Structural Basis for Activation of ZAP-70 by Phosphorylation of the SH2-Kinase Linker

PubMed Central

Yan, Qingrong; Barros, Tiago; Visperas, Patrick R.; Deindl, Sebastian; Kadlecek, Theresa A.; Weiss, Arthur

2013-01-01

Serial activation of the tyrosine kinases Lck and ZAP-70 initiates signaling downstream of the T cell receptor. We previously reported the structure of an autoinhibited ZAP-70 variant in which two regulatory tyrosine residues (315 and 319) in the SH2-kinase linker were replaced by phenylalanine. We now present a crystal structure of ZAP-70 in which Tyr 315 and Tyr 319 are not mutated, leading to the recognition of a five-residue sequence register error in the SH2-kinase linker of the original crystallographic model. The revised model identifies distinct roles for these two tyrosines. As seen in a recently reported structure of the related tyrosine kinase Syk, Tyr 315 of ZAP-70 is part of a hydrophobic interface between the regulatory apparatus and the kinase domain, and the integrity of this interface would be lost upon engagement of doubly phosphorylated peptides by the SH2 domains. Tyr 319 is not necessarily dislodged by SH2 engagement, which activates ZAP-70 only ∼5-fold in vitro. In contrast, phosphorylation by Lck activates ZAP-70 ∼100-fold. This difference is due to the ability of Tyr 319 to suppress ZAP-70 activity even when the SH2 domains are dislodged from the kinase domain, providing stringent control of ZAP-70 activity downstream of Lck. PMID:23530057

Structural basis for activation of ZAP-70 by phosphorylation of the SH2-kinase linker.

PubMed

Yan, Qingrong; Barros, Tiago; Visperas, Patrick R; Deindl, Sebastian; Kadlecek, Theresa A; Weiss, Arthur; Kuriyan, John

2013-06-01

Serial activation of the tyrosine kinases Lck and ZAP-70 initiates signaling downstream of the T cell receptor. We previously reported the structure of an autoinhibited ZAP-70 variant in which two regulatory tyrosine residues (315 and 319) in the SH2-kinase linker were replaced by phenylalanine. We now present a crystal structure of ZAP-70 in which Tyr 315 and Tyr 319 are not mutated, leading to the recognition of a five-residue sequence register error in the SH2-kinase linker of the original crystallographic model. The revised model identifies distinct roles for these two tyrosines. As seen in a recently reported structure of the related tyrosine kinase Syk, Tyr 315 of ZAP-70 is part of a hydrophobic interface between the regulatory apparatus and the kinase domain, and the integrity of this interface would be lost upon engagement of doubly phosphorylated peptides by the SH2 domains. Tyr 319 is not necessarily dislodged by SH2 engagement, which activates ZAP-70 only ∼5-fold in vitro. In contrast, phosphorylation by Lck activates ZAP-70 ∼100-fold. This difference is due to the ability of Tyr 319 to suppress ZAP-70 activity even when the SH2 domains are dislodged from the kinase domain, providing stringent control of ZAP-70 activity downstream of Lck.

Role of zinc-finger anti-viral protein in host defense against Sindbis virus

PubMed Central

Kozaki, Tatsuya; Takahama, Michihiro; Misawa, Takuma; Matsuura, Yoshiharu; Saitoh, Tatsuya

2015-01-01

Accumulating evidence indicates that type I interferon (IFN) mediates the host protective response to RNA viruses. However, the anti-viral effector molecules involved in this response have not been fully identified. Here, we show that zinc-finger anti-viral protein (ZAP), an IFN-inducible gene, plays a critical role in the elimination of Sindbis virus (SINV) in vitro and in vivo. The loss of ZAP greatly enhances the replication of SINV but does not inhibit type I IFN production in primary mouse embryonic fibroblasts (MEFs). ZAP binds and destabilizes SINV RNA, thereby suppressing the replication of SINV. Type I IFN fails to suppress SINV replication in ZAP-deficient MEFs, whereas the ectopic expression of ZAP is sufficient to suppress the replication of SINV in MEFs lacking the expression of type I IFN and the IFN-inducible genes. ZAP-deficient mice are highly susceptible to SINV infection, although they produce sufficient amounts of type I IFN. Therefore, ZAP is an RNA-sensing anti-viral effector molecule that mediates the type-I-IFN-dependent host defense against SINV. PMID:25758257

Identification of putative Z-ring-associated proteins, involved in cell division in human pathogenic bacteria Helicobacter pylori.

PubMed

Kamran, Mohammad; Sinha, Swati; Dubey, Priyanka; Lynn, Andrew M; Dhar, Suman K

2016-07-01

Cell division in bacteria is initiated by FtsZ, which forms a Z ring at the middle of the cell, between the nucleoids. The Z ring is stabilized by Z ring-associated proteins (Zaps), which crosslink the FtsZ filaments and provide strength. The deletion of Zaps leads to the elongation phenotype with an abnormal Z ring. The components of cell division in Helicobacter pylori are similar to other gram negative bacteria except for the absence of few components including Zaps. Here, we used HHsearch to identify homologs of the missing cell division proteins and got potential hits for ZapA and ZapB, as well as for few other cell division proteins. We further validated the function of the putative ZapA homolog by genetic complementation, immuno-colocalization and biochemical analysis. © 2016 Federation of European Biochemical Societies.

Zoonoses action plan Salmonella monitoring programme: an investigation of the sampling protocol.

PubMed

Snary, E L; Munday, D K; Arnold, M E; Cook, A J C

2010-03-01

The Zoonoses Action Plan (ZAP) Salmonella Programme was established by the British Pig Executive to monitor Salmonella prevalence in quality-assured British pigs at slaughter by testing a sample of pigs with a meat juice enzyme-linked immunosorbent assay for antibodies against group B and C(1) Salmonella. Farms were assigned a ZAP level (1 to 3) depending on the monitored prevalence, and ZAP 2 or 3 farms were required to act to reduce the prevalence. The ultimate goal was to reduce the risk of human salmonellosis attributable to British pork. A mathematical model has been developed to describe the ZAP sampling protocol. Results show that the probability of assigning a farm the correct ZAP level was high, except for farms that had a seroprevalence close to the cutoff points between different ZAP levels. Sensitivity analyses identified that the probability of assigning a farm to the correct ZAP level was dependent on the sensitivity and specificity of the test, the number of batches taken to slaughter each quarter, and the number of samples taken per batch. The variability of the predicted seroprevalence was reduced as the number of batches or samples increased and, away from the cutoff points, the probability of being assigned the correct ZAP level increased as the number of batches or samples increased. In summary, the model described here provided invaluable insight into the ZAP sampling protocol. Further work is required to understand the impact of the program for Salmonella infection in British pig farms and therefore on human health.

The Structural Basis for Activation and Inhibition of ZAP-70 Kinase Domain.

PubMed

Huber, Roland G; Fan, Hao; Bond, Peter J

2015-10-01

ZAP-70 (Zeta-chain-associated protein kinase 70) is a tyrosine kinase that interacts directly with the activated T-cell receptor to transduce downstream signals, and is hence a major player in the regulation of the adaptive immune response. Dysfunction of ZAP-70 causes selective T cell deficiency that in turn results in persistent infections. ZAP-70 is activated by a variety of signals including phosphorylation of the kinase domain (KD), and binding of its regulatory tandem Src homology 2 (SH2) domains to the T cell receptor. The present study investigates molecular mechanisms of activation and inhibition of ZAP-70 via atomically detailed molecular dynamics simulation approaches. We report microsecond timescale simulations of five distinct states of the ZAP-70 KD, comprising apo, inhibited and three phosphorylated variants. Extensive analysis of local flexibility and correlated motions reveal crucial transitions between the states, thus elucidating crucial steps in the activation mechanism of the ZAP-70 KD. Furthermore, we rationalize previously observed staurosporine-bound crystal structures, suggesting that whilst the KD superficially resembles an "active-like" conformation, the inhibitor modulates the underlying protein dynamics and restricts it in a compact, rigid state inaccessible to ligands or cofactors. Finally, our analysis reveals a novel, potentially druggable pocket in close proximity to the activation loop of the kinase, and we subsequently use its structure in fragment-based virtual screening to develop a pharmacophore model. The pocket is distinct from classical type I or type II kinase pockets, and its discovery offers promise in future design of specific kinase inhibitors, whilst mutations in residues associated with this pocket are implicated in immunodeficiency in humans.

ZAP-70 compared with immunoglobulin heavy-chain gene mutation status as a predictor of disease progression in chronic lymphocytic leukemia.

PubMed

Rassenti, Laura Z; Huynh, Lang; Toy, Tracy L; Chen, Liguang; Keating, Michael J; Gribben, John G; Neuberg, Donna S; Flinn, Ian W; Rai, Kanti R; Byrd, John C; Kay, Neil E; Greaves, Andrew; Weiss, Arthur; Kipps, Thomas J

2004-08-26

The course of chronic lymphocytic leukemia (CLL) is variable. In aggressive disease, the CLL cells usually express an unmutated immunoglobulin heavy-chain variable-region gene (IgV(H)) and the 70-kD zeta-associated protein (ZAP-70), whereas in indolent disease, the CLL cells usually express mutated IgV(H) but lack expression of ZAP-70. We evaluated the CLL B cells from 307 patients with CLL for ZAP-70 and mutations in the rearranged IgV(H) gene. We then investigated the association between the results and the time from diagnosis to initial therapy. We found that ZAP-70 was expressed above a defined threshold level in 117 of the 164 patients with an unmutated IgV(H) gene (71 percent), but in only 24 of the 143 patients with a mutated IgV(H) gene (17 percent, P<0.001). Among the patients with ZAP-70-positive CLL cells, the median time from diagnosis to initial therapy in those who had an unmutated IgV(H) gene (2.8 years) was not significantly different from the median time in those who had a mutated IgV(H) gene (4.2 years, P=0.07). However, the median time from diagnosis to initial treatment in each of these groups was significantly shorter than the time in patients with ZAP-70-negative CLL cells who had either mutated or unmutated IgV(H) genes (P<0.001). The median time from diagnosis to initial therapy among patients who did not have ZAP-70 was 11.0 years in those with a mutated IgV(H) gene and 7.1 years in those with an unmutated IgV(H) gene (P<0.001). Although the presence of an unmutated IgV(H) gene is strongly associated with the expression of ZAP-70, ZAP-70 is a stronger predictor of the need for treatment in B-cell CLL. Copyright 2004 Massachusetts Medical Society

Kolmogorov-Smirnov statistical test for analysis of ZAP-70 expression in B-CLL, compared with quantitative PCR and IgV(H) mutation status.

PubMed

Van Bockstaele, Femke; Janssens, Ann; Piette, Anne; Callewaert, Filip; Pede, Valerie; Offner, Fritz; Verhasselt, Bruno; Philippé, Jan

2006-07-15

ZAP-70 has been proposed as a surrogate marker for immunoglobulin heavy-chain variable region (IgV(H)) mutation status, which is known as a prognostic marker in B-cell chronic lymphocytic leukemia (CLL). The flow cytometric analysis of ZAP-70 suffers from difficulties in standardization and interpretation. We applied the Kolmogorov-Smirnov (KS) statistical test to make analysis more straightforward. We examined ZAP-70 expression by flow cytometry in 53 patients with CLL. Analysis was performed as initially described by Crespo et al. (New England J Med 2003; 348:1764-1775) and alternatively by application of the KS statistical test comparing T cells with B cells. Receiver-operating-characteristics (ROC)-curve analyses were performed to determine the optimal cut-off values for ZAP-70 measured by the two approaches. ZAP-70 protein expression was compared with ZAP-70 mRNA expression measured by a quantitative PCR (qPCR) and with the IgV(H) mutation status. Both flow cytometric analyses correlated well with the molecular technique and proved to be of equal value in predicting the IgV(H) mutation status. Applying the KS test is reproducible, simple, straightforward, and overcomes a number of difficulties encountered in the Crespo-method. The KS statistical test is an essential part of the software delivered with modern routine analytical flow cytometers and is well suited for analysis of ZAP-70 expression in CLL. (c) 2006 International Society for Analytical Cytology.

ZAPs: Using Interactive Programs for Learning Psychology

ERIC Educational Resources Information Center

Hulshof, Casper D.; Eysink, Tessa H. S.; Loyens, Sofie; de Jong, Ton

2005-01-01

ZAPs are short, self-contained computer programs that encourage students to experience psychological phenomena in a vivid, self-explanatory way, and that are meant to evoke enthusiasm about psychological topics. ZAPs were designed according to principles that originate from experiential and discovery learning theories. The interactive approach…

Phosphorylation of SLP-76 by the ZAP-70 protein-tyrosine kinase is required for T-cell receptor function.

PubMed

Bubeck Wardenburg, J; Fu, C; Jackman, J K; Flotow, H; Wilkinson, S E; Williams, D H; Johnson, R; Kong, G; Chan, A C; Findell, P R

1996-08-16

Two families of tyrosine kinases, the Src and Syk families, are required for T-cell receptor activation. While the Src kinases are responsible for phosphorylation of receptor-encoded signaling motifs and for up-regulation of ZAP-70 activity, the downstream substrates of ZAP-70 are unknown. Evidence is presented herein that the Src homology 2 (SH2) domain-containing leukocyte protein of 76 kDa (SLP-76) is a substrate of ZAP-70. Phosphorylation of SLP-76 is diminished in T cells that express a catalytically inactive ZAP-70. Moreover, SLP-76 is preferentially phosphorylated by ZAP-70 in vitro and in heterologous cellular systems. In T cells, overexpression of wild-type SLP-76 results in a hyperactive receptor, while expression of a SLP-76 molecule that is unable to be tyrosine-phosphorylated attenuates receptor function. In addition, the SH2 domain of SLP-76 is required for T-cell receptor function, although its role is independent of the ability of SLP-76 to undergo tyrosine phosphorylation. As SLP-76 interacts with both Grb2 and phospholipase C-gamma1, these data indicate that phosphorylation of SLP-76 by ZAP-70 provides an important functional link between the T-cell receptor and activation of ras and calcium pathways.

Trichostatin A down-regulates ZAP-70, LAT and SLP-76 content in Jurkat T cells.

PubMed

Januchowski, Radosław; Jagodzinski, Paweł P

2007-02-01

We exploited Jurkat leukemia T cell clone E6-1 as a model of Trichostatin A (TSA) effect on cellular levels of ZAP-70, LAT and SLP-76 molecules involved in the signal transduction pathway from T cell receptor to nucleus. Using reverse transcription real-time quantitative PCR and Western blotting analysis we observed that TSA resulted in ZAP-70, LAT and SLP-76 transcript and protein down-regulation in Jurkat leukemia T cells. We also found that TSA reduced half-life of ZAP-70, LAT and SLP-76 mRNAs from 4.8, 3.5, and 4.8 to approximately 2.3, 1.9 and 1.7 h, respectively. Employing the protein biosynthesis inhibitor cycloheximide, we demonstrated the involvement of RNase and/or mRNA stabilization protein in ZAP-70, LAT and SLP-76 mRNAs stabilization. The effect of TSA on ZAP-70, LAT and SLP-76 content in T cells confirms an immunosuppressive effect by TSA, and the usefulness of this histone deacetylase inhibitor in the treatment of autoimmune diseases.

ZapE Is a Novel Cell Division Protein Interacting with FtsZ and Modulating the Z-Ring Dynamics

PubMed Central

Marteyn, Benoit S.; Karimova, Gouzel; Fenton, Andrew K.; Gazi, Anastasia D.; West, Nicholas; Touqui, Lhousseine; Prevost, Marie-Christine; Betton, Jean-Michel; Poyraz, Oemer; Ladant, Daniel; Gerdes, Kenn; Sansonetti, Philippe J.; Tang, Christoph M.

2014-01-01

ABSTRACT Bacterial cell division requires the formation of a mature divisome complex positioned at the midcell. The localization of the divisome complex is determined by the correct positioning, assembly, and constriction of the FtsZ ring (Z-ring). Z-ring constriction control remains poorly understood and (to some extent) controversial, probably due to the fact that this phenomenon is transient and controlled by numerous factors. Here, we characterize ZapE, a novel ATPase found in Gram-negative bacteria, which is required for growth under conditions of low oxygen, while loss of zapE results in temperature-dependent elongation of cell shape. We found that ZapE is recruited to the Z-ring during late stages of the cell division process and correlates with constriction of the Z-ring. Overexpression or inactivation of zapE leads to elongation of Escherichia coli and affects the dynamics of the Z-ring during division. In vitro, ZapE destabilizes FtsZ polymers in an ATP-dependent manner. PMID:24595368

Variations in the detection of ZAP-70 in chronic lymphocytic leukemia: Comparison with IgV(H) mutation analysis.

PubMed

Sheikholeslami, M R; Jilani, I; Keating, M; Uyeji, J; Chen, K; Kantarjian, H; O'Brien, S; Giles, F; Albitar, M

2006-07-15

Lack of immunoglobulin heavy chain genes (IgV(H)) mutation in patients with chronic lymphocytic leukemia (CLL) is associated with rapid disease progression and shorter survival. The zeta-chain (T-cell receptor) associated protein kinase 70 kDa (ZAP-70) has been reported to be a surrogate marker for IgV(H) mutation status, and its expression in leukemic cells correlates with unmutated IgV(H). However, ZAP-70 detection by flow cytometry varies significantly dependant on the antibodies used, the method of performing the assay, and the condition of the cells in the specimen. The clinical value of ZAP-70 testing when samples are shipped under poorly controlled conditions is not known. Furthermore, testing in a research environment may differ from testing in a routine clinical laboratory. We validated an assay for ZAP-70 by comparing results with clinical outcome and the mutation status of the IgV(H). Using stored samples, we show significant correlation between ZAP-70 expression and clinical outcome as well as IgV(H) mutation at a cut-off point of 15%. While positive samples (>15% positivity) remain positive when kept in the laboratory environment for 48 h after initial testing, results obtained from samples from CLL patients tested after shipping at room temperature for routine testing showed no correlation with IgV(H) mutation status when 15% cut-off was used. In these samples, cut-point of 10% correlated with the IgV(H) mutation (P = 0.0001). This data suggests that although ZAP-70 positivity correlates with IgV(H) mutation status and survival, variations in sample handling and preparation may influence results. We show that IgV(H) mutation results, unlike ZAP-70 remain correlated with CD38 expression and beta-2 microglobulin in shipped samples, and ZAP-70 testing should not be used as the sole criterion for stratifying patients for therapy. (c) 2006 International Society for Analytical Cytology.

