility and a variety of physiological and disease-linked responses. Wild type Rac1 signaling entails dissociation of the GTPase from cytosolic Rac1-Rho GDP dissociation inhibitor (GDI) complexes, translocation to membranes, activation by exchange factors, effector binding, and activation of downstream signaling cascades. Out of those steps, membrane translocation is the less understood. Using transfections of a expression cDNA library in cells expressing a Rac1 bioreporter, we previously identified a cytoskeletal feedback loop nucleated by the F-actin binding protein coronin 1A (Coro1A) that promotes Rac1 translocation to the plasma membrane by facilitating the Pak-dependent dissociation of Rac1-Rho GDI complexes. This screening identified other potential regulators of this process, including WDR26, basigin, and TMEM8A. Here, we show that WDR26 promotes Rac1 translocation following a Coro1A-like and Coro1A-dependent mechanism. By contrast, basigin and TMEM8A stabilize Rac1 at the plasma membrane by inhibiting the internalization of caveolin-rich membrane subdomains. This latter pathway is F-actin-dependent but Coro1A-, Pak- and Rho GDI-independent.

  15. An evaluation of three commercial fecal transport systems for the recovery of enteric pathogens.

    PubMed

    Mundy, L S; Shanholtzer, C J; Willard, K E; Peterson, L R

    1991-09-01

    Twenty-five isolates, including six strains of Shigella species, six strains of Salmonella species, five strains of Yersinia enterocolitica, six strains of Campylobacter jejuni, and two strains of Vibrio parahaemolyticus, were inoculated at a concentration of 1.5 x 10(4) colony-forming units/mL into the following transport systems: Fekal Enteric Plus (Trend Scientific, Inc., St. Paul, MN), Cary Blair Transport Medium (Remel, Inc., Lenexa, KS), and Para-Pak C & S (Meridian Diagnostics, Inc., Cincinnati, OH). The Fekal Enteric Plus system showed a greater than or equal to 50% recovery of the original bacterial inoculum after 96 hours for all Salmonella, Shigella, and Yersinia strains tested and after as long as 72 hours for Vibrio strains. The Cary Blair Transport System showed greater than or equal to 50% recovery of the initial inoculum at 96 hours for five of six Salmonella strains, four of six Shigella strains, all Yersinia strains, and one of two Vibrio strains. With the use of the Para-Pak C & S, greater than or equal to 50% recovery of the original inoculum after 96 h was demonstrated for five of six Salmonella strains, four of six Shigella strains, all Yersinia strains, and one of two Vibrio strains. All three systems did not demonstrate even 10% recovery of the initial C. jejuni inoculum at 24 hours when held at room temperature.

  16. TFAP2C-mediated upregulation of TGFBR1 promotes lung tumorigenesis and epithelial–mesenchymal transition

    PubMed Central

    Kim, Wanyeon; Kim, EunGi; Lee, Sungmin; Kim, Daehoon; Chun, Jahyun; Park, Kang Hyun; Youn, HyeSook; Youn, BuHyun

    2016-01-01

    TFAP2C (transcription factor-activating enhancer-binding protein 2C) expression has been positively correlated with poor prognosis in patients with certain types of cancer, but the mechanisms underlying TFAP2C-mediated tumorigenesis in non-small-cell lung cancer (NSCLC) are still unknown. We previously performed a microarray analysis to identify TFAP2C regulation genes, and TGFBR1 (transforming growth factor-β receptor type 1) was found to be upregulated by TFAP2C. We observed that TFAP2C or TGFBR1 overexpression led to oncogenic properties, such as cell viability, proliferation and cell cycle progression. TGFBR1 upregulation induced by TFAP2C also promoted cell motility and migration, leading to malignant development. We also found that PAK1 (p21 protein (Cdc42/Rac)-activated kinase 1) signaling was involved in TFAP2C/TGFBR1-induced tumorigenesis. These results were confirmed by an in vivo xenograft model and patient tissue samples. This study shows that TFAP2C promoted tumor progression by upregulation of TGFBR1 and consequent activation of PAK1 signaling. PMID:27885255

  17. Next-Generation “-omics” Approaches Reveal a Massive Alteration of Host RNA Metabolism during Bacteriophage Infection of Pseudomonas aeruginosa

    PubMed Central

    Monot, Marc; Kogadeeva, Maria; Sauer, Uwe; Jorth, Peter; Whiteley, Marvin; Debarbieux, Laurent; Lavigne, Rob

    2016-01-01

    As interest in the therapeutic and biotechnological potentials of bacteriophages has grown, so has value in understanding their basic biology. However, detailed knowledge of infection cycles has been limited to a small number of model bacteriophages, mostly infecting Escherichia coli. We present here the first analysis coupling data obtained from global next-generation approaches, RNA-Sequencing and metabolomics, to characterize interactions between the virulent bacteriophage PAK_P3 and its host Pseudomonas aeruginosa. We detected a dramatic global depletion of bacterial transcripts coupled with their replacement by viral RNAs over the course of infection, eventually leading to drastic changes in pyrimidine metabolism. This process relies on host machinery hijacking as suggested by the strong up-regulation of one bacterial operon involved in RNA processing. Moreover, we found that RNA-based regulation plays a central role in PAK_P3 lifecycle as antisense transcripts are produced mainly during the early stage of infection and viral small non coding RNAs are massively expressed at the end of infection. This work highlights the prominent role of RNA metabolism in the infection strategy of a bacteriophage belonging to a new characterized sub-family of viruses with promising therapeutic potential. PMID:27380413

  18. Comparative phytohormone profiles, lipid kinase and lipid phosphatase activities in barley aleurone, coleoptile, and root tissues.

    PubMed

    Meringer, Maria V; Villasuso, Ana L; Pasquaré, Susana J; Giusto, Norma M; Machado, Estela E; Racagni, Graciela E

    2012-09-01

    We analyzed lipid kinase and lipid phosphatase activities and determined endogenous phytohormone levels by liquid chromatography-tandem mass spectrometry in root and coleoptile tissues following germination of barley (Hordeum vulgare) seeds. The enzymes showing highest activity in aleurone cells were diacylglycerol kinase (DAG-k, EC 2.7.1.107) and phosphatidate kinase (PA-k). The ratio of gibberellins (GAs) to abscisic acid (ABA) was 2-fold higher in aleurone than in coleoptile or root tissues. In coleoptiles, phosphatidylinositol 4-kinase (PI4-k, EC 2.7.1.67) showed the highest enzyme activity, and jasmonic acid (JA) level was higher than in aleurone. In roots, activities of PI4-k, DAG-k, and PA-k were similar, and salicylic acid (SA) showed the highest concentration. In the assays to evaluate the hydrolysis of DGPP (diacylglycerol pyrophosphate) and PA (phosphatidic acid) we observed that PA hydrolysis by LPPs (lipid phosphate phosphatases) was not modified; however, the diacylglycerol pyrophosphate phosphatase (DGPPase) was strikingly higher in coleoptile and root tissues than to aleurone. Relevance of these findings in terms of signaling responses and seedling growth is discussed.

  19. Comparison of two immunochromatographic assays and the indirect immunofluorescence antibody test for diagnosis of Trypanosoma cruzi infection in dogs in south central Louisiana.

    PubMed

    Nieto, Prixia D; Boughton, Roger; Dorn, Patricia L; Steurer, Frank; Raychaudhuri, Syamal; Esfandiari, Javan; Gonçalves, Edson; Diaz, James; Malone, John B

    2009-11-12

    Two rapid tests evaluated in dogs considered to be of high risk of infection with the Chagas parasite Trypanosoma cruzi using two immunochromatographic assays: Trypanosoma Detect for canine, InBios, Seattle, WA and CHAGAS STAT-PAK assay, Chembio Diagnostic Systems, Medford, NY, in south central Louisiana. For this purpose a serological survey was carried out in a total of 122 dogs and a serum bank was created. These 122 animals were first tested by IFAT that was used as the standard test. From the serum bank 50 samples were tested using the two rapid Chagas assays and results compared to the standard test IFAT. The serological survey using IFAT showed a prevalence of T. cruzi infection in 22.1% of the tested dogs. In the immunochromatographic assays, 13 and 11 animals were positive on rapid assay: Trypanosoma Detect for canine, InBios and CHAGAS STAT-PAK, Chembio Diagnostic Systems, respectively compared to 11 positive by IFAT. These two immunochromatographic tests have shown high susceptibility and specificity compared to our standard method IFAT. The rapid, easy and accurate screening assays used in conjunction with confirmatory tests, would be an excellent tool for veterinarians to diagnose T. cruzi infection. Early detection of T. cruzi infection may prevent complications through an effective treatment. Greater awareness by veterinarians of the risk, clinical findings, history along with diagnostic methods will contribute greatly to an understanding of the true prevalence of Chagas disease in dogs in Louisiana.

  20. Inca: a novel p21-activated kinase-associated protein required for cranial neural crest development.

    PubMed

    Luo, Ting; Xu, Yanhua; Hoffman, Trevor L; Zhang, Tailin; Schilling, Thomas; Sargent, Thomas D

    2007-04-01

    Inca (induced in neural crest by AP2) is a novel protein discovered in a microarray screen for genes that are upregulated in Xenopus embryos by the transcriptional activator protein Tfap2a. It has no significant similarity to any known protein, but is conserved among vertebrates. In Xenopus, zebrafish and mouse embryos, Inca is expressed predominantly in the premigratory and migrating neural crest (NC). Knockdown experiments in frog and fish using antisense morpholinos reveal essential functions for Inca in a subset of NC cells that form craniofacial cartilage. Cells lacking Inca migrate successfully but fail to condense into skeletal primordia. Overexpression of Inca disrupts cortical actin and prevents formation of actin "purse strings", which are required for wound healing in Xenopus embryos. We show that Inca physically interacts with p21-activated kinase 5 (PAK5), a known regulator of the actin cytoskeleton that is co-expressed with Inca in embryonic ectoderm, including in the NC. These results suggest that Inca and PAK5 cooperate in restructuring cytoskeletal organization and in the regulation of cell adhesion in the early embryo and in NC cells during craniofacial development.

  1. DO COMMERCIAL SEROLOGIC TESTS FOR TRYPANOSOMA CRUZI INFECTION DETECT MEXICAN STRAINS IN WOMEN AND NEWBORNS?

    PubMed Central

    Gamboa-León, Rubi; Gonzalez-Ramirez, Claudia; Padilla-Raygoza, Nicolas; Sosa-Estani, Sergio; Caamal-Kantun, Alejandra; Buekens, Pierre; Dumonteil, Eric

    2012-01-01

    We sought to determine the serological test that could be used for Trypanosoma cruzi seroprevalence studies in Mexico, where lineage I predominates. In a previous study among pregnant women and their newborns in the states of Yucatan and Guanajuato, we reported a 0.8–0.9% of prevalence for T. cruzi–specific antibodies by Stat-Pak and Wiener ELISA. We have expanded this study here by performing an additional non-commercial ELISA and confirming the seropositives with Western blot, using whole antigens of a local parasite strain. We found a seroprevalence of 0.6% (3/500) in Merida and 0.4% in Guanajuato (2/488). The 5 seropositive umbilical cord samples reacted to both non-commercial ELISA and Western blot tests, and only 1 of the maternal samples was not reactive to non-commercial ELISA. A follow-up of the newborns at 10 mo was performed in Yucatan to determine the presence of T. cruzi antibodies in children as evidence of congenital infection. None of the children was seropositive. One newborn from an infected mother died at 2 wk of age of cardiac arrest, but T. cruzi infection was not confirmed. The T. cruzi seroprevalence data obtained with both commercial tests (Stat-Pak and ELISA Wiener) are similar to those from non-commercial tests using a local Mexican strain of T. cruzi. PMID:21506787

  2. Control of mammary myoepithelial cell contractile function by α3β1 integrin signalling.

    PubMed

    Raymond, Karine; Cagnet, Stéphanie; Kreft, Maaike; Janssen, Hans; Sonnenberg, Arnoud; Glukhova, Marina A

    2011-05-18

    In the functionally differentiated mammary gland, basal myoepithelial cells contract to eject the milk produced by luminal epithelial cells from the body. We report that conditional deletion of a laminin receptor, α3β1 integrin, from myoepithelial cells leads to low rates of milk ejection due to a contractility defect but does not interfere with the integrity or functional differentiation of the mammary epithelium. In lactating mammary gland, in the absence of α3β1, focal adhesion kinase phosphorylation is impaired, the Rho/Rac balance is altered and myosin light-chain (MLC) phosphorylation is sustained. Cultured mammary myoepithelial cells depleted of α3β1 contract in response to oxytocin, but are unable to maintain the state of post-contractile relaxation. The expression of constitutively active Rac or its effector p21-activated kinase (PAK), or treatment with MLC kinase (MLCK) inhibitor, rescues the relaxation capacity of mutant cells, strongly suggesting that α3β1-mediated stimulation of the Rac/PAK pathway is required for the inhibition of MLCK activity, permitting completion of the myoepithelial cell contraction/relaxation cycle and successful lactation. This is the first study highlighting the impact of α3β1 integrin signalling on mammary gland function.

  3. Merlin/NF2 functions upstream of the nuclear E3 ubiquitin ligase CRL4DCAF1 to suppress oncogenic gene expression.

    PubMed

    Cooper, Jonathan; Li, Wei; You, Liru; Schiavon, Gaia; Pepe-Caprio, Angela; Zhou, Lu; Ishii, Ryohei; Giovannini, Marco; Hanemann, C Oliver; Long, Stephen B; Erdjument-Bromage, Hediye; Zhou, Pengbo; Tempst, Paul; Giancotti, Filippo G

    2011-08-23

    Integrin-mediated activation of PAK (p21-activated kinase) causes phosphorylation and inactivation of the FERM (4.1, ezrin, radixin, moesin) domain-containing protein Merlin, which is encoded by the NF2 (neurofibromatosis type 2) tumor suppressor gene. Conversely, cadherin engagement inactivates PAK, thus leading to accumulation of unphosphorylated Merlin. Current models imply that Merlin inhibits cell proliferation by inhibiting mitogenic signaling at or near the plasma membrane. We have recently shown that the unphosphorylated, growth-inhibiting form of Merlin accumulates in the nucleus and binds to the E3 ubiquitin ligase CRL4(DCAF1) to suppress its activity. Depletion of DCAF1 blocks the hyperproliferation caused by inactivation of Merlin. Conversely, expression of a Merlin-insensitive DCAF1 mutant counteracts the antimitogenic effect of Merlin. Expression of Merlin or silencing of DCAF1 in Nf2-deficient cells induce an overlapping, tumor-suppressive program of gene expression. Mutations present in some tumors from NF2 patients disrupt Merlin's ability to interact with or inhibit CRL4(DCAF1). Lastly, depletion of DCAF1 inhibits the hyperproliferation of Schwannoma cells isolated from NF2 patients and suppresses the oncogenic potential of Merlin-deficient tumor cell lines. Current studies are aimed at identifying the substrates and mechanism of action of CRL4(DCAF1) and examining its role in NF2-dependent tumorigenesis in mouse models. We propose that Merlin mediates contact inhibition and suppresses tumorigenesis by translocating to the nucleus to inhibit CRL4(DCAF1).

  4. Sepsis-induced elevation in plasma serotonin facilitates endothelial hyperpermeability

    PubMed Central

    Li, Yicong; Hadden, Coedy; Cooper, Anthonya; Ahmed, Asli; Wu, Hong; Lupashin, Vladimir V.; Mayeux, Philip R.; Kilic, Fusun

    2016-01-01

    Hyperpermeability of the endothelial barrier and resulting microvascular leakage are a hallmark of sepsis. Our studies describe the mechanism by which serotonin (5-HT) regulates the microvascular permeability during sepsis. The plasma 5-HT levels are significantly elevated in mice made septic by cecal ligation and puncture (CLP). 5-HT-induced permeability of endothelial cells was associated with the phosphorylation of p21 activating kinase (PAK1), PAK1-dependent phosphorylation of vimentin (P-vimentin) filaments, and a strong association between P-vimentin and ve-cadherin. These findings were in good agreement with the findings with the endothelial cells incubated in serum from CLP mice. In vivo, reducing the 5-HT uptake rates with the 5-HT transporter (SERT) inhibitor, paroxetine blocked renal microvascular leakage and the decline in microvascular perfusion. Importantly, mice that lack SERT showed significantly less microvascular dysfunction after CLP. Based on these data, we propose that the increased endothelial 5-HT uptake together with 5-HT signaling disrupts the endothelial barrier function in sepsis. Therefore, regulating intracellular 5-HT levels in endothelial cells represents a novel approach in improving sepsis-associated microvascular dysfunction and leakage. These new findings advance our understanding of the mechanisms underlying cellular responses to intracellular/extracellular 5-HT ratio in sepsis and refine current views of these signaling processes during sepsis. PMID:26956613

  5. Metal oxide coating of carbon supports for supercapacitor applications.

    SciTech Connect

    Boyle, Timothy J.; Tribby, Louis, J; Lakeman, Charles D. E.; Han, Sang M.; Lambert, Timothy N.; Fleig, Patrick F.

    2008-07-01

    The global market for wireless sensor networks in 2010 will be valued close to $10 B, or 200 M units. TPL, Inc. is a small Albuquerque based business that has positioned itself to be a leader in providing uninterruptible power supplies in this growing market with projected revenues expected to exceed $26 M in 5 years. This project focused on improving TPL, Inc.'s patent-pending EnerPak{trademark} device which converts small amounts of energy from the environment (e.g., vibrations, light or temperature differences) into electrical energy that can be used to charge small energy storage devices. A critical component of the EnerPak{trademark} is the supercapacitor that handles high power delivery for wireless communications; however, optimization and miniaturization of this critical component is required. This proposal aimed to produce prototype microsupercapacitors through the integration of novel materials and fabrication processes developed at New Mexico Technology Research Collaborative (NMTRC) member institutions. In particular, we focused on developing novel ruthenium oxide nanomaterials and placed them into carbon supports to significantly increase the energy density of the supercapacitor. These improvements were expected to reduce maintenance costs and expand the utility of the TPL, Inc.'s device, enabling New Mexico to become the leader in the growing global wireless power supply market. By dominating this niche, new customers were expected to be attracted to TPL, Inc. yielding new technical opportunities and increased job opportunities for New Mexico.

  6. The EOP Visualization Module Integrated into the Plasma On-Line Nuclear Power Plant Safety Monitoring and Assessment System

    SciTech Connect

    Hornaes, Arne; Hulsund, John Einar; Vegh, Janos; Major, Csaba; Horvath, Csaba; Lipcsei, Sandor; Kapocs, Gyoergy

    2001-08-15

    An ambitious project to replace the unit information systems (UISs) at the Hungarian Paks nuclear power plant was started in 1998-99. The basic aim of the reconstruction project is to install a modern, distributed UIS architecture on all four Paks VVER-440 units. The new UIS includes an on-line plant safety monitoring and assessment system (PLASMA), which contains a critical safety functions monitoring module and provides extensive operator support during the execution of the new, symptom-oriented emergency operating procedures (EOPs). PLASMA includes a comprehensive EOP visualization module, based on the COPMA-III procedure-handling software developed by the Organization for Economic Cooperation and Development, Halden Reactor Project. Intranet technology is applied for the presentation of the EOPs with the use of a standard hypertext markup language (HTML) browser as a visualization tool. The basic design characteristics of the system, with a detailed description of its user interface and functions of the new EOP display module, are presented.

  7. Antioxidative properties of defatted dabai pulp and peel prepared by solid phase extraction.

    PubMed

    Khoo, Hock Eng; Azlan, Azrina; Ismail, Amin; Abas, Faridah

    2012-08-14

    Solid phase extraction (SPE) using Sep-Pak® cartridges is one of the techniques used for fractionation of antioxidant compounds in waste of dabai oil extraction (defatted dabai parts). The aim of this study was to determine the phenolic compounds and antioxidant capacity in crude extracts and several SPE fractions from methanolic extract of defatted dabai pulp and peel. Based on SPE, Sep-Pak® cyanopropyl and C₁₈ cartridges were used to fractionate the antioxidant-rich crude extracts into water and methanolic fractions. Analyzed using LC-MS, flavonoids, anthocyanins, saponin derivatives and other unknown antioxidative compounds were detected in the defatted dabai crude extracts and their SPE fractions. Anthocyanins were the major phenolic compounds identified in the defatted dabai peel and detected in most of the SPE fractions. Methanolic fractions of defatted dabai parts embraced higher total phenolics and antioxidant capacity than water fractions. This finding also revealed the crude extracts of defatted dabai peel have the most significant antioxidant properties compared to the methanolic and water fractions studied. The crude extract of defatted dabai parts remain as the most potent antioxidant as it contains mixture of flavonoids, anthocyanins and other potential antioxidants.

  8. Morphology of root nodules and nodule-like structures formed by Rhizobium and Agrobacterium strains containing a Rhizobium meliloti megaplasmid

    PubMed Central

    1983-01-01

    We examined expression of the megaplasmid pRme41b of Rhizobium meliloti in two different Rhizobium sp. Strains and in Agrobacterium tumefaciens. Transfer of pRme41b into these bacteria was facilitated by insertion of a recombinant plasmid coding for mobilization functions of RP4 into the nif region (Kondorosi, A., E. Kondorosi, C.E. Pankhurst, W. J. Broughton, and Z. Banfalvi, 1982, Mol. Gen. Genet., 188:433-439). In all cases, transconjugants formed nodule-like structures on the roots of Medicago sativa. These structures were largely composed of meristematic cells but they were not invaded by bacteria. Bacteria were found only within infection threads in root hairs, and within intercellular spaces of the outermost cells of the structures. The donor strain of R. meliloti containing pAK11 or pAK12 in pRme41b initially produced nodules on M. sativa that did not fix nitrogen (Fix- ). In these nodules, bacteria were released from infection threads into the host cells but they did not multiply appreciably. Any bacteroids formed degenerated prematurely. In some cases, however, reversion to a Fix+ phenotype occurred after 4 to 6 wk. Bacteria released into newly infected cells in these nodules showed normal development into bacteriods. PMID:6885919

  9. Do commercial serologic tests for Trypanosoma cruzi infection detect Mexican strains in women and newborns?

    PubMed

    Gamboa-León, Rubi; Gonzalez-Ramirez, Claudia; Padilla-Raygoza, Nicolas; Sosa-Estani, Sergio; Caamal-Kantun, Alejandra; Buekens, Pierre; Dumonteil, Eric

    2011-04-01

    We sought to determine the serological test that could be used for Trypanosoma cruzi seroprevalence studies in Mexico, where lineage I predominates. In a previous study among pregnant women and their newborns in the states of Yucatan and Guanajuato, we reported a 0.8-0.9% of prevalence for T. cruzi -specific antibodies by Stat-Pak and Wiener ELISA. We have expanded this study here by performing an additional non-commercial ELISA and confirming the seropositives with Western blot, using whole antigens of a local parasite strain. We found a seroprevalence of 0.6% (3/500) in Merida and 0.4% in Guanajuato (2/488). The 5 seropositive umbilical cord samples reacted to both non-commercial ELISA and Western blot tests, and only 1 of the maternal samples was not reactive to non-commercial ELISA. A follow-up of the newborns at 10 mo was performed in Yucatan to determine the presence of T. cruzi antibodies in children as evidence of congenital infection. None of the children was seropositive. One newborn from an infected mother died at 2 wk of age of cardiac arrest, but T. cruzi infection was not confirmed. The T. cruzi seroprevalence data obtained with both commercial tests (Stat-Pak and ELISA Wiener) are similar to those from non-commercial tests using a local Mexican strain of T. cruzi.

  10. 3q29 microdeletion syndrome: clinical and molecular characterization of a new syndrome.

    PubMed

    Willatt, Lionel; Cox, James; Barber, John; Cabanas, Elisabet Dachs; Collins, Amanda; Donnai, Dian; FitzPatrick, David R; Maher, Eddy; Martin, Howard; Parnau, Josep; Pindar, Lesley; Ramsay, Jacqueline; Shaw-Smith, Charles; Sistermans, Erik A; Tettenborn, Michael; Trump, Dorothy; de Vries, Bert B A; Walker, Kate; Raymond, F Lucy

    2005-07-01

    We report the identification of six patients with 3q29 microdeletion syndrome. The clinical phenotype is variable despite an almost identical deletion size. The phenotype includes mild-to-moderate mental retardation, with only slightly dysmorphic facial features that are similar in most patients: a long and narrow face, short philtrum, and high nasal bridge. Autism, gait ataxia, chest-wall deformity, and long and tapering fingers were noted in at least two of six patients. Additional features--including microcephaly, cleft lip and palate, horseshoe kidney and hypospadias, ligamentous laxity, recurrent middle ear infections, and abnormal pigmentation--were observed, but each feature was only found once, in a single patient. The microdeletion is approximately 1.5 Mb in length, with molecular boundaries mapping within the same or adjacent bacterial artificial chromosome (BAC) clones at either end of the deletion in all patients. The deletion encompasses 22 genes, including PAK2 and DLG1, which are autosomal homologues of two known X-linked mental retardation genes, PAK3 and DLG3. The presence of two nearly identical low-copy repeat sequences in BAC clones on each side of the deletion breakpoint suggests that nonallelic homologous recombination is the likely mechanism of disease causation in this syndrome.

  11. Occurrence of paleozoic and early mesozoic radiolaria in Thailand (preliminary report)

    NASA Astrophysics Data System (ADS)

    Sashida, Katsuo; Igo, Hisayoshi; Hisafa, Ken-Ichiro; Nakornsri, Nikorn; Ampornmaha, Apsorn

    Paleozoic and Early Mesozoic radiolarians are newly recovered from chert and associated fine-grained clastic rocks in Thailand. This study clarifies the geologic age of these radiolarian rocks and their paleogeographic and geotectonic significance. Devonian, Early Carboniferous and Permian radiolarians were found in the "Fang Chert" which outcrops along the Chiang Mai-Fang Road, upper north Thailand, Early Carboniferous radiolarians were recovered from a sequence of tuffaceous shale and chert exposed in the Pak Chom area along the Mekong River, and well-preserved Late Devonian and Early Carboniferous radiolarians were also recovered from cherts exposed along the Pak Chom-Loei Road near Phu Laem, north of Loei, in northeast Thailand. These Devonian to Carboniferous radiolarian faunas are apparently identical with those reported from eastern and western Australia. Well-preserved Early Triassic conodonts and radiolarians were obtained from a limeston exposed near Patthalung, southern Thailand. Most of the radiolarian species of this fauna show close affinity with those reported from the Upper Paleozoic rocks, and are new species except for some spicule-type forms. Based on the above-mentioned newly obtained micropaleontological evidence, the geotectonic significance of these radiolarian rocks are briefly discussed in relation to the paleography of the Palaeo-Tethys Ocean, Sibumas and Indochina Terranes, and Australia during the Late Devonian to Middle Permian times.

  12. Next-Generation "-omics" Approaches Reveal a Massive Alteration of Host RNA Metabolism during Bacteriophage Infection of Pseudomonas aeruginosa.

    PubMed

    Chevallereau, Anne; Blasdel, Bob G; De Smet, Jeroen; Monot, Marc; Zimmermann, Michael; Kogadeeva, Maria; Sauer, Uwe; Jorth, Peter; Whiteley, Marvin; Debarbieux, Laurent; Lavigne, Rob

    2016-07-01

    As interest in the therapeutic and biotechnological potentials of bacteriophages has grown, so has value in understanding their basic biology. However, detailed knowledge of infection cycles has been limited to a small number of model bacteriophages, mostly infecting Escherichia coli. We present here the first analysis coupling data obtained from global next-generation approaches, RNA-Sequencing and metabolomics, to characterize interactions between the virulent bacteriophage PAK_P3 and its host Pseudomonas aeruginosa. We detected a dramatic global depletion of bacterial transcripts coupled with their replacement by viral RNAs over the course of infection, eventually leading to drastic changes in pyrimidine metabolism. This process relies on host machinery hijacking as suggested by the strong up-regulation of one bacterial operon involved in RNA processing. Moreover, we found that RNA-based regulation plays a central role in PAK_P3 lifecycle as antisense transcripts are produced mainly during the early stage of infection and viral small non coding RNAs are massively expressed at the end of infection. This work highlights the prominent role of RNA metabolism in the infection strategy of a bacteriophage belonging to a new characterized sub-family of viruses with promising therapeutic potential.

  13. BAD contributes to RAF-mediated proliferation and cooperates with B-RAF-V600E in cancer signaling.

    PubMed

    Polzien, Lisa; Baljuls, Angela; Albrecht, Marco; Hekman, Mirko; Rapp, Ulf R

    2011-05-20

    BAD (Bcl-2 antagonist of cell death) belongs to the proapoptotic BH3-only subfamily of Bcl-2 proteins. Physiological activity of BAD is highly controlled by phosphorylation. To further analyze the regulation of BAD function, we investigated the role of recently identified phosphorylation sites on BAD-mediated apoptosis. We found that in contrast to the N-terminal phosphorylation sites, the serines 124 and 134 act in an antiapoptotic manner because the replacement by alanine led to enhanced cell death. Our results further indicate that RAF kinases represent, besides PAK1, BAD serine 134 phosphorylating kinases. Importantly, in the presence of wild type BAD, co-expression of survival kinases, such as RAF and PAK1, leads to a strongly increased proliferation, whereas substitution of serine 134 by alanine abolishes this process. Furthermore, we identified BAD serine 134 to be strongly involved in survival signaling of B-RAF-V600E-containing tumor cells and found that phosphorylation of BAD at this residue is critical for efficient proliferation in these cells. Collectively, our findings provide new insights into the regulation of BAD function by phosphorylation and its role in cancer signaling.

  14. Use of Dimples to Suppress Boundary Layer Separation on a Low Pressure Turbine Blade

    NASA Astrophysics Data System (ADS)

    Rouser, Kurt P.

    2002-12-01

    Flow separation on a low pressure turbine blade is explored at Reynolds numbers of 25k, 45k and 100k, Experimental data is collected in a low-speed, draw-down wind tunnel using a cascade of eight Pak-B blades, Flow is examined from measurements of blade surface pressures, boundary layer parameters, exit velocities, and total pressure losses across the blade, Two recessed dimple shapes are assessed for suppressing flow separation and associated losses, One dimple is spherical, and the second is asymmetric, formed from a full dimple spanwise half-filled, A single row of each dimple shape is tested at 50%, 55% and 65% axial chord, Symmetric dimples reduce separation losses by as much as 28%, while asymmetric dimples reduce losses by as much as 23%, A complementary three-dimensional computational study is conducted to visualize local flow structure, Computational analysis uses Gridgen v13,3 as a mesh generator, Fluent v6,O as a flow solver and FIELDVIEW - v8,0 for graphic display and analysis, Computational results for Pak-B blades at a Reynolds number of 25k indicate that both dimple shapes cause a span-wise vortex to rollup within the dimple and provide a localized pressure drop,

  15. Extracts of Tsai Tai (Brassica chinensis): enhanced antioxidant activity and anti-aging effects both in vitro and in Caenorhabditis elegans.

    PubMed

    Chen, Jing; Zhang, Ju; Xiang, Yanxia; Xiang, Limin; Liu, Yongmei; He, Xiangjiu; Zhou, Xiaoju; Liu, Xin; Huang, Zebo

    2016-02-01

    Tsai Tai is one of the most widely consumed Brassica vegetables in Asian countries because of its good taste and its nutritional benefits. This study evaluated the antioxidant capacity and possible associated health benefits of 3 Tsai Tai (Brassica chinensis) varieties, namely, Hon Tsai Tai, Pak Choi and Choi Sum. The DPPH radical scavenging ability and reducing power assays were performed to evaluate the in vitro activities of the extracts. Caenorhabditis elegans was used as an in vivo model for evaluation of beneficial health effects, including antioxidant activity and delayed aging. In vitro, the Hon Tsai Tai extract exhibited higher antioxidant activities than Pak Choi and Choi Sum, and the total phenolic contents were significantly correlated with the DPPH and RP values. In vivo, the three assayed Tsai Tai extracts significantly increased resistance against paraquat-induced oxidative stress with an increase in survival rates from 15% to 28% compared with controls. However, only the extract from Hon Tsai Tai significantly prolonged the lifespan of Caenorhabditis elegans, with an 8% increase in the mean lifespan with respect to controls. Further evidence of antioxidant protection was obtained by assessing ROS production via the DCF assay. The analyses of intracellular SOD activity and MDA content confirmed the existence of an antioxidant protective effect. These results suggest that Tsai Tai might serve as a good source of natural antioxidants, and in particular, Hon Tsai Tai could be explored as a potential dietary supplement to retard aging.

  16. Characterization of Novel Molecular Mechanisms Favoring Rac1 Membrane Translocation

    PubMed Central

    Castro-Castro, Antonio; Muriel, Olivia; del Pozo, Miguel A.

    2016-01-01

    The Rac1 GTPase plays key roles in cytoskeletal organization, cell motility and a variety of physiological and disease-linked responses. Wild type Rac1 signaling entails dissociation of the GTPase from cytosolic Rac1-Rho GDP dissociation inhibitor (GDI) complexes, translocation to membranes, activation by exchange factors, effector binding, and activation of downstream signaling cascades. Out of those steps, membrane translocation is the less understood. Using transfections of a expression cDNA library in cells expressing a Rac1 bioreporter, we previously identified a cytoskeletal feedback loop nucleated by the F-actin binding protein coronin 1A (Coro1A) that promotes Rac1 translocation to the plasma membrane by facilitating the Pak-dependent dissociation of Rac1-Rho GDI complexes. This screening identified other potential regulators of this process, including WDR26, basigin, and TMEM8A. Here, we show that WDR26 promotes Rac1 translocation following a Coro1A-like and Coro1A-dependent mechanism. By contrast, basigin and TMEM8A stabilize Rac1 at the plasma membrane by inhibiting the internalization of caveolin-rich membrane subdomains. This latter pathway is F-actin-dependent but Coro1A-, Pak- and Rho GDI-independent. PMID:27835684

  17. Synthetic Peptide Vaccine Development: Designing Dual Epitopes into a Single Pilin Peptide Immunogen Generates Antibody Cross-reactivity between Two Strains of Pseudomonas aeruginosa

    PubMed Central

    Hackbarth, Clifton; Hodges, Robert S.

    2010-01-01

    One of the main challenges of Pseudomonas aeruginosa (P. aeruginosa) vaccine development is the design of an antigen that elicits cross-reactive antibodies against multiple virulent strains. Using a rational design approach, we have developed a single 17-residue peptide immunogen that generates antibodies that target the receptor binding domain (RBD) of the type IV pilus of more than one strain of P. aeruginosa. Using the RBD sequence, of native strain PAO as a template, we have systematically changed up to five residues in the PAO sequence of the peptide immunogen, into that of the PAK sequence. We show by indirect and competitive ELISA, that the mutant peptide immunogens elicit the development of polyclonal sera that is cross-reactive to both native strain PAO and PAK pilin. We further show that there are at least two separate antibody populations in the polyclonal sera that possess closely-related epitopes but which are each strain specific. Moreover, part of the epitope for the PAO specific antibodies consists of several residues outside the disulfide loop of the receptor binding domain. This allows us to create two unique epitopes within the same receptor binding domain sequence. PMID:20807222

  18. Characterization of antralin: an endogenous protein inhibitor of radioligand binding to (/sup 3/H)RO 5-4864 and (/sup 3/H)nitrendipine binding sites

    SciTech Connect

    Mantione, C.R.; Goldman, M.E.; Bolger, G.T.; Lueddens, H.W.M.; Paul, S.M.; Skolnick, P.

    1986-03-01

    The presence of substances isolated from rat antral stomach that inhibit the binding of (/sup 3/H)Ro 5-4864 (4'-chlorodiazepam) to peripheral-type benzodiazepine receptors (PBR), but do not affect the binding of (/sup 3/H)diazepam to brain-type benzodiazepine receptors have been described. These substances, now obtained by isolation on Sep-Pak C/sub 18/ cartridges, also had no effect on (/sup 3/)PK 11195 binding which labels an antagonist site associated with PBR. Neither was any significant inhibition of (/sup 3/H)Ro 15-1788, (/sup 3/H)..beta..CCM, or (/sup 3/H)dihydroalprenolol binding observed. However, the Sep-Pak eluates were found to produce a concentration-dependent inhibition of (/sup 3/H)nitrendipine binding equivalent to that seen with (/sup 3/H)Ro 5-4864. This material, termed antralin, reduced the apparent K/sub d/ of both radioligands without significantly affecting B/sub max/. Its activity was destroyed by both heat treatment and pronase, and partially reduced by trypsin. Furthermore, its activity was enhanced by Ca/sup + +/ (0.1 mM). Antralin levels were unevenly distributed in rat tissues. These findings suggest that antralin is a protein that may regulate both PBR and dihydropyridine Ca/sup + +/-channel antagonist binding sites in vivo.

  19. Burn injury characteristics: findings from Pakistan National Emergency Department Surveillance Study

    PubMed Central

    2015-01-01

    Background Burn injury is an important yet under-researched area in Pakistan. The objective of this study was to determine the characteristics and associated outcomes of burn injury patients presenting to major emergency departments in Pakistan. Methods Pakistan National Emergency Department Surveillance (Pak-NEDS) was a pilot active surveillance conducted between November 2010 and March 2011. Information related to patient demographics, mode of arrival, cause of burn injury, and outcomes was analyzed for this paper. Data were entered using Epi Info and analyzed using SPSS v.20. Ethical approval was obtained from all participating sites. Results There were 403 burn injury patients in Pak-NEDS, with a male to female ratio 2:1. About 48.9% of the burn injury patients (n = 199) were between 10 - 29 years of age. There was no statistically significant difference between unintentional and intentional burn injury patients except for body part injured (p-value 0.004) and ED disposition (p-value 0.025). Among 21 patients who died, most were between 40 - 49 years of age (61.9%) and suffered from fire burns (81%). Conclusion Burn injuries are a burden on emergency rooms in Pakistan. We were able to demonstrate the significant burden of burn injuries that is not addressed by specialized burn centers. PMID:26692165

  20. Selection of in vivo expressed genes of Escherichia coli O157:H7 strain EDL933 in ground meat under elevated temperature conditions.

    PubMed

    Kroj, Andrea; Schmidt, Herbert

    2012-10-01

    Enterohemorrhagic Escherichia coli O157:H7 strains are important foodborne pathogens that are often transmitted to humans by the ingestion of raw or undercooked meat of bovine origin. To investigate adaptation of this pathogen during persistence and growth in ground meat, we established an in vivo expression technology model to identify genes that are expressed during growth in this food matrix under elevated temperatures (42°C). To improve on the antibiotic-based selection method, we constructed the promoter trap vector pAK-1, containing a promoterless kanamycin resistance gene. A genomic library of E. coli O157:H7 strain EDL933 was constructed in pAK-1 and used for promoter selection in ground meat. The 20 in vivo expressed genes identified were associated with transport processes, metabolism, macromolecule synthesis, and stress response. For most of the identified genes, only hypothetical functions could be assigned. The results of our study provide the first insights into the complex response of E. coli O157:H7 to a ground meat environment under elevated temperatures and establish a suitable vector for promoter studies or selection of in vivo induced promoters in foods such as ground meat.

  1. Profiling the changes in signaling pathways in ascorbic acid/β-glycerophosphate-induced osteoblastic differentiation.

    PubMed

    Chaves Neto, Antonio Hernandes; Queiroz, Karla Cristiana; Milani, Renato; Paredes-Gamero, Edgar Julian; Justo, Giselle Zenker; Peppelenbosch, Maikel P; Ferreira, Carmen Veríssima

    2011-01-01

    Despite numerous reports on the ability of ascorbic acid and β-glycerophosphate (AA/β-GP) to induce osteoblast differentiation, little is known about the molecular mechanisms involved in this phenomenon. In this work, we used a peptide array containing specific consensus sequences (potential substrates) for protein kinases and traditional biochemical techniques to examine the signaling pathways modulated during AA/β-GP-induced osteoblast differentiation. The kinomic profile obtained after 7 days of treatment with AA/β-GP identified 18 kinase substrates with significantly enhanced or reduced phosphorylation. Peptide substrates for Akt, PI3K, PKC, BCR, ABL, PRKG1, PAK1, PAK2, ERK1, ERBB2, and SYK showed a considerable reduction in phosphorylation, whereas enhanced phosphorylation was observed in substrates for CHKB, CHKA, PKA, FAK, ATM, PKA, and VEGFR-1. These findings confirm the potential usefulness of peptide microarrays for identifying kinases known to be involved in bone development in vivo and in vitro and show that this technique can be used to investigate kinases whose function in osteoblastic differentiation is poorly understood.

  2. Tgf-beta induced Erk phosphorylation of smad linker region regulates smad signaling.

    PubMed

    Hough, Chris; Radu, Maria; Doré, Jules J E

    2012-01-01

    The Transforming Growth Factor-Beta (TGF-β) family is involved in regulating a variety of cellular processes such as apoptosis, differentiation, and proliferation. TGF-β binding to a Serine/Threonine kinase receptor complex causes the recruitment and subsequent activation of transcription factors known as smad2 and smad3. These proteins subsequently translocate into the nucleus to negatively or positively regulate gene expression. In this study, we define a second signaling pathway leading to TGF-β receptor activation of Extracellular Signal Regulated Kinase (Erk) in a cell-type dependent manner. TGF-β induced Erk activation was found in phenotypically normal mesenchymal cells, but not normal epithelial cells. By activating phosphotidylinositol 3-kinase (PI3K), TGF-β stimulates p21-activated kinase2 (Pak2) to phosphorylate c-Raf, ultimately resulting in Erk activation. Activation of Erk was necessary for TGF-β induced fibroblast replication. In addition, Erk phosphorylated the linker region of nuclear localized smads, resulting in increased half-life of C-terminal phospho-smad 2 and 3 and increased duration of smad target gene transcription. Together, these data show that in mesenchymal cell types the TGF-β/PI3K/Pak2/Raf/MEK/Erk pathway regulates smad signaling, is critical for TGF-β-induced growth and is part of an integrated signaling web containing multiple interacting pathways rather than discrete smad/non-smad pathways.

  3. A new functional role uncovered for RASGRF2 in control of nuclear migration in cone photoreceptors during postnatal retinal development.

    PubMed

    Jimeno, David; Santos, Eugenio

    2017-01-02

    Despite their homologous structure and central nervous system(CNS) expression patterns, the GRF1 and GRF2 guanine nucleotide exchange factors(GEF) appear to play distinct, non-overlapping functions in cellular excitability, synaptic plasticity or neuromodulation. We recently uncovered a new functional role of GRF2 controlling nuclear migration in cone photoreceptors during postnatal neuroepithelial differentiation of the mouse retina. Analyzing GRF2-KO mice, we detected the specific accumulation of abnormally located, "ectopic" cone photoreceptor nuclei in the photoreceptor segment(PS) layer of their retinas. This alteration was accompanied by defective electroretinograms(ERG) indicative of impaired cone-mediated visual function, and accumulation around the "ectopic" nuclei of signaling molecules known to be functionally relevant for intracellular organelle migration, cytoskeletal reorganization or cell polarity establishment including PAR3, PAR6, and the phosphorylated proteins pPAK, pMLC2 and pVASP. We propose a mechanism whereby the absence of a productive functional interaction between GRF2 and its downstream target CDC42 leads to altered formation/structure of PAR-containing, polarity-related macromolecular complexes and abnormal activation of downstream signaling mediated by activated, phosphorylated forms of PAK, VASP and MLC2. As cone photoreceptors are responsible for color vision and visual acuity, these observations are potentially relevant for degenerative diseases of the human retina, harboring almost double number of cones than mice.

  4. Analysis of 26-hydroxycholesterol in plasma by high performance liquid chromatography

    SciTech Connect

    Kou, I.L.; Holmes, R.P.

    1986-03-01

    A method utilizing HPLC has been developed for the analysis of 26-hydroxycholesterol in plasma. The method involves initially saponifying samples for estimation of the total sterol or extracting samples with acetonitrile to estimate the free sterol. This is followed by a C-18 Sep Pak fractionation of the extracts, further purification on a Nova Pak C-18 HPLC column, and finally by quantitation on a silicic acid HPLC column by measuring the absorbance at 210 nm. Losses of sterol during this multi-step procedure were determined by estimating the recovery of /sup 3/H-26-hydroxycholesterol added to samples before extraction or saponification. The reproducibility of the method was evaluated by repetitive analysis of one plasma sample. The coefficient of variation was 6.8% for six analyses. In agreement with the mass spectrometric determinations of Javitt et al. The authors observed total plasma levels in the range of 140-260 ng/ml in young adult humans, rats, and rabbits. In the plasma of older human patients undergoing coronary by-pass surgery levels ranged from 335-950 ng/ml, suggesting that an increase in the level of 26-hydroxycholesterol is either age-related or related to the development of acute atherosclerosis.

  5. Lipid profiling by electrospray ionization tandem mass spectrometry and the identification of lipid phosphorylation by kinases in potato stolons

    PubMed Central

    Cenzano, Ana M.; Cantoro, Renata; Teresa Hernandez-Sotomayor, S. M.; Abdala, Guillermina I.; Racagni, Graciela E.

    2013-01-01

    There is limited information about the involvement of lipids and esterified fatty acids in signaling pathways during plant development. The purpose of this study was to evaluate the lipid composition and molecular species of potato (Solanum tuberosum L., cv. Spunta) stolons and to identify phosphorylated lipids in the first two developmental stages of tuber formation. Lipid profiling was determined using ESI-MS/MS, a useful method for the determination of the biosynthesis and catabolism of lipids based on their fatty acid composition. The most prevalent compound identified in this study was phosphatidic acid (PA); digalactosyldiacylglycerol (DGDG) was the second most abundant compound. A 34:2 species was identified in PA, phosphatidylcholine (PC), phosphatidylinositol (PI), and phosphatidylethanolamine (PE). The identification of lipid phosphorylation by kinases was revealed by the presence of the phosphorylated lipids. PA was metabolized to diacylglycerol pyrophosphate (DGPP) by phosphatidic acid kinase (PAK). This work establishes a correlation between lipid fatty acid composition and lipid metabolism enzymes at the beginning of tuber formation and is the first report of PAK activity in the early events of potato tuber formation. PMID:22142228

  6. Nonvirus encoded proteins could be embedded into Bombyx mori cypovirus polyhedra.

    PubMed

    Zhang, Yi-Ling; Xue, Ren-Yu; Cao, Guang-Li; Meng, Xiang-Kun; Zhu, Yue-Xiong; Pan, Zhong-Hua; Gong, Cheng-Liang

    2014-01-01

    To explore whether the nonvirus encoded protein could be embedded into Bombyx mori cypovirus (BmCPV) polyhedra. The stable transformants of BmN cells expressing a polyhedrin (Polh) gene of BmCPV were constructed by transfection with a non-transposon derived vector containing a polh gene. The polyhedra were purified from the midguts of BmCPV-infected silkworms and the transformed BmN cells, respectively. The proteins embedded into polyhedra were determined by mass spectrometry analysis. Host derived proteins were detected in the purified polyhedra. Analysis of structure and hydrophilicity of embedded proteins indicated that the hydrophilic proteins, in structure, were similar to the left-handed structure of polyhedrin or the N-terminal domain of BmCPV structural protein VP3, which were easily embedded into the BmCPV polyhedra. The lysate of polyhedra purified from the infected transformation of BmN cells with modified B. mori baculovirus BmPAK6 could infect BmN cells, indicating that B. mori baculovirus could be embedded into BmCPV polyhedra. Both the purified polyhedra and its lysate could be coloured by X-gal, indicating that the β-galactosidase expressed by BmPAK6 could be incorporated into BmCPV polyhedra. These results suggested that some heterologous proteins and baculovirus could be embedded into polyhedra in an unknown manner.

  7. Probabilistic acute dietary exposure assessment of the Chinese population to cypermethrin residues.

    PubMed

    Sun, J-F; Liu, P; Li, C-Y; Li, J-X; Wang, C-N; Min, J; Hu, D; Wu, Y-N

    2011-01-01

    A probabilistic estimation of dietary exposure to cypermethrin residues for the Chinese population was performed. Cypermethrin residue data were obtained from the national food contamination monitoring program for 2001-2006, encompassing 14,096 samples from 36 commodities with a detection rate of 10.4%. Food consumption data were gathered from the national nutrition and health survey conducted in 2002, comprising 65,915 consumers aged 2-100 years and 3701 children of 2-6 years old. The whole country was roughly divided into six regions and the ranges of the median and of P99.9 exposure estimated for these regions were 0.018-0.026 and 3.131-7.095 µg kg(-1 )bw day(-1), respectively. Pak-choi and Chinese cabbage contributed 33.9 and 13.2%, respectively, to the cypermethrin intake for the general population, while pak-choi and citrus covered 30.7 and 22.5% of the total intake for children, respectively. The exposure of the rural population was higher than urban populations. Rural areas mainly located in the plains of central China had among the highest exposure of the six regions, accounting for 17.7% of the ARfD at P99.9, while the 99.99th percentile of exposure for children, especially rural children, far exceeded the ARfD, which is a cause for concern.

  8. Social learning and amygdala disruptions in Nf1 mice are rescued by blocking p21-activated kinase

    PubMed Central

    Molosh, Andrei I.; Johnson, Philip L.; Spence, John P.; Arendt, David; Federici, Lauren M.; Bernabe, Cristian; Janasik, Steven P.; Segu, Zaneer M.; Khanna, Rajesh; Goswami, Chirayu; Zhu, Weiguo; Park, Su-Jung; Li, Lang; Mechref, Yehia S.; Clapp, D. Wade; Shekhar, Anantha

    2014-01-01

    Children with Neurofibromatosis type 1 (NF1) are increasingly recognized to have high prevalence of social difficulties and autism spectrum disorders (ASD). We demonstrated selective social learning deficit in mice with deletion of a single Nf1 gene (Nf1+/−), along with greater activation of mitogen activated protein kinase pathway in neurons from amygdala and frontal cortex, structures relevant to social behaviors. The Nf1+/− mice showed aberrant amygdala glutamate/GABA neurotransmission; deficits in long-term potentiation; and specific disruptions in expression of two proteins associated with glutamate and GABA neurotransmission: a disintegrin and metalloprotease domain 22 (ADAM22) and heat shock protein 70 (HSP70), respectively. All of these amygdala disruptions were normalized by co-deletion of p21 protein-activated kinase (Pak1) gene. We also rescued the social behavior deficits in Nf1+/− mice with pharmacological blockade of Pak1 directly in the amygdala. These findings provide novel insights and therapeutic targets for NF1 and ASD patients. PMID:25242307

  9. Evaluation of Viscosity Characteristics of Spin-Coated UV Nanoimprint Resin

    NASA Astrophysics Data System (ADS)

    Atobe, Hidemasa; Hiroshima, Hiroshi; Wang, Qing

    2010-06-01

    The viscosity of a UV-curable resin used in UV nanoimprint is one of the key parameters to determine the process speed of resin filling in the recesses of a mold. We have developed an apparatus to measure the viscosity of a spin-coated thin liquid film on a wafer. With this viscosity measuring apparatus we examined the effect of the film thickness, exposure of resin to atmosphere, and temperature of resin on the viscosity of PAK-01, which is a UV-curable resin commonly employed in UV nanoimprint. The viscosity of this resin with its film thickness larger than 2.3 µm showed a constant value of 83 mPa·s, which is almost the same as that of the bulk PAK-01. At below 2.3 µm, the viscosity seemed to increase with decreasing film thickness. The viscosity of the spin-coated resin was also found to increase with the duration of its exposure to atmosphere; for a 30 min exposure to atmosphere, the viscosity reached up to 461 mPa·s. It is considered that during the prolonged exposure, the volatile component of the resin evaporated. When subjected to heat treatment, the viscosity of the UV-curable resin did not seem to depend on film thickness and maintained a steady value of 385 mPa·s. It was found that as the film thickness decreased the viscosity approached the value obtained by heat treatment.

  10. Evaluation of Viscosity Characteristics of Spin-Coated UV Nanoimprint Resin

    NASA Astrophysics Data System (ADS)

    Hidemasa Atobe,; Hiroshi Hiroshima,; Qing Wang,

    2010-06-01

    The viscosity of a UV-curable resin used in UV nanoimprint is one of the key parameters to determine the process speed of resin filling in the recesses of a mold. We have developed an apparatus to measure the viscosity of a spin-coated thin liquid film on a wafer. With this viscosity measuring apparatus we examined the effect of the film thickness, exposure of resin to atmosphere, and temperature of resin on the viscosity of PAK-01, which is a UV-curable resin commonly employed in UV nanoimprint. The viscosity of this resin with its film thickness larger than 2.3 μm showed a constant value of 83 mPa\\cdots, which is almost the same as that of the bulk PAK-01. At below 2.3 μm, the viscosity seemed to increase with decreasing film thickness. The viscosity of the spin-coated resin was also found to increase with the duration of its exposure to atmosphere; for a 30 min exposure to atmosphere, the viscosity reached up to 461 mPa\\cdots. It is considered that during the prolonged exposure, the volatile component of the resin evaporated. When subjected to heat treatment, the viscosity of the UV-curable resin did not seem to depend on film thickness and maintained a steady value of 385 mPa\\cdots. It was found that as the film thickness decreased the viscosity approached the value obtained by heat treatment.

  11. Src and FAK kinases cooperate to phosphorylate paxillin kinase linker, stimulate its focal adhesion localization, and regulate cell spreading and protrusiveness.

    PubMed

    Brown, Michael C; Cary, Leslie A; Jamieson, Jennifer S; Cooper, Jonathan A; Turner, Christopher E

    2005-09-01

    The ArfGAP paxillin kinase linker (PKL)/G protein-coupled receptor kinase-interacting protein (GIT)2 has been implicated in regulating cell spreading and motility through its transient recruitment of the p21-activated kinase (PAK) to focal adhesions. The Nck-PAK-PIX-PKL protein complex is recruited to focal adhesions by paxillin upon integrin engagement and Rac activation. In this report, we identify tyrosine-phosphorylated PKL as a protein that associates with the SH3-SH2 adaptor Nck, in a Src-dependent manner, after cell adhesion to fibronectin. Both cell adhesion and Rac activation stimulated PKL tyrosine phosphorylation. PKL is phosphorylated on tyrosine residues 286/392/592 by Src and/or FAK and these sites are required for PKL localization to focal adhesions and for paxillin binding. The absence of either FAK or Src-family kinases prevents PKL phosphorylation and suppresses localization of PKL but not GIT1 to focal adhesions after Rac activation. Expression of an activated FAK mutant in the absence of Src-family kinases partially restores PKL localization, suggesting that Src activation of FAK is required for PKL phosphorylation and localization. Overexpression of the nonphosphorylated GFP-PKL Triple YF mutant stimulates cell spreading and protrusiveness, similar to overexpression of a paxillin mutant that does not bind PKL, suggesting that failure to recruit PKL to focal adhesions interferes with normal cell spreading and motility.

  12. The Chemistry and Biological Activities of Mimosine: A Review.

    PubMed

    Nguyen, Binh Cao Quan; Tawata, Shinkichi

    2016-08-01

    Mimosine [β-[N-(3-hydroxy-4-oxypyridyl)]-α-aminopropionic acid] is a non-protein amino acid found in the members of Mimosoideae family. There are a considerable number of reports available on the chemistry, methods for estimation, biosynthesis, regulation, and degradation of this secondary metabolite. On the other hand, over the past years of active research, mimosine has been found to have various biological activities such as anti-cancer, antiinflammation, anti-fibrosis, anti-influenza, anti-virus, herbicidal and insecticidal activities, and others. Mimosine is a leading compound of interest for use in the development of RAC/CDC42-activated kinase 1 (PAK1)-specific inhibitors for the treatment of various diseases/disorders, because PAK1 is not essential for the growth of normal cells. Interestingly, the new roles of mimosine in malignant glioma treatment, regenerative dentistry, and phytoremediation are being emerged. These identified properties indicate an exciting future for this amino acid. The present review is focused on the chemistry and recognized biological activities of mimosine in an attempt to draw a link between these two characteristics. Copyright © 2016 John Wiley & Sons, Ltd.

  13. Signaling Cascades Governing Cdc42-Mediated Chondrogenic Differentiation and Mensenchymal Condensation.

    PubMed

    Wang, Jirong R; Wang, Chaojun J; Xu, Chengyun Y; Wu, Xiaokai K; Hong, Dun; Shi, Wei; Gong, Ying; Chen, Haixiao X; Long, Fanxin; Wu, Ximei M

    2016-03-01

    Endochondral ossification consists of successive steps of chondrocyte differentiation, including mesenchymal condensation, differentiation of chondrocytes, and hypertrophy followed by mineralization and ossification. Loss-of-function studies have revealed that abnormal growth plate cartilage of the Cdc42 mutant contributes to the defects in endochondral bone formation. Here, we have investigated the roles of Cdc42 in osteogenesis and signaling cascades governing Cdc42-mediated chondrogenic differentiation. Though deletion of Cdc42 in limb mesenchymal progenitors led to severe defects in endochondral ossification, either ablation of Cdc42 in limb preosteoblasts or knockdown of Cdc42 in vitro had no obvious effects on bone formation and osteoblast differentiation. However, in Cdc42 mutant limb buds, loss of Cdc42 in mesenchymal progenitors led to marked inactivation of p38 and Smad1/5, and in micromass cultures, Cdc42 lay on the upstream of p38 to activate Smad1/5 in bone morphogenetic protein-2-induced mesenchymal condensation. Finally, Cdc42 also lay on the upstream of protein kinase B to transactivate Sox9 and subsequently induced the expression of chondrocyte differential marker in transforming growth factor-β1-induced chondrogenesis. Taken together, by using biochemical and genetic approaches, we have demonstrated that Cdc42 is involved not in osteogenesis but in chondrogenesis in which the BMP2/Cdc42/Pak/p38/Smad signaling module promotes mesenchymal condensation and the TGF-β/Cdc42/Pak/Akt/Sox9 signaling module facilitates chondrogenic differentiation.

  14. Biological Effects of Trichoderma harzianum Peptaibols on Mammalian Cells

    PubMed Central

    Peltola, Joanna; Ritieni, Alberto; Mikkola, Raimo; Grigoriev, Pavel A.; Pócsfalvi, Gabriella; Andersson, Maria A.; Salkinoja-Salonen, Mirja S.

    2004-01-01

    Trichoderma species isolated from water-damaged buildings were screened for toxicity by using boar sperm cells as indicator cells. The crude methanolic cell extract from Trichoderma harzianum strain ES39 inhibited the boar sperm cell motility at a low exposure concentration (50% effective concentration, 1 to 5 μg [dry weight] ml of extended boar semen−1). The same exposure concentration depleted the boar sperm cells of NADH2. Inspection of the exposed boar sperm cells by transmission electron microscopy revealed damage to the plasma membrane. By using the black lipid membrane technique, it was shown that the semipurified metabolites (eluted from a SepPak C18 cartridge) of T. harzianum strain ES39 induced voltage-dependent conductivity. The high-performance liquid chromatography-purified metabolites of T. harzianum strain ES39 dissipated the mitochondrial membrane potential (Δψm) of human lung epithelial carcinoma cells (cell line A549). The semipurified metabolites (eluted from a SepPak C18 cartridge) of T. harzianum strain ES39 were analyzed by mass spectrometry (MS). Matrix-assisted laser desorption ionization and nanoflow electrospray ionization MS revealed five major peptaibols, each of which contained 18 residues and had a mass ranging from 1,719 to 1,775 Da. Their partial amino acid sequences were determined by collision-induced dissociation tandem MS. PMID:15294840

  15. Characterization of Pseudomonas aeruginosa Phage C11 and Identification of Host Genes Required for Virion Maturation

    PubMed Central

    Cui, Xiaoli; You, Jiajia; Sun, Li; Yang, Xiaojing; Zhang, Tian; Huang, Kechong; Pan, Xuewei; Zhang, Fenjiao; He, Yang; Yang, Hongjiang

    2016-01-01

    The underlying mechanisms of phage-host interactions largely remained to be elucidated. In this work, Pseudomonas aeruginosa phage C11 was first characterized as a Myoviridae virus having a linear dsDNA molecule of 94109 bp with 1173 bp identical terminal direct repeats (TDR). Then the mutants resistant to phage C11 were screened in a Tn5G transposon mutant library of P. aeruginosa PAK, including two mutants with decreased adsorption rates (DAR) and five mutants with wild-type adsorption rates (WAR). When the WAR mutants were incubated with phage C11, their growth rates were significantly inhibited; the replication of the phage genomic DNA was detected in all the WAR mutants with the real-time quantitative PCR analysis; and the synthesized phage genomic DNA was processed into monomers for packaging evidenced by the southern blot analysis. Moreover, with strain PAK as indicator, small quantities of phage C11 were synthesized in the WAR mutants. Taken together, these data suggested the identified genes of the WAR mutants are necessary for efficient synthesis of the infectious phage particles. Finally, the WAR mutants were detected sensitive to two other Pseudomonas phages closely related with C11, further implying the evolved diversity and complexity of the phage-host interactions in both sides. PMID:28000703

  16. Analysis of baseline and alternative air quality scenarios for Pakistan: an integrated approach.

    PubMed

    Mir, Kaleem Anwar; Purohit, Pallav; Goldstein, Gary A; Balasubramanian, Rajasekhar

    2016-11-01

    This study aims to assess the current and future air pollution and associated health impacts in Pakistan. In this study, the Pakistan Integrated Energy Model (Pak-IEM) is used to assess current and future energy consumption in Pakistan. To assess air pollution levels and associated health impacts, we used the Greenhouse gas and Air pollution INteractions and Synergies (GAINS) model. A linkage has been established between both the models to feed the energy outputs from Pak-IEM into GAINS for exploring different scenarios. Mainly, the emissions of three air pollutants (SO2, NOx, and PM2.5) as well as the associated health impacts of increased emissions are assessed. Baseline emission scenario (BES) shows a growth in emissions of SO2, NOx, and PM2.5 by a factor of 2.4, 2.2, and 2.5 between 2007 and 2030. In terms of health impacts, by 2030, annual mean concentrations of fine particles (PM2.5) would increase to more than 150 μg/m(3) in some parts of Punjab region of Pakistan, for which loss in statistical life expectancy is calculated to increase from 30 to 60 months in 2007 up to 60-100 months in 2030 on average.

  17. Sepsis-induced elevation in plasma serotonin facilitates endothelial hyperpermeability.

    PubMed

    Li, Yicong; Hadden, Coedy; Cooper, Anthonya; Ahmed, Asli; Wu, Hong; Lupashin, Vladimir V; Mayeux, Philip R; Kilic, Fusun

    2016-03-09

    Hyperpermeability of the endothelial barrier and resulting microvascular leakage are a hallmark of sepsis. Our studies describe the mechanism by which serotonin (5-HT) regulates the microvascular permeability during sepsis. The plasma 5-HT levels are significantly elevated in mice made septic by cecal ligation and puncture (CLP). 5-HT-induced permeability of endothelial cells was associated with the phosphorylation of p21 activating kinase (PAK1), PAK1-dependent phosphorylation of vimentin (P-vimentin) filaments, and a strong association between P-vimentin and ve-cadherin. These findings were in good agreement with the findings with the endothelial cells incubated in serum from CLP mice. In vivo, reducing the 5-HT uptake rates with the 5-HT transporter (SERT) inhibitor, paroxetine blocked renal microvascular leakage and the decline in microvascular perfusion. Importantly, mice that lack SERT showed significantly less microvascular dysfunction after CLP. Based on these data, we propose that the increased endothelial 5-HT uptake together with 5-HT signaling disrupts the endothelial barrier function in sepsis. Therefore, regulating intracellular 5-HT levels in endothelial cells represents a novel approach in improving sepsis-associated microvascular dysfunction and leakage. These new findings advance our understanding of the mechanisms underlying cellular responses to intracellular/extracellular 5-HT ratio in sepsis and refine current views of these signaling processes during sepsis.

  18. A novel role for RhoA GTPase in the regulation of airway smooth muscle contraction.

    PubMed

    Zhang, Wenwu; Huang, Youliang; Wu, Yidi; Gunst, Susan J

    2015-02-01

    Recent studies have demonstrated a novel molecular mechanism for the regulation of airway smooth muscle (ASM) contraction by RhoA GTPase. In ASM tissues, both myosin light chain (MLC) phosphorylation and actin polymerization are required for active tension generation. RhoA inactivation dramatically suppresses agonist-induced tension development and completely inhibits agonist-induced actin polymerization, but only slightly reduces MLC phosphorylation. The inhibition of MLC phosphatase does not reverse the effects of RhoA inactivation on contraction or actin polymerization. Thus, RhoA regulates ASM contraction through its effects on actin polymerization rather than MLC phosphorylation. Contractile stimulation of ASM induces the recruitment and assembly of paxillin, vinculin, and focal adhesion kinase (FAK) into membrane adhesion complexes (adhesomes) that regulate actin polymerization by catalyzing the activation of cdc42 GTPase by the G-protein-coupled receptor kinase-interacting target (GIT) - p21-activated kinase (PAK) - PAK-interacting exchange factor (PIX) complex. Cdc42 is a necessary and specific activator of the actin filament nucleation activator, N-WASp. The recruitment and activation of paxillin, vinculin, and FAK is prevented by RhoA inactivation, thus preventing cdc42 and N-WASp activation. We conclude that RhoA regulates ASM contraction by catalyzing the assembly and activation of membrane adhesome signaling modules that regulate actin polymerization, and that the RhoA-mediated assembly of adhesome complexes is a fundamental step in the signal transduction process in response to a contractile agonist.