MFI ratio estimation of ZAP-70 in B-CLL by flow cytometry can be improved by considering the isotype-matched antibody signal.

PubMed

Marquez, M-E; Deglesne, P-A; Suarez, G; Romano, E

2011-04-01

The IgV(H) mutational status of B-cell chronic lymphocytic leukemia (B-CLL) is of prognostic value. Expression of ZAP-70 in B-CLL is a surrogate marker for IgV(H) unmutated (UM). As determination of IgV(H) mutational status involves a methodology currently unavailable for most clinical laboratories, it is important to have available a reliable technique for ZAP-70 estimation in B-CLL. Flow cytometry (FC) is a convenient technique for this purpose. However, there is still no adequate way for data analysis, which would prevent the assignment of false positive or negative expression. We have modified the currently most accepted technique, which uses the ratio of the mean fluorescent index (MFI) of B-CLL to T cells. The MFI for parallel antibody isotype staining is subtracted from the ZAP-70 MFI of both B-CLL and T cells. We validated this technique comparing the results obtained for ZAP-70 expression by FC with those obtained with quantitative PCR for the same patients. We applied the technique in a series of 53 patients. With this modification, a better correlation between ZAP-70 expression and IgV(H) UM was obtained. Thus, the MFI ratio B-CLL/T cell corrected by isotype is a reliable analysis technique to estimate ZAP-70 expression in B-CLL. © 2010 Blackwell Publishing Ltd.

Modification by covalent reaction or oxidation of cysteine residues in the tandem-SH2 domains of ZAP-70 and Syk can block phosphopeptide binding.

PubMed

Visperas, Patrick R; Winger, Jonathan A; Horton, Timothy M; Shah, Neel H; Aum, Diane J; Tao, Alyssa; Barros, Tiago; Yan, Qingrong; Wilson, Christopher G; Arkin, Michelle R; Weiss, Arthur; Kuriyan, John

2015-01-01

Zeta-chain associated protein of 70 kDa (ZAP-70) and spleen tyrosine kinase (Syk) are non-receptor tyrosine kinases that are essential for T-cell and B-cell antigen receptor signalling respectively. They are recruited, via their tandem-SH2 (Src-homology domain 2) domains, to doubly phosphorylated immunoreceptor tyrosine-based activation motifs (ITAMs) on invariant chains of immune antigen receptors. Because of their critical roles in immune signalling, ZAP-70 and Syk are targets for the development of drugs for autoimmune diseases. We show that three thiol-reactive small molecules can prevent the tandem-SH2 domains of ZAP-70 and Syk from binding to phosphorylated ITAMs. We identify a specific cysteine residue in the phosphotyrosine-binding pocket of each protein (Cys39 in ZAP-70, Cys206 in Syk) that is necessary for inhibition by two of these compounds. We also find that ITAM binding to ZAP-70 and Syk is sensitive to the presence of H2O2 and these two cysteine residues are also necessary for inhibition by H2O2. Our findings suggest a mechanism by which the reactive oxygen species generated during responses to antigen could attenuate signalling through these kinases and may also inform the development of ZAP-70 and Syk inhibitors that bind covalently to their SH2 domains.

Modification by covalent reaction or oxidation of cysteine residues in the Tandem-SH2 Domains of ZAP-70 and Syk Can Block Phosphopeptide Binding

PubMed Central

Visperas, Patrick R.; Winger, Jonathan A.; Horton, Timothy M.; Shah, Neel H.; Aum, Diane J.; Tao, Alyssa; Barros, Tiago; Yan, Qingrong; Wilson, Christopher G.; Arkin, Michelle R.; Weiss, Arthur; Kuriyan, John

2015-01-01

Zeta-chain Associated Protein of 70kDa (ZAP-70) and Spleen tyrosine kinase (Syk) are non-receptor tyrosine kinases that are essential for T-cell and B-cell antigen receptor signaling, respectively. They are recruited, via their tandem-SH2 domains, to doubly-phosphorylated Immunoreceptor Tyrosine-based Activation Motifs (ITAMs) on invariant chains of immune antigen receptors. Because of their critical roles in immune signaling, ZAP-70 and Syk are targets for the development of drugs for autoimmune diseases. We show that three thiol-reactive small molecules can prevent the tandem-SH2 domains of ZAP-70 and Syk from binding to phosphorylated ITAMs. We identify a specific cysteine residue in the phosphotyrosine-binding pocket of each protein (Cys 39 in ZAP-70, Cys 206 in Syk) that is necessary for inhibition by two of these compounds. We also find that ITAM binding to ZAP-70 and Syk is sensitive to the presence of hydrogen peroxide, and these two cysteine residues are also necessary for inhibition by hydrogen peroxide. Our findings suggest a mechanism by which the generation of reactive oxygen species generated during responses to antigen could attenuate signaling through these kinases, and may also inform the development of ZAP-70 and Syk inhibitors that bind covalently to their SH2 domains. PMID:25287889

The ZAP Project: Designing Interactive Computer Tools for Learning Psychology

ERIC Educational Resources Information Center

Hulshof, Casper; Eysink, Tessa; de Jong, Ton

2006-01-01

In the ZAP project, a set of interactive computer programs called "ZAPs" was developed. The programs were designed in such a way that first-year students experience psychological phenomena in a vivid and self-explanatory way. Students can either take the role of participant in a psychological experiment, they can experience phenomena themselves,…

The Effect of Six Production Variables on Recall of Television Commercials during Fast Forward "Zapping."

ERIC Educational Resources Information Center

White, Sylvia E.; And Others

In recent years, broadcast advertisers have become concerned with the tendency of video cassette recorder (VCR) owners to "zap"--fast forward delete--commercials. However, research has shown that viewers retain some recall of commercials, even with fast forward zapping, and it is to the benefit of advertisers to discover ways of…

Silencing Inhibits Cre-Mediated Recombination of the Z/AP and Z/EG Reporters in Adult Cells

PubMed Central

Long, Michael A.; Rossi, Fabio M. V.

2009-01-01

Background The Cre-loxP system has been used to enable tissue specific activation, inactivation and mutation of many genes in vivo and has thereby greatly facilitated the genetic dissection of several cellular and developmental processes. In such studies, Cre-reporter strains, which carry a Cre-activated marker gene, are frequently utilized to validate the expression profile of Cre transgenes, to act as a surrogate marker for excision of a second allele, and to irreversibly label cells for lineage tracing experiments. Principal Findings We have studied three commonly used Cre-reporter strains, Z/AP, Z/EG and R26R-EYFP and have demonstrated that although each reporter can be reliably activated by Cre during early development, exposure to Cre in adult hematopoietic cells results in a much lower frequency of marker-positive cells in the Z/AP or Z/EG strains than in the R26R-EYFP strain. In marker negative cells derived from the Z/AP and Z/EG strains, the transgenic promoter is methylated and Cre-mediated recombination of the locus is inhibited. Conclusions These results show that the efficiency of Cre-mediated recombination is not only dependent on the genomic context of a given loxP-flanked sequence, but also on stochastic epigenetic mechanisms underlying transgene variegation. Furthermore, our data highlights the potential shortcomings of utilizing the Z/AP and Z/EG reporters as surrogate markers of excision or in lineage tracing experiments. PMID:19415111

Silencing inhibits Cre-mediated recombination of the Z/AP and Z/EG reporters in adult cells.

PubMed

Long, Michael A; Rossi, Fabio M V

2009-01-01

The Cre-loxP system has been used to enable tissue specific activation, inactivation and mutation of many genes in vivo and has thereby greatly facilitated the genetic dissection of several cellular and developmental processes. In such studies, Cre-reporter strains, which carry a Cre-activated marker gene, are frequently utilized to validate the expression profile of Cre transgenes, to act as a surrogate marker for excision of a second allele, and to irreversibly label cells for lineage tracing experiments. We have studied three commonly used Cre-reporter strains, Z/AP, Z/EG and R26R-EYFP and have demonstrated that although each reporter can be reliably activated by Cre during early development, exposure to Cre in adult hematopoietic cells results in a much lower frequency of marker-positive cells in the Z/AP or Z/EG strains than in the R26R-EYFP strain. In marker negative cells derived from the Z/AP and Z/EG strains, the transgenic promoter is methylated and Cre-mediated recombination of the locus is inhibited. These results show that the efficiency of Cre-mediated recombination is not only dependent on the genomic context of a given loxP-flanked sequence, but also on stochastic epigenetic mechanisms underlying transgene variegation. Furthermore, our data highlights the potential shortcomings of utilizing the Z/AP and Z/EG reporters as surrogate markers of excision or in lineage tracing experiments.

77 FR 56703 - Public Hearing to Determine Whether ZAP Has Met Notification and Remedy Requirements

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-13

...: NHTSA will hold a public hearing on whether ZAP,\\1\\ a publicly owned company based in Santa Rosa... those noncompliances in two recalls involving Model Year (MY) 2008 ZAP Xebra three-wheeled vehicles.../ . DATES: The public hearing will be held beginning at 10 a.m. ET on October 9, 2012 in the Oklahoma City...

The Touch and Zap method for in vivo whole-cell patch recording of intrinsic and visual responses of cortical neurons and glial cells.

PubMed

Schramm, Adrien E; Marinazzo, Daniele; Gener, Thomas; Graham, Lyle J

2014-01-01

Whole-cell patch recording is an essential tool for quantitatively establishing the biophysics of brain function, particularly in vivo. This method is of particular interest for studying the functional roles of cortical glial cells in the intact brain, which cannot be assessed with extracellular recordings. Nevertheless, a reasonable success rate remains a challenge because of stability, recording duration and electrical quality constraints, particularly for voltage clamp, dynamic clamp or conductance measurements. To address this, we describe "Touch and Zap", an alternative method for whole-cell patch clamp recordings, with the goal of being simpler, quicker and more gentle to brain tissue than previous approaches. Under current clamp mode with a continuous train of hyperpolarizing current pulses, seal formation is initiated immediately upon cell contact, thus the "Touch". By maintaining the current injection, whole-cell access is spontaneously achieved within seconds from the cell-attached configuration by a self-limited membrane electroporation, or "Zap", as seal resistance increases. We present examples of intrinsic and visual responses of neurons and putative glial cells obtained with the revised method from cat and rat cortices in vivo. Recording parameters and biophysical properties obtained with the Touch and Zap method compare favourably with those obtained with the traditional blind patch approach, demonstrating that the revised approach does not compromise the recorded cell. We find that the method is particularly well-suited for whole-cell patch recordings of cortical glial cells in vivo, targeting a wider population of this cell type than the standard method, with better access resistance. Overall, the gentler Touch and Zap method is promising for studying quantitative functional properties in the intact brain with minimal perturbation of the cell's intrinsic properties and local network. Because the Touch and Zap method is performed semi-automatically, this approach is more reproducible and less dependent on experimenter technique.

The role of MatP, ZapA and ZapB in chromosomal organization and dynamics in Escherichia coli

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mannik, Jaana; Castillo, Daniel E.; Yang, Da

Despite extensive research over several decades, a comprehensive view of how the Escherichia coli chromosome is organized within the nucleoid, and how two daughter chromosomes segregate has yet to emerge. Here we investigate the role of the MatP, ZapA and ZapB proteins in organizing the replication terminus (Ter) region and in the chromosomal segregation process. Quantitative image analysis of the fluorescently labeled Ter region shows that the replication terminus attaches to the divisome in a single segment along the perimeter of the cell in a MatP, ZapA and ZapB-dependent manner. The attachment does not significantly affect the bulk chromosome segregationmore » in slow growth conditions. With or without the attachment, two chromosomal masses separate from each other at a speed comparable to the cell growth. The separation starts even before the replication terminus region positions itself at the center of the nucleoid. Modeling of the segregation based on conformational entropy correctly predicts the positioning of the replication terminus region within the nucleoid. Furthermore, the model produces a distinctly different chromosomal density distribution than the experiment, indicating that the conformational entropy plays a limited role in segregating the chromosomes in the late stages of replication.« less

The role of MatP, ZapA and ZapB in chromosomal organization and dynamics in Escherichia coli

DOE PAGES

Mannik, Jaana; Castillo, Daniel E.; Yang, Da; ...

2016-01-13

Despite extensive research over several decades, a comprehensive view of how the Escherichia coli chromosome is organized within the nucleoid, and how two daughter chromosomes segregate has yet to emerge. Here we investigate the role of the MatP, ZapA and ZapB proteins in organizing the replication terminus (Ter) region and in the chromosomal segregation process. Quantitative image analysis of the fluorescently labeled Ter region shows that the replication terminus attaches to the divisome in a single segment along the perimeter of the cell in a MatP, ZapA and ZapB-dependent manner. The attachment does not significantly affect the bulk chromosome segregationmore » in slow growth conditions. With or without the attachment, two chromosomal masses separate from each other at a speed comparable to the cell growth. The separation starts even before the replication terminus region positions itself at the center of the nucleoid. Modeling of the segregation based on conformational entropy correctly predicts the positioning of the replication terminus region within the nucleoid. Furthermore, the model produces a distinctly different chromosomal density distribution than the experiment, indicating that the conformational entropy plays a limited role in segregating the chromosomes in the late stages of replication.« less

A new multivariate zero-adjusted Poisson model with applications to biomedicine.

PubMed

Liu, Yin; Tian, Guo-Liang; Tang, Man-Lai; Yuen, Kam Chuen

2018-05-25

Recently, although advances were made on modeling multivariate count data, existing models really has several limitations: (i) The multivariate Poisson log-normal model (Aitchison and Ho, ) cannot be used to fit multivariate count data with excess zero-vectors; (ii) The multivariate zero-inflated Poisson (ZIP) distribution (Li et al., 1999) cannot be used to model zero-truncated/deflated count data and it is difficult to apply to high-dimensional cases; (iii) The Type I multivariate zero-adjusted Poisson (ZAP) distribution (Tian et al., 2017) could only model multivariate count data with a special correlation structure for random components that are all positive or negative. In this paper, we first introduce a new multivariate ZAP distribution, based on a multivariate Poisson distribution, which allows the correlations between components with a more flexible dependency structure, that is some of the correlation coefficients could be positive while others could be negative. We then develop its important distributional properties, and provide efficient statistical inference methods for multivariate ZAP model with or without covariates. Two real data examples in biomedicine are used to illustrate the proposed methods. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A novel human autoimmune syndrome caused by combined hypomorphic and activating mutations in ZAP-70

PubMed Central

Chan, Alice Y.; Punwani, Divya; Kadlecek, Theresa A.; Cowan, Morton J.; Olson, Jean L.; Mathes, Erin F.; Sunderam, Uma; Man Fu, Shu; Srinivasan, Rajgopal; Kuriyan, John; Brenner, Steven E.; Weiss, Arthur

2016-01-01

A brother and sister developed a previously undescribed constellation of autoimmune manifestations within their first year of life, with uncontrollable bullous pemphigoid, colitis, and proteinuria. The boy had hemophilia due to a factor VIII autoantibody and nephrotic syndrome. Both children required allogeneic hematopoietic cell transplantation (HCT), which resolved their autoimmunity. The early onset, severity, and distinctive findings suggested a single gene disorder underlying the phenotype. Whole-exome sequencing performed on five family members revealed the affected siblings to be compound heterozygous for two unique missense mutations in the 70-kD T cell receptor ζ-chain associated protein (ZAP-70). Healthy relatives were heterozygous mutation carriers. Although pre-HCT patient T cells were not available, mutation effects were determined using transfected cell lines and peripheral blood from carriers and controls. Mutation R192W in the C-SH2 domain exhibited reduced binding to phosphorylated ζ-chain, whereas mutation R360P in the N lobe of the catalytic domain disrupted an autoinhibitory mechanism, producing a weakly hyperactive ZAP-70 protein. Although human ZAP-70 deficiency can have dysregulated T cells, and autoreactive mouse thymocytes with weak Zap-70 signaling can escape tolerance, our patients’ combination of hypomorphic and activating mutations suggested a new disease mechanism and produced previously undescribed human ZAP-70–associated autoimmune disease. PMID:26783323

Phosphorylation of a Tyrosine Residue on Zap70 by Lck and Its Subsequent Binding via an SH2 Domain May Be a Key Gatekeeper of T Cell Receptor Signaling In Vivo.

PubMed

Thill, Peter A; Weiss, Arthur; Chakraborty, Arup K

2016-09-15

The initiation of signaling in T lymphocytes in response to the binding of the T cell receptor (TCR) to cognate ligands is a key step in the emergence of adaptive immune responses. Conventional models posit that TCR signaling is initiated by the phosphorylation of receptor-associated immune receptor activation motifs (ITAMs). The cytoplasmic tyrosine kinase Zap70 binds to phosphorylated ITAMs, is subsequently activated, and then propagates downstream signaling. While evidence for such models is provided by experiments with cell lines, in vivo, Zap70 is bound to phosphorylated ITAMs in resting T cells. However, Zap70 is activated only upon TCR binding to cognate ligand. We report the results of computational studies of a new model for the initiation of TCR signaling that incorporates these in vivo observations. Importantly, the new model is shown to allow better and faster TCR discrimination between self-ligands and foreign ligands. The new model is consistent with many past experimental observations, and experiments that could further test the model are proposed. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

A novel potential/viscous flow coupling technique for computing helicopter flow fields

NASA Technical Reports Server (NTRS)

Summa, J. Michael; Strash, Daniel J.; Yoo, Sungyul

1993-01-01

The primary objective of this work was to demonstrate the feasibility of a new potential/viscous flow coupling procedure for reducing computational effort while maintaining solution accuracy. This closed-loop, overlapped velocity-coupling concept has been developed in a new two-dimensional code, ZAP2D (Zonal Aerodynamics Program - 2D), a three-dimensional code for wing analysis, ZAP3D (Zonal Aerodynamics Program - 3D), and a three-dimensional code for isolated helicopter rotors in hover, ZAPR3D (Zonal Aerodynamics Program for Rotors - 3D). Comparisons with large domain ARC3D solutions and with experimental data for a NACA 0012 airfoil have shown that the required domain size can be reduced to a few tenths of a percent chord for the low Mach and low angle of attack cases and to less than 2-5 chords for the high Mach and high angle of attack cases while maintaining solution accuracies to within a few percent. This represents CPU time reductions by a factor of 2-4 compared with ARC2D. The current ZAP3D calculation for a rectangular plan-form wing of aspect ratio 5 with an outer domain radius of about 1.2 chords represents a speed-up in CPU time over the ARC3D large domain calculation by about a factor of 2.5 while maintaining solution accuracies to within a few percent. A ZAPR3D simulation for a two-bladed rotor in hover with a reduced grid domain of about two chord lengths was able to capture the wake effects and compared accurately with the experimental pressure data. Further development is required in order to substantiate the promise of computational improvements due to the ZAPR3D coupling concept.