  19. Dynamic interplay between nitration and phosphorylation of tubulin cofactor B in the control of microtubule dynamics

    PubMed Central

    Rayala, Suresh K.; Martin, Emil; Sharina, Iraida G.; Molli, Poonam R.; Wang, Xiaoping; Jacobson, Raymond; Murad, Ferid; Kumar, Rakesh

    2007-01-01

    Tubulin cofactor B (TCoB) plays an important role in microtubule dynamics by facilitating the dimerization of α- and β-tubulin. Recent evidence suggests that p21-activated kinase 1 (Pak1), a major signaling nodule in eukaryotic cells, phosphorylates TCoB on Ser-65 and Ser-128 and plays an essential role in microtubule regrowth. However, to date, no upstream signaling molecules have been identified to antagonize the functions of TCoB, which might help in maintaining the equilibrium of microtubules. Here, we discovered that TCoB is efficiently nitrated, mainly on Tyr-64 and Tyr-98, and nitrated-TCoB attenuates the synthesis of new microtubules. In addition, we found that nitration of TCoB antagonizes signaling-dependent phosphorylation of TCoB, whereas optimal nitration of TCoB requires the presence of functional Pak1 phosphorylation sites, thus providing a feedback mechanism to regulate phosphorylation-dependent MT regrowth. Together these findings identified TCoB as the third cytoskeleton protein to be nitrated and suggest a previously undescribed mechanism, whereby growth factor signaling may coordinately integrate nitric oxide signaling in the regulation of microtubule dynamics. PMID:18048340

  20. Differential signalling by muscarinic receptors in smooth muscle: m2-mediated inactivation of myosin light chain kinase via Gi3, Cdc42/Rac1 and p21-activated kinase 1 pathway, and m3-mediated MLC20 (20 kDa regulatory light chain of myosin II) phosphorylation via Rho-associated kinase/myosin phosphatase targeting subunit 1 and protein kinase C/CPI-17 pathway.

    PubMed

    Murthy, Karnam S; Zhou, Huiping; Grider, John R; Brautigan, David L; Eto, Masumi; Makhlouf, Gabriel M

    2003-08-15

    Signalling via m3 and m2 receptors in smooth muscles involved activation of two G-protein-dependent pathways by each receptor. m2 receptors were coupled via Gbetagammai3 with activation of phospholipase C-beta3, phosphoinositide 3-kinase and Cdc42/Rac1 (where Cdc stands for cell division cycle) and p21-activated kinase 1 (PAK1), resulting in phosphorylation and inactivation of myosin light chain kinase (MLCK). Each step was inhibited by methoctramine and pertussis toxin. PAK1 activity was abolished in cells expressing both Cdc42-DN (where DN stands for dominant negative) and Rac1-DN. MLCK phosphorylation was inhibited by PAK1 antibody, and in cells expressing Cdc42-DN and Rac1-DN. m3 receptors were coupled via Galpha(q/11) with activation of phospholipase C-beta1 and via RhoA with activation of Rho-associated kinase (Rho kinase), phospholipase D and protein kinase C (PKC). Rho kinase and phospholipase D activities were inhibited by C3 exoenzyme and in cells expressing RhoA-DN. PKC activity was inhibited by bisindolylmaleimide, and in cells expressing RhoA-DN; PKC activity was also inhibited partly by Y27632 (44+/-5%). PKC-induced phosphorylation of PKC-activated 17 kDa inhibitor protein of type 1 phosphatase (CPI-17) at Thr38 was abolished by bisindolylmaleimide and inhibited partly by Y27632 (28+/-3%). Rho-kinase-induced phosphorylation of myosin phosphatase targeting subunit (MYPT1) and was abolished by Y27632. Sustained phosphorylation of 20 kDa regulatory light chain of myosin II (MLC20) and contraction were abolished by bisindolylmaleimide Y27632 and C3 exoenzyme and in cells expressing RhoA-DN. The results suggest that Rho-kinase-dependent phosphorylation of MYPT1 and PKC-dependent phosphorylation and enhancement of CPI-17 binding to the catalytic subunit of MLC phosphatase (MLCP) act co-operatively to inhibit MLCP activity, leading to sustained stimulation of MLC20 phosphorylation and contraction. Because Y27632 inhibited both Rho kinase and PKC activities

  1. Differential signalling by muscarinic receptors in smooth muscle: m2-mediated inactivation of myosin light chain kinase via Gi3, Cdc42/Rac1 and p21-activated kinase 1 pathway, and m3-mediated MLC20 (20 kDa regulatory light chain of myosin II) phosphorylation via Rho-associated kinase/myosin phosphatase targeting subunit 1 and protein kinase C/CPI-17 pathway.

    PubMed Central

    Murthy, Karnam S; Zhou, Huiping; Grider, John R; Brautigan, David L; Eto, Masumi; Makhlouf, Gabriel M

    2003-01-01

    Signalling via m3 and m2 receptors in smooth muscles involved activation of two G-protein-dependent pathways by each receptor. m2 receptors were coupled via Gbetagammai3 with activation of phospholipase C-beta3, phosphoinositide 3-kinase and Cdc42/Rac1 (where Cdc stands for cell division cycle) and p21-activated kinase 1 (PAK1), resulting in phosphorylation and inactivation of myosin light chain kinase (MLCK). Each step was inhibited by methoctramine and pertussis toxin. PAK1 activity was abolished in cells expressing both Cdc42-DN (where DN stands for dominant negative) and Rac1-DN. MLCK phosphorylation was inhibited by PAK1 antibody, and in cells expressing Cdc42-DN and Rac1-DN. m3 receptors were coupled via Galpha(q/11) with activation of phospholipase C-beta1 and via RhoA with activation of Rho-associated kinase (Rho kinase), phospholipase D and protein kinase C (PKC). Rho kinase and phospholipase D activities were inhibited by C3 exoenzyme and in cells expressing RhoA-DN. PKC activity was inhibited by bisindolylmaleimide, and in cells expressing RhoA-DN; PKC activity was also inhibited partly by Y27632 (44+/-5%). PKC-induced phosphorylation of PKC-activated 17 kDa inhibitor protein of type 1 phosphatase (CPI-17) at Thr38 was abolished by bisindolylmaleimide and inhibited partly by Y27632 (28+/-3%). Rho-kinase-induced phosphorylation of myosin phosphatase targeting subunit (MYPT1) and was abolished by Y27632. Sustained phosphorylation of 20 kDa regulatory light chain of myosin II (MLC20) and contraction were abolished by bisindolylmaleimide Y27632 and C3 exoenzyme and in cells expressing RhoA-DN. The results suggest that Rho-kinase-dependent phosphorylation of MYPT1 and PKC-dependent phosphorylation and enhancement of CPI-17 binding to the catalytic subunit of MLC phosphatase (MLCP) act co-operatively to inhibit MLCP activity, leading to sustained stimulation of MLC20 phosphorylation and contraction. Because Y27632 inhibited both Rho kinase and PKC activities

  2. Highly sensitive measurements of substrates and inhibitors on the basis of tyrosinase sensors and recycling systems

    NASA Astrophysics Data System (ADS)

    Streffer, Katrin

    2002-12-01

    compounds in river and sea water and the results could correlated very well with the corresponding DIN-test for the determination of phenolic compounds. An other developed tyrosinasesensor showed a very high sensitiveness for catecholamines, substances which are of special importance in the medical diagnostics. In addition, the investigations of two different tyrosinases, which were carried out also in the context of this thesis, have shown, that a special tyrosinase (tyrosinase from Streptomyces antibioticus) will be the better choice as tyrosinase from Agaricus bisporus, which is used in the area of biosensor research till now, if one wants to develop in future even more sensitive tyrosinasesensors. Furthermore, first successes became reached on a molecular biological field, the production of tyrosinasemutants with special, before well-considered features. These successes can be used to develop a new generation of tyrosinasesensors, tyrosinasesensors in which tyrosinase can be bound directionally both to the corresponding physical pickup or also to another enzyme. From this one expects to achieve ways minimized which the substance to be determined (or whose product) otherwise must cover. Finally, this should result in an clearly visible increase of sensitivity of the Biosensor. Analytische Chemie heute meint nicht länger nur die große Messtechnik, die zeit- und kostenintensiv ist, die außerdem nur von qualifiziertem Personal zu bedienen ist und deren Resultate nur durch dieses Personal auswertbar sind. Meist erfordert diese sagen wir 'klassische analytische Messtechnik' auch noch spezielle Räumlichkeiten und oft eine relative große Menge an speziell vorbereiteten Proben. Neben dieser klassischen analytischen Messtechnik hat sich besonders in den letzten Jahren eine auf bestimmte Stoffgruppen und Anforderungen zugeschnittene Messtechnik durchgesetzt, die oft auch durch einen Laien bedient werden kann. Meist sind es sehr kleine Geräte. Auch die ben

  3. HILI destabilizes microtubules by suppressing phosphorylation and Gigaxonin-mediated degradation of TBCB

    PubMed Central

    Tan, Hao; Liao, Hua; Zhao, Lianfang; Lu, Yilu; Jiang, Siyuan; Tao, Dachang; Liu, Yunqiang; Ma, Yongxin

    2017-01-01

    Human PIWIL2, aka HILI, is a member of PIWI protein family and overexpresses in various tumors. However, the underlying mechanisms of HILI in tumorigenesis remain largely unknown. TBCB has a critical role in regulating microtubule dynamics and is overexpressed in many cancers. Here we report that HILI inhibits Gigaxonin-mediated TBCB ubiquitination and degradation by interacting with TBCB, promoting the binding between HSP90 and TBCB, and suppressing the interaction between Gigaxonin and TBCB. Meanwhile, HILI can also reduce phosphorylation level of TBCB induced by PAK1. Our results showed that HILI suppresses microtubule polymerization and promotes cell proliferation, migration and invasion via TBCB for the first time, revealing a novel mechanism for HILI in tumorigenesis. PMID:28393858

  4. Life on a Surface in a Low-Reynolds-Number Flow

    NASA Astrophysics Data System (ADS)

    Stone, Howard A.

    2014-11-01

    There are many studies of the dynamics of swimming microorganisms. There are far fewer studies of how bacteria that attach to surfaces respond to motions of the surrounding fluid. Such motions can influence bacterial orientation, which can, in turn, impact bacterial colonization and motility on surfaces. Moreover, as biofilms develop, stresses from the flow on the surface can cause three-dimensional rearrangements of the soft biofilm in the form of ``streamers'' that can bridge the sides of a porous system; such suspended filaments trigger rapid clogging. Some of the underlying fluid dynamics will be discussed in the spirit of how the flow couples to the spatial and temporal evolution of these bacterial systems. Joint work with N. Autrusson, B. Bassler, K. Drescher, Z. Gitai, L. Guglielmini, F. Ingremeau, M.K. Kim, S. Lecuyer, O. Pak, A. Persat, R. Rusconi, Y. Shen, A. Siryaporn, N. Wingreen.

  5. The Cleaning of Aluminum Frame Assembly Units

    SciTech Connect

    Shen, T H

    2001-05-16

    The Brulin immersion and the precision cleaning experiments have shown that neither the Brulin solution nor the precision cleaning in AstroPak causes the smut formation on aluminum surfaces. The acid-bath cleaning in GTC is the primary source of the smut formation. The current GTC acid formulation etches the aluminum matrix quite aggressively, but does not appear to appreciably attack the Si particles. Therefore, this acid-bath cleaning will leave the cast-aluminum part surfaces with many protruded Si particles, which could potentially cause smut problems in the cleaning process down-stream. To ensure the removal of all loose Si particles from the cast-aluminum parts, it is necessary to physically hand-wipe and vigorously wash the acid-bath cleaned surfaces. Furthermore, the casting porosity in alloy A356 could be another source in causing high swipe readings in the FAU parts.

  6. Identification of anaerobes on the Minitek System, compared to a conventional system.

    PubMed

    Holloway, Y; Dankert, J

    1979-01-01

    The Minitek Miniaturized System (BBL) was compared to a conventional method (V.P.I.) for the indentification of anaerobes. Anaerobiosis was achieved by using the GasPak system (BBL) and a glove box. Gram staining, colonial morphology, gas-liquid chromatography and biochemical reactions as detailed in the Virginia Polytechnic Institute anaerobe laboratory manual were compared to the biochemical results obtained on the Minitek system. A total of 32 anaerobes was used. The percentage of correlation was an overall 98.2%. Discrepancies were found mostly in the production of indole, the reduction of nitrate, and the hydrolysis of aesculin. A further 95 anaerobes were tested on these three reactions, and found to give an overall agreement of 92.9%. It was found necessary to use some additional tests as well as gas-liquid chromatography for definitive identification. The microsystem was also evaluated as to time saved, cost and ease of use.

  7. Rapid method for measuring rotenone in water at piscicidal concentrations

    USGS Publications Warehouse

    Dawson, V.K.; Harman, P.D.; Schultz, D.P.; Allen, J.L.

    1983-01-01

    A high-performance liquid chromatography (HPLC) procedure that is rapid, specific, and sensitive (limit of detection <0.005 mg/liter) was developed for monitoring application and degradation rates of rotenone. For analysis, a water sample is buffered to pH 5 and injected through a Sep Pak(R) C18 disposable cartridge. The cartridge adsorbs and retains the rotenone which then can be eluted quantitatively from the cartridge with a small volume of methanol. This step effectively concentrates the sample and provides sample cleanup. The methanol extract is analyzed directly by HPLC on an MCH 10 reverse-phase column; methanol: water (75:25, volume : volume) is the mobile phase and flow rate is 1.5 ml/minute. The rotenone is detected by ultraviolet spectrophotometry at a wavelength of 295 nm.

  8. Photodegradation and flow-injection determination of simetryn herbicide by luminol chemiluminescence detection.

    PubMed

    Waseem, Amir; Yaqoob, Mohammad; Nabi, Abdul

    2008-08-01

    A novel and simple flow injection chemiluminescence method is reported for the determination of simetryn, a common herbicide. The method is based on the direct oxidation of luminol by the photoproducts of the simetryn in alkaline medium in the absence of catalyst/oxidant. The linear concentration range was 0.01 - 2 microg mL(-1) simetryn with a correlation coefficient (r(2)) of 0.9997 and relative standard deviations (RSD; n = 4) in the range of 0.9 - 2.3%. The limit of detection (S/N = 3) was 7.5 ng mL(-1) with a sample throughput of 100 h(-1). The proposed method has been applied to determine simetryn in natural waters using Sep-Pak C(18) cartridges for solid phase extraction (SPE) procedure. The recoveries were in the range of 97 +/- 1 to 104 +/- 2%. The mechanism of chemiluminescence reaction has also been discussed briefly.

  9. Application of the System Identification Technique to Goal-Directed Saccades.

    DTIC Science & Technology

    1984-07-30

    found in Appendix Al, yields 47 8 - 5.0938109 x 107 -.’ ’ o . - . -.. .... * . * . s - - * ** . - -. .. . . 63 P3 W 1.548 x 10 P = 3.4455 x 10 P2 P1 ...absolute value of displacement for subject 2.I Fig.S. he tme nteral romthe tar of he accae t th the ime t pak vlocty v abslut vale o • - " " "-’. di...AT FAG-B AG FANT) P 3 9 + P20 + p1 + p0e (13) where 6 = 57 .296K A/C4 p = I1 C1/C41 and P0 = C/C 4 State equations are usually written for ease in

  10. Detection of Lactobacillus acidophilus in Feces of Humans, Pigs, and Chickens1

    PubMed Central

    Gilliland, S. E.; Speck, M. L.; Morgan, C. G.

    1975-01-01

    Lactobacilli in fecal material from humans, pigs, and chickens were enumerated on lactobacillus selective agar (LBS). In all samples, higher numbers of lactobacilli were detected when plates were incubated in a system flushed with CO2 rather than in air. Much higher numbers of bacteria from human feces were detected when the LBS agar plates were incubated anaerobically in a hydrogen-carbon dioxide atmosphere (GasPak) than when incubated in CO2. The bacteria from human feces isolated on LBS agar incubated anaerobically were predominately bifidobacteria. Cultures from all three sources isolated on LBS agar incubated under CO2 were lactobacilli, including Lactobacillus acidophilus. Differences were observed in biochemical characteristics of some of the L. acidophilus isolated from all three sources. Guanine plus cytosine base ratios of deoxyribonucleic acid isolated from L. acidophilus cultures from humans were lower, in most cases, than those from pigs and chickens. PMID:811162

  11. KCC2 Gates Activity-Driven AMPA Receptor Traffic through Cofilin Phosphorylation.

    PubMed

    Chevy, Quentin; Heubl, Martin; Goutierre, Marie; Backer, Stéphanie; Moutkine, Imane; Eugène, Emmanuel; Bloch-Gallego, Evelyne; Lévi, Sabine; Poncer, Jean Christophe

    2015-12-02

    Expression of the neuronal K/Cl transporter KCC2 is tightly regulated throughout development and by both normal and pathological neuronal activity. Changes in KCC2 expression have often been associated with altered chloride homeostasis and GABA signaling. However, recent evidence supports a role of KCC2 in the development and function of glutamatergic synapses through mechanisms that remain poorly understood. Here we show that suppressing KCC2 expression in rat hippocampal neurons precludes long-term potentiation of glutamatergic synapses specifically by preventing activity-driven membrane delivery of AMPA receptors. This effect is independent of KCC2 transporter function and can be accounted for by increased Rac1/PAK- and LIMK-dependent cofilin phosphorylation and actin polymerization in dendritic spines. Our results demonstrate that KCC2 plays a critical role in the regulation of spine actin cytoskeleton and gates long-term plasticity at excitatory synapses in cortical neurons.

  12. Presence of immunoreactive atrial natriuretic peptide in follicular fluid, ovary and ovarian perfusates

    SciTech Connect

    Kim, Suhn Hee; Cho, Kyung Woo; Seul, Kyung Hwan; Ryu, Hoon; Koh, Gou Young )

    1989-01-01

    Immunoreactive atrial natriuretic peptide (ir-ANP) was measured in the follicular fluid of pig ovarian follicle, and rabbit ovarian homogenates and perfusates using a specific radioimmunoassay (RIA). Serial dilution curves made with the extracts of follicular fluid, ovarian homogenates and perfusates using Sep-Pak C18 cartridges were parallel with the RIA standard curve. On gel filtration chromatography and reverse phase HPLC, all extracted materials showed high and low molecular weight forms of ir-ANP. The amount of ir-ANP in rabbit ovary was 40.7{plus minus}0.39 pg/mg and that in follicular fluid of pig ovarian follicle was 18.88{plus minus}2.49 pg/ml.

  13. Determination of polycyclic aromatic hydrocarbons in diesel exhaust particulate matter and diesel fuel oil.

    PubMed

    Obuchi, A; Aoyama, H; Ohi, A; Ohuchi, H

    1984-11-16

    Clean-up procedures were developed for a method for determining the amount of polycyclic aromatic hydrocarbons (PAHs) in diesel exhaust particulate matter and in diesel fuel oils using reversed-phase high-performance liquid chromatography (HPLC). They were based mainly on the elimination of insoluble matter and aliphatic compounds that affect the performance of HPLC, from the dichloromethane extracts of particulate matter or from oils, with the aid of a disposable preparation column containing reversed-phase packings (Sep-Pak C18). Using these procedures, it is possible to detect 1 ng of benzo(a)pyrene in 30 mg of particulate matter with more than a 97% recovery or 0.5 ng in 50 microliters of oil with 91% recovery. Examples of analyses are given for particulate matter emitted from a diesel test engine and for diesel fuel oils, such as gas oil, residual oil and coal-liquefied oil.

  14. Symposium on Underwater Physiology (7th), July 5-10, 1980, Athens, Greece

    DTIC Science & Technology

    1980-07-10

    o Chairman of the Syyzou±= Governing Board O04T4 01 9. 109�" ONGANIZATION4 NAM& AND A00112111 Ia *NGAM £LEMEW PFIC 117, TASKr AREAA IWA UNI UM...CONTROWLUNG1 OPFICE NAM9 AND A010N1196 12. REPORIT DAY% Offiie of Naval Research 5 3 . l NUSR 7PGES 800 North quinoy StreetIs U1114aiPAK Ari~ on, VA 22217 96...were from, countries other than the U.S.A. I’I n .1 S, N 0102.. ... . .. O 6601 Unalasified ~~ SECURITY~ CL.ASS’ ICATION Olt TMIS PA@ 3 ?WSMug 0.60 Itilf

  15. A baseline study of benthic community associated with Amphioxus Sand in subtropical Hong Kong.

    PubMed

    Chen, Y; Cheung, S G; Shin, P K S

    2013-07-15

    An annual investigation on the seasonal changes of benthic community structure associated with Amphioxus Sand was conducted at two sites in the eastern waters of subtropical Hong Kong, where three species of amphioxus, Branchiostoma belcheri, B. japonicum and B. malayanum coexist. A total of 84 species and 4169 individuals were recorded at Tai Long Wan, whereas a total of 87 species and 3915 individuals were recorded at Pak Lap Wan. Benthic polychaetes were dominant, including high abundance of Onuphis eremita and Prionospio malmgreni. Results of cluster analysis showed significant community structures between the two areas because of difference in sediment granulometry. However, temporal changes within these Amphioxus Sand communities were minimal. In general, the Amphioxus Sand communities in Hong Kong showed higher species richness of Polychaeta as compared with similar studies elsewhere, possibly implying an increased level of organic pollution in Hong Kong waters.

  16. Assessment of computer codes for VVER-440/213-type nuclear power plants

    SciTech Connect

    Szabados, L.; Ezsol, Gy.; Perneczky

    1995-09-01

    Nuclear power plant of VVER-440/213 designed by the former USSR have a number of special features. As a consequence of these features the transient behaviour of such a reactor system should be different from the PWR system behaviour. To study the transient behaviour of the Hungarian Paks Nuclear Power Plant of VVER-440/213-type both analytical and experimental activities have been performed. The experimental basis of the research in the PMK-2 integral-type test facility , which is a scaled down model of the plant. Experiments performed on this facility have been used to assess thermal-hydraulic system codes. Four tests were selected for {open_quotes}Standard Problem Exercises{close_quotes} of the International Atomic Energy Agency. Results of the 4th Exercise, of high international interest, are presented in the paper, focusing on the essential findings of the assessment of computer codes.

  17. Packing in a tradition of recycling: Manufacturer-turned-recycler Free-Flow Packaging Corp. , Redwood City, Calif

    SciTech Connect

    White, K.M.

    1994-01-01

    Free-Flow Packaging Corp. recycles polystyrene. Loose-fill -- an industry name for expanded polystyrene (EPS) packaging modules, or what the public more commonly calls peanuts'' -- represents a material that can easily and economically be recycled over and over. The company manufactures a 100% recycled packaging peanut called FLO-PAK, as well as a variety of other EPS packaging products. Indeed, to date, Free-Flow Packaging has set up post-consumer EPS recycling operations at five of its 11 manufacturing facilities, both across the country and overseas. The corporation's original facility in Redwood City began this tradition when it first started processing industrial EPS scrap in 1978 and, later, pioneered the recycling of post-consumer EPS on site for use in its products in 1989. Now, only five years later, the result has produced a recycling operation that is truly successful, profitable, and closed-loop.

  18. Solid-phase extraction using hierarchical organosilicates for enhanced detection of nitroenergetic targets.

    PubMed

    Johnson, Brandy J; Melde, Brian J; Leska, Iwona A; Charles, Paul T; Hewitt, Alan D

    2011-05-01

    A novel porous organosilicate material was evaluated for application as a solid phase extraction sorbent for preconcentration of nitroenergetic targets from aqueous solution prior to HPLC analysis. The performance of the sorbent in spiked deionized water, groundwater, and surface water was evaluated. Targets considered included 2,4,6-trinitrotoluene, 2,4-dinitrotoluene, RDX, HMX, and nitroglycerin. The sorbent was shown to provide improved performance over Sep-Pak RDX. The impact of complex matrices on target preconcentration by the sorbent was also found to be less dramatic than that observed for LiChrolut EN. The impact of changes in pH on target preconcentration was considered. Aqueous soil extracts generated from samples collected at sites of ordnance testing were also used to evaluate the materials. The results presented here demonstrate the potential of this novel sorbent for application as a solid phase extraction material for the preconcentration of nitroenergetic targets from aqueous solutions.

  19. Scribble Scaffolds a Signalosome for Active Forgetting.

    PubMed

    Cervantes-Sandoval, Isaac; Chakraborty, Molee; MacMullen, Courtney; Davis, Ronald L

    2016-06-15

    Forgetting, one part of the brain's memory management system, provides balance to the encoding and consolidation of new information by removing unused or unwanted memories or by suppressing their expression. Recent studies identified the small G protein, Rac1, as a key player in the Drosophila mushroom bodies neurons (MBn) for active forgetting. We subsequently discovered that a few dopaminergic neurons (DAn) that innervate the MBn mediate forgetting. Here we show that Scribble, a scaffolding protein known primarily for its role as a cell polarity determinant, orchestrates the intracellular signaling for normal forgetting. Knocking down scribble expression in either MBn or DAn impairs normal memory loss. Scribble interacts physically and genetically with Rac1, Pak3, and Cofilin within MBn, nucleating a forgetting signalosome that is downstream of dopaminergic inputs that regulate forgetting. These results bind disparate molecular players in active forgetting into a single signaling pathway: Dopamine→ Dopamine Receptor→ Scribble→ Rac→ Cofilin.

  20. Immunoreactive prohormone atrial natriuretic peptides 1-30 and 31-67 - Existence of a single circulating amino-terminal peptide

    NASA Technical Reports Server (NTRS)

    Chen, Yu-Ming; Whitson, Peggy A.; Cintron, Nitza M.

    1990-01-01

    Sep-Pak C18 extraction of human plasma and radioimmunoassay using antibodies which recognize atrial natriuretic peptide (99-128) and the prohormone sequences 1-30 and 31-67 resulted in mean values from 20 normal subjects of 26.2 (+/- 9.2), 362 (+/- 173) and 368 (+/- 160) pg/ml, respectively. A high correlation coefficient between values obtained using antibodies recognizing prohormone sequences 1-30 and 31-67 was observed (R = 0.84). Extracted plasma immunoreactivity of 1-30 and 31-67 both eluted at 46 percent acetonitrile. In contrast, chromatographic elution of synthetic peptides 1-30 and 31-67 was observed at 48 and 39 percent acetonitrile, respectively. Data suggest that the radioimmunoassay of plasma using antibodies recognizing prohormone sequences 1-30 and 31-67 may represent the measurement of a unique larger amino-terminal peptide fragment containing antigenic sites recognized by both antisera.

  1. Golgi membrane fission requires the CtBP1-S/BARS-induced activation of lysophosphatidic acid acyltransferase δ.

    PubMed

    Pagliuso, Alessandro; Valente, Carmen; Giordano, Lucia Laura; Filograna, Angela; Li, Guiling; Circolo, Diego; Turacchio, Gabriele; Marzullo, Vincenzo Manuel; Mandrich, Luigi; Zhukovsky, Mikhail A; Formiggini, Fabio; Polishchuk, Roman S; Corda, Daniela; Luini, Alberto

    2016-07-12

    Membrane fission is an essential cellular process by which continuous membranes split into separate parts. We have previously identified CtBP1-S/BARS (BARS) as a key component of a protein complex that is required for fission of several endomembranes, including basolateral post-Golgi transport carriers. Assembly of this complex occurs at the Golgi apparatus, where BARS binds to the phosphoinositide kinase PI4KIIIβ through a 14-3-3γ dimer, as well as to ARF and the PKD and PAK kinases. We now report that, when incorporated into this complex, BARS binds to and activates a trans-Golgi lysophosphatidic acid (LPA) acyltransferase type δ (LPAATδ) that converts LPA into phosphatidic acid (PA); and that this reaction is essential for fission of the carriers. LPA and PA have unique biophysical properties, and their interconversion might facilitate the fission process either directly or indirectly (via recruitment of proteins that bind to PA, including BARS itself).

  2. Agrin as a Mechanotransduction Signal Regulating YAP through the Hippo Pathway.

    PubMed

    Chakraborty, Sayan; Njah, Kizito; Pobbati, Ajaybabu V; Lim, Ying Bena; Raju, Anandhkumar; Lakshmanan, Manikandan; Tergaonkar, Vinay; Lim, Chwee Teck; Hong, Wanjin

    2017-03-07

    The Hippo pathway effectors YAP and TAZ act as nuclear sensors of mechanical signals in response to extracellular matrix (ECM) cues. However, the identity and nature of regulators in the ECM and the precise pathways relaying mechanoresponsive signals into intracellular sensors remain unclear. Here, we uncover a functional link between the ECM proteoglycan Agrin and the transcriptional co-activator YAP. Importantly, Agrin transduces matrix and cellular rigidity signals that enhance stability and mechanoactivity of YAP through the integrin-focal adhesion- and Lrp4/MuSK receptor-mediated signaling pathways. Agrin antagonizes focal adhesion assembly of the core Hippo components by facilitating ILK-PAK1 signaling and negating the functions of Merlin and LATS1/2. We further show that Agrin promotes oncogenesis through YAP-dependent transcription and is clinically relevant in human liver cancer. We propose that Agrin acts as a mechanotransduction signal in the ECM.

  3. Environmental Impacts of Airport Policy Alternatives,

    DTIC Science & Technology

    1977-09-01

    NAl YSIS F U: C A S-E --- Am S A/(; AKR,/’P PA .’IC- CA kt’j IT 440*’t~ VuPi ) USO AIRPt.T TYPr CYCL’- LILATEI M10NOY. CAfccFJINS N ITRnc0E4 TA- I...AMES A/C ARR/09P PAR TIC - CARPrN HY0Rr1- nXIrs OFn_ _ A IRT TYPE CYCLES ULATES MONOX. CA;,)NS Tic.rN 18 13.99 A.68 232. 4 50.37 27-.14 20...52.09 26569.62 48196.86 73 TABLE 2.30 Emission Analysis for Case 29 (Pounds/Day) Ak’E% A/C AKR/r"P PAk,, TIC - CAP f)N HYDR- VX10cS OF A O , " To C

  4. Fray, a Drosophila serine/threonine kinase homologous to mammalian PASK, is required for axonal ensheathment

    NASA Technical Reports Server (NTRS)

    Leiserson, W. M.; Harkins, E. W.; Keshishian, H.

    2000-01-01

    Fray is a serine/threonine kinase expressed by the peripheral glia of Drosophila, whose function is required for normal axonal ensheathment. Null fray mutants die early in larval development and have nerves with severe swelling and axonal defasciculation. The phenotype is associated with a failure of the ensheathing glia to correctly wrap peripheral axons. When the fray cDNA is expressed in the ensheathing glia of fray mutants, normal nerve morphology is restored. Fray belongs to a novel family of Ser/Thr kinases, the PF kinases, whose closest relatives are the PAK kinases. Rescue of the Drosophila mutant phenotype with PASK, the rat homolog of Fray, demonstrates a functional homology among these proteins and suggests that the Fray signaling pathway is widely conserved.

  5. Application of TL dosemeters for dose distribution measurements at high temperatures in nuclear reactors.

    PubMed

    Osvay, M; Deme, S

    2006-01-01

    Al2O3:Mg,Y ceramic thermoluminescence dosemeters were developed at the Institute of Isotopes for high dose applications at room temperatures. The glow curve of Al2O3:Mg,Y exhibits two peaks--one at 250 degrees C (I) and another peak at approximately 400 degrees C (II). In order to extend the application of these dosemeters to high temperatures, the effect of irradiation temperature was investigated using temperature controlled heating system during high dose irradiation at various temperatures (20-100 degrees C). The new calibration and measuring method has been successfully applied for dose mapping within the hermetic zone of the Paks Nuclear Power Plant even at high temperature parts of blocks.

  6. [Determination method of ultra-high-intensity sweetener, advantame, in processed foods by HPLC and LC-MS/MS].

    PubMed

    Kobayashi, Miki; Terada, Hisaya; Nakajima, Masahiro

    2015-01-01

    A simple method using HPLC and LC-MS/MS was developed for the determination of ultra-high-intensity sweetener, advantame, in processed foods. Advantame was extracted by dialysis, and cleaned up on a Sep-Pak Plus C18 cartridge, then determined by HPLC and LC-MS/MS. The recoveries from 5 kinds of processed foods fortified at the levels of 0.001 g/kg and 0.01 g/kg were 64.1-89.9% (RSD 0.9-6.9%) by HPLC and 68.8-99.9% (RSD 0.8-4.9%) by LC-MS/MS. The quantitation limit was 0.0004 g/kg by HPLC and 0.00004 g/kg by LC-MS/MS.

  7. Analysis of fumonisin B1 in rodent feed by CE with fluorescence detection of the FMOC derivative.

    PubMed

    Holcomb, M; Thompson, H C

    1996-01-01

    A fast, simple, and environmentally friendly method has been developed for the analysis of fumonisin B1 (FB1) in rodent feed using CE. FB1 is the major fumonisin metabolite produced by the fungus Fusarium moniliforme and has been implicated in human and animal diseases. FB1 was extracted from rodent feed with acetonitrile/water (50/50) (vol/vol) and cleaned up with a C18 Sep-Pak Vac cartridge (Waters Corp., Milford, MA, U.S.A.). FB1 was quantitated after elution from the column using capillary electrophoresis with a 25-mM sodium borate buffer (pH 9.0) containing 10% acetonitrile and fluorescence detection of the (9-fluorenylmethyl) chloroformate (FMOC) derivative. The minimum detectable amount in rodent feed was 0.5 pmm. Recovery values in spiked rodent feed averaged more than 87% over the 2-20 ppm range.