Carbonaceous material production from vegetable residue and their use in the removal of textile dyes present in wastewater

NASA Astrophysics Data System (ADS)

Peláez-Cid, A. A.; Tlalpa-Galán, M. A.; Herrera-González, A. M.

2013-06-01

This paper presents the adsorption results of acid, basic, direct, vat, and reactive-type dyes on carbonaceous adsorbent materials prepared starting off vegetable residue such as Opuntia ficus indica and Casimiroa edulis fruit wastes. The adsorbents prepared from Opuntia ficus indica waste were designated: TunaAsh, CarTunaT, and CarTunaQ. The materials obtained from Casimiroa edulis waste were named: CenZAP, CarZAPT, and CarZAPQ. TunaAsh and CenZAP consist of ashes obtained at 550 °C CarTunaT and CarZAPT consist of the materials carbonized at 400 °C lastly, CarTunaQ and CarZAPQ consist of chemically activated carbons using H3PO4 at 400 °C. Only the chemically activated materials were washed with distilled water until a neutral pH was obtained after their carbonization. All materials were ground and sieved to obtain a particle size ranging from 0.25 to 0.84 mm. The static adsorption results showed that both ashes and chemically activated carbon are more efficient at dye removal for both vegetable residues. For TunaAsh and CarTunaQ, removal rates of up to 100% in the cases of basic, acid, and direct dyes were achieved. Regarding wastewater containing reactive dyes, the efficiency ranged from 60 to 100%. For vat effluents, it ranged from 42 to 52%. In the case of CenZAP and CarZAPQ, it was possible to treat reactive effluents with rates ranging between 63 and 91%. Regarding vat effluents, it ranged from 57 to 68%. The process of characterization for all materials was done using scanning electron microscopy and infrared spectroscopy.

EPA Registers the Wolbachia ZAP Strain in Live Male Asian Tiger Mosquitoes

EPA Pesticide Factsheets

EPA registered a new mosquito biopesticide – ZAP Males® - that can reduce local populations of the type of mosquito (Aedes albopictus, or Asian Tiger Mosquitoes) that can spread numerous diseases of significant human health concern, including the Zika viru

ZAP-70 staining in chronic lymphocytic leukemia.

PubMed

Villamor, Neus

2005-05-01

Chronic lymphocytic leukemia (CLL) is the most common chronic leukemia in Western countries. The disease has an extremely variable clinical course, and several prognostic features have been identified to assess individual risk. The configuration of the immunoglobulin variable heavy-chain gene (IgV(H)) is a strong predictor of the outcome. CLL patients with unmutated IgV(H) status have an aggressive clinical course and a short survival. Unfortunately, analysis of IgV(H) gene configuration is not available in most clinical laboratories. A small number of genes are differentially expressed between unmutated IgV(H) and mutated IgV(H) clinical forms of CLL. One of these genes is ZAP-70, which is detected in leukemic cells from patients with the unmutated IgV(H) form of CLL. Flow cytometry presents advantages over other methods to detect ZAP-70, and its quantification by flow cytometry has proved its predictive value. This unit focuses on protocols to quantify ZAP-70 by flow cytometry in CLL.

Progranulin Is a Novel Independent Predictor of Disease Progression and Overall Survival in Chronic Lymphocytic Leukemia

PubMed Central

Göbel, Maria; Eisele, Lewin; Möllmann, Michael; Hüttmann, Andreas; Johansson, Patricia; Scholtysik, René; Bergmann, Manuela; Busch, Raymonde; Döhner, Hartmut; Hallek, Michael; Seiler, Till; Stilgenbauer, Stephan; Klein-Hitpass, Ludger; Dührsen, Ulrich; Dürig, Jan

2013-01-01

Progranulin (Pgrn) is a 88 kDa secreted protein with pleiotropic functions including regulation of cell cycle progression, cell motility, wound repair and tumorigenesis. Using microarray based gene expression profiling we have recently demonstrated that the gene for Pgrn, granulin (GRN), is significantly higher expressed in aggressive CD38+ZAP-70+ as compared to indolent CD38−ZAP-70− chronic lymphocytic leukemia (CLL) cases. Here, we measured Pgrn plasma concentrations by enzyme-linked immunosorbent assay (ELISA) in the Essen CLL cohort of 131 patients and examined Pgrn for association with established prognostic markers and clinical outcome. We found that high Pgrn plasma levels were strongly associated with adverse risk factors including unmutated IGHV status, expression of CD38 and ZAP-70, poor risk cytogenetics (11q-, 17p-) as detected by flourescence in situ hybridization (FISH) and high Binet stage. Pgrn as well as the aforementioned risk factors were prognostic for time to first treatment and overall survival in this series. Importantly, these results could be confirmed in the independent multicentric CLL1 cohort of untreated Binet stage A patients (n = 163). Here, multivariate analysis of time to first treatment revealed that high risk Pgrn (HR = 2.06, 95%-CI = 1.13–3.76, p = 0.018), unmutated IGHV status (HR = 5.63, 95%-CI = 3.05–10.38, p<0.001), high risk as defined by the study protocol (HR = 2.06, 95%-CI = 1.09–3.89, p = 0.026) but not poor risk cytogenetics were independent prognostic markers. In summary our results suggest that Pgrn is a novel, robust and independent prognostic marker in CLL that can be easily measured by ELISA. PMID:24009671

Progranulin is a novel independent predictor of disease progression and overall survival in chronic lymphocytic leukemia.

PubMed

Göbel, Maria; Eisele, Lewin; Möllmann, Michael; Hüttmann, Andreas; Johansson, Patricia; Scholtysik, René; Bergmann, Manuela; Busch, Raymonde; Döhner, Hartmut; Hallek, Michael; Seiler, Till; Stilgenbauer, Stephan; Klein-Hitpass, Ludger; Dührsen, Ulrich; Dürig, Jan

2013-01-01

Progranulin (Pgrn) is a 88 kDa secreted protein with pleiotropic functions including regulation of cell cycle progression, cell motility, wound repair and tumorigenesis. Using microarray based gene expression profiling we have recently demonstrated that the gene for Pgrn, granulin (GRN), is significantly higher expressed in aggressive CD38(+)ZAP-70(+) as compared to indolent CD38(-)ZAP-70(-) chronic lymphocytic leukemia (CLL) cases. Here, we measured Pgrn plasma concentrations by enzyme-linked immunosorbent assay (ELISA) in the Essen CLL cohort of 131 patients and examined Pgrn for association with established prognostic markers and clinical outcome. We found that high Pgrn plasma levels were strongly associated with adverse risk factors including unmutated IGHV status, expression of CD38 and ZAP-70, poor risk cytogenetics (11q-, 17p-) as detected by flourescence in situ hybridization (FISH) and high Binet stage. Pgrn as well as the aforementioned risk factors were prognostic for time to first treatment and overall survival in this series. Importantly, these results could be confirmed in the independent multicentric CLL1 cohort of untreated Binet stage A patients (n = 163). Here, multivariate analysis of time to first treatment revealed that high risk Pgrn (HR = 2.06, 95%-CI = 1.13-3.76, p = 0.018), unmutated IGHV status (HR = 5.63, 95%-CI = 3.05-10.38, p<0.001), high risk as defined by the study protocol (HR = 2.06, 95%-CI = 1.09-3.89, p = 0.026) but not poor risk cytogenetics were independent prognostic markers. In summary our results suggest that Pgrn is a novel, robust and independent prognostic marker in CLL that can be easily measured by ELISA.

An electrostatic selection mechanism controls sequential kinase signaling downstream of the T cell receptor

PubMed Central

Shah, Neel H; Wang, Qi; Yan, Qingrong; Karandur, Deepti; Kadlecek, Theresa A; Fallahee, Ian R; Russ, William P; Ranganathan, Rama; Weiss, Arthur; Kuriyan, John

2016-01-01

The sequence of events that initiates T cell signaling is dictated by the specificities and order of activation of the tyrosine kinases that signal downstream of the T cell receptor. Using a platform that combines exhaustive point-mutagenesis of peptide substrates, bacterial surface-display, cell sorting, and deep sequencing, we have defined the specificities of the first two kinases in this pathway, Lck and ZAP-70, for the T cell receptor ζ chain and the scaffold proteins LAT and SLP-76. We find that ZAP-70 selects its substrates by utilizing an electrostatic mechanism that excludes substrates with positively-charged residues and favors LAT and SLP-76 phosphosites that are surrounded by negatively-charged residues. This mechanism prevents ZAP-70 from phosphorylating its own activation loop, thereby enforcing its strict dependence on Lck for activation. The sequence features in ZAP-70, LAT, and SLP-76 that underlie electrostatic selectivity likely contribute to the specific response of T cells to foreign antigens. DOI: http://dx.doi.org/10.7554/eLife.20105.001 PMID:27700984

The hypermorph FtsA* protein has an in vivo role in relieving the Escherichia coli proto-ring block caused by excess ZapC+

PubMed Central

Ortiz, Cristina; Casanova, Mercedes; Palacios, Pilar

2017-01-01

Assembly of the proto-ring, formed by the essential FtsZ, FtsA and ZipA proteins, and its progression into a divisome, are essential events for Escherichia coli division. ZapC is a cytoplasmic protein that belongs to a group of non-essential components that assist FtsZ during proto-ring assembly. Any overproduction of these proteins leads to faulty FtsZ-rings, resulting in a cell division block. We show that ZapC overproduction can be counteracted by an excess of the ZipA-independent hypermorph FtsA* mutant, but not by similar amounts of wild type FtsA+. An excess of FtsA+ allowed regular spacing of the ZapC-blocked FtsZ-rings, but failed to promote recruitment of the late-assembling proteins FtsQ, FtsK and FtsN and therefore, to activate constriction. In contrast, overproduction of FtsA*, besides allowing correct FtsZ-ring localization at midcell, restored the ability of FtsQ, FtsK and FtsN to be incorporated into active divisomes. PMID:28877250

Differential Requirements for Src-Family Kinases in SYK or ZAP70-Mediated SLP-76 Phosphorylation in Lymphocytes

PubMed Central

Fasbender, Frank; Claus, Maren; Wingert, Sabine; Sandusky, Mina; Watzl, Carsten

2017-01-01

In a synthetic biology approach using Schneider (S2) cells, we show that SLP-76 is directly phosphorylated at tyrosines Y113 and Y128 by SYK in the presence of ITAM-containing adapters such as CD3ζ, DAP12, or FcεRγ. This phosphorylation was dependent on at least one functional ITAM and a functional SH2 domain within SYK. Inhibition of Src-kinases by inhibitors PP1 and PP2 did not reduce SLP-76 phosphorylation in S2 cells, suggesting an ITAM and SYK dependent, but Src-kinase independent signaling pathway. This direct ITAM/SYK/SLP-76 signaling pathway therefore differs from previously described ITAM signaling. However, the SYK-family kinase ZAP70 required the additional co-expression of the Src-family kinases Fyn or Lck to efficiently phosphorylate SLP-76 in S2 cells. This difference in Src-family kinase dependency of SYK versus ZAP70-mediated ITAM-based signaling was further demonstrated in human lymphocytes. ITAM signaling in ZAP70-expressing T cells was dependent on the activity of Src-family kinases. In contrast, Src-family kinases were partially dispensable for ITAM signaling in SYK-expressing B cells or in natural killer cells, which express SYK and ZAP70. This demonstrates that SYK can signal using a Src-kinase independent ITAM-based signaling pathway, which may be involved in calibrating the threshold for lymphocyte activation. PMID:28736554

Differential Requirements for Src-Family Kinases in SYK or ZAP70-Mediated SLP-76 Phosphorylation in Lymphocytes.

PubMed

Fasbender, Frank; Claus, Maren; Wingert, Sabine; Sandusky, Mina; Watzl, Carsten

2017-01-01

In a synthetic biology approach using Schneider (S2) cells, we show that SLP-76 is directly phosphorylated at tyrosines Y113 and Y128 by SYK in the presence of ITAM-containing adapters such as CD3ζ, DAP12, or FcεRγ. This phosphorylation was dependent on at least one functional ITAM and a functional SH2 domain within SYK. Inhibition of Src-kinases by inhibitors PP1 and PP2 did not reduce SLP-76 phosphorylation in S2 cells, suggesting an ITAM and SYK dependent, but Src-kinase independent signaling pathway. This direct ITAM/SYK/SLP-76 signaling pathway therefore differs from previously described ITAM signaling. However, the SYK-family kinase ZAP70 required the additional co-expression of the Src-family kinases Fyn or Lck to efficiently phosphorylate SLP-76 in S2 cells. This difference in Src-family kinase dependency of SYK versus ZAP70-mediated ITAM-based signaling was further demonstrated in human lymphocytes. ITAM signaling in ZAP70-expressing T cells was dependent on the activity of Src-family kinases. In contrast, Src-family kinases were partially dispensable for ITAM signaling in SYK-expressing B cells or in natural killer cells, which express SYK and ZAP70. This demonstrates that SYK can signal using a Src-kinase independent ITAM-based signaling pathway, which may be involved in calibrating the threshold for lymphocyte activation.

Non-degradative Ubiquitination of Protein Kinases

PubMed Central

Ball, K. Aurelia; Johnson, Jeffrey R.; Lewinski, Mary K.; Guatelli, John; Verschueren, Erik; Krogan, Nevan J.; Jacobson, Matthew P.

2016-01-01

Growing evidence supports other regulatory roles for protein ubiquitination in addition to serving as a tag for proteasomal degradation. In contrast to other common post-translational modifications, such as phosphorylation, little is known about how non-degradative ubiquitination modulates protein structure, dynamics, and function. Due to the wealth of knowledge concerning protein kinase structure and regulation, we examined kinase ubiquitination using ubiquitin remnant immunoaffinity enrichment and quantitative mass spectrometry to identify ubiquitinated kinases and the sites of ubiquitination in Jurkat and HEK293 cells. We find that, unlike phosphorylation, ubiquitination most commonly occurs in structured domains, and on the kinase domain, ubiquitination is concentrated in regions known to be important for regulating activity. We hypothesized that ubiquitination, like other post-translational modifications, may alter the conformational equilibrium of the modified protein. We chose one human kinase, ZAP-70, to simulate using molecular dynamics with and without a monoubiquitin modification. In Jurkat cells, ZAP-70 is ubiquitinated at several sites that are not sensitive to proteasome inhibition and thus may have other regulatory roles. Our simulations show that ubiquitination influences the conformational ensemble of ZAP-70 in a site-dependent manner. When monoubiquitinated at K377, near the C-helix, the active conformation of the ZAP-70 C-helix is disrupted. In contrast, when monoubiquitinated at K476, near the kinase hinge region, an active-like ZAP-70 C-helix conformation is stabilized. These results lead to testable hypotheses that ubiquitination directly modulates kinase activity, and that ubiquitination is likely to alter structure, dynamics, and function in other protein classes as well. PMID:27253329

Pressurized grout remote backfilling at AML sites near Beulah and Zap, North Dakota

DOE Office of Scientific and Technical Information (OSTI.GOV)

Weiner, E.J.; Dodd, W.E.

1999-07-01

The Abandoned Mine Lands (AML) Division of the North Dakota Public Service Commission (PSC) is charged with the reclamation of hazardous abandoned mine sites in North Dakota. Several underground lignite coalmines were operated near the cities of Beulah and Zap, North Dakota, from the early 1900's until about 1955. Coal seams in this area were relatively thick and the overburden generally shallow. As these mines have deteriorated with time, deep collapse features, or sinkholes, have surfaced in many areas. These features are very dangerous, especially when they occur at or near residential and commercial areas and public roads. In themore » past five years, sinkholes have surfaced beneath a commercial building (boat dealership, lounge, and gas station) and beneath a nearby occupied mobile home north of Beulah. sinkholes have also surfaced near KHOL Radio Station in Beulah and in the right of way of a public road south of Zap. The AML Division has conducted several emergency sinkhole-filling projects in these areas. In 1995--97, the AML Division conducted exploratory drilling which confirmed the presence of collapsing underground mines at these sites. The remediation of these sites around Beulah/Zap will take place over several years and involve three or more separate contracts due to budget considerations. In 1997, the AML Division began reclamation at these sties utilizing pressurized grout remote backfilling. In this technique, a cementitious grout is pumped through cased drill holes directly into the mine cavities to fill them and thereby stabilize the surface from collapse. The successful contractor for Phase One of the project was The Concrete Doctor, Inc. (TCDI). This paper will concentrate on Phase One of this work performed from June through September 1997. This project is especially interesting because grout was pumped through holes drilled inside the occupied commercial building. Grout was also pumped through angled holes that intercepted mined workings directly beneath the structure. Several specialized monitoring techniques were used to alert contractor if any movement in the structures occurred during grouting activities. Informational meetings were conducted by TCDI and PDC held with landowners, business owners, residents and road authorities before, during and after the project.« less

H2A.Z Acidic Patch Couples Chromatin Dynamics to Regulation of Gene Expression Programs during ESC Differentiation

PubMed Central

Subramanian, Vidya; Mazumder, Aprotim; Surface, Lauren E.; Butty, Vincent L.; Fields, Paul A.; Alwan, Allison; Torrey, Lillian; Thai, Kevin K.; Levine, Stuart S.; Bathe, Mark; Boyer, Laurie A.