  8. DOCK8 regulates lymphocyte shape integrity for skin antiviral immunity.

    PubMed

    Zhang, Qian; Dove, Christopher G; Hor, Jyh Liang; Murdock, Heardley M; Strauss-Albee, Dara M; Garcia, Jordan A; Mandl, Judith N; Grodick, Rachael A; Jing, Huie; Chandler-Brown, Devon B; Lenardo, Timothy E; Crawford, Greg; Matthews, Helen F; Freeman, Alexandra F; Cornall, Richard J; Germain, Ronald N; Mueller, Scott N; Su, Helen C

    2014-12-15

    DOCK8 mutations result in an inherited combined immunodeficiency characterized by increased susceptibility to skin and other infections. We show that when DOCK8-deficient T and NK cells migrate through confined spaces, they develop cell shape and nuclear deformation abnormalities that do not impair chemotaxis but contribute to a distinct form of catastrophic cell death we term cytothripsis. Such defects arise during lymphocyte migration in collagen-dense tissues when DOCK8, through CDC42 and p21-activated kinase (PAK), is unavailable to coordinate cytoskeletal structures. Cytothripsis of DOCK8-deficient cells prevents the generation of long-lived skin-resident memory CD8 T cells, which in turn impairs control of herpesvirus skin infections. Our results establish that DOCK8-regulated shape integrity of lymphocytes prevents cytothripsis and promotes antiviral immunity in the skin.

  9. GIS, GPS, political history, and geo-demography of the Aramamisi Yanomamo expansion

    NASA Technical Reports Server (NTRS)

    Chagnon, Napoleon A.

    1991-01-01

    This year a joint US-Venezuelan research project was initiated to locate, identify, and begin long term research on the ecology of the Siapa Basin and the cultural adaptations of Yanomamo communities to this habit, a poorly mapped region of southern Venezuela. During two separate field trips, representing over 500 hours of flying time, local informants were asked to guide a helicopter to villages and other areas of anthropological interest. The location of each place was established with a GPS instrument. Both the Trimble Navigation TransPak and Magellan Nav 1000 Pro were used on these trips. A brief summary of the results and preliminary comparisons of both instruments will be presented.

  10. Separation and purification of astaxanthin from Phaffia rhodozyma by preparative high-speed counter-current chromatography.

    PubMed

    Du, Xiping; Dong, Congcong; Wang, Kai; Jiang, Zedong; Chen, Yanhong; Yang, Yuanfan; Chen, Feng; Ni, Hui

    2016-09-01

    An effective high-speed counter-current chromatography (HSCCC) method was established for the preparative isolation and purification of astaxanthin from Phaffia rhodozyma. With a two-phase solvent system composed of n-hexane-acetone-ethanol-water (1:1:1:1, v/v/v/v), 100mg crude extract of P. rhodozyma was separated to yield 20.6mg of astaxanthin at 92.0% purity. By further one step silica gel column chromatography, the purity reached 99.0%. The chemical structure of astaxanthin was confirmed by thin layer chromatography (TLC), UV spectroscopy scanning, high performance liquid chromatography with a ZORBAX SB-C18 column and a Waters Nova-pak C18 column, and ESI/MS/MS.

  11. Golgi membrane fission requires the CtBP1-S/BARS-induced activation of lysophosphatidic acid acyltransferase δ

    PubMed Central

    Pagliuso, Alessandro; Valente, Carmen; Giordano, Lucia Laura; Filograna, Angela; Li, Guiling; Circolo, Diego; Turacchio, Gabriele; Marzullo, Vincenzo Manuel; Mandrich, Luigi; Zhukovsky, Mikhail A.; Formiggini, Fabio; Polishchuk, Roman S.; Corda, Daniela; Luini, Alberto

    2016-01-01

    Membrane fission is an essential cellular process by which continuous membranes split into separate parts. We have previously identified CtBP1-S/BARS (BARS) as a key component of a protein complex that is required for fission of several endomembranes, including basolateral post-Golgi transport carriers. Assembly of this complex occurs at the Golgi apparatus, where BARS binds to the phosphoinositide kinase PI4KIIIβ through a 14-3-3γ dimer, as well as to ARF and the PKD and PAK kinases. We now report that, when incorporated into this complex, BARS binds to and activates a trans-Golgi lysophosphatidic acid (LPA) acyltransferase type δ (LPAATδ) that converts LPA into phosphatidic acid (PA); and that this reaction is essential for fission of the carriers. LPA and PA have unique biophysical properties, and their interconversion might facilitate the fission process either directly or indirectly (via recruitment of proteins that bind to PA, including BARS itself). PMID:27401954

  12. DOCK8 regulates lymphocyte shape integrity for skin antiviral immunity

    PubMed Central

    Zhang, Qian; Dove, Christopher G.; Hor, Jyh Liang; Murdock, Heardley M.; Strauss-Albee, Dara M.; Garcia, Jordan A.; Mandl, Judith N.; Grodick, Rachael A.; Jing, Huie; Chandler-Brown, Devon B.; Lenardo, Timothy E.; Crawford, Greg; Matthews, Helen F.; Freeman, Alexandra F.; Cornall, Richard J.; Germain, Ronald N.

    2014-01-01

    DOCK8 mutations result in an inherited combined immunodeficiency characterized by increased susceptibility to skin and other infections. We show that when DOCK8-deficient T and NK cells migrate through confined spaces, they develop cell shape and nuclear deformation abnormalities that do not impair chemotaxis but contribute to a distinct form of catastrophic cell death we term cytothripsis. Such defects arise during lymphocyte migration in collagen-dense tissues when DOCK8, through CDC42 and p21-activated kinase (PAK), is unavailable to coordinate cytoskeletal structures. Cytothripsis of DOCK8-deficient cells prevents the generation of long-lived skin-resident memory CD8 T cells, which in turn impairs control of herpesvirus skin infections. Our results establish that DOCK8-regulated shape integrity of lymphocytes prevents cytothripsis and promotes antiviral immunity in the skin. PMID:25422492

  13. [Assessment of the risk factors for urolithiasis in cosmonauts during long flights

    NASA Technical Reports Server (NTRS)

    Arzamazov, G. S.; Witson, P. A.; Larina, O. N.; Pastushkova, L. Kh; Pak, C. T.; Whitson, P. A. (Principal Investigator)

    1996-01-01

    Twelve cosmonauts flown aboard the Mir orbital station for 6-12 months and twelve candidates for cosmonauts were examined in an effort to determine a degree of urolithiasis risk. Prior to flight, on flight day 310 (in one cosmonaut) and after flight, the daily urinary excretion of the components influencing lithiasis formation was determined as well as a computer-aided calculation of urine saturation by lithogenic salts was performed (Ch. Pak, USA). In the in- and postflight periods, the greater number of indices under study were negatively changed in cosmonauts. Excretion dynamics of the lithogenesis inhibitors, i.e., citrates and magnesium, is of polar directionality. Frequency of deviations from the normal indices of urolithic risk in cosmonauts is primarily conditioned by low diuresis, urine supersaturation with calcium oxalate, undissociated uric acid, brushit, hypercalciuria, and changed pH.

  14. Seasonal variations of imposex indices and butyltin concentrations in the rock shell Thais clavigera collected from Hong Kong waters.

    PubMed

    Qiu, Jian-Wen; Chan, Ka Ming; Leung, Kenneth Mei Yee

    2011-01-01

    This study investigated butyltin contamination in Hong Kong waters using the rock shell Thais clavigera as a biomonitor. We collected rock shells from nine coastal sites in both winter 2005 and summer 2006 to examine the spatial and seasonal patterns. The relative penis size index (RPSI) and vas deferens sequence index (VDSI) were determined following a standard protocol, while the tissue concentrations of butyltins (i.e., mono-, di- and tri-butyltin) were measured using gas chromatography. The results showed that imposex was particularly severe in T. clavigera collected from Victoria Harbour, Pak Sha Wan and Sai Kung Pier where shipping activities were frequent. Both imposex indices had a positive correlation with tissue concentration of tributyltin in T. clavigera. While VDSI exhibited little seasonal variation, RPSI of winter specimens was significantly higher than that of summer specimens. Thus, sampling season should be standardised when using RPSI as a biomarker.

  15. Expression of feline recombinant interferon-gamma in baculovirus and demonstration of biological activity.

    PubMed

    Argyle, D J; Harris, M; Lawrence, C; McBride, K; Barron, R; McGillivray, C; Onions, D E

    1998-07-08

    We have previously reported the cloning of the coding sequence for feline-specific interferon-gamma. Here, we describe the expression of this sequence in a baculovirus system and demonstrate the biological activity of the recombinant protein. The coding sequence for feline interferon was directionally cloned into the baculovirus transfer vector pAcCL29-1. Transfer vector and linearized wild-type AcMNPV (BacPAK6) were used to co-transfect Sf9 cells by calcium phosphate coprecipitation. Subsequently, wild-type and recombinant viruses were separated by plaque assay. Recombinant plaques were expanded and a master stock of virus is produced. Production of biologically active interferon-gamma from infected Sf9 cells was demonstrated using a standard cytopathic effect reduction assay, utilising vesicular stomatitis virus (VSV), and an MHC class II induction assay.

  16. Analysis of organochlorine pesticides in human milk: preliminary results.

    PubMed

    Campoy, C; Jiménez, M; Olea-Serrano, M F; Moreno-Frías, M; Cañabate, F; Olea, N; Bayés, R; Molina-Font, J A

    2001-11-01

    In the face of evidence of human milk contamination by organochlorine pesticides, an analysis was performed on samples of milk obtained from healthy lactating women in the provinces of Granada and Almeria in Southern Spain. The samples were obtained by the Neonate Section of the Department of Pediatrics of Granada University Hospital (Neonatology Division) and by the Neonatal Service of Poniente Hospital in El Ejido, Almería. A liquid-liquid extraction procedure was performed. The cleaning of the sample before gas chromatography-mass spectrometry (GC-MS) used silica Sep-Pak. Among other pesticides, aldrin, dieldrin, DDT and its metabolites, lindane, methoxychlor and endosulfan were identified. The presence of these products was confirmed by mass spectrometry. The identification and quantification of these organochlorine molecules is important because they have estrogenic effects.

  17. [Mobile intensive-care-unit for transportation of premature and newborn babies at risc (author's transl)].

    PubMed

    Wille, L; Obladen, M; Schlunk, P; Weisser, J

    1975-02-01

    This is a technical report on a specially equipped ambulance for transportation of high-risk, seriously ill neonates. A mobile neonatal intensive-care-unit operating independently of the car utilized an Ohio-transport-incubator with 12V-DC portable power pak and collapsible stand, battery-operated ECG-monitor with optical and acoustical signal, a ECG-monitor with optical and acoustical signal, a battery-operated infusion pump, a Bird-respirator mark 8 with oxygen-blender, nebulizer and infant circuit with modification for PEEP as well as additional accessories. Ambulance-duty service is guaranteed by the German Red Cross (DRK) to facilitate transfer at any time, while skilled personal (physician, nurse) of the intensive care ward in on 24 hs call.

  18. A survey of compiler development aids. [concerning lexical, syntax, and semantic analysis

    NASA Technical Reports Server (NTRS)

    Buckles, B. P.; Hodges, B. C.; Hsia, P.

    1977-01-01

    A theoretical background was established for the compilation process by dividing it into five phases and explaining the concepts and algorithms that underpin each. The five selected phases were lexical analysis, syntax analysis, semantic analysis, optimization, and code generation. Graph theoretical optimization techniques were presented, and approaches to code generation were described for both one-pass and multipass compilation environments. Following the initial tutorial sections, more than 20 tools that were developed to aid in the process of writing compilers were surveyed. Eight of the more recent compiler development aids were selected for special attention - SIMCMP/STAGE2, LANG-PAK, COGENT, XPL, AED, CWIC, LIS, and JOCIT. The impact of compiler development aids were assessed some of their shortcomings and some of the areas of research currently in progress were inspected.

  19. An alternative method for monitoring carbonyls, and the development of a 24-port fully automated carbonyl sampler for PAMS program

    SciTech Connect

    Parmar, S.S.; Ugarova, L.; Fernandes, C.; Guyton, J.; Lee, C.P.

    1994-12-31

    The authors have investigated the possibility of collecting different aldehydes and ketones on different sorbents such as silica gel, molecular sieve and charcoal followed by solvent extraction, DNPH derivatization and HPLC/UV analysis. Carbonyl collection efficiencies for these sorbents were calculated relative to a DNPH coated C{sub 18} sep-pak cartridge. From a limited number of laboratory experiments, at various concentrations, it appears that silica gel tubes can be used for sampling aldehydes (collection efficiencies {approximately} 1), whereas charcoal tubes are suitable for collecting ketones. Molecular sieve was found to be unsuitable for collecting most of the carbonyl studied. The authors also report the development of a fully automated 24-port carbonyl sampler specially designed for EPA`s PAMS program.

  20. Artificial Intelligence In Computational Fluid Dynamics

    NASA Technical Reports Server (NTRS)

    Vogel, Alison Andrews

    1991-01-01

    Paper compares four first-generation artificial-intelligence (Al) software systems for computational fluid dynamics. Includes: Expert Cooling Fan Design System (EXFAN), PAN AIR Knowledge System (PAKS), grid-adaptation program MITOSIS, and Expert Zonal Grid Generation (EZGrid). Focuses on knowledge-based ("expert") software systems. Analyzes intended tasks, kinds of knowledge possessed, magnitude of effort required to codify knowledge, how quickly constructed, performances, and return on investment. On basis of comparison, concludes Al most successful when applied to well-formulated problems solved by classifying or selecting preenumerated solutions. In contrast, application of Al to poorly understood or poorly formulated problems generally results in long development time and large investment of effort, with no guarantee of success.

  1. Recent progress in the use of ‘omics technologies in brassicaceous vegetables

    PubMed Central

    Witzel, Katja; Neugart, Susanne; Ruppel, Silke; Schreiner, Monika; Wiesner, Melanie; Baldermann, Susanne

    2015-01-01

    Continuing advances in ‘omics methodologies and instrumentation is enhancing the understanding of how plants cope with the dynamic nature of their growing environment. ‘Omics platforms have been only recently extended to cover horticultural crop species. Many of the most widely cultivated vegetable crops belong to the genus Brassica: these include plants grown for their root (turnip, rutabaga/swede), their swollen stem base (kohlrabi), their leaves (cabbage, kale, pak choi) and their inflorescence (cauliflower, broccoli). Characterization at the genome, transcript, protein and metabolite levels has illustrated the complexity of the cellular response to a whole series of environmental stresses, including nutrient deficiency, pathogen attack, heavy metal toxicity, cold acclimation, and excessive and sub-optimal irradiation. This review covers recent applications of ‘omics technologies to the brassicaceous vegetables, and discusses future scenarios in achieving improvements in crop end-use quality. PMID:25926843

  2. Application of modified supercritical carbon dioxide extraction to microbial quinone analysis.

    PubMed

    Irvan; Hasanudin, Udin; Faisal, Muhammad; Daimon, Hiroyuki; Fujie, Koichi

    2006-01-01

    Supercritical carbon dioxide (scCO2) was applied to extract microbial quinones from activated sludge. Identification and analysis was then performed using high-performance liquid chromatography (HPLC) equipped with ultraviolet-visible (UV-Vis) detector and photodiode array detector (PDA). Extracted microbial quinones were trapped and separated as menaquinones (MK) and ubiquinones (Q) species using two Sep-Pak Plus Silica cartridges joined in series. Four ubiquinones and 12 menaquinones species were identified in 0.1 g dried activated sludge based on retention time and spectrum analysis. Among the tested various polar solvents, methanol showed to be the best modifier, based on the highest total quinone content extracted and the lowest dissimilarity index. The diversity index of quinone and the number of quinone species using methanol-modified scCO2 were similar to that of the conventional method (organic solvent extraction).

  3. Isolation and Characterization of Glycophorin from Carp Red Blood Cell Membranes

    PubMed Central

    Aoki, Takahiko; Chimura, Kenji; Nakao, Nobuhiro; Mizuno, Yasuko

    2014-01-01

    We isolated a high-purity carp glycophorin from carp erythrocyte membranes following extraction using the lithium diiodosalicylate (LIS)-phenol method and streptomycin treatment. The main carp glycophorin was observed to locate at the position of the carp and human band-3 proteins on an SDS-polyacrylamide gel. Only the N-glycolylneuraminic acid (NeuGc) form of sialic acid was detected in the carp glycophorin. The oligosaccharide fraction was separated into two components (P-1 and P-2) using a Glyco-Pak DEAE column. We observed bacteriostatic activity against five strains of bacteria, including two known fish pathogens. Fractions from the carp erythrocyte membrane, the glycophorin oligosaccharide and the P-1 also exhibited bacteriostatic activity; whereas the glycolipid fraction and the glycophorin fraction without sialic acid did not show the activity. The carp glycophorin molecules attach to the flagellum of V. anguillarum or the cell surface of M. luteus and inhibited bacterial growth. PMID:25110961

  4. A novel 47.2 Mb duplication on chromosomal bands Xq21.1-25 associated with mental retardation.

    PubMed

    Jin, Zhijuan; Yu, Li; Geng, Juan; Wang, Jian; Jin, Xingming; Huang, Hong

    2015-08-01

    We present array comparative genomic hybridization (aCGH) characterization of a novel Xq21.1-25 duplication in a 2-year-old girl with facial dysmorphism, mental retardation and short stature. Analysis of aCGH results revealed a 47,232kb duplication region that harbored 231 RefSeq genes, including 32 OMIM genes. Ten genes (i.e., ZNF711, SRPX2, RAB40AL, MID2, ACSL4, PAK3, UBE2A, UPF3B, CUL4B, and GRIA3) in the duplication interval have been associated with mental retardation. We discuss the genotype-phenotype correlation in this case. Our case provides evidence for an association of mental retardation with X chromosome duplication.

  5. A Stability Indicating HPLC Method for the Determination of Nitisinone in Capsules

    PubMed Central

    Souri, Effat; Lahiji, Farnaz R.; Nourhashemi, Tannaz; Jalalizadeh, H.

    2015-01-01

    In this study a simple and efficient stability-indicating HPLC method with short run time was developed for the determination of nitisinone. The stress degradation of nitisinone was studied in different acidic, basic, oxidative, thermal and photolytic conditions. The chromatographic separation was achieved on a Nova-Pak C18 column using a mixture of 50 mM NaH2PO4 (pH 2.5) and acetonitrile (45:55, v/v) as mobile phase. UV detection was performed at 280 nm. Good linearity was observed over the concentration range of 0.5-50 μg/ml with r2>0.999. The within-day and between-day precision values were less than 2%. The proposed method could be used for the determination of nitisinone in the presence of its degradation products and also dosage form excipients for the quality control purposes. PMID:26180282

  6. Molecular pathways: targeting the kinase effectors of RHO-family GTPases.

    PubMed

    Prudnikova, Tatiana Y; Rawat, Sonali J; Chernoff, Jonathan

    2015-01-01

    RHO GTPases, members of the RAS superfamily of small GTPases, are adhesion and growth factor-activated molecular switches that play important roles in tumor development and progression. When activated, RHO-family GTPases such as RAC1, CDC42, and RHOA, transmit signals by recruiting a variety of effector proteins, including the protein kinases PAK, ACK, MLK, MRCK, and ROCK. Genetically induced loss of RHO function impedes transformation by a number of oncogenic stimuli, leading to an interest in developing small-molecule inhibitors that either target RHO GTPases directly, or that target their downstream protein kinase effectors. Although inhibitors of RHO GTPases and their downstream signaling kinases have not yet been widely adopted for clinical use, their potential value as cancer therapeutics continues to facilitate pharmaceutical research and development and is a promising therapeutic strategy.

  7. Senescence-inducing stress promotes proteolysis of phosphoglycerate mutase via ubiquitin ligase Mdm2

    PubMed Central

    Mikawa, Takumi; Maruyama, Takeshi; Okamoto, Koji; Nakagama, Hitoshi; Lleonart, Matilde E.; Tsusaka, Takeshi; Hori, Kousuke; Murakami, Itsuo; Izumi, Taisuke; Takaori-Kondo, Akifumi; Yokode, Masayuki; Peters, Gordon; Beach, David

    2014-01-01

    Despite the well-documented clinical significance of the Warburg effect, it remains unclear how the aggressive glycolytic rates of tumor cells might contribute to other hallmarks of cancer, such as bypass of senescence. Here, we report that, during oncogene- or DNA damage–induced senescence, Pak1-mediated phosphorylation of phosphoglycerate mutase (PGAM) predisposes the glycolytic enzyme to ubiquitin-mediated degradation. We identify Mdm2 as a direct binding partner and ubiquitin ligase for PGAM in cultured cells and in vitro. Mutations in PGAM and Mdm2 that abrogate ubiquitination of PGAM restored the proliferative potential of primary cells under stress conditions and promoted neoplastic transformation. We propose that Mdm2, a downstream effector of p53, attenuates the Warburg effect via ubiquitination and degradation of PGAM. PMID:24567357

  8. Senescence-inducing stress promotes proteolysis of phosphoglycerate mutase via ubiquitin ligase Mdm2.

    PubMed

    Mikawa, Takumi; Maruyama, Takeshi; Okamoto, Koji; Nakagama, Hitoshi; Lleonart, Matilde E; Tsusaka, Takeshi; Hori, Kousuke; Murakami, Itsuo; Izumi, Taisuke; Takaori-Kondo, Akifumi; Yokode, Masayuki; Peters, Gordon; Beach, David; Kondoh, Hiroshi

    2014-03-03

    Despite the well-documented clinical significance of the Warburg effect, it remains unclear how the aggressive glycolytic rates of tumor cells might contribute to other hallmarks of cancer, such as bypass of senescence. Here, we report that, during oncogene- or DNA damage-induced senescence, Pak1-mediated phosphorylation of phosphoglycerate mutase (PGAM) predisposes the glycolytic enzyme to ubiquitin-mediated degradation. We identify Mdm2 as a direct binding partner and ubiquitin ligase for PGAM in cultured cells and in vitro. Mutations in PGAM and Mdm2 that abrogate ubiquitination of PGAM restored the proliferative potential of primary cells under stress conditions and promoted neoplastic transformation. We propose that Mdm2, a downstream effector of p53, attenuates the Warburg effect via ubiquitination and degradation of PGAM.

  9. Inhibitory effects of spices and herbs on iron availability.

    PubMed

    Tuntipopipat, Siriporn; Zeder, Christophe; Siriprapa, Pudsadee; Charoenkiatkul, Somsri

    2009-01-01

    Spices and herbs are extensively used in indigenous diets in tropical regions where prevalence of iron deficiency is still high. They are rich in polyphenolic compounds that are expected to inhibit iron absorption by forming iron complexes in the intestine, making dietary iron less available for absorption. The effects of six spices and herbs (chili pepper, garlic, 'Pak kyheng' (Thai leafy vegetable), shallot, tamarind, turmeric) and one mixture of spices (curry paste) on iron availability were determined by measuring the percentage dialyzable iron after addition of spices and herbs to a rice meal after simulated digestion. All tested spices and herbs contained from 0.5 to 33 mg polyphenol per meal and were potent inhibitors of iron availability (20-90%), reducing iron availability in a dose-dependent manner--with the exception of tamarind, which at 11 mg polyphenol per meal enhanced iron availability. Our findings demonstrate that culinary spices and herbs can play an important role in iron nutrition.

  10. CD66-mediated phagocytosis of Opa52 Neisseria gonorrhoeae requires a Src-like tyrosine kinase- and Rac1-dependent signalling pathway.

    PubMed

    Hauck, C R; Meyer, T F; Lang, F; Gulbins, E

    1998-01-15

    The interaction of Neisseria gonorrhoeae with human phagocytes is a hallmark of gonococcal infections. Recently, CD66 molecules have been characterized as receptors for Opa52-expressing gonococci on human neutrophils. Here we show that Opa52-expressing gonococci or Escherichia coli or F(ab) fragments directed against CD66, respectively, activate a signalling cascade from CD66 via Src-like protein tyrosine kinases, Rac1 and PAK to Jun-N-terminal kinase. The induced signal is distinct from Fcgamma-receptor-mediated signalling and is specific for Opa52, since piliated Opa- gonococci, commensal Neisseria cinerea or E.coli do not stimulate this signalling pathway. Inhibition of Src-like kinases or Rac1 prevents the uptake of Opa52 bacteria, demonstrating the crucial role of this signalling cascade for the opsonin-independent, Opa52/CD66-mediated phagocytosis of pathogenic Neisseria.

  11. The sediment and hydrographic characteristics of three horseshoe crab nursery beaches in hong kong

    NASA Astrophysics Data System (ADS)

    Chiu, Helen M. C.; Morton, Brian

    2003-04-01

    Horseshoe crab juveniles have been recorded from sand and sandy-mud nursery beaches at Pak Nai (western New Territories), San Tau and Shui Hau (Lantau Island), Hong Kong. In order to provide a better understanding of these beaches and to identify those plausible factors which have made them preferred by spawning horseshoe crabs, environmental parameters, including temperature, salinity, pH and dissolved oxygen content of the water, and particle size distribution and organic matter content of the sediments at the three sites, were determined and compared. The hydrographic, and sediment data obtained for the three study sites have revealed some common environmental features. The three nursery beaches are relatively remote, and far (in Hong Kong terms) from urbanized and densely populated areas. The beaches are generally well sheltered from strong wave action and inundated regularly by estuarine waters. Horseshoe crab adults tend to select these beaches for spawning as their protected features ensures the laid eggs are less likely to be washed out of the sand, and hatched juveniles can feed on the meiofauna and grow. Sediments of the three beaches largely comprise medium-sized sand particles and are moderately sorted, suggesting medium porosity and good water permeability. Such a sand type, with the generally high oxygen levels in incursing waters, may help create a well-oxygenated micro-environment for the normal development of horseshoe crab eggs, larvae and juveniles. Lantau Island beaches at San Tau and Shui Hau are relatively free from organic pollution, as reflected in generally high dissolved oxygen level, and low BOD5 and ammonia nitrogen values. Pak Nai is, however, more polluted.

  12. Bactericidal Performance of Flame-Sprayed Nanostructured Titania-Copper Composite Coatings

    NASA Astrophysics Data System (ADS)

    George, N.; Mahon, M.; McDonald, A.

    2010-09-01

    A large concern surrounding stainless steel surfaces is the ability of bacteria to grow and attach to them quite easily. One possible solution to destroy these pathogens is to coat surfaces with a biocidal agent. The photocatalytic effect of titanium dioxide (TiO2) is known to have a bactericidal effect. Coatings of TiO2 were prepared on 1010 low carbon steel substrates using an oxy-acetylene flame spray torch. TiO2 coatings containing 5 wt.% copper (Cu) were fabricated to increase the bactericidal effect of the coating. After deposition, the coatings were polished to an average roughness of 1 μm. Solutions of Pseudomonas aeruginosa (PAK) bacteria were placed onto the coating surface for periods of up to 3 h, and the amount of surviving bacteria were counted. Some samples were irradiated with white light and other samples were held in a dark chamber. In coatings of copper-free flame-sprayed TiO2, the high flame temperatures facilitated the conversion of the anatase phase to the rutile phase, which limited the photocatalytic destruction of the bacterial cells. However, TiO2-copper composite coatings showed a large bactericidal effect, killing approximately 75% of PAK bacterial cells after 3 h. Under the same conditions, the TiO2-copper composite coatings had the same bactericidal capabilities as pure copper surfaces, with the composite coatings showing improved bactericidal performance when exposed to light. It was proposed that increased concentrations of reactive oxide species produced due to TiO2 photocatalysis improved the performance of the irradiated TiO2-copper composite coatings.

  13. Dead on arrival in a low-income country: results from a multicenter study in Pakistan

    PubMed Central

    2015-01-01

    Background This study assessed the characteristics of dead on arrival (DOA) patients in Pakistan. Methods Data about the DOA patients were extracted from Pakistan National Emergency Department Surveillance study (Pak-NEDS). This study recruited all ED patients presenting to seven tertiary care hospitals during a four-month period between November 2010 and March 2011. This study included patients who were declared dead-on-arrival by the ED physician. Results A total of 1,557 DOA patients (7 per 1,000 visits) were included in the Pak-NEDS. Men accounted for two-thirds (64%) of DOA patients. Those aged 20-49 years accounted for about 46% of DOA patients. Nine percent (n = 72) of patients were brought by ambulance, and most patients presented at a public hospital (80%). About 11% of DOA patients had an injury. Factors significantly associated (p < 0.05) with ambulance use were men (adjusted odds ratio [aOR] = 2.72), brought to a private hospital (OR = 2.74), and being injured (aOR = 1.89). Cardiopulmonary resuscitation (CPR) was performed on 6% (n = 42) of patients who received treatment. Those brought to a private hospital were more likely to receive CPR (aOR = 2.81). Conclusion This study noted a higher burden of DOA patients in Pakistan compared to other resourceful settings (about 1 to 2 per 1,000 visits). A large proportion of patients belonging to productive age groups, and the low prevalence of ambulance and CPR use, indicate a need for improving the prehospital care and basic life support training in Pakistan. PMID:26689125

  14. [Deletion of IV a2 gene from adenoviral genome by lambda-Red recombinase system and packaging of the recombinant adenovirus].

    PubMed

    Liu, Yun-Fan; Yu, Chi-Jie; Wang, Gang; Tian, Wen-Hong; Lu, Yue; Liu, Xue-Rong; Dong, Xiao-Yan; Zheng, Gang; Shen, Wei; Wu, Xiao-Bing; Ruan, Li

    2011-05-01

    This investigation is to delete the most of the coding sequence (1104 bp) of the IV a2 gene in an adenovirus genome by a lambda-Red recombinase system-mediated PCR-targeting approach and rescue a recombinant adenovirus with IV a2 deletion. First, the template pAK of PCR targeting, containing kanamycin cassette, was constructed. Then, a linear fragment for PCR targeting, which had 39 bp homologous arms at both of its terminus, was amplified by PCR from the pAK. The pFG140 and the linear fragment were electroporated into E. coli BW25113/pIJ790 sequentially and the recombinant pFG140-deltaIV a2 (1104) was established by homologous recombination between the linear fragment and the pFG140 with aid of X-Red recombinase. The precise deletion of 1 104 bp fragment from IV a2 was confirmed by restriction endonucleases digestion and DNA sequencing. ORF of IV a2 was amplified by PCR from pFG140 and then cloned into the pAAV2neo vector. The recombinant adenovirus Ad5delta IV a2 (1104) was rescued by co-transfection of pFG140-deltaIV a2 (1104) and pAAV2neo-IV a2 into HEK293 cells. It was shown by Western Blot that IV a2 could not be detected in the Ad5deltaIV a2 (1104)- infected HEK293 cells. This study established a PCR-targeting strategy for manipulating adenovirus genome directly by a lambda-Red recombinase system, and a recombinant adenovirus with IV a2 deletion was obtained.

  15. The pediatric disease spectrum in emergency departments across Pakistan: data from a pilot surveillance system

    PubMed Central

    2015-01-01

    Background There is an increasing number of urgently ill and injured children being seen in emergency departments (ED) of developing countries. The pediatric disease burden in EDs across Pakistan is generally unknown. Our main objective was to determine the spectrum of disease and injury among children seen in EDs in Pakistan through a nationwide ED-based surveillance system. Methods Through the Pakistan National Emergency Department Surveillance (Pak-NEDS), data were collected from November 2010 to March 2011 in seven major tertiary care centers representing all provinces of Pakistan. These included five public and two private hospitals, with a collective annual census of over one million ED encounters. Results Of 25,052 children registered in Pak-NEDS (10% of all patients seen): 61% were male, 13% under 5 years, while almost 65% were between 10 to < 16 years. The majority (90%) were seen in public hospital EDs. About half the patients were discharged from the EDs, 9% admitted to hospitals and only 1.3% died in the EDs. Injury (39%) was the most common presenting complaint, followed by fever/malaise (19%) and gastrointestinal symptoms (18%). Injury was more likely in males vs. females (43% vs. 33%; p < 0.001), with a peak presentation in the 5-12 year age group (45%). Conclusions Pediatric patients constitute a smaller proportion among general ED users in Pakistan. Injury is the most common presenting complaint for children seen in the ED. These data will help in resource allocation for cost effective pediatric ED service delivery systems. Prospective longer duration surveillance is needed in more representative pediatric EDs across Pakistan. PMID:26691052

  16. Stress-induced response, localization, and regulation of the Pmk1 cell integrity pathway in Schizosaccharomyces pombe.