2013-01-01

The histone H2A variant H2A.Z is essential for embryonic development and for proper control of developmental gene expression programs in embryonic stem cells (ESCs). Divergent regions of amino acid sequence of H2A.Z likely determine its functional specialization compared to core histone H2A. For example, H2A.Z contains three divergent residues in the essential C-terminal acidic patch that reside on the surface of the histone octamer as an uninterrupted acidic patch domain; however, we know little about how these residues contribute to chromatin structure and function. Here, we show that the divergent amino acids Gly92, Asp97, and Ser98 in the H2A.Z C-terminal acidic patch (H2A.ZAP3) are critical for lineage commitment during ESC differentiation. H2A.Z is enriched at most H3K4me3 promoters in ESCs including poised, bivalent promoters that harbor both activating and repressive marks, H3K4me3 and H3K27me3 respectively. We found that while H2A.ZAP3 interacted with its deposition complex and displayed a highly similar distribution pattern compared to wild-type H2A.Z, its enrichment levels were reduced at target promoters. Further analysis revealed that H2A.ZAP3 was less tightly associated with chromatin, suggesting that the mutant is more dynamic. Notably, bivalent genes in H2A.ZAP3 ESCs displayed significant changes in expression compared to active genes. Moreover, bivalent genes in H2A.ZAP3 ESCs gained H3.3, a variant associated with higher nucleosome turnover, compared to wild-type H2A.Z. We next performed single cell imaging to measure H2A.Z dynamics. We found that H2A.ZAP3 displayed higher mobility in chromatin compared to wild-type H2A.Z by fluorescent recovery after photobleaching (FRAP). Moreover, ESCs treated with the transcriptional inhibitor flavopiridol resulted in a decrease in the H2A.ZAP3 mobile fraction and an increase in its occupancy at target genes indicating that the mutant can be properly incorporated into chromatin. Collectively, our work suggests that the divergent residues in the H2A.Z acidic patch comprise a unique domain that couples control of chromatin dynamics to the regulation of developmental gene expression patterns during lineage commitment. PMID:23990805

B-cell subsets in the joint compartments of seropositive and seronegative rheumatoid arthritis (RA) and No-RA arthritides express memory markers and ZAP70 and characterize the aggregate pattern irrespectively of the autoantibody status.

PubMed

Michelutti, Alessandro; Gremese, Elisa; Morassi, Francesca; Petricca, Luca; Arena, Vincenzo; Tolusso, Barbara; Alivernini, Stefano; Peluso, Giusy; Bosello, Silvia Laura; Ferraccioli, Gianfranco

2011-01-01

The aim of the present study was to determine whether different subsets of B cells characterize synovial fluid (SF) or synovial tissue (ST) of seropositive or seronegative rheumatoid arthritis (RA) with respect to the peripheral blood (PB). PB, SF and ST of 14 autoantibody (AB)-positive (rheumatoid factor [RF]-IgM, RF-IgA, anti-citrullinated peptide [CCP]), 13 negative RA and 13 no-RA chronic arthritides were examined for B-cell subsets (Bm1-Bm5 and IgD-CD27 classifications), zeta-associated protein kinase-70 (ZAP70) expression on B cells and cytokine levels (interleukin [IL]-1β, tumor necrosis factor [TNF]-α, IL-6, IL-8 and monocyte chemotactic protein [MCP]-1). Synovial tissues were classified as aggregate and diffuse patterns. No differences were found in B-cell percentages or in subsets in PB and SF between AB(+) and AB(-) RA and no-RA. In both AB(+) and AB(-) RA (and no-RA), the percentage of CD19(+)/ZAP70(+) was higher in SF than in PB (AB(+): P = 0.03; AB(-): P = 0.01; no-RA: P = 0.01). Moreover, SF of both AB(+) and AB(-) RA (and no-RA) patients was characterized by a higher percentage of IgD-CD27(+) and IgD-CD27(-) B cells and lower percentage of IgD(+)CD27(-) (P < 0.05) B cells compared to PB. In SF, ZAP70 positivity is more represented in B cell CD27(+)/IgD(-)/CD38(-). The aggregate synovitis pattern was characterized by higher percentages of Bm5 cells in SF compared with the diffuse pattern (P = 0.05). These data suggest that no difference exists between AB(+) and AB(-) in B-cell subset compartmentalization. CD27(+)/IgD(-)/ZAP70(+) memory B cells accumulate preferentially in the joints of RA, suggesting a dynamic maturation of the B cells in this compartment.

Effects of pH during liquid storage of goat semen on sperm viability and fertilizing potential.

PubMed

Liu, Chang-He; Dong, Hai-Bo; Ma, Dong-Li; Li, You-Wei; Han, Dong; Luo, Ming-Jiu; Chang, Zhong-Le; Tan, Jing-He

2016-01-01

A specific problem in goat semen preservation is the detrimental effect of seminal plasma on sperm viability in extenders containing yolk or milk. Thus, the use of chemically defined extenders will have obvious advantages. Although previous studies indicate that the initial pH of an extender is crucial to sustain high sperm motility, changes in extender pH during long-term semen storage have not been observed. Monitoring extender pH at different times of semen storage and modeling its variation according to nonlinear models is thus important for protocol optimization for long-term liquid semen preservation. The present results showed that during long-term liquid storage of goat semen, both sperm motility and semen pH decreased gradually, and a strong correlation was observed between the two. Whereas increasing the initial extender pH from 6.04 to 6.25 or storage with stabilized pH improved, storage with artificially lowered pH impaired sperm motility. Extender renewal improved sperm motility by maintaining a stable pH. Sperm coating with chicken (Gallus gallus) egg yolk improved motility by increasing tolerance to pH decline. A new extender (n-mZAP) with a higher buffering capacity was formulated, and n-mZAP maintained higher sperm motility, membrane integrity and acrosome intactness than the currently used mZAP extender did. Goat semen liquid-stored for 12 d in n-mZAP produced pregnancy and kidding rates similar to those obtained with freshly collected semen following artificial insemination. In conclusion, maintenance of a stable pH during liquid semen storage dramatically improved sperm viability and fertilizing potential. Copyright © 2015 Elsevier B.V. All rights reserved.

INT WFC pre-discovery detection of the nova M31N 2017-11a (AT2017hvi, PTSS-17zap, TCP J00425419+4130425)

NASA Astrophysics Data System (ADS)

Mantero-Castaneda, E. A.; Hernandez-Sanchez, M.; Arce-Tord, C.; Esteban-Gutierrez, A.; Garcia-Broock, E.; Garcia-Rivas, M.; Gonzalez-Cuesta, L.; Hermosa-Munoz, L.; Jimenez-Gallardo, A.; Lopez-Navas, E.; Otero-Santos, J.; Prendin, M. G.; Rodriguez-Sanchez, M.; Perez-Fournon, I.

2017-12-01

We report the pre-discovery detection of the nova M31N 2017-11a (AT2017hvi, PTSS-17zap, TCP J00425419+4130425) with the Isaac Newton Telescope* Wide Field Camera on the night of 29 October 2017, 4.845 days before the discovery observations on 2017/11/04.695 by the PMO-Tsinghua Supernova Survey (PTSS).

The Touch and Zap Method for In Vivo Whole-Cell Patch Recording of Intrinsic and Visual Responses of Cortical Neurons and Glial Cells

PubMed Central

Schramm, Adrien E.; Marinazzo, Daniele; Gener, Thomas; Graham, Lyle J.

2014-01-01

Whole-cell patch recording is an essential tool for quantitatively establishing the biophysics of brain function, particularly in vivo. This method is of particular interest for studying the functional roles of cortical glial cells in the intact brain, which cannot be assessed with extracellular recordings. Nevertheless, a reasonable success rate remains a challenge because of stability, recording duration and electrical quality constraints, particularly for voltage clamp, dynamic clamp or conductance measurements. To address this, we describe “Touch and Zap”, an alternative method for whole-cell patch clamp recordings, with the goal of being simpler, quicker and more gentle to brain tissue than previous approaches. Under current clamp mode with a continuous train of hyperpolarizing current pulses, seal formation is initiated immediately upon cell contact, thus the “Touch”. By maintaining the current injection, whole-cell access is spontaneously achieved within seconds from the cell-attached configuration by a self-limited membrane electroporation, or “Zap”, as seal resistance increases. We present examples of intrinsic and visual responses of neurons and putative glial cells obtained with the revised method from cat and rat cortices in vivo. Recording parameters and biophysical properties obtained with the Touch and Zap method compare favourably with those obtained with the traditional blind patch approach, demonstrating that the revised approach does not compromise the recorded cell. We find that the method is particularly well-suited for whole-cell patch recordings of cortical glial cells in vivo, targeting a wider population of this cell type than the standard method, with better access resistance. Overall, the gentler Touch and Zap method is promising for studying quantitative functional properties in the intact brain with minimal perturbation of the cell's intrinsic properties and local network. Because the Touch and Zap method is performed semi-automatically, this approach is more reproducible and less dependent on experimenter technique. PMID:24875855

Lck is a relevant target in chronic lymphocytic leukaemia cells whose expression variance is unrelated to disease outcome.

PubMed

Till, Kathleen J; Allen, John C; Talab, Fatima; Lin, Ke; Allsup, David; Cawkwell, Lynn; Bentley, Alison; Ringshausen, Ingo; Duckworth, Andrew D; Pettitt, Andrew R; Kalakonda, Nagesh; Slupsky, Joseph R

2017-12-01

Pathogenesis of chronic lymphocytic leukaemia (CLL) is contingent upon antigen receptor (BCR) expressed by malignant cells of this disease. Studies on somatic hypermutation of the antigen binding region, receptor expression levels and signal capacity have all linked BCR on CLL cells to disease prognosis. Our previous work showed that the src-family kinase Lck is a targetable mediator of BCR signalling in CLL cells, and that variance in Lck expression associated with ability of BCR to induce signal upon engagement. This latter finding makes Lck similar to ZAP70, another T-cell kinase whose aberrant expression in CLL cells also associates with BCR signalling capacity, but also different because ZAP70 is not easily pharmacologically targetable. Here we describe a robust method of measuring Lck expression in CLL cells using flow cytometry. However, unlike ZAP70 whose expression in CLL cells predicts prognosis, we find Lck expression and disease outcome in CLL are unrelated despite observations that its inhibition produces effects that biologically resemble the egress phenotype taken on by CLL cells treated with idelalisib. Taken together, our findings provide insight into the pathobiology of CLL to suggest a more complex relationship between expression of molecules within the BCR signalling pathway and disease outcome.

Carbon and Metal Transformations Near the Continent-Ocean Margin Quantified with the ZAPS Instrument Package

DTIC Science & Technology

1999-09-30

Rudnicki M.D. and Klinkhammer G.P., (1999) A segment-scale survey of the Broken Spur hydrothermal plume , Deep-Sea Res. I, 46, 701-714. Guay C.K...We used our sled this year to explore the Bransfield Strait, Antarctica for hydrothermal vent sites (NSF -Office of Polar Programs; OPP-9725872). We...used the real time sensing capability to locate a vent field and map hydrothermal signals at several locations. We used the ZAPS package to investigate

Zapping Rocks on Mars

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wiens, Roger

Better understanding Mars means better understanding its geology. That’s why, sitting atop NASA’s Curiosity rover, is ChemCam, an instrument built by Los Alamos National Laboratory that shoots lasers at Martian rocks and analyzes the data. After nearly 1,500 rock zaps, ChemCam has uncovered some surprising facts about the Red Planet, including the discovery of igneous rocks. Soon, a new Los Alamos-built instrument—the SuperCam—will ride aboard the Mars 2020 rover and bring with it enhanced capabilities to unlock new secrets about the planet.

Effects of Pasteurella haemolytica leukotoxic culture supernatant on bovine neutrophil aggregation.

PubMed

Conlon, P; Gervais, M; Chaudhari, S; Conlon, J

1992-07-01

Pasteurella haemolytica A1 leukotoxic culture supernatant was evaluated for its ability to cause aggregation of bovine peripheral neutrophils. Neutrophils were isolated by a hypotonic lysis method and incubated with zymosan-activated plasma (ZAP), leukotoxic culture supernatant, antileukotoxin serum, calcium and magnesium-free media, p-bromophenacyl bromide and protein kinase C inhibitors. Aggregation was evaluated by changes in infrared light transmittance. Leukotoxic culture supernatant caused neutrophils to aggregate, and this effect was significantly removed by preincubation with antileukotoxin serum. Aggregation to ZAP and leukotoxin was dependent on the presence of extra-cellular calcium. Activation of protein kinase C by phorbol myristate acetate induced aggregation which was reduced by staurosporine; however, aggregation to leukotoxin did not involve protein kinase C activation. Phospholipase A2 inhibition did not alter the aggregation response to ZAP or to leukotoxin. The in vitro measurement of neutrophil aggregation induced by the leukotoxin of P. haemolytica reflects cytoskeletal and other activation events that may contribute to the intense inflammatory process which this organism induces in the lungs of cattle.

A unique proteomic profile on surface IgM ligation in unmutated chronic lymphocytic leukemia

PubMed Central

Perrot, Aurore; Pionneau, Cédric; Nadaud, Sophie; Davi, Frédéric; Leblond, Véronique; Jacob, Frédéric; Merle-Béral, Hélène; Herbrecht, Raoul; Béné, Marie-Christine; Gribben, John G.; Vallat, Laurent

2011-01-01

Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course with 2 extreme subsets: indolent, ZAP70− and mutated immunoglobulin heavy chain gene (M-CLL); and aggressive, ZAP70+ and unmutated immunoglobulin heavy chain (UM-CLL). Given the long-term suspicion of antigenic stimulation as a primum movens in the disease, the role of the B-cell receptor has been extensively studied in various experimental settings; albeit scarcely in a comparative dynamic proteomic approach. Here we use a quantitative 2-dimensional fluorescence difference gel electrophoresis technology to compare 48 proteomic profiles of the 2 CLL subsets before and after anti-IgM ligation. Differentially expressed proteins were subsequently identified by mass spectrometry. We show that unstimulated M- and UM-CLL cells display distinct proteomic profiles. Furthermore, anti-IgM stimulation induces a specific proteomic response, more pronounced in the more aggressive CLL. Statistical analyses demonstrate several significant protein variations according to stimulation conditions. Finally, we identify an intermediate form of M-CLL cells, with an indolent profile (ZAP70−) but sharing aggressive proteomic profiles alike UM-CLL cells. Collectively, this first quantitative and dynamic proteome analysis of CLL further dissects the complex molecular pathway after B-cell receptor stimulation and depicts distinct proteomic profiles, which could lead to novel molecular stratification of the disease. PMID:21602524

Inhibitory mechanism of five natural flavonoids against murine norovirus.

PubMed

Seo, Dong Joo; Choi, Changsun

2017-07-01

Human noroviruses (HuNoV), which are responsible for acute gastroenteritis, are becoming a serious public health concern worldwide. Since no effective antiviral drug or vaccine for HuNoV has been developed yet, some natural extracts and their active components have been investigated for their ability to inhibit noroviruses. However, their exact antiviral mechanisms have not been investigated. This study was performed to investigate the expression of interferon (IFN)-α, IFN-λ, tumor necrosis factor-α (TNF-α), Mx, and zinc finger CCCH type antiviral protein 1 (ZAP), 2'-5' oligo (A) synthetase (OAS), and inducible nitric oxide synthase (iNOS) in RAW 264.7 cells pre-treated with fisetin, daidzein, quercetin, epigallocatechin gallate (EGCG), and epicatechin gallate (ECG) that have anti-noroviral activity. Based on the antiviral activity of the five flavonoids, recently reported by our group, the expression of antiviral factors such as IFN-α, IFN-λ, TNF-α, IL-1β, IL-6, Mx, ZAP, OAS, and iNOS was investigated in RAW 264.7 cells pre-treated with these flavonoids. Anti-noroviral effect was determined by performing a plaque assay on cells treated with the flavonoid. RAW 264.7 cells were treated with fisetin, daidzein, quercetin, EGCG, and ECG. Then, mRNA of IFN-α, IFN-λ, TNF-α, IL-1β, IL-6, Mx, ZAP, OAS, and iNOS were measured by real-time RT-PCR. IFN-α, TNF-α, IL-1β, and IL-6 proteins were measured by ELISA. Pre-treatment with fisetin (50μM), fisetin (100μM), EGCG (100μM), quercetin (100μM), daidzein (50μM), and ECG (150μM) significantly reduced MNoV by 50.00±7.14 to 60.67±9.26%. The mRNA levels of IFN-α, IFN-λ, TNF-α, Mx, and ZAP were upregulated in RAW 264.7 cells pre-treated with fisetin, quercetin, and daidzein, but not in those pre-treated with EGCG or ECG. Regarding protein levels, IFN-α was significantly induced in cells pre-treated with fisetin, quercetin, and daidzein, whereas TNF-α was significantly induced only in cells pre-treated with daidzein. Pre-treatment of RAW 264.7 cells with the five flavonoids inhibited MNoV by upregulating the expression of antiviral cytokines (IFN-α, IFN-λ, and TNF-α) and interferon-stimulating genes (Mx and ZAP). Copyright © 2017 Elsevier GmbH. All rights reserved.

ZAP-70 Restoration in Mice by In Vivo Thymic Electroporation

PubMed Central

Kissenpfennig, Adrien; Poulin, Lionel Franz; Leserman, Lee; Marche, Patrice N.; Jouvin-Marche, Evelyne; Berger, François; Nguyen, Catherine

2008-01-01

Viral and non-viral vectors have been developed for gene therapy, but their use is associated with unresolved problems of efficacy and safety. Efficient and safe methods of DNA delivery need to be found for medical application. Here we report a new monopolar system of non-viral electro-gene transfer into the thymus in vivo that consists of the local application of electrical pulses after the introduction of the DNA. We assessed the proof of concept of this approach by correcting ZAP-70 deficient severe combined immunodeficiency (SCID) in mice. The thymic electro-gene transfer of the pCMV-ZAP-70-IRES-EGFP vector in these mice resulted in rapid T cell differentiation in the thymus with mature lymphocytes detected by three weeks in secondary lymphoid organs. Moreover, this system resulted in the generation of long-term functional T lymphocytes. Peripheral reconstituted T cells displayed a diversified T cell receptor (TCR) repertoire, and were responsive to alloantigens in vivo. This process applied to the thymus could represent a simplified and effective alternative for gene therapy of T cell immunodeficiencies. PMID:18446234

Effects of Pasteurella haemolytica leukotoxic culture supernatant on bovine neutrophil aggregation.

PubMed Central

Conlon, P; Gervais, M; Chaudhari, S; Conlon, J

1992-01-01

Pasteurella haemolytica A1 leukotoxic culture supernatant was evaluated for its ability to cause aggregation of bovine peripheral neutrophils. Neutrophils were isolated by a hypotonic lysis method and incubated with zymosan-activated plasma (ZAP), leukotoxic culture supernatant, antileukotoxin serum, calcium and magnesium-free media, p-bromophenacyl bromide and protein kinase C inhibitors. Aggregation was evaluated by changes in infrared light transmittance. Leukotoxic culture supernatant caused neutrophils to aggregate, and this effect was significantly removed by preincubation with antileukotoxin serum. Aggregation to ZAP and leukotoxin was dependent on the presence of extra-cellular calcium. Activation of protein kinase C by phorbol myristate acetate induced aggregation which was reduced by staurosporine; however, aggregation to leukotoxin did not involve protein kinase C activation. Phospholipase A2 inhibition did not alter the aggregation response to ZAP or to leukotoxin. The in vitro measurement of neutrophil aggregation induced by the leukotoxin of P. haemolytica reflects cytoskeletal and other activation events that may contribute to the intense inflammatory process which this organism induces in the lungs of cattle. PMID:1423054

Zapping Rocks on Mars

ScienceCinema

Wiens, Roger

2018-01-16

Better understanding Mars means better understanding its geology. That’s why, sitting atop NASA’s Curiosity rover, is ChemCam, an instrument built by Los Alamos National Laboratory that shoots lasers at Martian rocks and analyzes the data. After nearly 1,500 rock zaps, ChemCam has uncovered some surprising facts about the Red Planet, including the discovery of igneous rocks. Soon, a new Los Alamos-built instrument—the SuperCam—will ride aboard the Mars 2020 rover and bring with it enhanced capabilities to unlock new secrets about the planet.

Audience Selectivity and Involvement in the Newer Media Environment.

ERIC Educational Resources Information Center

Perse, Elizabeth M.

1990-01-01

Examines cable television viewer selectivity and involvement. Concludes through study of 342 cable subscribers who owned remote control devices that "zapping" (changing television channels during commercials) is an indication of ritualistic television viewing. Suggests that more salient motives are linked to the preexposure activities of…

Serum TK levels in CLL identify Binet stage A patients within biologically defined prognostic subgroups most likely to undergo disease progression.

PubMed

Matthews, Christine; Catherwood, Mark A; Morris, T C M; Kettle, Paul J; Drake, Mary B; Gilmore, William S; Alexander, H Denis

2006-10-01

Serum thymidine kinase (TK) levels have been shown to be correlated with survival in many malignancies, including chronic lymphocytic leukaemia (CLL). This study was designed to investigate associations between TK levels and other prognostic markers, in newly and previously diagnosed Binet stage A patients. Furthermore, the use of serum TK measurement to identify subcategories of disease within those defined by IgV(H) mutational status, gene usage and chromosomal aberrations was investigated. Ninety-one CLL patients were enrolled. Serum TK levels were measured using a radioenzyme assay. IgV(H) mutational status and V(H) gene usage were determined using BIOMED-2 primers and protocol. Recurring chromosomal abnormalities were detected by interphase fluorescent in situ hybridisation (FISH). Flow cytometry and reverse transcriptase polymerase chain reaction (RT-PCR) determined CD38 and Zap-70 expression, respectively. Significantly higher serum TK levels were found in IgV(H) unmutated, compared with IgV(H) mutated, patients (P < 0.001). Elevated TK levels were also found in patients with CD38 and Zap-70 positivity (P = 0.004, P < 0.001, respectively), short lymphocyte doubling time (LDT) (P = 0.044) and poor or intermediate prognosis chromosomal aberrations (P < 0.001). A TK level of >8.5 U/L best identified patients with progressive disease. Elevated TK levels could identify patients categorised, at diagnosis, into good prognosis subgroups by the various biological markers (mutated IgV(H), good prognosis chromosomal aberrations, Zap-70(-) and CD38(-)) who subsequently showed disease progression. Additionally, patients with V(H)3-21 gene usage showed high TK levels, irrespective of mutational status, and serum TK measurement retained predictive power as disease progressed in all subcategories studied.

THE RESPONSES OF PRAIRIE DEER MICE TO A FIELD SO2 GRADIENT

EPA Science Inventory

A capture-mark-release study of deer mice (Peromyscus) was conducted on two 10-acre grassland areas (Zonal Air Pollution Systems or ZAPS) at monthly intervals from April to September 1976. Both areas were subdivided into four in-line experimental plots, with three smaller plots a...

Regulation of coal polymer degradation by fungi. Tenth Quartery report, October 1996--December 1996

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvine, R.L.; Bumpus, J.A.

1997-01-28

It has long been known that low rank coal such as leonardite can be solubilized by strong base (>pH 12). Recent discoveries have also shown that leonardite is solubilized by Lewis bases at considerably lower pH values and by fungi that secrete certain Lewis bases (i.e., oxalate ion). During the current reporting period we have studied the ability of a strong base (sodium hydroxide, pH 12), and two fungi, Phanerochaete chrysosporium and Trametes versicolor, to solubilize Argonne Premium Coals. In general, Argonne Premium Coals were relatively resistant to base mediated solubilization. However, when these coals were preoxidized (150{degrees}C for sevenmore » days), substantial amounts of several coals were solubilized. Most affected were the Lewiston-Stockton bituminous coal, the Beulah-Zap lignite, the Wyodak-Anderson subbituminous coal and the Blind Canyon bituminous coal. Argonne Premium Coals were previously shown by us to be relatively resistant to solubilization by sodium oxalate. When preoxidized coals were treated with sodium oxalate, only the Beulah-Zap lignite was substantially solubilized. Although very small amounts of the other preoxidized coals were solubilized by treatment with oxalate, the small amount of solubilization that did take place was generally increased relative to that observed for coals that were not preoxidized. None of the Argonne Premium Coals were solubilized by P. chrysosporium or T. versicolor. Of considerable interest, however, is the observation that P. chrysosporium and T. versicolor mediated extensive solubilization of Lewiston-Stockton bituminous coal, the Beulah-Zap lignite and the Wyodak-Anderson subbituminous coal.« less

ZAP! Adapted: Incorporating design in the introductory electromagnetism lab

NASA Astrophysics Data System (ADS)

McNeil, J. A.

2002-04-01

In the last decade the Accreditation Board of Engineering and Technology(ABET) significantly reformed the criteria by which engineering programs are accredited. The new criteria are called Engineering Criteria 2000 (EC2000). Not surprisingly, engineering design constitutes an essential component of these criteria. The Engineering Physics program at the Colorado School of Mines (CSM) underwent an ABET general review and site visit in the fall of 2000. In preparation for this review and as part of a campus-wide curriculum reform the Physics Department was challenged to include elements of design in its introductory laboratories. As part of the background research for this reform, several laboratory programs were reviewed including traditional and studio modes as well as a course used by Cal Tech and MIT called "ZAP!" which incorporates design activities well-aligned with the EC2000 criteria but in a nontraditional delivery mode. CSM has adapted several ZAP! experiments to a traditional laboratory format while attempting to preserve significant design experiences. The new laboratory forms an important component of the reformed course which attempts to respect the psychological principles of learner-based education. This talk reviews the reformed introductory electromagnetism course and how the laboratories are integrated into the pedagogy along with design activities. In their new form the laboratories can be readily adopted by physics departments using traditional delivery formats.

SLP-76 mediates and maintains activation of the Tec family kinase ITK via the T cell antigen receptor-induced association between SLP-76 and ITK.

PubMed

Bogin, Yaron; Ainey, Carmit; Beach, Dvora; Yablonski, Deborah

2007-04-17

ITK (IL-2-inducible T cell kinase), a Tec family protein tyrosine kinase (PTK), is one of three PTKs required for T cell antigen receptor (TCR)-induced activation of phospholipase C-gamma1 (PLC-gamma1). Like Src and Abl family PTKs, ITK adopts an inactive, "closed" conformation, and its conversion to the active conformation is not well understood, nor have its direct substrates been identified. In a side-by-side comparison of ITK and ZAP-70 (zeta chain-associated protein kinase of 70 kDa), ITK efficiently phosphorylated Y(783) and Y(775) of PLC-gamma1, two phosphorylation sites that are critical for its activation, whereas ZAP-70 did not. SLP-76 (SH2-domain-containing leukocyte protein of 76 kDa), an adaptor required for TCR-induced activation of PLC-gamma1, was required for the phosphorylation of both PLC-gamma1 sites in intact cells. Furthermore, this event depended on the N-terminal tyrosines of SLP-76. Likewise, SLP-76, particularly its N-terminal tyrosines, was required for TCR-induced tyrosine phosphorylation and activation of ITK but was not required for the phosphorylation or activation of ZAP-70. Both ZAP-70 and ITK phosphorylated SLP-76 in vitro; thus, both PTKs are potential regulators of SLP-76, but only ITK is regulated by SLP-76. Upon TCR stimulation, a small fraction of ITK bound to SLP-76. This fraction, however, encompassed most of the catalytically active ITK. Catalytic activity was lost upon mild elution of ITK from the SLP-76-nucleated complex but was restored upon reconstitution of the complex. We propose that SLP-76 is required for ITK activation; furthermore, an ongoing physical interaction between SLP-76 and ITK is required to maintain ITK in an active conformation.

Activation of the Yeast UBI4 Polyubiquitin Gene by Zap1 Transcription Factor via an Intragenic Promoter Is Critical for Zinc-deficient Growth*

PubMed Central

MacDiarmid, Colin W.; Taggart, Janet; Jeong, Jeeyon; Kerdsomboon, Kittikhun; Eide, David J.

2016-01-01

Stability of many proteins requires zinc. Zinc deficiency disrupts their folding, and the ubiquitin-proteasome system may help manage this stress. In Saccharomyces cerevisiae, UBI4 encodes five tandem ubiquitin monomers and is essential for growth in zinc-deficient conditions. Although UBI4 is only one of four ubiquitin-encoding genes in the genome, a dramatic decrease in ubiquitin was observed in zinc-deficient ubi4Δ cells. The three other ubiquitin genes were strongly repressed under these conditions, contributing to the decline in ubiquitin. In a screen for ubi4Δ suppressors, a hypomorphic allele of the RPT2 proteasome regulatory subunit gene (rpt2E301K) suppressed the ubi4Δ growth defect. The rpt2E301K mutation also increased ubiquitin accumulation in zinc-deficient cells, and by using a ubiquitin-independent proteasome substrate we found that proteasome activity was reduced. These results suggested that increased ubiquitin supply in suppressed ubi4Δ cells was a consequence of more efficient ubiquitin release and recycling during proteasome degradation. Degradation of a ubiquitin-dependent substrate was restored by the rpt2E301K mutation, indicating that ubiquitination is rate-limiting in this process. The UBI4 gene was induced ∼5-fold in low zinc and is regulated by the zinc-responsive Zap1 transcription factor. Surprisingly, Zap1 controls UBI4 by inducing transcription from an intragenic promoter, and the resulting truncated mRNA encodes only two of the five ubiquitin repeats. Expression of a short transcript alone complemented the ubi4Δ mutation, indicating that it is efficiently translated. Loss of Zap1-dependent UBI4 expression caused a growth defect in zinc-deficient conditions. Thus, the intragenic UBI4 promoter is critical to preventing ubiquitin deficiency in zinc-deficient cells. PMID:27432887

Elevated IL-17A and IL-22 regulate expression of inducible CD38 and Zap-70 in chronic lymphocytic leukemia.

PubMed

Kouzegaran, Samaneh; Siroosbakht, Soheila; Farsad, Bahram Fariborz; Rezakhaniha, Bijan; Dormanesh, Banafshe; Behnod, Vahid; Tanha, Amir Saber

2018-01-01

In this study, we investigated the role and expression of interleukin (IL)-17A and IL-22 in chronic lymphocytic leukemia. We evaluated the expression of markers above on CLL by ELISA, qRT-PCR, flow cytometric analysis and nonparametric Kruskal-Wallis test. Quantitative RT-PCR revealed that the mRNA levels of IL-17A and IL-22 in PBMCs of CLL patients were upregulated compared with those from healthy subjects (mean ± SD: 1.96 ± 0.232 vs.0.72 ± 0.15, P < 0.001 and mean ± SD: 2.45 ± 0.534 vs.0.81 ± 0.26, P < 0.001, respectivily). In addition, findings showed that the IL-17A and IL-22 plasma level was significantly elevated than that from healthy control group (P  < 0.001). The median IL-17A and IL-22 in CLL patients and healthy control group were 48.28 ± 17.2 pg mL -1 ; 20.01 ± 11.16 pg mL -1 and 58.68 ± 23.4 pg mL -1 ;16.47 ± 10.31 P < 0.001, respectively. The levels of IL-17A and IL-22 was not significantly associated with the different stages of disease (Rai stages; Kruskal-Wallis test P > 0.05).No significant relationship was found between expression of CD38 and higher median serum levels of IL-17A in patients, but patients with negative expression of ZAP-70 showed a significant association with higher median serum levels of IL-17A compared with healthy subjects. (57.84 pg mL -1 vs. 31.67 pg mL -1 ; P = 0.016). IL-22 is elevated and associated with CD38 and Zap-70 expression in patients with CLL. No significant correlation was found between expression of CD38 and increased levels of IL-17A, negative expression of ZAP-70 showed a significant association with increased levels of IL-17A. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

Clinical and molecular predictors of disease severity and survival in chronic lymphocytic leukemia.

PubMed

Weinberg, J Brice; Volkheimer, Alicia D; Chen, Youwei; Beasley, Bethany E; Jiang, Ning; Lanasa, Mark C; Friedman, Daphne; Vaccaro, Gina; Rehder, Catherine W; Decastro, Carlos M; Rizzieri, David A; Diehl, Louis F; Gockerman, Jon P; Moore, Joseph O; Goodman, Barbara K; Levesque, Marc C

2007-12-01

Several parameters may predict disease severity and overall survival in chronic lymphocytic leukemia (CLL). The purpose of our study of 190 CLL patients was to compare immunoglobulin heavy chain variable region (IgV(H)) mutation status, cytogenetic abnormalities, and leukemia cell CD38 and Zap-70 to older, traditional parameters. We also wanted to construct a simple, inexpensive prognosis score that would significantly predict TTT and survival in patients at the time of diagnosis and help practicing clinicians. In univariate analyses, patients with higher clinical stage, higher leukocyte count at diagnosis, shorter leukocyte doubling time, elevated serum lactate dehydrogenase (LDH), unmutated immunoglobulin heavy chain variable region (IgV(H)) genes, and higher CD38 had a shorter overall survival and time-to-treatment (TTT). CLL cell Zap-70 expression was higher in patients with unmutated IgV(H), and those with higher Zap-70 tended to have shorter survival. IgV(H)4-34 or IgV(H)1-69 was the most common IgV(H) genes used (16 and 12%, respectively). Of those with IgV(H)1-69, 86% had unmutated IgV(H) and had a significantly shorter TTT. A cytogenetic abnormality was noted in 71% of the patients tested. Patients with 11q22 del and 17p13 del or complex abnormalities were significantly more likely to have unmutated IgV(H). We found that a prognostic score constructed using modified Rai stage, cellular CD38, and serum LDH (parameters easily obtained clinically) significantly predicted TTT and survival in patients at the time of diagnosis and performed as well or better than models using the newer markers.

Level of PAX5 in differential diagnosis of non-Hodgkin's lymphoma

PubMed Central

Bharti, Brij; Shukla, Sachin; Tripathi, Ratnakar; Mishra, Suman; Kumar, Mohan; Pandey, Manoj; Mishra, Rajnikant

2016-01-01

Background & objectives: The PAX5, a paired box transcription factor and B-cell activator protein (BSAP), activates B-cell commitment genes and represses non-B-cell lineage genes. About 14 transcript variants of PAX5 have been observed in human. Any alteration in its expression pattern leads to lymphogenesis or associated diseases and carcinogenesis in non-lymphoid tissues. Its mechanisms of function in pathophysiology of non-Hodgkin's lymphoma (NHL) are unclear. This study was intended to explore influence of PAX5 in cascade of NHL pathogenesis and diagnosis. Methods: Samples of 65 patients were evaluated by immunohistochemical staining for cellular localization of PAX5, CD19, CD3, cABL, p53, Ras and Raf and by TUNEL assay, RNA-isolation and reverse transcriptase (RT)-PCR, Western blot analysis, and lactate dehydrogenase (LDH) specific staining. Results: B-cell type NHL patients were positive for PAX5, p53, Ras, CD19, Raf and CD3. All of them showed TUNEL-positive cells. The differential expression pattern of PAX5, CD19, p53, CD3, ZAP70, HIF1α, Ras, Raf and MAPK (mitogen-activated protein kinase) at the levels of transcripts and proteins was observed. The LDH assay showed modulation of LDH4 and LDH5 isoforms in the lymph nodes of NHL patients. Interpretation & conclusions: The histological observations suggested that the patients represent diverse cases of NHL like mature B-cell type, mature T-cell type and high grade diffuse B-cell type NHL. The findings indicate that patients with NHL may also be analyzed for status of PAX5, CD19 and ZAP70, and their transcriptional and post-translational variants for the differential diagnosis of NHL and therapy. PMID:27748274

Into the Curriculum. Art: The Z Was Zapped [and] Art: Friendly Plastic [and] Music: American Composers [and] Reading/Language Arts: Chocolate Day [and] Science: Moose [and] Social Studies: Women's History Project.

ERIC Educational Resources Information Center

Rossi, Marie; And Others

1995-01-01

A group of six articles describes activities for art, music, reading/language arts, science, and social studies. Each article includes library media skills objectives, curriculum objectives, grade levels, resources, instructional roles, activity and procedures for completion, evaluation, and follow-up. (AEF)

Occurrence and morphology of carbonate concretions in the Beulah-Zap coal bed, Williston basin, North Dakota

USGS Publications Warehouse

Keighin, C.W.M.; Flores, R.M.; Rowland, T.

1996-01-01

Carbonate concretionary bodies were encountered during mining of the Beulah-Zap lignite seam in the Coteau Properties' Freedom mine, Mercer County, North Dakota. Preliminary studies show that areal and vertical distribution of the concretions are variable. All concretions examined are composed almost entirely of calcite. They occur as thin tabular bodies, as more or less elliptical forms, or as tear shaped bodies, and may occur individually or as clusters of buff-colored, poorly consolidated to solidly crystalline material. The carbonate masses vary in size from a few millimeters to tens of centimeters. Bedding in the lignite may display some compactional folding over dense spheroidal to elliptical concretions, indicating formation of the concretions prior to compaction. Internal morphology of the concretions is complex, and includes cone-in-cone structure, cross-cutting calcite veinlets, and multiple generations of calcite. Carbon isotope values suggest the concretions are composed of biogenic carbonate, probably related to early diagenesis and decomposition of organic matter (peat); oxygen isotope values are light, and consistent with a freshwater origin.

Use of MP3 players to increase asthma knowledge in inner-city African-American adolescents.