    PubMed

    Madrid, Marisa; Soto, Teresa; Khong, Hou Keat; Franco, Alejandro; Vicente, Jero; Pérez, Pilar; Gacto, Mariano; Cansado, José

    2006-01-27

    Mitogen-activated protein kinase (MAPK) signaling pathways are critical for the sensing and response of eukaryotic cells to extracellular changes. In Schizosaccharomyces pombe, MAPK Pmk1/Spm1 has been involved in cell wall construction, morphogenesis, cytokinesis, and ion homeostasis, as part of the so-called cell integrity pathway together with MAPK kinase kinase Mkh1 and MAPK kinase Pek1. We show that Pmk1 is activated in multiple stress situations, including hyper- or hypotonic stress, glucose deprivation, presence of cell wall-damaging compounds, and oxidative stress induced by hydrogen peroxide or pro-oxidants. The stress-induced activation of Pmk1 was completely dependent on Mkh1 and Pek1 function, supporting a nonbranched pathway in the regulation of MAPK activation. Fluorescence microscopy revealed that Mkh1, Pek1, and Pmp1 (a protein phosphatase that inactivates Pmk1) are cytoplasmic proteins. Mkh1 and Pek1 were also found at the septum, whereas Pmk1 localized in both cytoplasm and nucleus as well as in the mitotic spindle and septum during cytokinesis. Interestingly, Pmk1 subcellular localization was unaffected by stress or the absence of Mkh1 and Pek1, suggesting that its activation by the Mkh1-Pek1 cascade takes place at the cytoplasm and/or septum and that the active and inactive forms of this kinase cross the nuclear membrane. Cdc42 GTPase and its effectors, p21-activated kinases Pak2 and Pak1, are not upstream elements controlling the basal level or the stress-induced activation of Pmk1. However, Sty1 MAPK was essential for proper Pmk1 deactivation after hypertonic stress in a process regulated by Atf1 transcription factor. These results provide the first evidence for the existence of cross-talk between two MAPK cascades during the stress response in fission yeast.

  17. Study on determination of iron, cobalt, nickel, copper, zinc and manganese in drinking water by solid-phase extraction and RP-HPLC with 2-(2-quinolinylazo)-5-diethylaminophenol as precolumn derivatizing reagent.

    PubMed

    Hu, Qiufen; Yang, Guanyu; Yang, Jihong; Yin, Jiayuan

    2002-12-01

    A new method for the determination of iron, cobalt, nickel, copper, zinc and manganese in drinking water by the reversed-phase high-performance liquid chromatography (RP-HPLC) with 2-(2-quinolinylazo)-5-diethylaminophenol (QADEAP) as precolumn derivatizing reagent was studied in this paper. The iron, cobalt, nickel, copper, zinc, and manganese ions react with QADEAP to form color chelates in the presence of cetyl trimethylammonium bromide (CTMAB) and acetic acid-sodium acetic buffer solution medium of pH 4.0. These chelates were enriched by solid-phase extraction with a Waters Nova-Pak C18 cartridge and eluted the retained chelates from the cartridge with tetrahydrofuran (THF). The enrichment factor of 100 was achieved. Then the chelates were separated on a Waters Nova-Pak C18 column (3.9 x 150 mm, 5 microm) by gradient elution with methanol (containing 0.2% of acetic acid and 0.1% of CTMAB) and 0.05 mol L(-1) acetic acid-sodium acetic buffer solution (containing 0.1% of CTMAB) (pH 4.0) as mobile phase at a flow rate of 0.5 ml min(-1), and monitored with a photodiode array detector from 450 approximately 700 nm. The detection limits (S/N = 3) of iron, cobalt, nickel, copper, zinc and manganese are 0.8, 1.1, 0.9, 1.1, 1.5 and 2.0 ng L(-1), respectively, in the original sample. This method can be applied to determination at the microg L(-1) level of iron, cobalt, nickel, copper, zinc and manganese in drinking water with good results.

  18. Molecular Epidemiology of Hemoglobinopathies in Cambodia.

    PubMed

    Munkongdee, Thongperm; Tanakulmas, Jatuporn; Butthep, Punnee; Winichagoon, Pranee; Main, Barbara; Yiannakis, Miriam; George, Joby; Devenish, Robyn; Fucharoen, Suthat; Svasti, Saovaros

    2016-06-01

    Determining the magnitude of the thalassemia problem in a country is important for implementing a national prevention and control program. In order to acquire accurate thalassemia prevalence data, the gene frequency of α- and β-thalassemia (α- and β-thal) in different regions of a country should be determined. The molecular basis of thalassemia in Cambodia was performed by polymerase chain reaction (PCR)-based techniques in a community-based cross-sectional survey of 1631 unrelated individuals from three regions, Battambang, Preah Vihear and Phnom Penh. Thalassemia mutations were detected in 62.7% of the three studied population of Cambodia. Hb E (HBB: c.79G > A) was the most common β-globin gene mutation with a frequency ranging from 0.139 to 0.331, while the most frequent α-globin gene mutation was the -α(3.7) (rightward) deletion (0.098-0.255). The other frequencies were 0.001-0.003 for β-thal, 0.008-0.011 for α-thal-1 (- -(SEA)), 0.003-0.008 for α-thal-2 [-α(4.2) (leftward deletion)], 0.021-0.044 for Hb Constant Spring (Hb CS, HBA2: c.427T > C) and 0.009-0.036 for Hb Paksé (HBA2: c.429A > T). A regional specific thalassemia gene frequency was observed. Preah Vihear had the highest prevalence of Hb E (55.9%), α-thal-2 (24.0%) and nondeletional α-thal (15.1%), whereas Phnom Penh had the lowest frequency of thalassemia genes. Interestingly, in Preah Vihear, the frequency of Hb Paksé was extremely high (0.036), almost equivalent to that of Hb CS (0.044). Our results indicate the importance of micromapping and epidemiology studies of thalassemia, which will assist in establishing the national prevention and control program in Cambodia.

  19. Isolation and identification of androstanediol glucuronide from human plasma.

    PubMed

    Rao, P N; Burdett, J E; Moore, P H; Horton, R

    1987-11-01

    [3H]Dihydrotestosterone (50 microCi) was infused into normal men and women for 8 h. It was previously shown that this was sufficient time for this material to reach a steady state. Venous plasma was obtained at 6 and 8 h, pooled, and the unconjugated steroids removed by ether extraction. The remaining plasma was adjusted to pH 4.9 and the steroid conjugate was extracted first with ethyl acetate and then with an ether-ethanol mixture. The extracts were combined and taken to dryness. Steroid sulfates were solvolyzed using dioxane, and the mixture partitioned between ether and 1% NaOH. The aqueous phase was acidified and added to an XAD-2 column, washed with water, and the glucuronide fraction eluted with methanol. The solvent was concentrated and the methanol extract was passed through a C18 Sep-Pak, filtered through an Acrodisc CR and then subjected to gradient high performance liquid chromatography [HPLC] (Nova-Pak C18, KH2PO4, pH 3, and methanol). The fractions containing steroid glucuronides were collected and esterified with diazomethane and then acetylated with acetic anhydride in pyridine. The glucuronide triacetyl methyl ester (GAME) derivatives were then run in a second HPLC system (3 Lichrosorb 5 mu columns, 4 mm x 25 cm) using a gradient of ethanol-heptane and heptane. We clearly established that this system separates 3 alpha-diol GAME conjugated at the 17 and 3 positions (44 vs 50 min) with authentic samples previously synthesized in our laboratory. We concluded that the pooled plasma contained only the 17-GAME conjugate. No significant activity of the 3-glucuronide was detected. The natural compound in circulation, therefore, is 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide.

  20. Analysis of organophosphorus pesticides in whole blood by GC-MS-μECD with forensic purposes.

    PubMed

    Valente, Nuno I P; Tarelho, Sónia; Castro, André L; Silvestre, Armando; Teixeira, Helena M

    2015-07-01

    In the present work, two multi-residue methods for the determination of ten organophosphorus pesticides (OPs), namely chlorfenvinphos, chlorpyrifos, diazinon, dimethoate, fenthion, malathion, parathion, phosalone, pirimiphos-methyl and quinalphos, in post-mortem whole blood samples are presented. The adopted procedure uses GC-MS for screening and quantitation, and GC-μECD (electron capture detector) for compound confirmation. Three different Solid Phase Extraction (SPE) procedures for OPs with Oasis(®) hydrophilic lipophilic balanced (HLB) and Sep-Pak(®) C18 cartridges were tested, and followed by GC-μECD and GC-MS analysis. The Sep-Pak(®) C18 cartridges extraction procedure was selected since it generated analytical signals 5 times higher than those obtained with the two different Oasis(®) HLB cartridges extraction procedures. The method has shown to be selective for the isolation of selected OPs as well as to the chosen internal standard (ethion) in postmortem blood samples. Calibration curves between 50 and 5000 ng/mL were prepared using weighted linear regression models (1/x(2)). It was not possible to establish a working range for fenthion by GC-μECD due to the lower sensitivity of the detector to this compound, whereas for pirimiphos-methyl it was set between 500 and 5000 ng/mL. The limit of quantitation was established at 50 ng/mL for all analytes, except for pirimiphos-methyl by GC-μECD analysis (500 ng/mL). The average extraction efficiency ranged from 72 to 102%. The developed methods were considered robust and fit for the purpose, and had already been adopted in the laboratory routine analysis.

  1. Rab7 Regulates CDH1 Endocytosis, Circular Dorsal Ruffles Genesis, and Thyroglobulin Internalization in a Thyroid Cell Line.

    PubMed

    Mascia, Anna; Gentile, Flaviana; Izzo, Antonella; Mollo, Nunzia; De Luca, Maria; Bucci, Cecilia; Nitsch, Lucio; Calì, Gaetano

    2016-08-01

    Rab7 regulates the biogenesis of late endosomes, lysosomes, and autophagosomes. It has been proposed that a functional and physical interaction exists between Rab7 and Rac1 GTPases in CDH1 endocytosis and ruffled border formation. In FRT cells over-expressing Rab7, increased expression and activity of Rac1 was observed, whereas a reduction of Rab7 expression by RNAi resulted in reduced Rac1 activity, as measured by PAK1 phosphorylation. We found that CDH1 endocytosis was extremely reduced only in Rab7 over-expressing cells but was unchanged in Rab7 silenced cells. In Rab7 under or over-expressing cells, Rab7 and LC3B-II co-localized and co-localization in large circular structures occurred only in Rab7 over-expressing cells. These large circular structures occurred in about 10% of the cell population; some of them (61%) showed co-localization of Rab7 with cortactin and f-actin and were identified as circular dorsal ruffles (CDRs), the others as mature autophagosomes. We propose that the over-expression of Rab7 is sufficient to induce CDRs. Furthermore, in FRT cells, we found that the expression of the insoluble/active form of Rab7, rather than Rab5, or Rab8, was inducible by cAMP and that cAMP-stimulated FRT cells showed increased PAK1 phosphorylation and were no longer able to endocytose CDH1. Finally, we demonstrated that Rab7 over-expressing cells are able to endocytose exogenous thyroglobulin via pinocytosis/CDRs more efficiently than control cells. We propose that the major thyroglobulin endocytosis described in thyroid autonomous adenomas due to Rab7 increased expression, occurs via CDRs. J. Cell. Physiol. 231: 1695-1708, 2016. © 2015 Wiley Periodicals, Inc.

  2. A case study on fuel oil contamination in a mangrove swamp in Hong Kong.

    PubMed

    Tam, Nora F Y; Wong, Teresa W Y; Wong, Y S

    2005-01-01

    Mangroves commonly found along tropical and subtropical coastlines are susceptible to oil pollution. In December 2000, around 500 1 m tall Kandelia candel saplings at the age of 3-5 years old located at the foreshore region of Sheung Pak Nai swamp, Hong Kong SAR, were found to be damaged by oil pollution. More than 80% of the saplings were either dead or washed away and leaving less than 5% healthy saplings with dense green leaves. Elevated concentrations of light n-alkanes (ranging from n-C14 to n-C20), pristane and phytane were recorded in surface sediments collected in December 2000. The ratio between light and total n-alkanes was 0.4. The total petroleum hydrocarbons (60-80 microg g(-1) TPH) and unresolved complex mixtures (60-70 microg g(-1) UCM) were higher than the background values of other mangrove sediments in Hong Kong, which were 40 and 20 microg g(-1), respectively. In certain root zone sediments, TPH concentrations were above 1000 microg g(-1). These results suggest that surface sediments in Sheung Pak Nai were contaminated by petroleum oil, most likely by illegal discharge of fuel oil which occurred between 1998 and 2002. One year later, in December 2001, unhealthy saplings had recovered and re-grown. The concentrations of TPH and UCM in sediments declined to around 40 microg g(-1), pristane and phytane dropped by 80%, and the ratio of light to total n-alkanes was 0.15, suggesting that residual oil in sediments was weathered leading to a remarkable recovery of the unhealthy saplings.

  3. Identification of polymerase gene mutations that affect viral replication in H5N1 influenza viruses isolated from pigeons.

    PubMed

    Elgendy, Emad Mohamed; Arai, Yasuha; Kawashita, Norihito; Daidoji, Tomo; Takagi, Tatsuya; Ibrahim, Madiha Salah; Nakaya, Takaaki; Watanabe, Yohei

    2017-01-01

    Highly pathogenic avian influenza virus H5N1 infects a wide range of host species, with a few cases of sporadic pigeon infections reported in the Middle East and Asia. However, the role of pigeons in the ecology and evolution of H5N1 viruses remains unclear. We previously reported two H5N1 virus strains, isolated from naturally infected pigeons in Egypt, that have several unique mutations in their viral polymerase genes. Here, we investigated the effect of these mutations on H5N1 polymerase activity and viral growth and identified three mutations that affected viral polymerase activity. The results showed that the PB1-V3D mutation significantly decreased polymerase activity and viral growth in both mammalian and avian cells. In contrast, the PB2-K627E and PA-K158R mutations had moderate effects: PB2-K627E decreased and PA-K158R increased polymerase activity. Structural homology modelling indicated that the PB1-V3D residue was located in the PB1 core region that interacts with PA, predicting that the PB1 mutation would produce a stronger interaction between PB1 and PA that results in decreased replication of pigeon-derived H5N1 viruses. Our results identified several unique mutations responsible for changes in polymerase activity in H5N1 virus strains isolated from infected pigeons, emphasizing the importance of avian influenza surveillance in pigeons and in studying the possible role of pigeon-derived H5N1 viruses in avian influenza virus evolution.

  4. [Isolation and determination of the seeds of Pachyrrhizus errosus protein by high performance gel filtration chromatography (GFC)].

    PubMed

    Wu, H; Hao, B; Tang, G; Lin, Y

    1997-03-01

    From the seeds of Pachyrrhizus errosus, three protein constituents, namel PE1, PE2 and PE3, have been isolated and purified by extraction with 5mmol/L phosphate saline (0.9% NaCl) buffer (PB) at pH 7.2, and S-Sepharose Fast Flow Column (2.6cm x 15cm) chromatography which eluted with 5mmol/L phosphate buffer (pH 7.0) containing 1mmol/L NaCl. Three proteins were burther separated on two connected Protein-Pak 60+Protein-Pak 125 [7.5mm x 39cm, 10microm] columns with mobile phase of 0.2mol/L phosphate buffer (pH 6.5). The flow rate was kept constant at 0.8mL/min by YSB-2 type high press pump. The effluent was monitored at a wavelength of 280nm on photodiode array detector. These three proteins are proved to be homogeneous by SDS-PAGE, IEF and HPGFC experiments, and all present the typical absorption spectra in ultraviolet region. The moleculer weights of the three proteins are approxiamtely 33000D, 14500D and 14000D respectively by SDS-PAGE. But as using HPGFC analysis, the MW value of PE2 is 28000D. This indicates PE2 may be composed of two chains joined by disulfide bond, which is further proved from the latter amino acid composition analysis. The isoelectric points of three proteins are 4.5, 6.5 and 7.5 respectively by using IEF. The amion acids compositions of the three proteins were determined with OPA post-column derivatization/fluorescence detection.

  5. Improved Method for Quantifying Nonvolatile Residues on Surfaces and in Liquids

    SciTech Connect

    Benkovich, M.G.

    2004-03-30

    The objective of the project was to develop an improved method to quantify nonvolatile residues on surfaces and in liquids. The project accomplishments are summarized below: (1) ERA Systems, Inc., The MESERAN Company has designed, developed, built, evaluated, tested, and delivered MicroSolventEvaporator systems to KCP, Astro Pak, and Lockheed Martin, that automatically deposit and evaporate successive small quantities (5-10 microliters) of solvents onto clean reference surfaces. (2) ERA Systems, Inc., The MESERAN Company, and KCP have designed, procured, and evaluated stainless steel disks with specific machined grooves to be used as reference surfaces with the MicroSolventEvaporator and MESERAN Analyzers. (3) KCP evaluated various cleaning processes to easily clean the reference surfaces to acceptable levels. Even though some methods (or a combination of methods) may have worked better than others, an easy method that most companies could use to acceptably clean the disks was desired. Aqueous ultrasonic cleaning with Dirl Lum 603 (30 g per liter concentration) followed by flowing DI water and ultrasonic DI water rinses, nitrogen blow drying, and baking in a HEPA filtered oven at 220 F for 30 minutes proved to be a relatively simple method that most companies could use. (4) KCP developed calibrations of several contaminants on the chosen reference substrates so the MESERAN data could be converted into quantifiable amounts of contamination. (5) KCP performed a prove-in of the MicroSolventEvaporator in conjunction with the MESERAN Analyzer and compared the results to gravimetric data. The method worked and was able to provide data at low contamination levels that can not be reliably obtained gravimetrically. (6) Astro Pak and Lockheed Martin have done limited testing at their facilities. The MESERAN Company has agreed to let them keep the units for awhile so they can continue to perform tests to prove-in the process at their facilities even after the CRADA is

  6. The role of drinking water sources, consumption of vegetables and seafood in relation to blood arsenic concentrations of Jamaican children with and without Autism Spectrum Disorders.

    PubMed

    Rahbar, Mohammad H; Samms-Vaughan, Maureen; Ardjomand-Hessabi, Manouchehr; Loveland, Katherine A; Dickerson, Aisha S; Chen, Zhongxue; Bressler, Jan; Shakespeare-Pellington, Sydonnie; Grove, Megan L; Bloom, Kari; Wirth, Julie; Pearson, Deborah A; Boerwinkle, Eric

    2012-09-01

    Arsenic is a toxic metal with harmful effects on human health, particularly on cognitive function. Autism Spectrum Disorders (ASDs) are lifelong neurodevelopmental and behavioral disorders manifesting in infancy or early childhood. We used data from 130 children between 2 and 8 years (65 pairs of ASD cases with age- and sex-matched control), to compare the mean total blood arsenic concentrations in children with and without ASDs in Kingston, Jamaica. Based on univariable analysis, we observed a significant difference between ASD cases and controls (4.03 μg/L for cases vs. 4.48 μg/L for controls, P<0.01). In the final multivariable General Linear Model (GLM), after controlling for car ownership, maternal age, parental education levels, source of drinking water, consumption of "yam, sweet potato, or dasheen", "carrot or pumpkin", "callaloo, broccoli, or pak choi", cabbage, avocado, and the frequency of seafood consumption per week, we did not find a significant association between blood arsenic concentrations and ASD status (4.36 μg/L for cases vs. 4.65 μg/L for controls, P=0.23). Likewise, in a separate final multivariable GLM, we found that source of drinking water, eating avocado, and eating "callaloo, broccoli, or pak choi" was significantly associated with higher blood arsenic concentrations (all three P<0.05). Based on our findings, we recommend assessment of arsenic levels in water, fruits, and vegetables, as well as increased awareness among the Jamaican population regarding potential risks for various exposures to arsenic.

  7. Downregulation of ACE2/Ang-(1-7)/Mas axis promotes breast cancer metastasis by enhancing store-operated calcium entry.

    PubMed

    Yu, Changhui; Tang, Wei; Wang, Yuhao; Shen, Qiang; Wang, Bin; Cai, Chunqing; Meng, Xiaojing; Zou, Fei

    2016-07-01

    The renin-angiotensin system (RAS) is an important component of the tumor microenvironment and plays a key role in promoting cancer cell proliferation, angiogenesis, metabolism, migration and invasion. Meanwhile, the arm of angiotensin-converting enzyme (ACE)2/angiotensin-(1-7) [Ang-(1-7)]/Mas axis in connection with RAS is associated with anti-proliferative, vasodilatory and anti-metastatic properties. Previous studies have shown that Ang-(1-7) reduces the proliferation of orthotopic human breast tumor growth by inhibiting cancer-associated fibroblasts. However, the role of ACE/Ang-(1-7)/Mas axis in the metastasis of breast cancer cells is still unknown. In the present study, we found that ACE2 protein level is negatively correlated with the metastatic ability of breast cancer cells and breast tumor grade. Upregulation of ACE2/Ang-(1-7)/Mas axis inhibits breast cancer cell migration and invasion in vivo and in vitro. Mechanistically, ACE2/Ang-(1-7)/Mas axis activation inhibits store-operated calcium entry (SOCE) and PAK1/NF-κB/Snail1 pathways, and induces E-cadherin expression. In summary, our results demonstrate that downregulation of ACE2/Ang-(1-7)/Mas axis stimulates breast cancer metastasis through the activation of SOCE and PAK1/NF-κB/Snail1 pathways. These results provide new mechanisms by which breast cancer develop metastasis and shed light on developing novel anti-metastasis therapeutics for metastatic breast cancer by modulating ACE2/Ang-(1-7)/Mas axis.

  8. Two years’ study of Tetanus cases in a Paediatric Intensive Care Unit

    PubMed Central

    Naseem, Faizia; Mahar, Imtiaz Ahmad; Arif, Fehmina

    2016-01-01

    Objective: To study the demographic and clinical features, outcome, complications and treatment cost of tetanus patients admitted in Paediatirc Intensive Care Unit (PICU) of Civil Hospital Karachi (CHK). Methods: It is a descriptive observational study conducted at Civil Hospital Karachi from July 2013 to June 2015. Patients of tetanus admitted in PICU during the study period were enrolled. Data was collected from the file records of patients and included the demographic profile, clinical presentation, grade of severity, length of stay, complications and outcome. It also included the cost of treatment. Descriptive statistics were applied to describe the results. Results: During the study period, 23 cases of tetanus were admitted in P.I.C.U. twelve were male and 11 female. Majority of cases (13) belonged to age group 2-6 years. Seventeen cases were unvaccinated and 6 had received only BCG & OPV. None was appropriately vaccinated for age. There were 9 cases of post injury tetanus, 6 of them were males, 5 cases of otogenic tetanus and 9 cases had no clinically identifiable portal of entry. Eleven cases belonged to grade III severity of Ablett classification and 6 had grade IV severity. Mortality in our case series was 26%. Autonomic instability was seen in 17 patients and all of them needed ionotropic support. The estimated cost of per day treatment of a tetanus patient with mechanical ventilation was approximatly 31, 979/Pak Rs and without mechanical ventilation was 20,000/Pak Rs. Conclusion: Tetanus is an entirely preventable disease with a high mortality. Treatment is very costly as compared to vaccination which is free of cost. Complete vaccination and proper wound care is the only option to reduce the ongoing burden of tetanus. PMID:27375706

  9. A novel role for atypical MAPK kinase ERK3 in regulating breast cancer cell morphology and migration

    PubMed Central

    Al-Mahdi, Rania; Babteen, Nouf; Thillai, Kiruthikah; Holt, Mark; Johansen, Bjarne; Wetting, Hilde Ljones; Seternes, Ole-Morten; Wells, Claire M

    2015-01-01

    ERK3 is an atypical Mitogen-activated protein kinase (MAPK6). Despite the fact that the Erk3 gene was originally identified in 1991, its function is still unknown. MK5 (MAP kinase- activated protein kinase 5) also called PRAK is the only known substrate for ERK3. Recently, it was found that group I p21 protein activated kinases (PAKs) are critical effectors of ERK3. PAKs link Rho family of GTPases to actin cytoskeletal dynamics and are known to be involved in the regulation of cell adhesion and migration. In this study we demonstrate that ERK3 protein levels are elevated as MDA-MB-231 breast cancer cells adhere to collagen I which is concomitant with changes in cellular morphology where cells become less well spread following nascent adhesion formation. During this early cellular adhesion event we observe that the cells retain protrusive activity while reducing overall cellular area. Interestingly exogenous expression of ERK3 delivers a comparable reduction in cell spread area, while depletion of ERK3 expression increases cell spread area. Importantly, we have detected a novel specific endogenous ERK3 localization at the cell periphery. Furthermore we find that ERK3 overexpressing cells exhibit a rounded morphology and increased cell migration speed. Surprisingly, exogenous expression of a kinase inactive mutant of ERK3 phenocopies ERK3 overexpression, suggesting a novel kinase independent function for ERK3. Taken together our data suggest that as cells initiate adhesion to matrix increasing levels of ERK3 at the cell periphery are required to orchestrate cell morphology changes which can then drive migratory behavior. PMID:26588708

  10. Separation of aliphatic carboxylic acids and benzenecarboxylic acids by ion-exclusion chromatography with various cation-exchange resin columns and sulfuric acid as eluent.

    PubMed

    Ohta, Kazutoku; Ohashi, Masayoshi; Jin, Ji-Ye; Takeuchi, Toyohide; Fujimoto, Chuzo; Choi, Seong-Ho; Ryoo, Jae-Jeong; Lee, Kwang-Pill

    2003-05-16

    The application of various hydrophilic cation-exchange resins for high-performance liquid chromatography (sulfonated silica gel: TSKgel SP-2SW, carboxylated silica gel: TSKgel CM-2SW, sulfonated polymethacrylate resin: TSKgel SP-5PW, carboxylated polymethacrylate resins: TSKgel CM-5PW and TSKgel OA-Pak A) as stationary phases in ion-exclusion chromatography for C1-C7 aliphatic carboxylic acids (formic, acetic, propionic, butyric, isovaleric, valeric, isocaproic, caproic, 2-methylhexanoic and heptanoic acids) and benzenecarboxylic acids (pyromellitic, trimellitic, hemimellitic, o-phthalic, m-phthalic, p-phthalic, benzoic, salicylic acids and phenol) was carried out using diluted sulfuric acid as the eluent. Silica-based cation-exchange resins (TSKgel SP-2SW and TSKgel CM-2SW) were very suitable for the ion-exclusion chromatographic separation of these benzenecarboxylic acids. Excellent simultaneous separation of these benzenecarboxylic acids was achieved on a TSKgel SP-2SW column (150 x 6 mm I.D.) in 17 min using a 2.5 mM sulfuric acid at pH 2.4 as the eluent. Polymethacrylate-based cation-exchange resins (TSKgel SP-5PW, TSKgel CM-5PW and TSKgel OA-Pak A) acted as advanced stationary phases for the ion-exclusion chromatographic separation of these C1-C7 aliphatic carboxylic acids. Excellent simultaneous separation of these C1-C7 acids was achieved on a TSKgel CM-5PW column (150 x 6 mm I.D.) in 32 min using a 0.05 mM sulfuric acid at pH 4.0 as the eluent.

  11. Solid Waste Composition Study at Taman Universiti, Parit Raja, Batu Pahat

    NASA Astrophysics Data System (ADS)

    Kadir, A. A.; Sani, M. S. A. M.

    2016-07-01

    Solid waste management is recognised as one of the most challenging issues confronted by both the developed and developing countries. The problems rise due to growing population in current years which results in increased generation of waste with various compositions. The aim of this study was to determine the waste compositions at Taman Universiti. Taman Universiti is a mix residential and commercial area which a preferred residential location amongst students and lecturers due to its proximate location to UTHM main campus. The waste collection was carried out for 50 houses on a daily basis. The collection and sorting out method was conducted according to Malaysian Standard MS 2505:2012 and the data was collected and recorded The result showed that the average generation rate of household waste at Taman Universiti was 0.16kg/person/day and the moisture content was approximately ranging from 61%-68%. Household wastes collected were categorized and it consisted of food and organic, paper, rigid plastics, plastics film, baby diapers, glass, tetra pak, household hazardous waste, metal, rubber, textiles, garden waste and leather. The proportion of each wastes were approximately 64.67%, 9.36%, 9.22%, 5.33%, 3.51%, 2.53%, 1.37%, 1.05%, 0.84%, 0.85%, 0.80%, 0.27%, and 0.23%, respectively. Results from the analyses indicated that the food and organic waste are the major composition of household waste at Taman Universiti followed by the paper, rigid plastics, and plastic film. Meanwhile, the proportion of baby diapers, glass, tetra pak, household hazardous waste, metal, rubber, textiles, and garden decreasing accordingly. In addition, leather was recognized as the least category that contributed to the household waste.

  12. Mechanisms of HIV-1 Nef Function and Intracellular Signaling

    PubMed Central

    Foster, John L.; Denial, Sarah J.; Temple, Brenda R. S.; Garcia, J. Victor

    2013-01-01

    Advances in the last several years have enhanced mechanistic understanding of Nef induced CD4 and MHCI downregulation and have suggested a new paradigm for analyzing Nef function. In both of these cases, Nef acts by forming ternary complexes with significant contributions to stability imparted by non-canonical interactions. The mutational analyses and binding assays that have led to these conclusions are discussed. The recent progress has been dependent on conservative mutations and multi-protein binding assays. The poorly understood Nef functions of p21 activated protein kinase (PAK2) activation, enhancement of virion infectivity, and inhibition of immunoglobulin class switching are also likely to involve ternary complexes and non-canonical interactions. Hence, investigation of these latter Nef functions should benefit from a similar approach. Six historically used alanine substitutions for determining structure-function relationships of Nef are discussed. These are M20A, E62A/E63A/E64A/E65A (AAAA), P72A/P75A (AXXA), R106A, L164A/L165A, and D174A/D175A. Investigations of less disruptive mutations in place of AAAA and AXXA have led to different interpretations of mechanism. Two recent examples of this alternate approach applied to PAK2 activation F191 and critical residue D123 are presented. The implications of the new findings and the resulting new paradigm for Nef structure-function are discussed with respect to creating a map of Nef functions on the protein surface. We report the results of a PPI-Pred analysis for protein-protein interfaces. There are three predicted patches produced by the analysis which describe regions consistent with the currently known mutational analyses of Nef function. PMID:21336563

  13. YES, a Src Family Kinase, Is a Proximal Glucose-specific Activator of Cell Division Cycle Control Protein 42 (Cdc42) in Pancreatic Islet β Cells*

    PubMed Central

    Yoder, Stephanie M.; Dineen, Stacey L.; Wang, Zhanxiang; Thurmond, Debbie C.

    2014-01-01

    Second-phase insulin secretion sustains insulin release in the face of hyperglycemia associated with insulin resistance, requiring the continued mobilization of insulin secretory granules to the plasma membrane. Cdc42, the small Rho family GTPase recognized as the proximal glucose-specific trigger to elicit second-phase insulin secretion, signals downstream to activate the p21-activated kinase (PAK1), which then signals to Raf-1/MEK/ERK to induce filamentous actin (F-actin) remodeling, to ultimately mobilize insulin granules to the plasma membrane. However, the steps required to initiate Cdc42 activation in a glucose-specific manner in β cells have remained elusive. Toward this, we identified the involvement of the Src family kinases (SFKs), based upon the ability of SFK inhibitors to block glucose-stimulated Cdc42 and PAK1 activation events as well as the amplifying pathway of glucose-stimulated insulin release, in MIN6 β cells. Indeed, subsequent studies performed in human islets revealed that SFK phosphorylation was induced only by glucose and within 1 min of stimulation before the activation of Cdc42 at 3 min. Furthermore, pervanadate treatment validated the phosphorylation event to be tyrosine-specific. Although RT-PCR showed β cells to express five different SFK proteins, only two of these, YES and Fyn kinases, were found localized to the plasma membrane, and of these two, only YES kinase underwent glucose-stimulated tyrosine phosphorylation. Immunodetection and RNAi analyses further established YES kinase as a proximal glucose-specific signal in the Cdc42-signaling cascade. Identification of YES kinase provides new insight into the mechanisms underlying the sustainment of insulin secretion via granule mobilization/replenishment and F-actin remodeling. PMID:24610809

  14. Diosgenin inhibits superoxide generation in FMLP-activated mouse neutrophils via multiple pathways.

    PubMed

    Lin, Y; Jia, R; Liu, Y; Gao, Y; Zeng, X; Kou, J; Yu, B

    2014-12-01

    Diosgenin possesses anti-inflammatory and anticancer properties. Activated neutrophils produce high concentrations of the superoxide anion which is involved in the pathophysiology of inflammation-related diseases and cancer. In the present study, the inhibitory effect and possible mechanisms of diosgenin on superoxide generation were investigated in mouse bone marrow neutrophils. Diosgenin potently and concentration-dependently inhibited the extracellular and intracellular superoxide anion generation in Formyl-Met-Leu-Phe (FMLP)- activated neutrophils, with IC50 values of 0.50 ± 0.08 μM and 0.66 ± 0.13 μM, respectively. Such inhibition was not mediated by scavenging the superoxide anion or by a cytotoxic effect. Diosgenin inhibited the phosphorylation of p47phox and membrane translocation of p47phox and p67phox, and thus blocking the assembly of nicotinamide adenine dinucleotide phosphate oxidase. Moreover, cellular cyclic adenosine monophosphate (cAMP) levels and protein kinase A (PKA) expression were also effectively increased by diosgenin. It attenuated FMLP-induced increase of phosphorylation of cytosolic phospholipase A (cPLA2), p21-activated kinase (PAK), Akt, p38 mitogen-activated protein kinase (p38MAPK), extracellular signal-regulated kinase (ERK1/2), and c-Jun N-terminal kinase (JNK). Our data indicate that diosgenin exhibits inhibitory effects on superoxide anion production through the blockade of cAMP, PKA, cPLA2, PAK, Akt and MAPKs signaling pathways. The results may explain the clinical implications of diosgenin in the treatment of inflammation-related disorders.