PubMed

Mosnaim, Giselle S; Cohen, Marc S; Rhoads, Christopher H; Rittner, Sarah Stuart; Powell, Lynda H

2008-01-01

Low-income African-American adolescents suffer a disproportionate burden of asthma morbidity. To evaluate the ability of our intervention, the Adolescents' Disease Empowerment and Persistency Technology (ADEPT) for asthma, to increase asthma knowledge in our target population. This was a 14-week (2-week run-in and 12-week treatment) randomized, double-blind, placebo-controlled pilot study in which 28 inner-city African-American adolescents with asthma, between 10 and 18 years of age, were randomized to receive (1) celebrity asthma messages (experimental group), or (2) general health messages (control group) between music tracks on an MP3 player. The asthma messages were recorded by famous athletes, musicians, and other celebrities popular among this group of teenagers. Asthma knowledge, assessed by the ZAP Asthma Knowledge instrament, was collected pre- and post-intervention. Mean improvement in ZAP score was significantly higher in the experimental group (8.1%, SD 7.2%) than the control group (0.4%, SD 7.2%) (p = 0.05). These findings suggest that this may be an innovative and promising new approach to improving asthma outcomes in this difficult-to-reach population.

MSL Animation EDL and Sky Crane

NASA Image and Video Library

2011-11-07

Animation of Mars Science Laboratory (MSL), also known as the Curiosity rover, from cruise stage to EDL (entry, descent and landing), roving around the planet, zapping rocks with its laser and drilling into rocks.

Curiosity First Rock Star, Up-Close

NASA Image and Video Library

2012-08-17

This close-up image shows the first target NASA Curiosity rover aims to zap with its Chemistry and Camera ChemCam instrument. The instrument will analyze that spark with a telescope and identify the chemical elements in the target.

Cryptochrome-1 expression: a new prognostic marker in B-cell chronic lymphocytic leukemia.

PubMed

Lewintre, Eloisa Jantus; Martín, Cristina Reinoso; Ballesteros, Carlos García; Montaner, David; Rivera, Rosa Farrás; Mayans, José Ramón; García-Conde, Javier

2009-02-01

Chronic lymphocytic leukemia is an adult-onset leukemia with a heterogeneous clinical behavior. When chronic lymphocytic leukemia cases were divided on the basis of IgV(H) mutational status, widely differing clinical courses were revealed. Since IgV(H) sequencing is difficult to perform in a routine diagnostic laboratory, finding a surrogate for IgV(H) mutational status seems an important priority. In the present study, we proposed the use of Cryptochrome-1 as a new prognostic marker in early-stage chronic lymphocytic leukemia. Seventy patients (Binet stage A, without treatment) were included in the study. We correlated Cryptochrome-1 mRNA with well established prognostic markers such as IgV(H) mutations, ZAP70, LPL or CD38 expression and chromosomal abnormalities. High Cryptochrome-1 expression correlated with IgV(H) unmutated samples. In addition, Cryptochrome-1 was a valuable predictor of disease progression in early-stage chronic lymphocytic leukemia, therefore it can be introduced in clinical practice with the advantage of a simplified method of quantification.

Zinc starvation induces autophagy in yeast

PubMed Central

Kawamata, Tomoko; Horie, Tetsuro; Matsunami, Miou; Sasaki, Michiko; Ohsumi, Yoshinori

2017-01-01

Zinc is an essential nutrient for all forms of life. Within cells, most zinc is bound to protein. Because zinc serves as a catalytic or structural cofactor for many proteins, cells must maintain zinc homeostasis under severely zinc-deficient conditions. In yeast, the transcription factor Zap1 controls the expression of genes required for uptake and mobilization of zinc, but to date the fate of existing zinc-binding proteins under zinc starvation remains poorly understood. Autophagy is an evolutionarily conserved cellular degradation/recycling process in which cytoplasmic proteins and organelles are sequestered for degradation in the vacuole/lysosome. In this study, we investigated how autophagy functions under zinc starvation. Zinc depletion induced non-selective autophagy, which is important for zinc-limited growth. Induction of autophagy by zinc starvation was not directly related to transcriptional activation of Zap1. Instead, TORC1 inactivation directed zinc starvation-induced autophagy. Abundant zinc proteins, such as Adh1, Fba1, and ribosomal protein Rpl37, were degraded in an autophagy-dependent manner. But the targets of autophagy were not restricted to zinc-binding proteins. When cellular zinc is severely depleted, this non-selective autophagy plays a role in releasing zinc from the degraded proteins and recycling zinc for other essential purposes. PMID:28264932

Herpes zoster-associated severity and duration of pain, health-related quality of life, and healthcare utilization in Taiwan: a prospective observational study.

PubMed

Tsai, Tsen-Fang; Yao, Chien-An; Yu, Hsin-Su; Lan, Cheng-Che; Chao, Sheau-Chiou; Yang, Jen-Hung; Yang, Kuo-Chia; Chen, Ching-Yu; White, Ron R; Psaradellis, Eliofotisti; Rampakakis, Emmanouil; Kawai, Kosuke; Acosta, Camilo J; Sampalis, John S

2015-01-01

To assess the severity and duration of herpes zoster (HZ)-associated pain (ZAP) and its impact on quality of life (QoL) and healthcare utilization (HCRU) from a patient perspective in routine care in Taiwan. A prospective, observational, single-cohort study was conducted in five centers across Taiwan. Patients were recruited at different time points during their HZ episode and were followed for ≤180 days. ZAP was assessed with the Initial Zoster Impact Questionnaire and the Zoster Brief Pain Inventory, QoL with the EQ-5D, and HCRU with a simple questionnaire. A total of 150 patients were included with a mean age of 64.9 years and mean time since rash onset of 18.8 days. Prodromal pain was experienced by 64.7% of patients, of whom 91.8% reported moderate-to-severe pain. At enrollment, 98.0% of patients experienced ZAP. Mean ± SD worst pain score decreased from 5.95 ± 3.09 at enrollment to 2.65 ± 2.98 at 30 days and 0.28 ± 0.83 at 180 days. Postherpetic neuralgia was observed in 20.7% of patients. Mean ± SD EQ-5D score significantly decreased (P < 0.001) from 0.91 ± 0.16 before rash onset to 0.67 ± 0.18 after rash onset, showing significant QoL deterioration up to 60 days. The impact of HZ on QoL and pain severity was similar across age groups. Significant HCRU was observed including visits to the doctor (83.3% of patients), specialist (30.7%), emergency department (24.7%), physiotherapist (23.3%), and hospitalizations (20.7%). Severe morbidity and significant HCRU are associated with HZ in Taiwan, supporting the need for early intervention and preventive strategies to reduce the HZ-associated burden of illness. © 2014 The International Society of Dermatology.

Curiosity First Rock Star

NASA Image and Video Library

2012-08-17

This mosaic image shows the first target NASA Curiosity rover aims to zap ChemCam instrument. ChemCam will be firing a laser at this rock, provisionally named N165, and analyzing the glowing, ionized gas, called plasma, that the laser excites.

Software Reviews: "Pow! Zap! Ker-plunk! The Comic Book Maker" (Pelican Software).

ERIC Educational Resources Information Center

Porter, Bernajean

1990-01-01

Reviews the newest addition to Pelican's Creative Writing Series of instructional software, which uses the comic book format to provide a unique writing environment for satire, symbolism, sequencing, and combining text and graphics to communicate ideas. (SR)

Zapping Those Zits: Helping Teens Handle Acne.

ERIC Educational Resources Information Center

Martin, Paul

1992-01-01

Describes five types of acne and stresses the importance of appropriate treatment. Some dermatologists believe diet is critical in improving acne. Other treatments include the use of drying lotions and soaps, astringents, abrasive cleansers, prescription drugs, face peels, and dermabrasion. (SM)

Children's Online Privacy.

ERIC Educational Resources Information Center

Aidman, Amy

2000-01-01

The first federal Internet privacy law (the Children's Online Privacy Protection Act) provides safeguards for children by regulating collection of their personal information. Unfortunately, teens are not protected. Legislation is pending to protect children from online marketers such as ZapMe! Interactive technologies require constant vigilance.…

Genetics Home Reference: ZAP70-related severe combined immunodeficiency

MedlinePlus

... involved in the activation of helper T cells (CD4+ T cells). These cells direct and assist the functions of ... CD8+ T cells and an excess of inactive CD4+ T cells. The resulting shortage of active T cells causes ...

Zap! Bam! Flop!

ERIC Educational Resources Information Center

Case, Doug

1976-01-01

Too many teachers have misguided ideas about the principles of the oral approach to teaching EFL. They are either too sure of themselves and of the theory and become complacent, or their confidence is shaken by the failure of a technique they thought to be infallible. (SCC)

Clonal evolution in chronic lymphocytic leukemia: analysis of correlations with IGHV mutational status, NOTCH1 mutations and clinical significance.

PubMed

López, Cristina; Delgado, Julio; Costa, Dolors; Villamor, Neus; Navarro, Alba; Cazorla, Maite; Gómez, Cándida; Arias, Amparo; Muñoz, Concha; Cabezas, Sandra; Baumann, Tycho; Rozman, María; Aymerich, Marta; Colomer, Dolors; Pereira, Arturo; Cobo, Francesc; López-Guillermo, Armando; Campo, Elías; Carrió, Ana

2013-10-01

Chronic lymphocytic leukemia (CLL) is a lymphoproliferative disorder characterized with highly variable clinical course. The most common chromosomal abnormalities in CLL, using conventional and molecular cytogenetics, are trisomy 12, del(13)(q14), del(11)(q22-23), del(17)(p13), and del(6)(q21). Whereas the prognostic marker such as IGHV mutational status remains stable during course of the diseases, chromosomal aberrations may be acquired over time. The aim of this study was to determine the incidence, and biological significance of clonal evolution (CE) using conventional and molecular cytogenetics and its relationship with prognostic markers such as CD38, ZAP70, and the mutational status of IGHV and NOTCH1. One hundred and forty-three untreated CLL patients were included in the study. The median time interval between analyses was 32 months (range 6-156 months). Forty-seven patients (33%) had CE as evidenced by detection of new cytogenetic abnormalities during follow-up. CE was not correlated with high expression of ZAP70, unmutated IGHV genes or NOTCH1 mutations. Multivariate analysis revealed that CE and IGHV mutation status had a significant impact on TFS. The combination of conventional and molecular cytogenetics increased the detection of CE, this phenomenon probably being a reflection of genomic instability and conferring a more aggressive clinical course. Copyright © 2013 Wiley Periodicals, Inc.

Zinc starvation induces autophagy in yeast.

PubMed

Kawamata, Tomoko; Horie, Tetsuro; Matsunami, Miou; Sasaki, Michiko; Ohsumi, Yoshinori

2017-05-19

Zinc is an essential nutrient for all forms of life. Within cells, most zinc is bound to protein. Because zinc serves as a catalytic or structural cofactor for many proteins, cells must maintain zinc homeostasis under severely zinc-deficient conditions. In yeast, the transcription factor Zap1 controls the expression of genes required for uptake and mobilization of zinc, but to date the fate of existing zinc-binding proteins under zinc starvation remains poorly understood. Autophagy is an evolutionarily conserved cellular degradation/recycling process in which cytoplasmic proteins and organelles are sequestered for degradation in the vacuole/lysosome. In this study, we investigated how autophagy functions under zinc starvation. Zinc depletion induced non-selective autophagy, which is important for zinc-limited growth. Induction of autophagy by zinc starvation was not directly related to transcriptional activation of Zap1. Instead, TORC1 inactivation directed zinc starvation-induced autophagy. Abundant zinc proteins, such as Adh1, Fba1, and ribosomal protein Rpl37, were degraded in an autophagy-dependent manner. But the targets of autophagy were not restricted to zinc-binding proteins. When cellular zinc is severely depleted, this non-selective autophagy plays a role in releasing zinc from the degraded proteins and recycling zinc for other essential purposes. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Peculiar Expression of CD3-Epsilon in Kidney of Ginbuna Crucian Carp.

PubMed

Miyazawa, Ryuichiro; Murata, Norifumi; Matsuura, Yuta; Shibasaki, Yasuhiro; Yabu, Takeshi; Nakanishi, Teruyuki

2018-01-01

TCR/CD3 complex is composed of the disulfide-linked TCR-αβ heterodimer that recognizes the antigen as a peptide presented by the MHC, and non-covalently paired CD3γε- and δε-chains together with disulfide-linked ζ-chain homodimers. The CD3 chains play key roles in T cell development and T cell activation. In the present study, we found nor or extremely lower expression of CD3ε in head- and trunk-kidney lymphocytes by flow cytometric analysis, while CD3ε was expressed at the normal level in lymphocytes from thymus, spleen, intestine, gill, and peripheral blood. Furthermore, CD4-1 + and CD8α + T cells from kidney express Zap-70, but not CD3ε, while the T cells from other tissues express both Zap-70 and CD3ε, although expression of CD3ε was low. Quantitative analysis of mRNA expression revealed that the expression level of T cell-related genes including tcrb, cd3 ε, zap-70 , and lck in CD4-1 + and CD8α + T cells was not different between kidney and spleen. Western blot analysis showed that CD3ε band was detected in the cell lysates of spleen but not kidney. To be interested, CD3ε-positive cells greatly increased after 24 h in in vitro culture of kidney leukocytes. Furthermore, expression of CD3ε in both transferred kidney and spleen leukocytes was not detected or very low in kidney, while both leukocytes expressed CD3ε at normal level in spleen when kidney and spleen leukocytes were injected into the isogeneic recipient. Lower expression of CD3ε was also found in kidney T lymphocytes of goldfish and carp. These results indicate that kidney lymphocytes express no or lower level of CD3ε protein in the kidney, although the mRNA of the gene was expressed. Here, we discuss this phenomenon from the point of function of kidney as reservoir for T lymphocytes in teleost, which lacks lymph node and bone marrow.

High energy density Z-pinch plasmas using flow stabilization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shumlak, U., E-mail: shumlak@uw.edu; Golingo, R. P., E-mail: shumlak@uw.edu; Nelson, B. A., E-mail: shumlak@uw.edu

The ZaP Flow Z-Pinch research project[1] at the University of Washington investigates the effect of sheared flows on MHD instabilities. Axially flowing Z-pinch plasmas are produced that are 100 cm long with a 1 cm radius. The plasma remains quiescent for many radial Alfvén times and axial flow times. The quiescent periods are characterized by low magnetic mode activity measured at several locations along the plasma column and by stationary visible plasma emission. Plasma evolution is modeled with high-resolution simulation codes – Mach2, WARPX, NIMROD, and HiFi. Plasma flow profiles are experimentally measured with a multi-chord ion Doppler spectrometer. Amore » sheared flow profile is observed to be coincident with the quiescent period, and is consistent with classical plasma viscosity. Equilibrium is determined by diagnostic measurements: interferometry for density; spectroscopy for ion temperature, plasma flow, and density[2]; Thomson scattering for electron temperature; Zeeman splitting for internal magnetic field measurements[3]; and fast framing photography for global structure. Wall stabilization has been investigated computationally and experimentally by removing 70% of the surrounding conducting wall to demonstrate no change in stability behavior.[4] Experimental evidence suggests that the plasma lifetime is only limited by plasma supply and current waveform. The flow Z-pinch concept provides an approach to achieve high energy density plasmas,[5] which are large, easy to diagnose, and persist for extended durations. A new experiment, ZaP-HD, has been built to investigate this approach by separating the flow Z-pinch formation from the radial compression using a triaxial-electrode configuration. This innovation allows more detailed investigations of the sheared flow stabilizing effect, and it allows compression to much higher densities than previously achieved on ZaP by reducing the linear density and increasing the pinch current. Experimental results and scaling analyses will be presented. In addition to studying fundamental plasma science and high energy density physics, the ZaP and ZaP-HD experiments can be applied to laboratory astrophysics.« less

Organization and sequence of four flagellin-encoding genes of Edwardsiella icataluri

USDA-ARS?s Scientific Manuscript database

Edwardsiella ictaluri, the cause of enteric septicemia in channel catfish (Ictalurus punctatus), is motile by means of peritrichous flagella. We determined the complete flagellin gene sequences and their organization in E. ictaluri by sequencing genomic segments selected from a lambda-ZAP phage gen...

First Imaging of Laser-Induced Spark on Mars

NASA Image and Video Library

2014-07-16

NASA Curiosity Mars rover used the Mars Hand Lens Imager MAHLI camera on its arm to catch the first images of sparks produced by the rover laser being shot at a rock on Mars. The left image is from before the laser zapped this rock, called Nova.

Noncolocated Structural Vibration Suppression Using Zero Annihilation Periodic Control

NASA Technical Reports Server (NTRS)

Bayard, David S.; Boussalis, Dhemetrios

1993-01-01

The Zero Annihilation Periodic (ZAP) controller is applied to the problem of vibration control of a noncolocated flexible structure. It is shown that even though the transfer function is nonminimum-phase, a plant inverse controller can be designed which elicits a deadbeat closed-loop response.

78 FR 36545 - Amendment of an Experimental Use Permit

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-18

...., on behalf of James Mains, Ph.D., Mosquito Mate, Inc., 1122 Oak Hill Drive, Lexington, KY 40505- 3322... original application specified the release of 30,000 mosquitoes in Kentucky (0.7 acre). The applicant now intends to release 100,000 male Aedes albopictus mosquitoes infected with the Wolbachia pipientis ZAP...

Zap! Pow! Wham!: Comics, Graphic Novels, and Education.

ERIC Educational Resources Information Center

Barron, Daniel D.

1991-01-01

Discussion of the use of comic books and graphic novels in school and public libraries highlights the role of school library media programs; how to use comics and graphic novels in teaching; the Japanese use of comic books; their use in literacy education; and the influence of comics on reading habits. (LRW)

Inhibition of polymorphonuclear leukocyte function by Legionella pneumophila exoproducts.

PubMed

Sahney, N N; Lambe, B C; Summersgill, J T; Miller, R D

1990-08-01

Total exoproducts (relative molecular mass greater than 10,000) from wild-type strains of Legionella pneumophila markedly inhibited human polymorphonuclear leukocyte (PMN) superoxide anion generation, at sub-lethal concentrations, in response to four stimuli [1.7, 0, 0.6 and 3.4% of control for zymosan activated particles (ZAP), phorbol myristate acetate (PMA), calcium ionophore (A 23187), and formyl-methionyl-leucyl-phenylalanine (fMLP), respectively]. PMN chemotaxis towards fMLP and spontaneous migration, were also dramatically inhibited (2.8 and 2.9% of buffer-treated controls, respectively). In contrast, total exoproducts from the cas-1 strain of L. pneumophila, a protease-deficient mutant generated by ethyl methane sulfonate mutagenesis, failed to inhibit PMN superoxide production in response to ZAP and PMA and only partially inhibited PMN response to A 23187 and fMLP. PMN spontaneous migration was unaffected by treatment with total exoproducts from the mutant, while directed chemotaxis was partially inhibited (51.4%). These data demonstrated that L. pneumophila total exoproducts, primarily protease had significant inhibitory effects on normal PMN function and may play an important contributory role in the pathogenesis of legionnaire's disease.

Impact of glutathione metabolism on zinc homeostasis in Saccharomyces cerevisiae.

PubMed

Steiger, Matthias G; Patzschke, Anett; Holz, Caterina; Lang, Christine; Causon, Tim; Hann, Stephan; Mattanovich, Diethard; Sauer, Michael

2017-06-01

Zinc is a crucial mineral for all organisms as it is an essential cofactor for the proper function of a plethora of proteins and depletion of zinc causes oxidative stress. Glutathione is the major redox buffering agent in the cell and therefore important for mitigation of the adverse effects of oxidative stress. In mammalian cells, zinc deficiency is accompanied by a glutathione depletion. In the yeast Saccharomyces cerevisiae, the opposite effect is observed: under low zinc conditions, an elevated glutathione concentration is found. The main regulator to overcome zinc deficiency is Zap1p. However, we show that Zap1p is not involved in this glutathione accumulation phenotype. Furthermore, we found that in glutathione-accumulating strains also the metal ion-binding phytochelatin-2, which is an oligomer of glutathione, is accumulated. This increased phytochelatin concentration correlates with a lower free zinc level in the vacuole. These results suggest that phytochelatin is important for zinc buffering in S. cerevisiae and thus explains how zinc homeostasis is connected with glutathione metabolism. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

T cell receptor-driven transendothelial migration of human effector memory CD4 T cells involves Vav, Rac and Myosin IIA

PubMed Central

Manes, Thomas D.; Pober, Jordan S.

2013-01-01

Human effector memory (EM) CD4 T cells may be recruited from the blood into a site of inflammation in response either to inflammatory chemokines displayed on or specific antigen presented by venular endothelial cells (ECs), designated as chemokine-driven or TCR-driven transendothelial migration (TEM), respectively. We have previously described differences in the morphological appearance of transmigrating T cells as well as in the molecules that mediate T cell-EC interactions distinguishing these two pathways. Here we report that TCR-driven TEM requires ZAP-70-dependent activation of a pathway involving Vav, Rac and myosin IIA. Chemokine-driven TEM also utilizes ZAP-70, albeit in a quantitatively and spatially different manner of activation, and is independent of Vav, Rac and mysosin IIA, depending instead on an as yet unidentified GTP exchange factor that activates Cdc42. The differential use of small Rho family GTPases to activate the cytoskeleton is consistent with the morphological differences observed in T cells that undergo TEM in response to these distinct recruitment signals. PMID:23420881

CD45-mediated signaling pathway is involved in Rhizoctonia bataticola lectin (RBL)-induced proliferation and Th1/Th2 cytokine secretion in human PBMC

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pujari, Radha; Eligar, Sachin M.; Kumar, Natesh

2012-03-23

Highlights: Black-Right-Pointing-Pointer RBL, a potent mitogenic and complex N-glycan specific lectin binds to CD45 on PBMC. Black-Right-Pointing-Pointer RBL triggers CD45-mediated signaling involved in activation of p38MAPK and STAT-5. Black-Right-Pointing-Pointer Inhibition of CD45 PTPase signaling blocks RBL-induced ZAP70 phosphorylation. Black-Right-Pointing-Pointer RBL-CD45 mediated signaling is crucial for RBL-induced immunodulatory activities. -- Abstract: We earlier reported the mitogenic and immunostimulatory activities of Rhizoctonia bataticola lectin (RBL), purified from phytopathogenic fungus R. bataticola in human PBMC. The lectin demonstrates specificity towards glycoproteins containing complex N-glycans. Since CD45-protein tyrosine phosphatase that abundantly expresses N-glycans is important in T-cell signaling, the study aimed to investigate themore » involvement of CD45 in the immunomodulatory activities of RBL. Flowcytometry and confocal microscopy studies revealed that RBL exhibited binding to PBMC and colocalized with CD45. The binding was comparable in cells expressing different CD45 isoforms-RA, -RB and -RO. CD45 blocking antibody reduced the binding and proliferation of PBMC induced by RBL. CD45-PTPase inhibitor dephostatin inhibited RBL-induced proliferation, expression of CD25 and pZAP-70. RBL-induced secretion of Th1/Th2 cytokines were significantly inhibited in presence of dephostatin. Also, dephostatin blocked phosphorylation of p38MAPK and STAT-5 that was crucial for the biological functions of RBL. The study demonstrates the involvement of CD45-mediated signaling in RBL-induced PBMC proliferation and Th1/Th2 cytokine secretion through activation of p38MAPK and STAT-5.« less

Molecular mechanism of the Syk activation switch.

PubMed

Tsang, Emily; Giannetti, Anthony M; Shaw, David; Dinh, Marie; Tse, Joyce K Y; Gandhi, Shaan; Ho, Hoangdung; Wang, Sandra; Papp, Eva; Bradshaw, J Michael

2008-11-21

Many immune signaling pathways require activation of the Syk tyrosine kinase to link ligation of surface receptors to changes in gene expression. Despite the central role of Syk in these pathways, the Syk activation process remains poorly understood. In this work we quantitatively characterized the molecular mechanism of Syk activation in vitro using a real time fluorescence kinase assay, mutagenesis, and other biochemical techniques. We found that dephosphorylated full-length Syk demonstrates a low initial rate of substrate phosphorylation that increases during the kinase reaction due to autophosphorylation. The initial rate of Syk activity was strongly increased by either pre-autophosphorylation or binding of phosphorylated immune tyrosine activation motif peptides, and each of these factors independently fully activated Syk. Deletion mutagenesis was used to identify regions of Syk important for regulation, and residues 340-356 of the SH2 kinase linker region were identified to be important for suppression of activity before activation. Comparison of the activation processes of Syk and Zap-70 revealed that Syk is more readily activated by autophosphorylation than Zap-70, although both kinases are rapidly activated by Src family kinases. We also studied Syk activity in B cell lysates and found endogenous Syk is also activated by phosphorylation and immune tyrosine activation motif binding. Together these experiments show that Syk functions as an "OR-gate" type of molecular switch. This mechanism of switch-like activation helps explain how Syk is both rapidly activated after receptor binding but also sustains activity over time to facilitate longer term changes in gene expression.

Happy Zapping in the Classroom: Enhancing Teaching and Learning with Electronic Voting Systems

ERIC Educational Resources Information Center

Read, David

2010-01-01

The use of electronic voting systems (EVS) in teaching at all levels is increasing as the technology becomes cheaper and easier to use. Although many educators initially take a cynical view of the educational value of such technology, many of those who use EVS express a very positive view of their experiences. This article aims to share these…

Integrating the Schoolhouse and the Marketplace: A Preliminary Assessment of the Emerging Role of Electronic Technology.

ERIC Educational Resources Information Center

Molnar, Alex

This paper focuses on how, by reinforcing and exploiting the emphasis on electronic technologies in school reform, marketers are integrating school children into the advertising and marketing system. The ZapMe! program is a for-profit venture that proposes to derive its profits from activities that link the academic activities of schools and the…

AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) TESTING OF TWO NUTRIENT ANALYZERS AT AN INDUSTRIAL PLANT: SCHIMADZU SCIENTIFIC INSTRUMENTS, INC. TNPC-4110(C) AND A ZAPS TECHNOLOGIES, INC. MULTI-PARAMETER (MP-1)

EPA Science Inventory

The Environmental Technology Verification (ETV) Program, beginning as an initiative of the U.S. Environmental Protection Agency (EPA) in 1995, verifies the performance of commercially available, innovative technologies that can be used to measure environmental quality. The ETV p...

RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain and is required for ubiquitination.

PubMed

Choudhury, Nila Roy; Heikel, Gregory; Trubitsyna, Maryia; Kubik, Peter; Nowak, Jakub Stanislaw; Webb, Shaun; Granneman, Sander; Spanos, Christos; Rappsilber, Juri; Castello, Alfredo; Michlewski, Gracjan

2017-11-08

TRIM25 is a novel RNA-binding protein and a member of the Tripartite Motif (TRIM) family of E3 ubiquitin ligases, which plays a pivotal role in the innate immune response. However, there is scarce knowledge about its RNA-related roles in cell biology. Furthermore, its RNA-binding domain has not been characterized. Here, we reveal that the RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain, which we postulate to be a novel RNA-binding domain. Using CLIP-seq and SILAC-based co-immunoprecipitation assays, we uncover TRIM25's endogenous RNA targets and protein binding partners. We demonstrate that TRIM25 controls the levels of Zinc Finger Antiviral Protein (ZAP). Finally, we show that the RNA-binding activity of TRIM25 is important for its ubiquitin ligase activity towards itself (autoubiquitination) and its physiologically relevant target ZAP. Our results suggest that many other proteins with the PRY/SPRY domain could have yet uncharacterized RNA-binding potential. Together, our data reveal new insights into the molecular roles and characteristics of RNA-binding E3 ubiquitin ligases and demonstrate that RNA could be an essential factor in their enzymatic activity.

New insights into the Vav1 activation cycle in lymphocytes.

PubMed

Barreira, María; Rodríguez-Fdez, Sonia; Bustelo, Xosé R

2018-05-01

Vav1 is a hematopoietic-specific Rho GDP/GTP exchange factor and signaling adaptor. Although these activities are known to be stimulated by direct Vav1 phosphorylation, little information still exists regarding the regulatory layers that influence the overall Vav1 activation cycle. Using a collection of cell models and activation-mimetic Vav1 mutants, we show here that the dephosphorylated state of Vav1 in nonstimulated T cells requires the presence of a noncatalytic, phospholipase Cγ1-Slp76-mediated inhibitory pathway. Upon T cell stimulation, Vav1 becomes rapidly phosphorylated via the engagement of Lck and, to a much lesser extent, other Src family kinases and Zap70. In this process, Lck, Zap70 and the adaptor protein Lat contribute differently to the dynamics and amplitude of the Vav1 phosphorylated pool. Consistent with a multiphosphosite activation mechanism, the optimal stimulation of Vav1 can only be recapitulated by the combination of several activation-mimetic phosphosite mutants. The analysis of these mutants has also unveiled the presence of different Vav1 signaling competent states that are influenced by phosphosites present in the N- and C-terminal domains of the protein. Copyright © 2018 Elsevier Inc. All rights reserved.

Zap 'Em with Assistive Technology: Notetaking, Modified Materials, Assistive Writing Tools, References, Organizational Tools, Cognitive Assistance, Adapted Access.

ERIC Educational Resources Information Center

Lahm, Elizabeth A.; Morrissette, Sandra K.

This collection of materials describes different types of computer applications and software that can help students with disabilities. It contains information on: (1) Easy Access, a feature of the systems software on every Macintosh computer that allows use of the keypad instead of the mouse, options for slow keys, and options for sticky keys; (2)…

Defense Data Network/TOPS-20 Tutorial. An Interative Computer Program.

DTIC Science & Technology

1985-12-01

33 XI. ELECTRONIC MAIL HOST ( EMH )-------------------------- 34 XII. EMACS...contents of text buffer to a file X EXCHANGE , used to search for and replace text Z ZAP, puts your entire file into the print buffer 23 - -° SWITCH...prompt USC-ISIE.ARPA> Sample foreign host command level prompt FTP COMMAND LEVEL COMMANDS COMMAND USE(S) FTP Invokes the FTP protocol CONNECT Connects

LDOC1 mRNA is differentially expressed in chronic lymphocytic leukemia and predicts overall survival in untreated patients

PubMed Central

Duzkale, Hatice; Schweighofer, Carmen D.; Coombes, Kevin R.; Barron, Lynn L.; Ferrajoli, Alessandra; O'Brien, Susan; Wierda, William G.; Pfeifer, John; Majewski, Tadeusz; Czerniak, Bogdan A.; Jorgensen, Jeffrey L.; Medeiros, L. Jeffrey; Freireich, Emil J; Keating, Michael J.

2011-01-01

We previously identified LDOC1 as one of the most significantly differentially expressed genes in untreated chronic lymphocytic leukemia (CLL) patients with respect to the somatic mutation status of the immunoglobulin heavy-chain variable region genes. However, little is known about the normal function of LDOC1, its contribution to the pathophysiology of CLL, or its prognostic significance. In this study, we have investigated LDOC1 mRNA expression in a large cohort of untreated CLL patients, as well as in normal peripheral blood B-cell (NBC) subsets and primary B-cell lymphoma samples. We have confirmed that LDOC1 is dramatically down-regulated in mutated CLL cases compared with unmutated cases, and have identified a new splice variant, LDOC1S. We show that LDOC1 is expressed in NBC subsets (naive > memory), suggesting that it may play a role in normal B-cell development. It is also expressed in primary B-cell lymphoma samples, in which its expression is associated with somatic mutation status. In CLL, we show that high levels of LDOC1 correlate with biomarkers of poor prognosis, including cytogenetic markers, unmutated somatic mutation status, and ZAP70 expression. Finally, we demonstrate that LDOC1 mRNA expression is an excellent predictor of overall survival in untreated CLL patients. PMID:21310924

miR-150 influences B-cell receptor signaling in chronic lymphocytic leukemia by regulating expression of GAB1 and FOXP1

PubMed Central

Mraz, Marek; Chen, Liguang; Rassenti, Laura Z.; Ghia, Emanuela M.; Li, Hongying; Jepsen, Kristen; Smith, Erin N.; Messer, Karen; Frazer, Kelly A.; Kipps, Thomas J.

2014-01-01

We examined the microRNAs (miRNAs) expressed in chronic lymphocytic leukemia (CLL) and identified miR-150 as the most abundant, but with leukemia cell expression levels that varied among patients. CLL cells that expressed ζ-chain–associated protein of 70 kDa (ZAP-70) or that used unmutated immunoglobulin heavy chain variable (IGHV) genes, each had a median expression level of miR-150 that was significantly lower than that of ZAP-70–negative CLL cells or those that used mutated IGHV genes. In samples stratified for expression of miR-150, CLL cells with low-level miR-150 expressed relatively higher levels of forkhead box P1 (FOXP1) and GRB2-associated binding protein 1 (GAB1), genes with 3′ untranslated regions having evolutionary-conserved binding sites for miR-150. High-level expression of miR-150 could repress expression of these genes, which encode proteins that enhance B-cell receptor signaling, a putative CLL-growth/survival signal. Also, high-level expression of miR-150 was a significant independent predictor of longer treatment-free survival or overall survival, whereas an inverse association was observed for high-level expression of GAB1 or FOXP1 for overall survival. This study demonstrates that expression of miR-150 can influence the relative expression of GAB1 and FOXP1 and the signaling potential of the B-cell receptor, thereby possibly accounting for the noted association of expression of miR-150 and disease outcome. PMID:24787006

miR-150 influences B-cell receptor signaling in chronic lymphocytic leukemia by regulating expression of GAB1 and FOXP1.

PubMed

Mraz, Marek; Chen, Liguang; Rassenti, Laura Z; Ghia, Emanuela M; Li, Hongying; Jepsen, Kristen; Smith, Erin N; Messer, Karen; Frazer, Kelly A; Kipps, Thomas J

2014-07-03

We examined the microRNAs (miRNAs) expressed in chronic lymphocytic leukemia (CLL) and identified miR-150 as the most abundant, but with leukemia cell expression levels that varied among patients. CLL cells that expressed ζ-chain-associated protein of 70 kDa (ZAP-70) or that used unmutated immunoglobulin heavy chain variable (IGHV) genes, each had a median expression level of miR-150 that was significantly lower than that of ZAP-70-negative CLL cells or those that used mutated IGHV genes. In samples stratified for expression of miR-150, CLL cells with low-level miR-150 expressed relatively higher levels of forkhead box P1 (FOXP1) and GRB2-associated binding protein 1 (GAB1), genes with 3' untranslated regions having evolutionary-conserved binding sites for miR-150. High-level expression of miR-150 could repress expression of these genes, which encode proteins that enhance B-cell receptor signaling, a putative CLL-growth/survival signal. Also, high-level expression of miR-150 was a significant independent predictor of longer treatment-free survival or overall survival, whereas an inverse association was observed for high-level expression of GAB1 or FOXP1 for overall survival. This study demonstrates that expression of miR-150 can influence the relative expression of GAB1 and FOXP1 and the signaling potential of the B-cell receptor, thereby possibly accounting for the noted association of expression of miR-150 and disease outcome. © 2014 by The American Society of Hematology.

Sequential phosphorylation of SLP-76 at tyrosine 173 is required for activation of T and mast cells

PubMed Central

Sela, Meirav; Bogin, Yaron; Beach, Dvora; Oellerich, Thomas; Lehne, Johanna; Smith-Garvin, Jennifer E; Okumura, Mariko; Starosvetsky, Elina; Kosoff, Rachelle; Libman, Evgeny; Koretzky, Gary; Kambayashi, Taku; Urlaub, Henning; Wienands, Jürgen; Chernoff, Jonathan; Yablonski, Deborah

2011-01-01

Cooperatively assembled signalling complexes, nucleated by adaptor proteins, integrate information from surface receptors to determine cellular outcomes. In T and mast cells, antigen receptor signalling is nucleated by three adaptors: SLP-76, Gads and LAT. Three well-characterized SLP-76 tyrosine phosphorylation sites recruit key components, including a Tec-family tyrosine kinase, Itk. We identified a fourth, evolutionarily conserved SLP-76 phosphorylation site, Y173, which was phosphorylated upon T-cell receptor stimulation in primary murine and Jurkat T cells. Y173 was required for antigen receptor-induced phosphorylation of phospholipase C-γ1 (PLC-γ1) in both T and mast cells, and for consequent downstream events, including activation of the IL-2 promoter in T cells, and degranulation and IL-6 production in mast cells. In intact cells, Y173 phosphorylation depended on three, ZAP-70-targeted tyrosines at the N-terminus of SLP-76 that recruit and activate Itk, a kinase that selectively phosphorylated Y173 in vitro. These data suggest a sequential mechanism whereby ZAP-70-dependent priming of SLP-76 at three N-terminal sites triggers reciprocal regulatory interactions between Itk and SLP-76, which are ultimately required to couple active Itk to its substrate, PLC-γ1. PMID:21725281

Sequential phosphorylation of SLP-76 at tyrosine 173 is required for activation of T and mast cells.