  15. YES, a Src family kinase, is a proximal glucose-specific activator of cell division cycle control protein 42 (Cdc42) in pancreatic islet β cells.

    PubMed

    Yoder, Stephanie M; Dineen, Stacey L; Wang, Zhanxiang; Thurmond, Debbie C

    2014-04-18

    Second-phase insulin secretion sustains insulin release in the face of hyperglycemia associated with insulin resistance, requiring the continued mobilization of insulin secretory granules to the plasma membrane. Cdc42, the small Rho family GTPase recognized as the proximal glucose-specific trigger to elicit second-phase insulin secretion, signals downstream to activate the p21-activated kinase (PAK1), which then signals to Raf-1/MEK/ERK to induce filamentous actin (F-actin) remodeling, to ultimately mobilize insulin granules to the plasma membrane. However, the steps required to initiate Cdc42 activation in a glucose-specific manner in β cells have remained elusive. Toward this, we identified the involvement of the Src family kinases (SFKs), based upon the ability of SFK inhibitors to block glucose-stimulated Cdc42 and PAK1 activation events as well as the amplifying pathway of glucose-stimulated insulin release, in MIN6 β cells. Indeed, subsequent studies performed in human islets revealed that SFK phosphorylation was induced only by glucose and within 1 min of stimulation before the activation of Cdc42 at 3 min. Furthermore, pervanadate treatment validated the phosphorylation event to be tyrosine-specific. Although RT-PCR showed β cells to express five different SFK proteins, only two of these, YES and Fyn kinases, were found localized to the plasma membrane, and of these two, only YES kinase underwent glucose-stimulated tyrosine phosphorylation. Immunodetection and RNAi analyses further established YES kinase as a proximal glucose-specific signal in the Cdc42-signaling cascade. Identification of YES kinase provides new insight into the mechanisms underlying the sustainment of insulin secretion via granule mobilization/replenishment and F-actin remodeling.

  16. Determination of arsenic species in fish, crustacean and sediment samples from Thailand using high performance liquid chromatography (HPLC) coupled with inductively coupled plasma mass spectrometry (ICP-MS).

    PubMed

    Rattanachongkiat, S; Millward, G E; Foulkes, M E

    2004-04-01

    Suitable techniques have been developed for the extraction of arsenic species in a variety of biological and environmental samples from the Pak Pa-Nang Estuary and catchment, located in Southern Thailand, and for their determination using HPLC directly coupled with ICP-MS. The estuary catchment comprises a tin mining area and inhabitants of the region can suffer from various stages of arsenic poisoning. The important arsenic species, AsB, DMA, MMA, and inorganic arsenic (As III and V) have been determined in fish and crustacean samples to provide toxicological information on those fauna which contribute to the local diet. A Hamilton PRP-X100 anion-exchange HPLC system employing a step elution has been used successfully to achieve separation of the arsenic species. A nitric acid microwave digestion procedure, followed by carrier gas nitrogen addition- (N2)-ICP-MS analysis was used to measure total arsenic in sample digests and extracts. The arsenic speciation of the biological samples was preserved using a Trypsin enzymatic extraction procedure. Extraction efficiencies were high, with values of 82-102%(As) for fish and crustacean samples. Validation for these procedures was carried out using certified reference materials. Fish and crustacean samples from the Pak Pa-Nang Estuary showed a range for total arsenic concentration, up to 17 microg g(-1) dry mass. The major species of arsenic in all fauna samples taken was AsB, together with smaller quantities of DMA and, more importantly, inorganic As. For sediment samples, arsenic species were determined following phosphoric acid (1 M H3PO4) extraction in an open focused microwave system. A phosphate-based eluant, pH 6-7.5, with anion exchange HPLC coupled with ICP-MS was used for separation and detection of AsIII, AsV, MMA and DMA. The optimum conditions, identified using an estuarine sediment reference material (LGC), were achieved using 45 W power and a 20 minute heating period for extraction of 0.5 g sediment. The

  17. Chemotaxis and Binding of Pseudomonas aeruginosa to Scratch-Wounded Human Cystic Fibrosis Airway Epithelial Cells

    PubMed Central

    Schwarzer, Christian; Fischer, Horst; Machen, Terry E.

    2016-01-01

    Confocal imaging was used to characterize interactions of Pseudomonas aeruginosa (PA, expressing GFP or labeled with Syto 11) with CF airway epithelial cells (CFBE41o-, grown as confluent monolayers with unknown polarity on coverglasses) in control conditions and following scratch wounding. Epithelia and PAO1-GFP or PAK-GFP (2 MOI) were incubated with Ringer containing typical extracellular salts, pH and glucose and propidium iodide (PI, to identify dead cells). PAO1 and PAK swam randomly over and did not bind to nonwounded CFBE41o- cells. PA migrated rapidly (began within 20 sec, maximum by 5 mins) and massively (10–80 fold increase, termed “swarming”), but transiently (random swimming after 15 mins), to wounds, particularly near cells that took up PI. Some PA remained immobilized on cells near the wound. PA swam randomly over intact CFBE41o- monolayers and wounded monolayers that had been incubated with medium for 1 hr. Expression of CFTR and altered pH of the media did not affect PA interactions with CFBE41o- wounds. In contrast, PAO1 swarming and immobilization along wounds was abolished in PAO1 (PAO1ΔcheYZABW, no expression of chemotaxis regulatory components cheY, cheZ, cheA, cheB and cheW) and greatly reduced in PAO1 that did not express amino acid receptors pctA, B and C (PAO1ΔpctABC) and in PAO1 incubated in Ringer containing a high concentration of mixed amino acids. Non-piliated PAKΔpilA swarmed normally towards wounded areas but bound infrequently to CFBE41o- cells. In contrast, both swarming and binding of PA to CFBE41o- cells near wounds were prevented in non-flagellated PAKΔfliC. Data are consistent with the idea that (i) PA use amino acid sensor-driven chemotaxis and flagella-driven swimming to swarm to CF airway epithelial cells near wounds and (ii) PA use pili to bind to epithelial cells near wounds. PMID:27031335

  18. Tropical Pacific Mg/Ca paleo-thermometry: A contribution to the core top calibration

    NASA Astrophysics Data System (ADS)

    Clark, S.; Mekik, F.; Kienast, M.; Groeneveld, J.

    2009-12-01

    The magnesium to calcium ratio in foraminifera is widely used as a proxy to estimate ocean temperatures in Earth’s past. However, studies have shown that both dissolution within the sediments and seawater salinity influence the Mg/Ca in the shells of tropical foraminifera and skew paleo-temperature estimates. We measured Mg/Ca and d18O in shells of mixed layer-dwelling species (Globigernoides ruber sensu stricto, Globigernoides ruber sensu lato, and Globigerinoides sacculifer) from a series of core tops in the eastern equatorial Pacific. We specifically chose these three taxa because they are most commonly used for sea surface temperature estimates, particularly in down core work. We compared our results with global core top data from which paleo-temperature equations have already been derived and published (e.g. Brown and Elderfield, 1996; Lea et al., 2000; Dekens et al., 2002). We find that at the same surface ocean temperature, Mg/Ca values can range as much as 1.5 mmol/mol, and we find no influence of salinity on outliers from the expected exponential relationship between Mg/Ca and temperature. Nor do we see dissolution as the cause of this variation. Most samples used for core top calibration work are well above the calcite saturation horizon and show no evidence of dissolution. We demonstrate this both with carbonate dissolution proxies, like data using the Globorotalia menardii Fragmentation Index (MFI) (Mekik et al., 2002) from our same core tops, and with estimates of bottom water carbonate ion concentration. Potential other influences which cause these variations could include seasonality, differences in analytical methods, and potential differences among morphotypes of G. ruber. Brown S. and H. Elderfield, 1996. Variations in Mg/Ca in Sr/Ca ratios of planktonic foraminifera caused by postdepositional dissolution: Evidence of shallow Mg-dependant dissolution, Paleoceanography, 11,5,543-551. Dekens P., D. Lea, D. Pak, and H. Spero, 2002. Core top

  19. A portrait of tissue phosphoprotein stability in the clinical tissue procurement process.

    PubMed

    Espina, Virginia; Edmiston, Kirsten H; Heiby, Michael; Pierobon, Mariaelena; Sciro, Manuela; Merritt, Barbara; Banks, Stacey; Deng, Jianghong; VanMeter, Amy J; Geho, David H; Pastore, Lucia; Sennesh, Joel; Petricoin, Emanuel F; Liotta, Lance A

    2008-10-01

    inhibitors alone (ATF-2 Thr-71, SAPK/JNK Thr-183/Tyr-185, STAT1 Tyr-701, JAK1 Tyr-1022/1023, and PAK1/PAK2 Ser-199/204/192/197). This time course study 1) establishes the dynamic nature of specific phosphoproteins in excised tissue, 2) demonstrates augmented phosphorylation in the presence of phosphatase inhibitors, 3) shows that kinase inhibitors block the upsurge in phosphorylation of phosphoproteins, 4) provides a rational strategy for room temperature preservation of proteins, and 5) constitutes a foundation for developing evidence-based tissue procurement guidelines.

  20. Ambulance use in Pakistan: an analysis of surveillance data from emergency departments in Pakistan

    PubMed Central

    2015-01-01

    Background The utilization of ambulances in low- and middle-income countries is limited. The aim of this study was to ascertain frequency of ambulance use and characteristics of patients brought into emergency departments (EDs) through ambulance and non-ambulance modes of transportation. Methods The Pakistan National Emergency Departments Surveillance (Pak-NEDS) was a pilot active surveillance conducted in seven major tertiary-care EDs in six main cities of Pakistan between November 2010 and March 2011. Univariate and multivariate logistic regression was performed to investigate the factors associated with ambulance use. Results Out of 274,436 patients enrolled in Pak-NEDS, the mode of arrival to the ED was documented for 94. 9% (n = 260,378) patients, of which 4.1% (n = 10,546) came to EDs via ambulances. The mean age of patients in the ambulance group was significantly higher compared to the mean age of the non-ambulance group (38 ± 18.4 years versus 32.8 ± 14.9 years, p-value < 0.001). The most common presenting complaint in the ambulance group was head injury (12%) while among non-ambulance users it was fever (12%). Patients of all age groups were less likely to use an ambulance compared to those >45 years of age (p-value < 0.001) adjusted for gender, cities, hospital type, presenting complaint group and disposition. The adjusted odds ratio of utilizing ambulances for those with injuries was 3.5 times higher than those with non-injury complaints (p-value < 0.001). Patients brought to the ED by ambulance were 7.2 times more likely to die in the ED than non-ambulance patients after adjustment for other variables in the model. Conclusion Utilization of ambulances is very low in Pakistan. Ambulance use was found to be more among the elderly and those presenting with injuries. Patients presenting via ambulances were more likely to die in the ED. PMID:26689242

  1. Numerical Investigation of Thermal Stratification in the Primary Circuit of VVER-440 Type Reactors

    SciTech Connect

    Boros, Ildiko; Aszodi, Attila; Legradi, Gabor

    2006-07-01

    Thermal stratification in the primary loops and in the connected pipes can limit the lifetime of the piping, or lead to penetrating cracks due to the stresses caused by the temperature differences and the cyclic temperature changes. Therefore it is essential to determine the thermal hydraulic parameters of the stratified flow. The determination of the affected pipes can be based on the international operational experience and on engineering consideration. The most affected pipes in PWRs are the pressurizer surge line, the injection pipe of the emergency core cooling systems and the feedwater injection pipe of the steam generators. CFD codes can provide an appropriate tool for the examination of the development and the breaking up of the stratification and the determination of the temperature distribution. However, the challenge of the uncertainty of the boundary conditions has to be faced because of the unknown flow circumstances. According to an extensive evaluation, performed in 1998 by the VEIKI, in the VVER-440/213 units of Paks NPP the most affected pipe is the pressurizer surge line [1]. To find out the possible thermal stratification in the surge line, a temperature monitoring system was installed on the YP20 leg of the surge line of the Unit 1 of the Paks NPP in 2000. The measurements showed that during the heat-up period there is a thermal stratification almost all time in the surge line [2]. The maximum temperature differences reach 140 K (140 deg. C). The surge line has been modeled with the CFD code CFX-5.7. The performed transient simulations confirmed the existence of a thermal stratification in the surge line, but showed permanent recirculation of colder coolant in the lower layer, caused by the asymmetric arrangement of the surge line legs and the asymmetric connection of the two legs to the main loop. In this paper, the surge line model and the results of the transient simulations are presented. The CFD model of the injection pipe of the high

  2. Dissolved Gases in Seawater and Sediments (Paper 7R0315)

    NASA Astrophysics Data System (ADS)

    Key, R. M.

    1987-07-01

    the photooxidative daylight loss of oxygen from near-surface tropical waters (Gieskes and Kraay, 1982), isotopic fractionation between fresh and seawater and the atmosphere (Benson and Krause, 1984) , edge effects on chemistry in the 02 minimum zone (Mullins et al., 1985), and the relationship between oxygen and other biogeochemical properties (Pak, 1984; Blizard and Pak, 1984; Lewitus and Broenkow, 1985).

  3. Patterns of domestic exposure to carbon monoxide and particulate matter in households using biomass fuel in Janakpur, Nepal.

    PubMed

    Bartington, S E; Bakolis, I; Devakumar, D; Kurmi, O P; Gulliver, J; Chaube, G; Manandhar, D S; Saville, N M; Costello, A; Osrin, D; Hansell, A L; Ayres, J G

    2017-01-01

    Household Air Pollution (HAP) from biomass cooking fuels is a major cause of morbidity and mortality in low-income settings worldwide. In Nepal the use of open stoves with solid biomass fuels is the primary method of domestic cooking. To assess patterns of domestic air pollution we performed continuous measurement of carbon monoxide (CO) and particulate Matter (PM2.5) in 12 biomass fuel households in Janakpur, Nepal. We measured kitchen PM2.5 and CO concentrations at one-minute intervals for an approximately 48-h period using the TSI DustTrak II 8530/SidePak AM510 (TSI Inc, St. Paul MN, USA) or EL-USB-CO data logger (Lascar Electronics, Erie PA, USA) respectively. We also obtained information regarding fuel, stove and kitchen characteristics and cooking activity patterns. Household cooking was performed in two daily sessions (median total duration 4 h) with diurnal variability in pollutant concentrations reflecting morning and evening cooking sessions and peak concentrations associated with fire-lighting. We observed a strong linear relationship between PM2.5 measurements obtained by co-located photometric and gravimetric monitoring devices, providing local calibration factors of 4.9 (DustTrak) and 2.7 (SidePak). Overall 48-h average CO and PM2.5 concentrations were 5.4 (SD 4.3) ppm (12 households) and 417.6 (SD 686.4) μg/m(3) (8 households), respectively, with higher average concentrations associated with cooking and heating activities. Overall average PM2.5 concentrations and peak 1-h CO concentrations exceeded WHO Indoor Air Quality Guidelines. Average hourly PM2.5 and CO concentrations were moderately correlated (r = 0.52), suggesting that CO has limited utility as a proxy measure for PM2.5 exposure assessment in this setting. Domestic indoor air quality levels associated with biomass fuel combustion in this region exceed WHO Indoor Air Quality standards and are in the hazardous range for human health.

  4. Southern plains woodrats (Neotoma micropus) from southern Texas are important reservoirs of two genotypes of Trypanosoma cruzi and host of a putative novel Trypanosoma species.

    PubMed

    Charles, Roxanne A; Kjos, Sonia; Ellis, Angela E; Barnes, John C; Yabsley, Michael J

    2013-01-01

    Trypanosoma cruzi, the causative agent of Chagas' disease, is an important public health and veterinary pathogen. Although human cases are rare in the United States, infections in wildlife, and in some areas domestic dogs, are common. In 2008 and 2010, we investigated T. cruzi prevalence in possible vertebrate reservoirs in southern Texas, with an emphasis on southern plains woodrats (Neotoma micropus). Infection status was determined using a combination of culture isolation, polymerase chain reaction (PCR), and serologic testing. Based on PCR and/or culture, T. cruzi was detected in 35 of 104 (34%) woodrats, 3 of 4 (75%) striped skunks (Mephitis mephitis), 12 of 20 (60%) raccoons (Procyon lotor), and 5 of 28 (18%) other rodents including a hispid cotton rat (Sigmodon hispidus), rock squirrel (Otospermophilus variegatus), black rat (Rattus rattus), and two house mice (Mus musculus). Additionally, another Trypanosoma species was detected in 41 woodrats, of which 27 were co-infected with T. cruzi. Genetic characterization of T. cruzi revealed that raccoon, rock squirrel, and cotton rat isolates were genotype TcIV, while woodrats and skunks were infected with TcI and TcIV. Based on the Chagas Stat-Pak assay, antibodies were detected in 27 woodrats (26%), 13 raccoons (65%), 4 skunks (100%), and 5 other rodents (18%) (two white-ankled mice [Peromyscus pectoralis laceianus], two house mice, and a rock squirrel). Seroprevalence based on indirect immunofluorescence antibody testing was higher for both woodrats (37%) and raccoons (90%), compared with the Chagas Stat-Pak. This is the first report of T. cruzi in a hispid cotton rat, black rat, rock squirrel, and white-ankled mouse. These data indicate that based on culture and PCR testing, the prevalence of T. cruzi in woodrats is comparable with other common reservoirs (i.e., raccoons and opossums) in the United States. However, unlike raccoons and opossums, which tend to be infected with a particular genotype, southern

  5. Sequence stratigraphic analysis of Eocene Rock Strata, Offshore Indus, southwest Pakistan

    NASA Astrophysics Data System (ADS)

    Khan, Natasha; Rehman, Khaista; Ahmad, Sajjad; Khokher, Jamil; Hajana, M. Iqbal; Hanif, M.

    2016-09-01

    In this study, seismic data from two wells (Pak G2-1 and Indus Marine-1C) and age diagnostic larger benthic foraminifera (LBF) within drill cuttings has been used for the first time to identify depositional sequences within the carbonates in the Offshore Indus Basin, Pakistan. The Offshore Indus is tectonically categorized as a passive continental margin where carbonates occur as shelf carbonates in the near offshore and on volcanic seamounts in deeper waters. Seismic data analysis has indicated the presence of minor faults and carbonate buildups above the igneous basement in the south. Patterns of the seismic reflections enabled definition of three seismic facies units identified as: Unit 1 basement, represented by chaotic, moderate amplitude reflection configuration; while parallel bedding and the drape of overlying strata is typical character of Unit 2, carbonate mound facies. The younger Miocene channels represent Unit 3. The diagnosis of Alveolina vredenburgi/cucumiformis biozone confirmed the Ilerdian (55-52 Ma) stage constituting a second order cycle of deposition for the Eocene carbonates (identified as Unit 2). The carbonate succession has been mainly attributed to an early highstand system tract (HST). The environmental conditions remained favorable leading to the development of keep-up carbonates similar to pinnacle buildups as a result of aggradation during late transgressive system tract and an early HST. The carbonate sequence in the south (Pak G2-1) is thicker and fossiliferous representing inner to middle shelf depths based on fauna compared to the Indus Marine-1C in the north, which is devoid of fossils. Three biozones (SBZ 5, SBZ 6 and SBZ 8) were identified based on the occurrence of LBF. The base of the SBZ 5 zone marks the larger foraminifera turnover and the Paleocene-Eocene (P-E) boundary. The LBF encountered in this study coincides with earlier findings for the P-E boundary. Our findings indicate that the entire Ilerdian stage ranges from 55

  6. Effect of non-thermal air atmospheric pressure plasma jet treatment on gingival wound healing

    NASA Astrophysics Data System (ADS)

    Lee, Jung-Hwan; Choi, Eun-Ha; Kim, Kwang-Mahn; Kim, Kyoung-Nam

    2016-02-01

    Non-thermal atmospheric pressure plasmas have been applied in the biomedical field for the improvement of various cellular activities. In dentistry, the healing of gingival soft tissue plays an important role in health and aesthetic outcomes. While the biomedical application of plasma has been thoroughly studied in dentistry, a detailed investigation of plasma-mediated human gingival fibroblast (HGF) migration for wound healing and its underlying biological mechanism is still pending. Therefore, the aim of this study is to apply a non-thermal air atmospheric pressure plasma jet (NTAAPPJ) to HGF to measure the migration and to reveal the underlying biological mechanisms involved in the migration. After the characterization of NTAAPPJ by optical emission spectroscopy, the adherent HGF was treated with NTAAPPJ or air with a different flow rate. Cell viability, lipid peroxidation, migration, intracellular reactive oxygen species (ROS), and the expression of migration-related genes (EGFR, PAK1, and MAPK3) were investigated. The level of statistical significance was set at 0.05. NTAAPPJ and air treatment with a flow rate of 250-1000 standard cubic centimetres per minute (sccm) for up to 30 s did not induce significant decreases in cell viability or membrane damage. A significant increase in the migration of mitomycin C-treated HGF was observed after 30 s of NTAAPPJ treatment compared to 30 s air-only treatment, which was induced by high levels of intracellular reactive oxygen species (ROS). An increase in migration-related gene expression and EGFR activation was observed following NTAAPPJ treatment in an air flow rate-dependent manner. This is the first report that NTAAPPJ treatment induces an increase in HGF migration without changing cell viability or causing membrane damage. HGF migration was related to an increase in intracellular ROS, changes in the expression of three of the migration-related genes (EGFR, PAK1, and MAPK1), and EGFR activation. Therefore

  7. Pattern of presenting complaints recorded as near-drowning events in emergency departments: a national surveillance study from Pakistan

    PubMed Central

    2015-01-01

    Background Drowning is a heavy burden on the health systems of many countries, including Pakistan. To date, no effective large-scale surveillance has been in place to estimate rates of drowning and near-drowning in Pakistan. The Pakistan National Emergency Department Surveillance (Pak-NEDS) study aimed to fill this gap. Methods Patients who presented with a complaint of "near-drowning" were analyzed to explore patterns of true near-drowning (unintentional) and intentional injuries that led to the "near-drowning" complaint. Bivariate analysis was done to establish patterns among patients treated in emergency departments, including socio-demographic information, injury-related information, accompanying injuries, and emergency department resource utilization. Results A total of 133 patients (0.2% of all injury patients) with "near-drowning" as presenting complaints were recorded by the Pak-NEDS system. True near-drowning (50.0%) and intentional injuries that led to "near-drowning" complaints (50.0%) differed in nature of injuries. The highest proportion of true near-drowning incidents occurred among patients aged between 25-44 years (47.5%), and among males (77.5%). True near-drowning patients usually had other accompanying complaints, such as lower limb injury (40.0%). Very few patients were transported by ambulance (5.0%), and triage was done for 15% of patients. Eleven (27.5%) true near-drowning patients received cardiopulmonary resuscitation. Conclusion There was major under-reporting of drowning and near-drowning cases in the surveillance study. The etiology of near-drowning cases should be further studied. Patients who experienced non-fatal drownings were more commonly sent for medical care due to other accompanying conditions, rather than near-drowning event itself. There is also need for recognizing true near-drowning incidents. The results of this study provide information on data source selection, site location, emergency care standardization, and multi

  8. Identification of the mucin-binding adhesin of Pseudomonas cepacia isolated from patients with cystic fibrosis.

    PubMed Central

    Sajjan, S U; Forstner, J F

    1992-01-01

    In previous experiments, we have shown that isolates of Pseudomonas cepacia from sputa of patients with cystic fibrosis (CF), particularly those with severe lung infection, exhibited specific binding to purified respiratory or intestinal mucins (U. Sajjan, M. Corey, M. Karmali, and J. Forstner, J. Clin. Invest. 89:648-656, 1992). The present report describes the identification of the adhesin as a protein located on fimbriae of mucin-binding P. cepacia. From a total of 53 isolates available (from 22 patients with CF), we used three mucin-binding and three non-mucin-binding isolates for our experiments. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of crude P. cepacia homogenates was performed, the separated proteins were blotted onto nitrocellulose and overlaid with purified mucin, and mucin-binding components were detected with an antimucin antibody and then a second-antibody-alkaline phosphatase conjugate system. Only mucin-binding isolates exhibited a positively stained band at an Mr of 22,000. The 22-kDa protein was purified, and a polyclonal antibody specific for it was developed in rabbits. By electron microscopy and immunogold labelling, both the antibody and mucin (separately) were localized to pili present over the entire surface of the bacterial cells. Non-mucin-binding isolates did not have (or had very few) pili and did not stain with either mucin or the antibody to the 22-kDa protein. The purified 22-kDa protein and its antibody were each able to inhibit piliated P. cepacia binding to mucin. The amino acid composition of the 22-kDa protein was dissimilar to those of the major pilin proteins of Escherichia coli (type 1 pilus) and P. aeruginosa (PAK and PAO1 strains). Both the pili of P. aeruginosa PAK and PAO1 and antibodies to these pili failed to inhibit P. cepacia binding to mucin. Thus, P. cepacia adhesion to mucin is mediated by a pilin-associated 22-kDa protein which differs from epithelial-cell-binding pilin proteins of P. aeruginosa

  9. Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

    PubMed Central

    Weng, Ying-Jan; Husebekk, Anne; Skogen, Björn; Kjaer, Mette; Lin, Liang-Tzung; Burnouf, Thierry

    2016-01-01

    Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a severe disease that is caused by maternal alloantibodies generated during pregnancy or at delivery as a result of incompatibility between maternal and fetal human platelet antigens (HPAs) inherited from the father. Antibody-mediated immune suppression using anti-HPA-1a immunoglobulins is thought to be able to prevent FNAIT caused by HPA-1a. A fractionation process to prepare anti-HPA-1a immunoglobulin (Ig) G (IgG) from human plasma was therefore developed. Anti-HPA-1a plasma was obtained from volunteer mothers who underwent alloimmunization against HPA-1a during a previous pregnancy. Plasma was cryoprecipitated and the supernatant treated with caprylic acid and solvent/detergent (S/D), purified by chromatography, nanofiltered, concentrated, and sterile-filtered. The anti-HPA-1a immunoglobulin fraction was characterized for purity and safety. PAK12 and quantitative monoclonal antibody immobilization of platelet antigen (MAIPA) assays were used to detect anti-HPA-1a IgG. Hepatitis C virus (HCV) removal during nanofiltration was assessed by spiking experiments, using cell culture-derived reporter HCV and luciferase analysis. The caprylic acid treatment precipitated non-Ig proteins yielding a 90% pure Ig supernatant. S-HyperCel chromatography of the S/D-treated supernatant followed by HyperCel STAR AX provided high IgG recovery (>80%) and purity (>99.5%), and efficient IgA and IgM removal. Concentrations of complement factors C3 and C4 were < 0.5 and < 0.4 mg/dL, respectively. The final IgG could be nanofiltered on Planova 20N under conditions removing more than 3 log HCV infectivity to baseline mock infection level, and concentrated to ca. 30 g/L. Proteolytic activity and thrombin generation were low in the final fraction. The Pak12 and MAIPA assays showed good recovery of anti-HPA-1a throughout the process. Clinical-grade HPA-1a IgG can be prepared using a process compliant with current quality requirements

  10. Endocytosis and serpentine filopodia drive blebbishield-mediated resurrection of apoptotic cancer stem cells

    PubMed Central

    Jinesh, G G; Kamat, A M

    2016-01-01

    The blebbishield emergency program helps to resurrect apoptotic cancer stem cells (CSCs) themselves. Understanding the mechanisms behind this program is essential to block resurrection of CSCs during cancer therapy. Here we demonstrate that endocytosis drives serpentine filopodia to construct blebbishields from apoptotic bodies and that a VEGF-VEGFR2-endocytosis-p70S6K axis governs subsequent transformation. Disengagement of RalGDS from E-cadherin initiates endocytosis of RalGDS and its novel interaction partners cdc42, VEGFR2, cleaved β-catenin, and PKC-ζ as well as its known interaction partner K-Ras. We also report novel interactions of p45S6K (cleaved p70S6K) and PKM-ζ with PAK-1 filopodia-forming machinery specifically in blebbishields. Thus, a RalGDS-endocytosis-filopodia-VEGFR2-K-Ras-p70S6K axis drives the blebbishield emergency program, and therapeutic targeting of this axis might prevent resurrection of CSCs during cancer therapy. PMID:27226900

  11. Detection of 8,9-dihydro-(7'-guanyl)-9-hydroxyaflatoxin B1 in human urine

    SciTech Connect

    Autrup, H.; Wakhisi, J.; Vahakangas, K.; Wasunna, A.; Harris, C.C.

    1985-10-01

    A possible role of aflatoxin B1 (AFB) in the etiology of human liver cancer has been suggested from several epidemiological studies. This has been based upon the association between consumption of AFB-contaminated food and the liver cancer incidence in different parts of the world. To further establish the role of AFB as a major factor, we initiated a pilot study in three different districts of Kenya to determine the number of individuals exposed to significant amounts of AFB as measured by the urinary excretion of 8,9-dihydro-8-(7-guanyl)-9-hydroxyaflatoxin B1 (AFB-Gua), an adduct formed between the ultimate carcinogenic form of AFB and nucleic acids. This product has previously been detected in urine from rats treated with AFB. Urine collected at the outpatient clinics at the district hospitals were concentrated on C18 Sep-Pak columns and analyzed by high-pressure liquid chromatography under two different chromatographic conditions. The chemical identity of the samples showing a positive response in both chromatographic systems was verified by synchronous scanning fluorescence spectrophotometry. The highest number of individuals with detectable urinary AFB-Gua lived in either Murang'a district or the neighboring Meru and Embu districts. In Murang'a district a rate of 12% was observed in the January-March period, while only 1 of 32 patients (3%) had a detectable exposure in July-August.

  12. Computed tomographic examination of South American sea lions (Otaria flavescens) with suspected Mycobacterium pinnipedii infection.

    PubMed

    Jurczynski, K; Scharpegge, J; Ley-Zaporozhan, J; Ley, S; Cracknell, J; Lyashchenko, K; Greenwald, R; Schenk, J P

    2011-12-03

    Ten South American sea lions (Otaria flavescens) were presented for clinical evaluation and diagnosis of tuberculosis following known exposure to Mycobacterium pinnipedii. CT was used to determine whether foci of calcification in mediastinal lymph nodes, typically associated with pinniped tuberculosis, could be detected and whether CT was a useful diagnostic modality, in conjunction with other tests, for the diagnosis of tuberculosis in this species. Blood was collected from the caudal gluteal vein of each animal for serological testing using commercially available serological tests (ElephantTB STAT-PAK and DPP Vet; Chembio Diagnostic Systems) and a multiantigen print immunoassay (MAPIA), carried out at Chembio to verify the in-house kits. In four of nine animals that underwent CT scanning, lesions consistent with pinniped tuberculosis were apparent and these were confirmed at subsequent postmortem examination. The five remaining animals did not show any abnormalities on CT, with three being negative on serological tests, which were considered to be normal and potentially used as reference images for healthy sea lions. One animal could not be CT scanned due to its large size and weight (510 kg).

  13. The effect of American cranberry (Vaccinium macrocarpon) constituents on the growth inhibition, membrane integrity, and injury of Escherichia coli O157:H7 and Listeria monocytogenes in comparison to Lactobacillus rhamnosus.

    PubMed

    Lacombe, Alison; McGivney, Christine; Tadepalli, Shravani; Sun, Xiaohong; Wu, Vivian C H

    2013-06-01

    The antimicrobial properties of the American cranberry were studied against Escherichia coli O157:H7, Listeria monocytogenes, and Lactobacillus rhamnosus to determine the effects on growth inhibition, membrane permeability, and injury. Cranberry powder was separated using a C-18 Sep-Pak cartridge into sugars plus organic acids (F1), monomeric phenolics (F2), and anthocyanins plus proanthocyanidins (F3). Fraction 3 was further separated into anthocyanins (F4) and proanthocyanidins (F5) using an LH-20 Sephadex column. Each fraction was diluted in the brain heart infusion (BHI) broth to determine the minimum inhibitory/bactericidal concentrations (MIC/MBC). L. monocytogenes was the most susceptible to cranberry fraction treatment with the lowest MIC/MBC for each treatment, followed by E. coli O157:H7 and L. rhamnosus. Membrane permeability and potential was studied using LIVE/DEAD viability assay and using Bis (1, 3-dibutylbarbituric acid) trimethine oxonol (DiBAC4), respectively. L. rhamnosus demonstrated the highest permeability followed by E. coli O157:H7, and L. monocytogenes. L. rhamnosus demonstrated the highest recovery followed by E. coli O157:H7, and L. monocytogenes. Each cranberry fraction demonstrated membrane hyperpolarization at their native pH, while F2, F3, and F5 demonstrated membrane depolarization at neutral pH. With this knowledge cranberry compounds may be used to prevent maladies and potentially substitute for synthetic preservatives and antibiotics.

  14. Thiostrepton-induced gene expression in Streptomyces lividans.

    PubMed Central

    Murakami, T; Holt, T G; Thompson, C J

    1989-01-01

    Thiostrepton induced the expression of four proteins (17, 19, 30, and 56 kilodaltons) of unknown function in Streptomyces lividans. The chromosomal gene which encoded the 19-kilodalton protein (tipA) was cloned and sequenced. Transcription of the tipA promoter was induced at least 200-fold by thiostrepton. The tipA 200-fold by thiostrepton. The tipA transcriptional start site (located by S1 mapping and primer extension experiments) was preceded by a 45-base-pair imperfect inverted-repeat sequence which included the -10 and -35 regions of the promoter. Under noninducing conditions in vivo, this might form a cruciform structure which is not recognized by RNA polymerase. A 143-base-pair fragment including this region was cloned into a promoter probe vector, pIJ486. In this plasmid, pAK114, the thiostrepton-inducible tipA promoter controlled the expression of a kanamycin resistance gene encoding an aminoglycoside phosphotransferase. As little as 1 ng of thiostrepton spotted on a lawn of S. lividans(pAK114) induced kanamycin-resistant growth. Other thiostreptonlike antibiotics also induced tipA, but structurally unrelated antibiotics which inhibit translation had no effect. In S. lividans, the promoter could be induced by thiostrepton during either growth or stationary phase. The tipA promoter should be a valuable tool for expression studies in streptomycetes. Images PMID:2537819

  15. Determination of ortho-phenylphenol residues in lemon rind by high-performance liquid chromatography with electrochemical detection using a microbore column.

    PubMed

    Yang, Lan; Kotani, Akira; Hakamata, Hideki; Kusu, Fumiyo

    2004-01-01

    A simple and highly sensitive method has been developed for determining ortho-phenylphenol (OPP) in lemon rind by high-performance liquid chromatography with electrochemical detection using a microbore column (microHPLC-ECD). Based on the voltammetric behavior of OPP, microHPLC-ECD was established using a CAPCELL PAK C-18 UG 120 microbore ODS column, 17 mM acetic acid-sodium acetate buffer (pH 4.0)/acetonitrile (60/40, v/v) as a mobile phase and an applied potential at +0.9 V vs. Ag/AgCl. The current peak height was found to be linearly related to the amount of OPP injected from 3.4 pg to 1.7 ng (r > 0.999). The detection limit (S/N = 3) was 3.4 pg (20 fmol), which was 100 times greater in terms of sensitivity when compared to conventional HPLC with UV detection. Standard OPP at 0.425 ng was detected with a relative standard deviation (RSD) of 1.9% (n = 10). The OPP contents in several lemon samples were determined by the present method. The recoveries of OPP from lemon rind exceeded 98% with an RSD (n = 5) of less than 3.01%.

  16. Recovery of nosocomial fecal flora from frozen stool specimens and rectal swabs: comparison of preservatives for epidemiological studies.

    PubMed

    Bonten, M J; Nathan, C; Weinstein, R A

    1997-04-01

    The recovery of antibiotic-susceptible and -resistant aerobic Gram-negative bacilli from stool specimens and from mock rectal swabs after freezing (-20 degrees C) for as long as 4 weeks was studied using three preservatives: Cary-Blair (CB) transport medium, buffered glycerol saline (BGS), and Para Pak C&S solution (CS). In addition, the recovery of enterococci from rectal swabs was investigated after storage of swabs in Stuart's transport media at 4 degrees C as long as 4 weeks. The log10 decreases in bacterial counts from seeded stool suspensions frozen in BGS were 0.64 (i.e., fourfold) and 1.16 after 1 and 4 weeks, respectively, which were significantly less (p < .05) than 1 and 4 week decreases following freezing in CB (1.57 and 2.85) or in CS (1.50 and 2.45). The recovery of Gram-negative bacilli from patients' rectal swabs preserved in BGS was consistent with the results of the experiments with seeded stool suspensions. There was no detectable decrease in recovery of enterococci from rectal swabs stored at 4 degrees C. BGS performed well as a preservative for freezing stool specimens or rectal swabs for later recovery of nosocomial Gram-negative bacilli; enterococci survived well in refrigerated rectal swab specimens.

  17. Toward the Distribution of Orbital Parameters of Nearby Major Galaxy Mergers

    NASA Astrophysics Data System (ADS)

    Mortazavi Karvani, Seyed Alireza

    2016-01-01

    In this thesis project our goal is to measure the initial conditions of a sample of ~20 local disk-disk major galaxy mergers. Measuring the orbital parameters is possible by findingthe most similar galaxy merger simulation to the morphology and kinematics of the data.We have developed an automated modeling method based on the Identikit software package,which also estimates the uncertainty of the measured initial conditions. We tested our modeling method using an independent set of GADGET simulations, and we acquired reliable results onprograde merger systems. We observed the Hα kinematics of our sample using SparsePak IFU on the WIYN telescope at KPNO, and DIS on the 3.5m telescope at APO. For the few merger systems in our sample with archival HI data available, we compare the use of HI vs Hα as the kinematic tracer. This work lays the ground-work for the analysis of larger statistical samples of mergers from on-going IFU galaxy survey such as MaNGA.

  18. CDK5RAP3 is a novel repressor of p14ARF in hepatocellular carcinoma cells.

    PubMed

    Mak, Grace Wing-Yan; Lai, Wai-Lung; Zhou, Yuan; Li, Mingtao; Ng, Irene Oi-Lin; Ching, Yick-Pang

    2012-01-01

    CDK5 regulatory subunit associated protein 3 (CDK5RAP3) is a novel activator of PAK4 and processes important pro-metastatic function in hepatocarcinogenesis. However, it remains unclear if there are other mechanisms by which CDK5RAP3 promotes HCC metastasis. Here, we showed that in CDK5RAP3 stable knockdown SMMC-7721 HCC cells, p14(ARF) tumor suppressor was upregulated at protein and mRNA levels, and ectopic expression of CDK5RAP3 was found to repress the transcription of p14(ARF). Using chromatin immunoprecipitation assay, we demonstrated that CDK5RAP3 bound to p14(ARF) promoter in vivo. Furthermore, knockdown of p14(ARF) in CDK5RAP3 stable knockdown HCC cells reversed the suppression of HCC cell invasiveness mediated by knockdown of CDK5RAP3. Taken together, our findings provide the new evidence that overexpression of CDK5RAP3 promotes HCC metastasis via downregulation of p14(ARF).

  19. CDK5RAP3 Is a Novel Repressor of p14ARF in Hepatocellular Carcinoma Cells

    PubMed Central

    Mak, Grace Wing-Yan; Li, Mingtao; Ng, Irene Oi-Lin; Ching, Yick-Pang

    2012-01-01

    CDK5 regulatory subunit associated protein 3 (CDK5RAP3) is a novel activator of PAK4 and processes important pro-metastatic function in hepatocarcinogenesis. However, it remains unclear if there are other mechanisms by which CDK5RAP3 promotes HCC metastasis. Here, we showed that in CDK5RAP3 stable knockdown SMMC-7721 HCC cells, p14ARF tumor suppressor was upregulated at protein and mRNA levels, and ectopic expression of CDK5RAP3 was found to repress the transcription of p14ARF. Using chromatin immunoprecipitation assay, we demonstrated that CDK5RAP3 bound to p14ARF promoter in vivo. Furthermore, knockdown of p14ARF in CDK5RAP3 stable knockdown HCC cells reversed the suppression of HCC cell invasiveness mediated by knockdown of CDK5RAP3. Taken together, our findings provide the new evidence that overexpression of CDK5RAP3 promotes HCC metastasis via downregulation of p14ARF. PMID:22860085

  20. CIP4 controls CCL19-driven cell steering and chemotaxis in chronic lymphocytic leukemia.

    PubMed

    Malet-Engra, Gema; Viaud, Julien; Ysebaert, Loïc; Farcé, Manon; Lafouresse, Fanny; Laurent, Guy; Gaits-Iacovoni, Frédérique; Scita, Giorgio; Dupré, Loïc

    2013-06-01

    Solid tumor dissemination relies on the reprogramming of molecular pathways controlling chemotaxis. Whether the motility of nonsolid tumors such as leukemia depends on the deregulated expression of molecules decoding chemotactic signals remains an open question. We identify here the membrane remodeling F-BAR adapter protein Cdc42-interacting protein 4 (CIP4) as a key regulator of chemotaxis in chronic lymphocytic leukemia (CLL). CIP4 is expressed at abnormally high levels in CLL cells, where it is required for CCL19-induced chemotaxis. Upon CCL19 stimulation of CLL cells, CIP4 associates with GTP-bound Cdc42 and is recruited to the rear of the lamellipodium and along microspikes radiating through the lamellipodium. Consistent with its cellular distribution, CIP4 removal impairs both the assembly of the polarized lamellipodium and directional migration along a diffusible CCL19 gradient. Furthermore, CIP4 depletion results in decreased activation of WASP, but increased activation of PAK1 and p38 mitogen-activated protein kinase (MAPK). Notably, p38 MAPK inhibition results in impaired lamellipodium assembly and loss of directional migration. This suggests that CIP4 modulates both the WASP and p38 MAPK pathways to promote lamellipodium assembly and chemotaxis. Overall, our study reveals a critical role of CIP4 in mediating chemotaxis of CLL cells by controlling the dynamics of microspike-containing protrusions and cell steering.

  1. An improved liquid chromatographic method for the analysis of tylosin and its impurities.

    PubMed

    Ashenafi, Dunge; Hoogmartens, Jos; Adams, Erwin

    2011-10-01

    A selective reversed-phase (RP) liquid chromatographic (LC) method coupled with UV for the determination of tylosin and its related substances is described. The gradient method uses a Capcell pak C18 ACR column (25 cm×4.6 mm id, 5 μm) maintained at a temperature of 60°C. The mobile phases consist of acetonitrile, phosphate buffer pH 5.5 and water: (A; 27.5:10:62.5 v/v/v) and (B; 50:10:40 v/v/v). The flow rate is 1.0 mL/min and UV detection is performed at 280 nm. It allows the separation of all known and 22 other unknown related substances (≥0.02%) from the main compound and from one another. The method shows good precision, sensitivity, linearity (between 0.2 μg/mL and 1.25 mg/mL) and robustness. The limit of quantification is 0.2 μg/mL, corresponding to 0.020%. Seven bulk tylosin samples containing a large number of impurities were examined using this method.

  2. Characterization of the variation of carbonyl compounds concentrations before, during, and after the renovation of an apartment at Niterói, Brazil.

    PubMed

    Ochs, Soraya de Mendonça; Furtado, Leonardo de Almeida; Cerqueira, Wildson Vieira; Pereira Netto, Annibal Duarte

    2016-08-01

    The present work reports the variation of 31 carbonyl compounds (CC) in an apartment located at Niterói City, Rio de Janeiro State, Brazil. Eight sampling campaigns were conducted through a 1-year period, and three areas (living room, kitchen, and bedroom) were evaluated before, during, and after the renovation activities and reoccupation of the apartment. Samples were collected using SEP-PAK cartridges impregnated with 2,4-dinitrophenylhydrazine, and the hydrazones were analyzed using rapid resolution liquid chromatography with UV detection. The lowest total concentration of CC (19.0 ± 1.5 μg m(-3)) was found before the renovation when the apartment was empty, but door varnishing resulted in highest contamination of the apartment (1386 ± 384 μg m(-3)); however, an important dispersion of CC was observed in the subsequent sampling (148 ± 1.8 μg m(-3)). After apartment reoccupation, the indoor contamination seemed to depend on the routine activities taken there, such as household product use and cooking activities, but apparently, local temperature increase favored the vaporization of the volatile CC from the building materials in the apartment. As far as we are concerned, this is the first study comparing the concentrations of 31 CC in residential areas before, during, and after renovation activities taken in Brazil.

  3. Epidermal growth factor-induced mobilization of a ganglioside-specific sialidase (NEU3) to membrane ruffles

    SciTech Connect

    Yamaguchi, Kazunori; Hata, Keiko; Wada, Tadashi; Moriya, Setsuko; Miyagi, Taeko . E-mail: miyagi-ta173@pref.miyagi.jp

    2006-07-28

    Human ganglioside-specific sialidase, NEU3, localized at cell membranes is thought to regulate various biological processes at cell surfaces. We here explored functional subcellular localization of the sialidase by immunofluorescence and found accumulation at leading edges of cell membranes in the presence of serum in culture. In response to EGF, the sialidase redistributed rapidly to ruffling cell membranes of squamous carcinoma A431 cells and co-localized with Rac-1. NEU3 overexpression enhanced Rac-1 activation and cell migration as compared with controls in HeLa cells as well as in A431 cells. Consistent with co-localization with Rac-1 by immunofluorescence, NEU3 was found to co-precipitate with activated Rac bound to GST-PAK-1 fusion protein. NEU3 silencing by siRNA, in contrast, resulted in inhibition of Rac-1 activation. These results indicate that NEU3 is able to mobilize to membrane ruffles in response to growth stimuli and activate the Rac-1 signaling by co-localization with Rac-1, leading to increased cell motility.

  4. Role of endopeptidase 3.4.24.16 in the catabolism of neurotensin, in vivo, in the vascularly perfused dog ileum.

    PubMed

    Barelli, H; Fox-Threlkeld, J E; Dive, V; Daniel, E E; Vincent, J P; Checler, F

    1994-05-01

    1. The degradation of tritiated and unlabelled neurotensin (NT) following close intra-arterial infusion of the peptides in ileal segments of anaesthetized dogs was examined. 2. Intact NT and its catabolites recovered in the venous effluents were purified by chromatography on Sep-Pak columns followed by reverse-phase h.p.l.c. and identified by their retention times or by radioimmunoassay. 3. The half-life of neurotensin was estimated to be between 2 and 6 min. Four labelled catabolites, corresponding to free tyrosine, neurotensin (1-8), neurotensin (1-10) and neurotensin (1-11), were detected. 4. Neurotensin (1-11) was mainly generated by a phosphoramidon-sensitive cleavage, probably elicited by endopeptidase 24-11. 5. Two endopeptidase 3.4.24.16 inhibitors, phosphodiepryl 03 and the dipeptide Pro-Ile, dose-dependently potentiated the recovery of intact neurotensin. Furthermore, both agents inhibited the formation of neurotensin (1-10), the product that results from the hydrolysis of neurotensin by purified endopeptidase 3.4.24.16. In contrast, the endopeptidase 3.4.24.15 inhibitor Cpp-AAY-pAB neither protected neurotensin from degradation nor modified the production of neurotensin (1-10). 6. Our study is the first evidence to indicate that endopeptidase 3.4.24.16 contributes to the catabolism of neurotensin, in vivo, in the dog intestine.

  5. Development of a microwave-assisted-extraction-based method for the determination of aflatoxins B1, G1, B2, and G2 in grains and grain products.

    PubMed

    Chen, Si; Zhang, Hong

    2013-02-01

    This article describes the use of microwave-assisted extraction (MAE) as a pretreatment technique for the determination of aflatoxins B(1), G(1), B(2), and G(2) in grains and grain products. The optimal operation parameters, including extraction solvent, temperature, and time, were identified to be acetonitrile as the extraction solvent at 80 °C with 15 min of MAE. The extracts were cleaned up using solid-phase extraction followed by derivatization with trifluoroacetic acid and were determined by liquid chromatography-fluorescence detection. A Sep-Pak cartridge was chosen over Oasis HLB and Bond Elut cartridges. By the use of aflatoxin M(1) as an internal standard, relative recoveries of the aflatoxins ranged from 90.7 to 105.7 % for corn and from 88.1 to 103.4 % for wheat, with relative standard deviations between 2.5 and 8.7 %. A total of 36 samples from local markets were analyzed, and aflatoxin B(1) was found to be the predominant toxin, with concentrations ranging from 0.42 to 3.41 μg/kg.

  6. Insights into the cellular response triggered by silver nanoparticles using quantitative proteomics.

    PubMed

    Verano-Braga, Thiago; Miethling-Graff, Rona; Wojdyla, Katarzyna; Rogowska-Wrzesinska, Adelina; Brewer, Jonathan R; Erdmann, Helmut; Kjeldsen, Frank

    2014-03-25

    The use of nanoparticles in foods, materials, and clinical treatments has increased dramatically in the past decade. Because of the possibility of human exposure to nanoparticles, there is an urgent need to investigate the molecular mechanisms underlying the cellular responses that might be triggered. Such information is necessary to assess potential health risks arising from the use of nanoparticles, and for developing new formulations of next generation nanoparticles for clinical treatments. Using mass spectrometry-based proteomic technologies and complementary techniques (e.g., Western blotting and confocal laser scanning microscopy), we present insights into the silver nanoparticle-protein interaction in the human LoVo cell line. Our data indicate that some unique cellular processes are driven by the size. The 100 nm nanoparticles exerted indirect effects via serine/threonine protein kinase (PAK), mitogen-activated protein kinase (MAPK), and phosphatase 2A pathways, and the 20 nm nanoparticles induced direct effects on cellular stress, including generation of reactive oxygen species and protein carbonylation. In addition, we report that proteins involved in SUMOylation were up-regulated after exposure to 20 nm silver nanoparticles. These results were further substantiated by the observation of silver nanoparticles entering the cells; however, data indicate that this was determined by the size of the nanoparticles, since 20 nm particles entered the cells while 100 nm particles did not.

  7. Global Analysis of Serine-Threonine Protein Kinase Genes in Neurospora crassa ▿ †

    PubMed Central

    Park, Gyungsoon; Servin, Jacqueline A.; Turner, Gloria E.; Altamirano, Lorena; Colot, Hildur V.; Collopy, Patrick; Litvinkova, Liubov; Li, Liande; Jones, Carol A.; Diala, Fitz-Gerald; Dunlap, Jay C.; Borkovich, Katherine A.

    2011-01-01

    Serine/threonine (S/T) protein kinases are crucial components of diverse signaling pathways in eukaryotes, including the model filamentous fungus Neurospora crassa. In order to assess the importance of S/T kinases to Neurospora biology, we embarked on a global analysis of 86 S/T kinase genes in Neurospora. We were able to isolate viable mutants for 77 of the 86 kinase genes. Of these, 57% exhibited at least one growth or developmental phenotype, with a relatively large fraction (40%) possessing a defect in more than one trait. S/T kinase knockouts were subjected to chemical screening using a panel of eight chemical treatments, with 25 mutants exhibiting sensitivity or resistance to at least one chemical. This brought the total percentage of S/T mutants with phenotypes in our study to 71%. Mutants lacking apg-1, an S/T kinase required for autophagy in other organisms, possessed the greatest number of phenotypes, with defects in asexual and sexual growth and development and in altered sensitivity to five chemical treatments. We showed that NCU02245/stk-19 is required for chemotropic interactions between female and male cells during mating. Finally, we demonstrated allelism between the S/T kinase gene NCU00406 and velvet (vel), encoding a p21-activated protein kinase (PAK) gene important for asexual and sexual growth and development in Neurospora. PMID:21965514

  8. In vivo biosynthesis of L-(/sup 35/S)Cys-arginine vasopressin, -oxytocin, and -somatostatin: rapid estimation using reversed phase high pressure liquid chromatography. [Rats

    SciTech Connect

    Franco-Bourland, R.E.; Fernstrom, J.D.

    1981-01-01

    L(/sup 35/S)Cys-arginine vasopressin, -oxytocin, and -somatostatin were purified from hypothalami and neurohypophyses 4 h after rats received L(/sup 35/S)Cys via the third ventricle. After acetic acid extraction, Sephadex G-25 filtration, and chemoadsorption to C18-silica (Sep-Pak cartridges), the labeled peptides were rapidly separated by gradient elution, reversed phase, high pressure liquid chromatography (HPLC). The identity and isotopic purity of the labeled peptides were determined by several reversed phase HPLC procedures in conjunction with chemical modification. The labeled peptide fractions were at least 50% radiochemically pure. Using this HPLC isolation procedure, incorporation of L-(/sup 35/S)Cys into each peptide was determined in hydrated and dehydrated rats. Label incorporation into arginine vasopressin and oxytocin in the hypothalamus and the neurohypophysis of dehydrated rats was 2-3 times greater than that in hydrated rats. Incorporation of label into hypothalamic and neurohypophyseal somatostatin was unaffected by the hydration state of the animal. This procedure thus provides a very rapid, but sensitive, set of techniques for studying the control of small peptide biosynthesis in the brain.

  9. HPLC-UV method for simultaneous determination of MK-1775 and AZD-7762 in both acetonitrile-aqueous solution and mouse plasma.

    PubMed

    Ebeid, Kareem; Ho, Giang N; Salem, Aliasger K

    2017-02-15

    A sensitive and precise method is described for the simultaneous determination of two small molecule kinase inhibitors: MK-1775 (MK) and AZD-7762 (AZD), in acetonitrile (ACN)-aqueous solution and in mouse plasma. A Nova-Pak C18 reversed phase column (3.9mm×150mm, 4μm, 60Å) was utilized in the separation using an isocratic mobile phase of 0.1% v/v triethylamine in phosphate buffer (pH=7.4): acetonitrile (ACN) (60:40, v/v), at a flow rate of 0.8mL/min. Detection wavelength was set at 310nm for both MK and AZD, and 431nm for the internal standard sunitinib (SUN). The developed method was validated following the ICH guidelines and it was shown to be accurate, precise and linear in the range of 41ng/mL to 8333ng/mL for both drugs in the ACN-aqueous solution and from 83ng/mL to 8333ng/mL for both drugs in mouse plasma samples. For the first time, the presented data suggest the suitability of this method for the simultaneous separation and quantification of MK and AZD in both ACN aqueous solution as well as in mouse plasma samples.

  10. Precision prediction of the log power spectrum

    NASA Astrophysics Data System (ADS)

    Repp, A.; Szapudi, I.

    2017-01-01

    At translinear scales, the log power spectrum captures significantly more cosmological information than the standard power spectrum. At high wavenumbers k, the Fisher information in the standard power spectrum P(k) fails to increase in proportion to k, in part due to correlations between large- and small-scale modes. As a result, P(k) suffers from an information plateau on these translinear scales, so that analysis with the standard power spectrum cannot access the information contained in these small-scale modes. The log power spectrum PA(k), on the other hand, captures the majority of this otherwise lost information. Until now there has been no means of predicting the amplitude of the log power spectrum apart from cataloging the results of simulations. We here present a cosmology-independent prescription for the log power spectrum; this prescription displays accuracy comparable to that of Smith et al., over a range of redshifts and smoothing scales, and for wavenumbers up to 1.5 h Mpc-1.

  11. Experiments with an EVA Assistant Robot

    NASA Technical Reports Server (NTRS)

    Burridge, Robert R.; Graham, Jeffrey; Shillcutt, Kim; Hirsh, Robert; Kortenkamp, David

    2003-01-01

    Human missions to the Moon or Mars will likely be accompanied by many useful robots that will assist in all aspects of the mission, from construction to maintenance to surface exploration. Such robots might scout terrain, carry tools, take pictures, curate samples, or provide status information during a traverse. At NASA/JSC, the EVA Robotic Assistant (ERA) project has developed a robot testbed for exploring the issues of astronaut-robot interaction. Together with JSC's Advanced Spacesuit Lab, the ERA team has been developing robot capabilities and testing them with space-suited test subjects at planetary surface analog sites. In this paper, we describe the current state of the ERA testbed and two weeks of remote field tests in Arizona in September 2002. A number of teams with a broad range of interests participated in these experiments to explore different aspects of what must be done to develop a program for robotic assistance to surface EVA. Technologies explored in the field experiments included a fuel cell, new mobility platform and manipulator, novel software and communications infrastructure for multi-agent modeling and planning, a mobile science lab, an "InfoPak" for monitoring the spacesuit, and delayed satellite communication to a remote operations team. In this paper, we will describe this latest round of field tests in detail.

  12. [Chemical profiles of methylpyrazines contained in commercially available natto].

    PubMed

    Kanuma, Midori; Yamada, Chie; Oka, Kitaro

    2004-01-01

    Structures and amounts of methylpyrazines contained in commercial natto, a fermented soybean food in Japan, were determined using HPLC equipped with an acid-resistant reversed phase column, Capcell Pak C18 ACR (Shiseido). Mobile phase solvent mixtures consisted of acidic phosphate buffer solution (pH 2.0) containing 2% acetonitrile gave satisfactory results with baseline separation of the authentic specimens, such as naked pyrazine, monomethylpyrazine, 2,3-, 2,5-, and 2,6-dimethylpyrazine, trimethylpyrazine, and tetramethylpyrazine. We used the mobile-phase solvent with a flow rate of 1 ml/min at 15.0 degrees C. Before HPLC, commercial natto samples were treated with water to prepare diluted suspensions of surface mucous materials. The suspensions were treated on Sep-Pac C18 Cartridges (Waters) with phosphate buffer solutions containing 2-7% acetonitrile. The extracts were then injected into the analytical column to obtain chromatograms that were used to determine the structures and amounts of methylpyrazines. The results showed that a commercialiy packed natto contains a considerable amount of 2,5-dimethylpyrazine instead of the tetrametyl- and trimethylpyrazines in the traditional products. This may be a result of recent efforts of natto makers whose interests have been focused on new methods for preparing odorless products.

  13. Micelle-mediated extraction of dibenzocyclooctadiene lignans from Schisandra chinensis with analysis by high-performance liquid chromatography.

    PubMed

    Lee, Ki Yong; Shin, Yong-Jun; Kim, Dae Hyun; Park, Jin-Ho; Kim, Seung Hyun; Han, Sang Beom; Sung, Sang Hyun

    2014-08-01

    Micelle-mediated extraction offers a convenient alternative to conventional extraction systems. A new method based on micelle-mediated extraction was developed for the separation and determination of dibenzocyclooctadiene lignans from Schisandra chinensis by high-performance liquid chromatography with photodiode array detection. Various experimental conditions using the micelle-mediated method were investigated to evaluate the extraction process. Ethylene glycol monoalkyl ether (Genapol X-080), a non-ionic surfactant oligoethylene glycol monoalkyl ether, was chosen as the extract solvent. The chromatographic separation was accomplished on a Shiseido Capcell Pak C18 analytical column (250 × 4.6mm i.d., 5 µm particle diameter), detected by ultraviolet absorption at 254 nm. The isocratic elution was achieved with a mobile phase composed of water-acetonitrile-formic acid (70:30:0.1) at a flow rate of 0.6 mL/min. The method was optimized and fully validated against dibenzocyclooctadiene lignans (schizandrin, gomisin A and gomisin N). With 15% Genapol X-080, a liquid to solid ratio of 100:1 (mL/g) and ultrasonic-assisted extraction for 60 min, the extraction percentage of total dibenzocyclooctadiene lignans reached the highest value. The non-ionic surfactant Genapol X-080 solution is an effective alternative for the extraction of bioactive lignans from S. chinensis.

  14. Nuclear Rings in Galaxies - A Kinematic Perspective

    NASA Technical Reports Server (NTRS)

    Mazzuca, Lisa M.; Swaters, Robert A.; Knapen, Johan H.; Veilleux, Sylvain

    2011-01-01

    We combine DensePak integral field unit and TAURUS Fabry-Perot observations of 13 nuclear rings to show an interconnection between the kinematic properties of the rings and their resonant origin. The nuclear rings have regular and symmetric kinematics, and lack strong non-circular motions. This symmetry, coupled with a direct relationship between the position angles and ellipticities of the rings and those of their host galaxies, indicate the rings are in the same plane as the disc and are circular. From the rotation curves derived, we have estimated the compactness (v(sup 2)/r) up to the turnover radius, which is where the nuclear rings reside. We find that there is evidence of a correlation between compactness and ring width and size. Radially wide rings are less compact, and thus have lower mass concentration. The compactness increases as the ring width decreases. We also find that the nuclear ring size is dependent on the bar strength, with weaker bars allowing rings of any size to form.

  15. HGF-independent regulation of MET and GAB1 by nonreceptor tyrosine kinase FER potentiates metastasis in ovarian cancer

    PubMed Central

    Fan, Gaofeng; Zhang, Siwei; Gao, Yan; Greer, Peter A.; Tonks, Nicholas K.

    2016-01-01

    Ovarian cancer cells disseminate readily within the peritoneal cavity, which promotes metastasis, and are often resistant to chemotherapy. Ovarian cancer patients tend to present with advanced disease, which also limits treatment options; consequently, new therapies are required. The oncoprotein tyrosine kinase MET, which is the receptor for hepatocyte growth factor (HGF), has been implicated in ovarian tumorigenesis and has been the subject of extensive drug development efforts. Here, we report a novel ligand- and autophosphorylation-independent activation of MET through the nonreceptor tyrosine kinase feline sarcoma-related (FER). We demonstrated that the levels of FER were elevated in ovarian cancer cell lines relative to those in immortalized normal surface epithelial cells and that suppression of FER attenuated the motility and invasive properties of these cancer cells. Furthermore, loss of FER impaired the metastasis of ovarian cancer cells in vivo. Mechanistically, we demonstrated that FER phosphorylated a signaling site in MET: Tyr1349. This enhanced activation of RAC1/PAK1 and promoted a kinase-independent scaffolding function that led to recruitment and phosphorylation of GAB1 and the specific activation of the SHP2–ERK signaling pathway. Overall, this analysis provides new insights into signaling events that underlie metastasis in ovarian cancer cells, consistent with a prometastatic role of FER and highlighting its potential as a novel therapeutic target for metastatic ovarian cancer. PMID:27401557

  16. A new species of Bryoleuca Hampson, 1908 from Afghanistan (Lepidoptera, Noctuidae, Bryophilinae).

    PubMed

    Pekarsky, Oleg

    2015-11-13

    A new species of the raptricula species-group, Bryoleuca pljushtchi sp. n. is described. A diagnostic comparison is made with Bryoleuca raptricula ([Denis & Schiffermüller], 1775), Bryoleuca felina (Eversmann, 1852), Bryoleuca orthogramma (Boursin, 1954), Bryoleuca volodia An, Choi & Ronkay, 2013 and Bryoleuca nahnybidai Pekarsky, 2014. Adults and female genitalia of the new species and its closest relatives are illustrated. Modern taxonomic studies of the raptricula-group started by Fibiger et al. (2009) and got progress by the clarification of the taxonomic status of B. felina (Pekarsky & Ronkay 2010). Subsequently, further two newly recognised species were described in the last two years, B. volodia and B. nahnybidai. The first attempt to prepare the checklist of the subfamily was made by Hacker (1990) and, later, European check list was given in the Noctuidae Europaeae, Vol. 11 (Fibiger et al. 2009). As for the fauna of Afghanistan, Koçak and Kemal (2012) listed twelve Bryophilinae species one of which, B. raptricula, belongs to the genus Bryoleuca. The taxonomic situation of the raptricula species-complex is far not completely known yet. Present paper contains the description of a new Bryoleuca species from Afghanistan. The single female of this new species was collected by Ukrainian entomologists Igor Pljushtch and Oleg Pak in Central Afghanistan in 2013.

  17. X linked mental retardation: a clinical guide.

    PubMed

    Raymond, F L

    2006-03-01

    Mental retardation is more common in males than females in the population, assumed to be due to mutations on the X chromosome. The prevalence of the 24 genes identified to date is low and less common than expansions in FMR1, which cause Fragile X syndrome. Systematic screening of all other X linked genes in X linked families with mental retardation is currently not feasible in a clinical setting. The phenotypes of genes causing syndromic and non-syndromic mental retardation (NLGN3, NLGN4, RPS6KA3(RSK2), OPHN1, ATRX, SLC6A8, ARX, SYN1, AGTR2, MECP2, PQBP1, SMCX, and SLC16A2) are first discussed, as these may be the focus of more targeted mutation analysis. Secondly, the relative prevalence of genes causing only non-syndromic mental retardation (IL1RAPL1, TM4SF2, ZNF41, FTSJ1, DLG3, FACL4, PAK3, ARHGEF6, FMR2, and GDI) is summarised. Thirdly, the problem of recurrence risk where a molecular genetics diagnosis has not been made and what proportion of the male excess of mental retardation is due to monogenic disorders of the X chromosome are discussed.

  18. Syndapin Promotes Formation of a Postsynaptic Membrane System in Drosophila

    PubMed Central

    Fricke, Robert; Bhar, Debjani; Reddy-Alla, Suneel; Krishnan, K. S.; Bogdan, Sven

    2009-01-01

    Syndapins belong to the F-BAR domain protein family whose predicted functions in membrane tubulation remain poorly studied in vivo. At Drosophila neuromuscular junctions, syndapin is associated predominantly with a tubulolamellar postsynaptic membrane system known as the subsynaptic reticulum (SSR). We show that syndapin overexpression greatly expands this postsynaptic membrane system. Syndapin can expand the SSR in the absence of dPAK and Dlg, two known regulators of SSR development. Syndapin's N-terminal F-BAR domain, required for membrane tubulation in cultured cells, is required for SSR expansion. Consistent with a model in which syndapin acts directly on postsynaptic membrane, SSR expansion requires conserved residues essential for membrane binding in vitro. However, syndapin's Src homology (SH) 3 domain, which negatively regulates membrane tubulation in cultured cells, is required for synaptic targeting and strong SSR induction. Our observations advance knowledge of syndapin protein function by 1) demonstrating the in vivo relevance of membrane remodeling mechanisms suggested by previous in vitro and structural analyses, 2) showing that SH3 domains are necessary for membrane expansion observed in vivo, and 3) confirming that F-BAR proteins control complex membrane structures. PMID:19244343

  19. A second generation SNP and SSR integrated linkage map and QTL mapping for the Chinese mitten crab Eriocheir sinensis

    PubMed Central

    Qiu, Gao-Feng; Xiong, Liang-Wei; Han, Zhi-Ke; Liu, Zhi-Qiang; Feng, Jian-Bin; Wu, Xu-Gan; Yan, Yin-Long; Shen, Hong; Huang, Long; Chen, Li

    2017-01-01

    The Chinese mitten crab Eriocheir sinensis is the most economically important cultivated crab species in China, and its genome has a high number of chromosomes (2n = 146). To obtain sufficient markers for construction of a dense genetic map for this species, we employed the recently developed specific-locus amplified fragment sequencing (SLAF-seq) method for large-scale SNPs screening and genotyping in a F1 full-sib family of 149 individuals. SLAF-seq generated 127,677 polymorphic SNP markers, of which 20,803 valid markers were assigned into five segregation types and were used together with previous SSR markers for linkage map construction. The final integrated genetic map included 17,680 SNP and 629 SSR markers on the 73 linkage groups (LG), and spanned 14,894.9 cM with an average marker interval of 0.81 cM. QTL mapping localized three significant growth-related QTL to a 1.2 cM region in LG53 as well as 146 sex-linked markers in LG48. Genome-wide QTL-association analysis further identified four growth-related QTL genes named LNX2, PAK2, FMRFamide and octopamine receptors. These genes are involved in a variety of different signaling pathways including cell proliferation and growth. The map and SNP markers described here will be a valuable resource for the E. sinensis genome project and selective breeding programs. PMID:28045132

  20. LIM Kinase 1 Modulates Cortical Actin and CXCR4 Cycling and Is Activated by HIV-1 to Initiate Viral Infection*

    PubMed Central

    Vorster, Paul J.; Guo, Jia; Yoder, Alyson; Wang, Weifeng; Zheng, Yanfang; Xu, Xuehua; Yu, Dongyang; Spear, Mark; Wu, Yuntao

    2011-01-01

    Almost all viral pathogens utilize a cytoskeleton for their entry and intracellular transport. In HIV-1 infection, binding of the virus to blood resting CD4 T cells initiates a temporal course of cortical actin polymerization and depolymerization, a process mimicking the chemotactic response initiated from chemokine receptors. The actin depolymerization has been suggested to promote viral intracellular migration through cofilin-mediated actin treadmilling. However, the role of the virus-mediated actin polymerization in HIV infection is unknown, and the signaling molecules involved remain unidentified. Here we describe a pathogenic mechanism for triggering early actin polymerization through HIV-1 envelope-mediated transient activation of the LIM domain kinase (LIMK), a protein that phosphorylates cofilin. We demonstrate that HIV-mediated LIMK activation is through gp120-triggered transient activation of the Rack-PAK-LIMK pathway, and that knockdown of LIMK through siRNA decreases filamentous actin, increases CXCR4 trafficking, and diminishes viral DNA synthesis. These results suggest that HIV-mediated early actin polymerization may directly regulate the CXCR4 receptor during viral entry and is involved in viral DNA synthesis. Furthermore, we also demonstrate that in resting CD4 T cells, actin polymerization can be triggered through transient treatment with a pharmacological agent, okadaic acid, that activates LIMK and promotes HIV latent infection of resting CD4 T cells. Taken together, our results suggest that HIV hijacks LIMK to control the cortical actin dynamics for the initiation of viral infection of CD4 T cells. PMID:21321123

  1. Influence of variation in soil copper on the yield and nutrition of carrots grown in microplots on two organic soils. [Daucus carota

    SciTech Connect

    Mathur, S.P.; Belanger, A.

    1987-01-01

    Carrots (Daucus carota L. cv. Gold Pak 128) were grown in microplots of two organic soils at site A (a peat), and site B (a muck) in the summer of 1984. The soil surface (0 to 20 cm) varied in total Cu from 13 to 1659, and 81 to 1745 ..mu..g/g at sites A and B, respectively, mainly due to three levels of applications of CuSO/sub 4/ x 5H/sub 2/O in 1978. Neither the yield nor the nutrition (P, K, Ca, Mg, Fe, Mn, Zn, Cu, Mo and B) of the carrot crop was significantly adversely affected by the copper applications or the resultant total soil copper levels at both sites A and B. There were decreases in some foliar nutrient levels due to dilution effects attributable to the significant positive correlations between both root and leaf yields and total soil copper at site B. At both sites A and B, the copper additions appeared to have increased the availability of soil Mn, in accord with earlier evidence.

  2. Chorein sensitivity of cytoskeletal organization and degranulation of platelets.

    PubMed

    Schmidt, Eva-Maria; Schmid, Evi; Münzer, Patrick; Hermann, Andreas; Eyrich, Ann-Kathrin; Russo, Antonella; Walker, Britta; Gu, Shuchen; vom Hagen, Jennifer Müller; Faggio, Caterina; Schaller, Martin; Föller, Michael; Schöls, Ludger; Gawaz, Meinrad; Borst, Oliver; Storch, Alexander; Stournaras, Christos; Lang, Florian

    2013-07-01

    Chorea-acanthocytosis (ChAc), a lethal disease caused by defective chorein, is characterized by neurodegeneration and erythrocyte acanthocytosis. The functional significance of chorein in other cell types remained ill-defined. The present study revealed chorein expression in blood platelets. As compared to platelets from healthy volunteers, platelets from patients with ChAc displayed a 47% increased globular/filamentous actin ratio, indicating actin depolymerization. Moreover, phosphoinositide-3-kinase subunit p85 phosphorylation, p21 protein-activated kinase (PAK1) phosphorylation, as well as vesicle-associated membrane protein 8 (VAMP8) expression were significantly reduced in platelets from patients with ChAc (by 17, 22, and 39%, respectively) and in megakaryocytic (MEG-01) cells following chorein silencing (by 16, 54, and 11%, respectively). Activation-induced platelet secretion from dense granules (ATP release) and α granules (P-selectin exposure) were significantly less (by 55% after stimulation with 1 μg/ml CRP and by 33% after stimulation with 5 μM TRAP, respectively) in ChAc platelets than in control platelets. Furthermore, platelet aggregation following stimulation with different platelet agonists was significantly impaired. These observations reveal a completely novel function of chorein, i.e., regulation of secretion and aggregation of blood platelets.

  3. Health risk assessment of heavy metals and bacterial contamination in drinking water sources: a case study of Malakand Agency, Pakistan.

    PubMed

    Nawab, Javed; Khan, Sardar; Ali, Sharafat; Sher, Hassan; Rahman, Ziaur; Khan, Kifayatullah; Tang, Jianfeng; Ahmad, Aziz

    2016-05-01

    Human beings are frequently exposed to pathogens and heavy metals through ingestion of contaminated drinking water throughout the world particularly in developing countries. The present study aimed to assess the quality of water used for drinking purposes in Malakand Agency, Pakistan. Water samples were collected from different sources (dug wells, bore wells, tube wells, springs, and hand pumps) and analyzed for different physico-chemical parameters and bacterial pathogens (fecal coliform bacteria) using standard methods, while heavy metals were analyzed using atomic absorption spectrophotometry (AAS-PEA-700). In the study area, 70 % of water sources were contaminated with F. coliform representing high bacterial contamination. The heavy metals, such as Cd (29 and 8 %), Ni (16 and 78 %), and Cr (7 %), exceeded their respective safe limits of WHO (2006) and Pak-EPA (2008), respectively, in water sources, while Pb (9 %) only exceeded from WHO safe limit. The risk assessment tools such as daily intake of metals (DIMs) and health risk indexes (HRIs) were used for health risk estimation and were observed in the order of Ni > Cr > Mn > Pb > Cd and Cd > Ni > Pb > Mn > Cr, respectively. The HRI values of heavy metals for both children and adults were <1, showing lack of potential health risk to the local inhabitants of the study area.

  4. [Determination of poppy ingredients in chafing dish materials by isotopic internal standard coupled with multiple reaction monitoring and online full scan mass spectrometry].

    PubMed

    Zhu, Weixia; Sun, Zhuanlian; Yuan, Ping; Yang, Jizhou; Liu, Yafeng; Sun, Wuyong

    2014-12-01

    A confirmative method was developed for determining five poppy alkaloids including morphine, codeine, papaverine, tibane, noscapine in chafing dish ingredients by high performance liquid chromatography coupled with triple quadrupole linear ion trap mass spectrometry (HPLC-Q Trap MS). The sample was extracted with dilute HCl solution under heating condition. The removal of lipid procedure was performed with hexane. The purification was carried out on a mixed-cation solid-phase extraction column (MCX) and ethyl acetate-methanol containing 5% aqueous ammonia was used for elution. A PAK ST column was used to separate the analytes, and 5 mmol/L ammonium acetate methanol and 10 mmol/L ammonium acetate (pH 3. 6) were used as mobile phases. The five alkaloids was detected in the positive mode simultaneously by multiple reaction monitoring (MRM) and online enhanced product ion full scan (EPI). The LODs were 0.05-0.5 µg/kg and the LOQs were 0. 2-2 µg/kg for the five poppy alkaloids. The overall recoveries of the method varied from 64. 2% to 110. 6%, and the RSD were between 4. 2% and 12. 5%. The EPI mass spectra of positive samples were searched through standard library for qualitative confirmation. The detection of real hot pot material samples showed this method can be used for the simple and accurate determination of the five poppy alkaloid residues in chafing dish.

  5. Issues in interpreting the in vivo activity of Aurora-A inhibitors

    PubMed Central

    Shagisultanova, Elena; Dunbrack, Roland L.; Golemis, Erica A.

    2014-01-01

    Introduction Based on its role as a mitotic regulatory kinase, overexpressed and associated with aneuploidy in cancer, small molecule inhibitors have been developed for Aurora-A (AURKA) kinase. In preclinical and clinical assessments, these agents have shown efficacy in inducing stable disease or therapeutic response. In optimizing the use of Aurora-A inhibitors, it is critical to have robust capacity to measure the kinase activity of Aurora-A in tumors. Areas covered we provide an overview of molecular mechanisms of mitotic and non-mitotic activation of Aurora-A kinase, and interaction of Aurora-A with its regulatory partners. Typically, Aurora-A activity is measured by use of phospho-antibodies targeting an auto-phosphorylated T288 epitope. However, recent studies have identified alternative means of Aurora-A activation control, including allosteric regulation by partners, phosphorylation on alternative activating residues (S51, S98), dephosphorylation on inhibitory sites (S342), and T288 phosphorylation by alternative kinases such as Pak enzymes. Additional work has shown that the relative abundance of Aurora-A partners can affect the activity of Aurora-A inhibitors, and that Aurora-A activation also occurs in interphase cells. Expert opinion Taken together, this work suggests the need for comprehensive analysis of Aurora-A activity and expression of Aurora-A partners in order to stratify patients for likely therapeutic response. PMID:25384454

  6. A Novel Biosurfactant Produced by Aureobasidium pullulans L3-GPY from a Tiger Lily Wild Flower, Lilium lancifolium Thunb.

    PubMed Central

    Kim, Jong Shik; Lee, In Kyoung; Yun, Bong Sik

    2015-01-01

    Yeast biosurfactants are important biotechnological products in the food industry, and they have medical and cosmeceutical applications owing to their specific modes of action, low toxicity, and applicability. Thus, we have isolated and examined biosurfactant-producing yeast for various industrial and medical applications. A rapid and simple method was developed to screen biosurfactant-producing yeasts for high production of eco-friendly biosurfactants. Using this method, several potential niches of biosurfactant-producing yeasts, such as wild flowers, were investigated. We successfully selected a yeast strain, L3-GPY, with potent surfactant activity from a tiger lily, Lilium lancifolium Thunb. Here, we report the first identification of strain L3-GPY as the black yeast Aureobasidium pullulans. In addition, we isolated a new low-surface-tension chemical, designated glycerol-liamocin, from the culture supernatant of strain L3-GPY through consecutive chromatography steps, involving an ODS column, solvent partition, silica gel, Sephadex LH-20, and an ODS Sep-Pak cartridge column. The chemical structure of glycerol-liamocin, determined by mass spectrometry and nuclear magnetic resonance spectroscopy, indicates that it is a novel compound with the molecular formula C33H62O12. Furthermore, glycerol-liamocin exhibited potent biosurfactant activity (31 mN/m). These results suggest that glycerol-liamocin is a potential novel biosurfactantfor use in various industrial applications. PMID:25849549

  7. A Systems Biology Comparison of Ovarian Cancers Implicates Putative Somatic Driver Mutations through Protein-Protein Interaction Models

    PubMed Central

    Yang, Mary Qu; Elnitski, Laura

    2016-01-01

    Ovarian carcinomas can be aggressive with a high mortality rate (e.g., high-grade serous ovarian carcinomas, or HGSOCs), or indolent with much better long-term outcomes (e.g., low-malignant-potential, or LMP, serous ovarian carcinomas). By comparing LMP and HGSOC tumors, we can gain insight into the mechanisms underlying malignant progression in ovarian cancer. However, previous studies of the two subtypes have been focused on gene expression analysis. Here, we applied a systems biology approach, integrating gene expression profiles derived from two independent data sets containing both LMP and HGSOC tumors with protein-protein interaction data. Genes and related networks implicated by both data sets involved both known and novel disease mechanisms and highlighted the different roles of BRCA1 and CREBBP in the two tumor types. In addition, the incorporation of somatic mutation data revealed that amplification of PAK4 is associated with poor survival in patients with HGSOC. Thus, perturbations in protein interaction networks demonstrate differential trafficking of network information between malignant and benign ovarian cancers. The novel network-based molecular signatures identified here may be used to identify new targets for intervention and to improve the treatment of invasive ovarian cancer as well as early diagnosis. PMID:27788148

  8. Effect of additives on tribochemical stability of greases with easter base oil

    SciTech Connect

    Nikonorov, E.M.; Demina, L.V.; Skryabina, T.G.; Sosulina, L.N.

    1984-03-01

    This article investigates the possibility of improving the tribochemical stability of soap-base grease formulated with polyol ester oils, by the use of additives of the phenol and amine types. Model greases formulated with a pentaerythritol ester (PET) and the polyester PAK, taken in a 9:1 ratio, were examined. The tribochemical stability of the greases was evaluated according to the rate of conversion of their dispersion medium in a rolling friction zone. The rate of additive exhaustion was evaluated by analogy with the rate of triboconversion of the dispersion medium. The additives used were phenyl-..cap alpha..-naphthylamine (PAN), diphenylamine (DPA), 2, 6-di-tert-butyl-p-cresol (ionol), a technical-grade dialkylated diphenylamine (DAT), and mixtures of the inhibitors. It is determined that the greatest effect in increasing the tribochemical stability is achieved by the introduction of combinations of additives: DAT + ionol (in Li and cCa greases) and PAN + DAT (in cCa grease).

  9. Characterization of toxins from the broad-banded water snake Helicops angulatus (Linnaeus, 1758): isolation of a cysteine-rich secretory protein, Helicopsin.

    PubMed

    Estrella, Amalid; Sánchez, Elda E; Galán, Jacob A; Tao, W Andy; Guerrero, Belsy; Navarrete, Luis F; Rodríguez-Acosta, Alexis

    2011-04-01

    Helicops angulatus (broad-banded water snake) according to recent proposals is presently cited in the family Dipsadidae, subfamily Xenodontinae, forming the tribe Hydropsini along with the genera Hydrops and Pseudoeryx. The current work characterizes the proteolytic and neurotoxic activities of H. angulatus crude toxins from salivary excretion (SE) and describes the isolation and identification of a cysteine-rich secretory protein (CRISP) called helicopsin. The SE lethal dose (LD₅₀) was 5.3 mg/kg; however, the SE did not contain hemorrhagic activity. Helicopsin was purified using activity-guided, Superose 12 10/300 GL molecular exclusion, Mono Q10 ion exchange, and Protein Pak 60 molecular exclusion. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a highly purified band of approximately 20 kDa. The minimal lethal dose for helicopsin was 0.4 mg/kg. Liquid chromatography mass spectrometry (LC-MS/MS) analysis identified 2 unique peptides MEWYPEAAANAER and YTQIVWYK, representing a protein sequence (deleted homology) belonging to cysteine-rich secretory proteins, which are conserved in snake venoms (CRISPs). CRISPs are a large family of cysteine-rich secretory proteins found in various organisms and participate in diverse biological processes. Helicopsin exhibited robust neurotoxic activity as evidenced by immediate death (~8 min) due to respiratory paralysis in NIH mice. These observations for helicopsin purified from H. angulatus provide further evidence of the extensive distribution of highly potent neurotoxins in the Colubroidea superfamily of snakes than previously described.

  10. Mnk kinase pathway: Cellular functions and biological outcomes

    PubMed Central

    Joshi, Sonali; Platanias, Leonidas C

    2014-01-01

    The mitogen-activated protein kinase (MAPK) interacting protein kinases 1 and 2 (Mnk1 and Mnk2) play important roles in controlling signals involved in mRNA translation. In addition to the MAPKs (p38 or Erk), multiple studies suggest that the Mnk kinases can be regulated by other known kinases such as Pak2 and/or other unidentified kinases by phosphorylation of residues distinct from the sites phosphorylated by the MAPKs. Several studies have established multiple Mnk protein targets, including PSF, heterogenous nuclear ribonucleoprotein A1, Sprouty 2 and have lead to the identification of distinct biological functions and substrate specificity for the Mnk kinases. In this review we discuss the pathways regulating the Mnk kinases, their known substrates as well as the functional consequences of engagement of pathways controlled by Mnk kinases. These kinases play an important role in mRNA translation via their regulation of eukaryotic initiation factor 4E (eIF4E) and their functions have important implications in tumor biology as well as the regulation of drug resistance to anti-oncogenic therapies. Other studies have identified a role for the Mnk kinases in cap-independent mRNA translation, suggesting that the Mnk kinases can exert important functional effects independently of the phosphorylation of eIF4E. The role of Mnk kinases in inflammation and inflammation-induced malignancies is also discussed. PMID:25225600

  11. Comparison of new Ampac bags and FireDebrisPAK® bags as packaging for fire debris analysis.

    PubMed

    Borusiewicz, Rafal

    2012-07-01

    The FireDebrisPAK(®) bags that were produced by Kapak were considered to be one of the best containers for fire debris. Kapak bags were discontinued; however, from July 2010, Ampac is offering a new packaging material. The aim of the presented research was to compare the properties (durability, background interferences, and permeability) of Kapak bags and packaging material offered by Ampac. The analysis was conducted by passive adsorption from the headspace with subsequent thermal desorption and analysis by GC-MS. The results proved that the properties of the compared materials are similar. Their greatest advantage is that they are impermeable for components of flammable liquids, so there is no danger of losing analytes or cross-contamination. Their one significant drawback is that they should not be exposed to temperatures above 80°C. At this temperature, they become soft, their inner layer is compromised (becomes sticky), and they emit some volatile organic compounds. Among them, there are compounds that constitute the components of some of flammable liquids.

  12. The Photometric and Kinematic Structure of Face-on Disk Galaxies. III. Kinematic Inclinations from Hα Velocity Fields

    NASA Astrophysics Data System (ADS)

    Andersen, David R.; Bershady, Matthew A.

    2013-05-01

    Using the integral field unit DensePak on the WIYN 3.5 m telescope we have obtained Hα velocity fields of 39 nearly face-on disks at echelle resolutions. High-quality, uniform kinematic data and a new modeling technique enabled us to derive accurate and precise kinematic inclinations with mean i kin = 23° for 90% of these galaxies. Modeling the kinematic data as single, inclined disks in circular rotation improves upon the traditional tilted-ring method. We measure kinematic inclinations with a precision in sin i of 25% at 20° and 6% at 30°. Kinematic inclinations are consistent with photometric and inverse Tully-Fisher inclinations when the sample is culled of galaxies with kinematic asymmetries, for which we give two specific prescriptions. Kinematic inclinations can therefore be used in statistical "face-on" Tully-Fisher studies. A weighted combination of multiple, independent inclination measurements yield the most precise and accurate inclination. Combining inverse Tully-Fisher inclinations with kinematic inclinations yields joint probability inclinations with a precision in sin i of 10% at 15° and 5% at 30°. This level of precision makes accurate mass decompositions of galaxies possible even at low inclination. We find scaling relations between rotation speed and disk-scale length identical to results from more inclined samples. We also observe the trend of more steeply rising rotation curves with increased rotation speed and light concentration. This trend appears to be uncorrelated with disk surface brightness.

  13. Simultaneous determination of methamphetamine and its metabolite, amphetamine, in urine using a high performance liquid chromatography column-switching method.

    PubMed

    Kumihashi, Mitsuru; Ameno, Kiyoshi; Shibayama, Takayuki; Suga, Keisuke; Miyauchi, Hiroshi; Jamal, Mostofa; Wang, Weihuan; Uekita, Ikuo; Ijiri, Iwao

    2007-01-01

    We describe here a simple, precise, and highly sensitive method for the simultaneous determination of methamphetamine (MA) and amphetamine (AM) in urine using a high performance liquid chromatography (HPLC) column-switching method. A PK-2A (Shodex) column was used for extraction and deproteinization, and a CAPCELL PAK SCX semi-micro, polymer-coated cation-exchange column was employed for separation. The urine sample was mixed with an equal volume of borate buffer (0.1M, pH 9.4), and then 100 microl of the mixture was injected into the HPLC column. The column was switched for 6 min, and then 10 min later detection was performed at 210 nm. Recovery yields of the MA and AM spiked in the urine were 93.0-100.4% with a coefficient of variation of less than 1%. The calibration curves of MA and AM were in the range of 0.1-10 microg/ml with good linearity (r(2)=0.999), with the limit of qualification being 0.005 microg/ml. This method of using HPLC with column-switching can be used for both qualification and quantification of MA and its metabolite, AM, in urine, especially in forensic cases.

  14. Determination of pesticide residues in coconut water by liquid-liquid extraction and gas chromatography with electron-capture plus thermionic specific detection and solid-phase extraction and high-performance liquid chromatography with ultraviolet detection.

    PubMed

    Brito, N M; Navickiene, S; Polese, L; Jardim, E F G; Abakerli, R B; Ribeiro, M L

    2002-05-31

    Two simple methods were developed to determine 11 pesticides in coconut water, a natural isotonic drink rich in salts, sugars and vitamins consumed by the people and athletes. The first procedure involves solid-phase extraction using Sep-Pak Vac C18 disposable cartridges with methanol for elution. Isocratic analysis was carried out by means of high-performance liquid chromatography with ultraviolet detection at 254 nm to analyse captan, chlorothalonil, carbendazim, lufenuron and diafenthiuron. The other procedure is based on liquid-liquid extraction with hexane-dichloromethane (1:1, v/v), followed by gas chromatographic analysis with effluent splitting to electron-capture detection for determination of endosulfan, captan, tetradifon and trichlorfon and thermionic specific detection for determination of malathion, parathion-methyl and monocrotophos. The methods were validated with fortified samples at different concentration levels (0.01-12.0 mg/kg). Average recoveries ranged from 75 to 104% with relative standard deviations between 1.4 and 11.5%. Each recovery analysis was repeated at least five times. Limits of detection ranged from 0.002 to 2.0 mg/kg. The analytical procedures were applied to 15 samples and no detectable amounts of the pesticides were found in any samples under the conditions described.

  15. Simultaneous determination of 1-(2'-deoxy-2'-fluoro-β-D-arabinofuranosyl) uracil (FAU) and 1-(2'-deoxy-2'-fluoro-β-D-arabinofuranosyl) 5-methyluracil (FMAU) in human plasma by liquid chromatography/tandem mass spectrometry.

    PubMed

    Wiegand, Richard; Wu, Jianmei; Shields, Anthony F; Lorusso, Patricia; Li, Jing

    2012-04-01

    A liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) assay was developed and validated for simultaneous determination of 1-(2'-deoxy-2'-fluoro-β-D-arabinofuranosyl) uracil (FAU) and its active metabolite 1-(2'-deoxy-2'-fluoro-β-D-arabinofuranosyl) 5-methyluracil (FMAU) in human plasma. FAU and FMAU were extracted from plasma samples using solid-phase extraction with Waters Sep-Pak® Vac C₁₈ cartridge. Chromatographic separation was achieved on a Waters Atlantis T3 C₁₈ column with a gradient mobile phase consisting of methanol and water with 0.45% formic acid (v/v) running at a flow rate of 0.2 ml/min. The analytes were monitored by triple quadrupole mass spectrometer under positive ionization mode. The lower limit of quantitation (LLOQ) was 10 and 2 ng/ml for FAU and FMAU in plasma, respectively. Calibration curves were linear over FAU and FMAU plasma concentration range of 10-2000 and 2-1000 ng/ml, respectively. The intra-day and inter-day accuracy and precision were within the generally accepted criteria for bioanalytical method (<15%). The method has been successfully employed to characterize the plasma pharmacokinetics of FAU and FMAU in cancer patients receiving 1-h intravenous infusion of FAU 50 mg/m².

  16. Diacylglycerol kinase ζ regulates RhoA activation via a kinase-independent scaffolding mechanism.

    PubMed

    Ard, Ryan; Mulatz, Kirk; Abramovici, Hanan; Maillet, Jean-Christian; Fottinger, Alexandra; Foley, Tanya; Byham, Michèle-Renée; Iqbal, Tasfia Ahmed; Yoneda, Atsuko; Couchman, John R; Parks, Robin J; Gee, Stephen H

    2012-10-01

    Rho GTPases share a common inhibitor, Rho guanine nucleotide dissociation inhibitor (RhoGDI), which regulates their expression levels, membrane localization, and activation state. The selective dissociation of individual Rho GTPases from RhoGDI ensures appropriate responses to cellular signals, but the underlying mechanisms are unclear. Diacylglycerol kinase ζ (DGKζ), which phosphorylates diacylglycerol to yield phosphatidic acid, selectively dissociates Rac1 by stimulating PAK1-mediated phosphorylation of RhoGDI on Ser-101/174. Similarly, phosphorylation of RhoGDI on Ser-34 by protein kinase Cα (PKCα) selectively releases RhoA. Here we show DGKζ is required for RhoA activation and Ser-34 phosphorylation, which were decreased in DGKζ-deficient fibroblasts and rescued by wild-type DGKζ or a catalytically inactive mutant. DGKζ bound directly to the C-terminus of RhoA and the regulatory arm of RhoGDI and was required for efficient interaction of PKCα and RhoA. DGKζ-null fibroblasts had condensed F-actin bundles and altered focal adhesion distribution, indicative of aberrant RhoA signaling. Two targets of the RhoA effector ROCK showed reduced phosphorylation in DGKζ-null cells. Collectively our findings suggest DGKζ functions as a scaffold to assemble a signaling complex that functions as a RhoA-selective, GDI dissociation factor. As a regulator of Rac1 and RhoA activity, DGKζ is a critical factor linking changes in lipid signaling to actin reorganization.

  17. Partial purification of endogenous digitalis-like compound(s) in cord blood

    SciTech Connect

    Balzan, S.; Ghione, S.; Biver, P.; Gazzetti, P.; Montali, U. )

    1991-02-01

    Increasing evidence indicates the presence of endogenous digitalis-like compound(s) in human body fluids. In this preliminary report, we describe a study of the partial purification by HPLC of these compounds in the plasma of neonates (who have particularly high concentrations of this substance) and adults. Plasma samples from neonates (cord blood) and adults, lyophilized and extracted with methanol, were applied on a 300 x 3.9 mm C18 Nova Pak column and eluted with a mobile phase of acetonitrile/methanol/water (17/17/66 or 14/14/72 by vol) and, after 30 min, with 100% methanol. We assayed eluted fractions for inhibitory activity of 86Rb uptake and for digoxin-like immunoreactivity. The elution profile revealed a first peak of inhibitory activity of 86Rb uptake at the beginning of the chromatography; another peak was eluted with the 100% methanol. The two peaks also cross-reacted with antidigoxin antibodies. Because the second peak could possibly reflect the nonspecific interference of various lipophilic compounds, we focused our attention on the first peak. For these fractions dose-response curves for 86Rb uptake and for displacement of digoxin were parallel, respectively, to those of ouabain and digoxin, suggesting similarities of digoxin-like immunoreactive substance to cardiac glycosides. Similar chromatographic profiles were also obtained for plasma from adults, suggesting that the endogenous glycoside-like compound(s) in the neonate may be the same as those in the adult.

  18. MLST-Based Population Genetic Analysis in a Global Context Reveals Clonality amongst Cryptococcus neoformans var. grubii VNI Isolates from HIV Patients in Southeastern Brazil

    PubMed Central

    Ferreira-Paim, Kennio; Andrade-Silva, Leonardo; Fonseca, Fernanda M.; Ferreira, Thatiana B.; Mora, Delio J.; Andrade-Silva, Juliana; Khan, Aziza; Dao, Aiken; Reis, Eduardo C.; Almeida, Margarete T. G.; Maltos, Andre; Junior, Virmondes R.; Trilles, Luciana; Rickerts, Volker; Chindamporn, Ariya; Sykes, Jane E.; Cogliati, Massimo; Nielsen, Kirsten; Boekhout, Teun; Fisher, Matthew; Kwon-Chung, June; Engelthaler, David M.; Lazéra, Marcia; Meyer, Wieland; Silva-Vergara, Mario L.

    2017-01-01

    Cryptococcosis is an important fungal infection in immunocompromised individuals, especially those infected with HIV. In Brazil, despite the free availability of antiretroviral therapy (ART) in the public health system, the mortality rate due to Cryptococcus neoformans meningitis is still high. To obtain a more detailed picture of the population genetic structure of this species in southeast Brazil, we studied 108 clinical isolates from 101 patients and 35 environmental isolates. Among the patients, 59% had a fatal outcome mainly in HIV-positive male patients. All the isolates were found to be C. neoformans var. grubii major molecular type VNI and mating type locus alpha. Twelve were identified as diploid by flow cytometry, being homozygous (AαAα) for the mating type and by PCR screening of the STE20, GPA1, and PAK1 genes. Using the ISHAM consensus multilocus sequence typing (MLST) scheme, 13 sequence types (ST) were identified, with one being newly described. ST93 was identified from 81 (75%) of the clinical isolates, while ST77 and ST93 were identified from 19 (54%) and 10 (29%) environmental isolates, respectively. The southeastern Brazilian isolates had an overwhelming clonal population structure. When compared with populations from different continents based on data extracted from the ISHAM-MLST database (mlst.mycologylab.org) they showed less genetic variability. Two main clusters within C. neoformans var. grubii VNI were identified that diverged from VNB around 0.58 to 4.8 million years ago. PMID:28099434

  19. Development of the software tool for generation and visualization of the finite element head model with bone conduction sounds

    NASA Astrophysics Data System (ADS)

    Nikolić, Dalibor; Milošević, Žarko; Saveljić, Igor; Filipović, Nenad

    2015-12-01

    Vibration of the skull causes a hearing sensation. We call it Bone Conduction (BC) sound. There are several investigations about transmission properties of bone conducted sound. The aim of this study was to develop a software tool for easy generation of the finite element (FE) model of the human head with different materials based on human head anatomy and to calculate sound conduction through the head. Developed software tool generates a model in a few steps. The first step is to do segmentation of CT medical images (DICOM) and to generate a surface mesh files (STL). Each STL file presents a different layer of human head with different material properties (brain, CSF, different layers of the skull bone, skin, etc.). The next steps are to make tetrahedral mesh from obtained STL files, to define FE model boundary conditions and to solve FE equations. This tool uses PAK solver, which is the open source software implemented in SIFEM FP7 project, for calculations of the head vibration. Purpose of this tool is to show impact of the bone conduction sound of the head on the hearing system and to estimate matching of obtained results with experimental measurements.

  20. Use of a new tandem cation/anion exchange system with clinical-scale generators provides high specific volume solutions of technetium-99m and rhenium-188

    SciTech Connect

    Knapp, F.R. Jr.; Beets, A.L.; Mirzadeh, S.; Guhlke, S. |

    1998-03-01

    In this paper the authors describe the first application of a simple and inexpensive post elution tandem cation-anion exchange column system which is based on generator elution with salts of weak acids such as ammonium acetate instead of saline solution to provide very high specific volume solutions of technetium-99m and rhenium-188 from clinical scale molybdenum-99/technetium-99m generator prepared from low specific activity (n,y) molybdenum-99, and tungsten-188/rhenium-188 generators, respectively. Initial passage of the bolus through a strong cation exchange cartridge converts the ammonium acetate to acetic acid which is essentially not ionized at the acidic pH, allowing specific subsequent amine type (QMA SepPak{trademark}) anion exchange cartridge column trapping of the microscopic levels of the pertechnetate or perrhenate. Subsequent elution of the anion cartridge with a small volume (< 1 mL) of saline then provides high specific volume solutions of technetium-99m by concentration of the high eluant volumes obtained by elution of clinical-scale (1 Ci) generators. This new approach also works very effectively to obtain high specific volume solutions of rhenium-188 (> 500 mCi/mL) from the alumina-based tungsten-188/rhenium-188 generator.