PubMed

Sela, Meirav; Bogin, Yaron; Beach, Dvora; Oellerich, Thomas; Lehne, Johanna; Smith-Garvin, Jennifer E; Okumura, Mariko; Starosvetsky, Elina; Kosoff, Rachelle; Libman, Evgeny; Koretzky, Gary; Kambayashi, Taku; Urlaub, Henning; Wienands, Jürgen; Chernoff, Jonathan; Yablonski, Deborah

2011-07-01

Cooperatively assembled signalling complexes, nucleated by adaptor proteins, integrate information from surface receptors to determine cellular outcomes. In T and mast cells, antigen receptor signalling is nucleated by three adaptors: SLP-76, Gads and LAT. Three well-characterized SLP-76 tyrosine phosphorylation sites recruit key components, including a Tec-family tyrosine kinase, Itk. We identified a fourth, evolutionarily conserved SLP-76 phosphorylation site, Y173, which was phosphorylated upon T-cell receptor stimulation in primary murine and Jurkat T cells. Y173 was required for antigen receptor-induced phosphorylation of phospholipase C-γ1 (PLC-γ1) in both T and mast cells, and for consequent downstream events, including activation of the IL-2 promoter in T cells, and degranulation and IL-6 production in mast cells. In intact cells, Y173 phosphorylation depended on three, ZAP-70-targeted tyrosines at the N-terminus of SLP-76 that recruit and activate Itk, a kinase that selectively phosphorylated Y173 in vitro. These data suggest a sequential mechanism whereby ZAP-70-dependent priming of SLP-76 at three N-terminal sites triggers reciprocal regulatory interactions between Itk and SLP-76, which are ultimately required to couple active Itk to its substrate, PLC-γ1.

ZaP-HD: High Energy Density Z-Pinch Plasmas using Sheared Flow Stabilization

NASA Astrophysics Data System (ADS)

Golingo, R. P.; Shumlak, U.; Nelson, B. A.; Claveau, E. L.; Doty, S. A.; Forbes, E. G.; Hughes, M. C.; Kim, B.; Ross, M. P.; Weed, J. R.

2015-11-01

The ZaP-HD flow Z-pinch project investigates scaling the flow Z-pinch to High Energy Density Plasma, HEDP, conditions by using sheared flow stabilization. ZaP used a single power supply to produce 100 cm long Z-pinches that were quiescent for many radial Alfven times and axial flow-through times. The flow Z-pinch concept provides an approach to achieve HED plasmas, which are dimensionally large and persist for extended durations. The ZaP-HD device replaces the single power supply from ZaP with two separate power supplies to independently control the plasma flow and current in the Z-pinch. Equilibrium is determined by diagnostic measurements of the density with interferometry and digital holography, the plasma flow and temperature with passive spectroscopy, the magnetic field with surface magnetic probes, and plasma emission with optical imaging. The diagnostics fully characterize the plasma from its initiation in the coaxial accelerator, through the pinch, and exhaust from the assembly region. The plasma evolution is modeled with high resolution codes: Mach2, WARPX, and NIMROD. Experimental results and scaling analyses are presented. This work is supported by grants from the U.S. Department of Energy and the U.S. National Nuclear Security Administration.

A Reactor Development Scenario for the FuZE Sheared-Flow Stabilized Z-pinch

NASA Astrophysics Data System (ADS)

McLean, Harry S.; Higginson, D. P.; Schmidt, A.; Tummel, K. K.; Shumlak, U.; Nelson, B. A.; Claveau, E. L.; Forbes, E. G.; Golingo, R. P.; Stepanov, A. D.; Weber, T. R.; Zhang, Y.

2017-10-01

We present a conceptual design, scaling calculations, and development path for a pulsed fusion reactor based on a flow-stabilized Z-pinch. Experiments performed on the ZaP and ZaP-HD devices have largely demonstrated the basic physics of sheared-flow stabilization at pinch currents up to 100 kA. Initial experiments on the FuZE device, a high-power upgrade of ZaP, have achieved 20 usec of stability at pinch current 100-200 kA and pinch diameter few mm for a pinch length of 50 cm. Scaling calculations based on a quasi-steady-state power balance show that extending stable duration to 100 usec at a pinch current of 1.5 MA and pinch length of 50 cm, results in a reactor plant Q 5. Future performance milestones are proposed for pinch currents of: 300 kA, where Te and Ti are calculated to exceed 1-2 keV; 700 kA, where DT fusion power would be expected to exceed pinch input power; and 1 MA, where fusion energy per pulse exceeds input energy per pulse. This work funded by USDOE ARPA-E and performed under the auspices of Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. LLNL-ABS-734770.

High-Pressure Neutron Diffraction Studies for Materials Sciences and Energy Sciences

NASA Astrophysics Data System (ADS)

Zhao, Y.; Los Alamos High Pressure Materials Research Team

2013-05-01

The development of neutron diffraction under extreme pressure (P) and temperature (T) conditions is highly valuable to condensed matter physics, crystal chemistry, materials sciences, as well as earth and planetary sciences. We have incorporated a 500-ton press TAP-98 into the HiPPO diffractometer at LANSCE to conduct in situ high P-T neutron diffraction experiments. We have worked out a large gem-crystal anvil cell, ZAP, to conduct neutron diffraction experiments at high-P and low-T. The ZAP cell can be used to integrate multiple experimental techniques such as neutron diffraction, laser spectroscopy, and ultrasonic interferometery. Recently, we have developed high-P low-T gas/fluid cells in conjunction with neutron diffraction and inelastic neutron scattering instruments. These techniques enable in-situ and real-time examination of gas uptake/release processes and allow high-resolution time-dependent determination of changes in crystal structure and related reaction kinetics. We have successfully used these techniques to study the equation of state, structural phase transition, and thermo-mechanical properties of metals, ceramics, and minerals. We have conducted researches on the formation of methane and hydrogen clathrates, and hydrogen adsorption of the inclusion compounds such as the recently discovered metal-organic frameworks (MOFs). The aim of our research is to accurately map phase diagram, lattice parameters, thermal parameters, bond lengths, bond angles, neighboring atomic environments, and phase stability in P-T-X space. We are currently developing further high P-T technology with a new "true" triaxial loading press, TAP_6x, to compress cubic sample package to achieve pressures up to 20 GPa and temperatures up to 2000 K in routine experiments. The implementation of TAP_6x300 with high-pressure neutron beamlines is underway for simultaneous high P-T neutron diffraction, ultrasonic, calorimetry, radiography, and tomography studies. Studies based on high-pressure neutron diffraction are important for multidisciplinary science, particularly for the theoretical/computational modeling/simulations.;

Towards Modernizing the Electrical Grid

DTIC Science & Technology

2011-05-01

Project is building four t t b t th t illpro o ype ro o s a w someday operate on extraterrestrial surfaces. The project coordinates four NASA ...Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to a penalty for failing to comply with a...Rivers’s PBRT (Proton Beam Radiation Therapy) system zaps tumors with accelerated protons. The treatment must be continuous for 30-40 days; downtime

Del(20q) in patients with chronic lymphocytic leukemia: a therapy-related abnormality involving lymphoid or myeloid cells.

PubMed

Yin, C Cameron; Tang, Guilin; Lu, Gary; Feng, Xiaoli; Keating, Michael J; Medeiros, L Jeffrey; Abruzzo, Lynne V

2015-08-01

Deletion 20q (Del(20q)), a common cytogenetic abnormality in myeloid neoplasms, is rare in chronic lymphocytic leukemia. We report 64 patients with chronic lymphocytic leukemia and del(20q), as the sole abnormality in 40, a stemline abnormality in 21, and a secondary abnormality in 3 cases. Fluorescence in situ hybridization (FISH) analysis revealed an additional high-risk abnormality, del(11q) or del(17p), in 25/64 (39%) cases. In most cases, the leukemic cells showed atypical cytologic features, unmutated IGHV (immunoglobulin heavy-chain variable region) genes, and ZAP70 positivity. The del(20q) was detected only after chemotherapy in all 27 cases with initial karyotypes available. With a median follow-up of 90 months, 30 patients (47%) died, most as a direct consequence of chronic lymphocytic leukemia. Eight patients developed a therapy-related myeloid neoplasm, seven with a complex karyotype. Combined morphologic and FISH analysis for del(20q) performed in 12 cases without morphologic evidence of a myeloid neoplasm localized the del(20q) to the chronic lymphocytic leukemia cells in 5 (42%) cases, and to myeloid/erythroid cells in 7 (58)% cases. The del(20q) was detected in myeloid cells in all 4 cases of myelodysplastic syndrome. In aggregate, these data indicate that chronic lymphocytic leukemia with del(20q) acquired after therapy is heterogeneous. In cases with morphologic evidence of dysplasia, the del(20q) likely resides in the myeloid lineage. However, in cases without morphologic evidence of dysplasia, the del(20q) may represent clonal evolution and disease progression. Combining morphologic analysis with FISH for del(20q) or performing FISH on immunomagnetically selected sub-populations to localize the cell population with this abnormality may help guide patient management.

Del(20q) in patients with chronic lymphocytic leukemia: A therapy-related abnormality involving lymphoid or myeloid cells

PubMed Central

Yin, C. Cameron; Tang, Guilin; Lu, Gary; Feng, Xiaoli; Keating, Michael J.; Medeiros, L. Jeffrey; Abruzzo, Lynne V.

2015-01-01

Del(20q), a common cytogenetic abnormality in myeloid neoplasms, is rare in chronic lymphocytic leukemia. We report 64 patients with chronic lymphocytic leukemia and del(20q), as the sole abnormality in 40, a stemline abnormality in 21, and a secondary abnormality in 3 cases. FISH analysis revealed an additional high-risk abnormality, del(11q) or del(17p), in 27/64 (42%) cases. In most cases, the leukemic cells showed atypical cytologic features, unmutated IGHV genes and ZAP70 positivity. The del(20q) was detected only after chemotherapy in all 27 cases with initial karyotypes available. With a median follow-up of 90 months, 30 patients (47%) died, most as a direct consequence of chronic lymphocytic leukemia. Eight patients developed a therapy-related myeloid neoplasm, seven with a complex karyotype. Combined morphologic and FISH analysis for del(20q) performed in 12 cases without morphologic evidence of a myeloid neoplasm localized the del(20q) to the chronic lymphocytic leukemia cells in 5 (42%) cases, and to myeloid/erythroid cells in 7 (58)% cases. The del(20q) was detected in myeloid cells in all 4 cases of myelodysplastic syndrome. In aggregate, these data indicate that chronic lymphocytic leukemia with del(20q) acquired after therapy is heterogeneous. In cases with morphologic evidence of dysplasia, the del(20q) likely resides in the myeloid lineage. However, in cases without morphologic evidence of dysplasia, the del(20q) may represent clonal evolution and disease progression. Combining morphologic analysis with FISH for del(20q) or performing FISH on immunomagnetically-selected subpopulations to localize the cell population with this abnormality may help guide patient management. PMID:25953391

Identification of cytosolic phosphodiesterases in the erythrocyte: A possible role for PDE5

PubMed Central

Adderley, Shaquria P.; Thuet, Kelly M.; Sridharan, Meera; Bowles, Elizabeth A.; Stephenson, Alan H.; Ellsworth, Mary L.; Sprague, Randy S.

2011-01-01

Summary Background Within erythrocytes (RBCs), cAMP levels are regulated by phosphodiesterases (PDEs). Increases in cAMP and ATP release associated with activation of β-adrenergic receptors (βARs) and prostacyclin receptors (IPRs) are regulated by PDEs 2, 4 and PDE 3, respectively. Here we establish the presence of cytosolic PDEs in RBCs and determine a role for PDE5 in regulating levels of cGMP. Material/Methods Purified cytosolic proteins were obtained from isolated human RBCs and western analysis was performed using antibodies against PDEs 3A, 4 and 5. Rabbit RBCs were incubated with dbcGMP, a cGMP analog, to determine the effect of cGMP on cAMP levels. To determine if cGMP affects receptor-mediated increases in cAMP, rabbit RBCs were incubated with dbcGMP prior to addition of isoproterenol (ISO), a βAR receptor agonist. To demonstrate that endogenous cGMP produces the same effect, rabbit and human RBCs were incubated with SpNONOate (SpNO), a nitric oxide donor, and YC1, a direct activator of soluble guanylyl cyclase (sGC), in the absence and presence of a selective PDE5 inhibitor, zaprinast (ZAP). Results Western analysis identified PDEs 3A, 4D and 5A. dbcGMP produced a concentration dependent increase in cAMP and ISO-induced increases in cAMP were potentiated by dbcGMP. In addition, incubation with YC1 and SpNO in the presence of ZAP potentiated βAR-induced increases in cAMP. Conclusions PDEs 2, 3A and 5 are present in the cytosol of human RBCs. PDE5 activity in RBCs regulates cGMP levels. Increases in intracellular cGMP augment cAMP levels. These studies suggest a novel role for PDE5 in erythrocytes. PMID:21525805

IP-FCM measures physiologic protein-protein interactions modulated by signal transduction and small-molecule drug inhibition.

PubMed

Smith, Stephen E P; Bida, Anya T; Davis, Tessa R; Sicotte, Hugues; Patterson, Steven E; Gil, Diana; Schrum, Adam G

2012-01-01

Protein-protein interactions (PPI) mediate the formation of intermolecular networks that control biological signaling. For this reason, PPIs are of outstanding interest in pharmacology, as they display high specificity and may represent a vast pool of potentially druggable targets. However, the study of physiologic PPIs can be limited by conventional assays that often have large sample requirements and relatively low sensitivity. Here, we build on a novel method, immunoprecipitation detected by flow cytometry (IP-FCM), to assess PPI modulation during either signal transduction or pharmacologic inhibition by two different classes of small-molecule compounds. First, we showed that IP-FCM can detect statistically significant differences in samples possessing a defined PPI change as low as 10%. This sensitivity allowed IP-FCM to detect a PPI that increases transiently during T cell signaling, the antigen-inducible interaction between ZAP70 and the T cell antigen receptor (TCR)/CD3 complex. In contrast, IP-FCM detected no ZAP70 recruitment when T cells were stimulated with antigen in the presence of the src-family kinase inhibitor, PP2. Further, we tested whether IP-FCM possessed sufficient sensitivity to detect the effect of a second, rare class of compounds called SMIPPI (small-molecule inhibitor of PPI). We found that the first-generation non-optimized SMIPPI, Ro-26-4550, inhibited the IL-2:CD25 interaction detected by IP-FCM. This inhibition was detectable using either a recombinant CD25-Fc chimera or physiologic full-length CD25 captured from T cell lysates. Thus, we demonstrate that IP-FCM is a sensitive tool for measuring physiologic PPIs that are modulated by signal transduction and pharmacologic inhibition.

Chronic Lymphocytic Leukemia in Chornobyl Cleanup Workers.

PubMed

Bazyka, Dimitry; Gudzenko, Natalya; Dyagil, Iryna; Goroh, Eugeny; Polyschuk, Oksana; Trotsuk, Natalya; Babkina, Nataly; Romanenko, Anatoly

2016-08-01

This paper describes the chronic lymphocytic leukemia (CLL) incidence in a cohort of 110,645 (enlarged later to 152,520) male Ukrainian cleanup workers of the Chornobyl (Chernobyl) accident who were exposed to a range of radiation doses over the 1986-1990 time period. The standardized incidence rates are presented for a 27-y period after the exposure. For 2007-2012 period, the authors have identified the incident CLL cases in an enlarged cohort of 152,520 persons by linkage of the cohort file with the Ukrainian National Cancer Registry (NCRU). CLL data for the previous period (1987-2006) were identified in a frame of the Ukrainian-American leukemia study in the original cohort of 110,645 male clean-up workers. A significant CLL incidence excess was shown for the entire study period 1987-2012, with more prominent levels for the earliest years (1987-1996) when the standardized incidence rate (SIR) value was estimated to be 3.61 with 95% confidence interval from 2.32 to 4.91. In 2007-2012, the CLL incidence decreased substantially but still exceeded the national level although not significantly. In parallel, the several studies were performed at the National Research Center for Radiation Medicine (NRCRM) to explore if any clinical and cytogenetic features of CLL existed in the clean-up workers. The clinical study included 80 exposed and 70 unexposed CLL cases. Among the major clinical differences of the CLL course in the clean-up workers were a shorter period of white blood cells (WBC) doubling (10.7 vs. 18.0; p<0.001), frequent infectious episodes, lymphoadenopathy and hepatosplenomegaly (37 vs. 16), higher expression for CD38, and lower expression for ZAP-70 antigen.

ChemCam for Mars Science Laboratory rover, undergoing pre-flight testing

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

2011-10-20

Los Alamos National Laboratory and partners developed a laser instrument, ChemCam, that will ride on the elevated mast of the Mars Science Laboratory rover Curiosity. The system allows Curiosity to "zap" rocks from a distance, reading their chemical composition through spectroscopic analysis. In this video, laboratory shaker-table testing of the instrument ensures that all of its components are solidly attached and resistant to damage from the rigors of launch, travel and landing.

ChemCam for Mars Science Laboratory rover, undergoing pre-flight testing

ScienceCinema

None

2018-06-06

Los Alamos National Laboratory and partners developed a laser instrument, ChemCam, that will ride on the elevated mast of the Mars Science Laboratory rover Curiosity. The system allows Curiosity to "zap" rocks from a distance, reading their chemical composition through spectroscopic analysis. In this video, laboratory shaker-table testing of the instrument ensures that all of its components are solidly attached and resistant to damage from the rigors of launch, travel and landing.

A LANL Scientist's Dream Takes Off to Zap Rocks on Mars

ScienceCinema

Wiens, Roger

2018-05-11

Roger Wiens, with a team of 40 people at Los Alamos National Laboratory and the collaboration of the French space institute IRAP, created ChemCam, a laser spectrometer and telescope device aboard the Curiosity rover. ChemCam will blast rocks from as far as 7 meters, vaporize bits of their surfaces, and spectroscopically determine their chemical composition, aiding in the search for life on Mars, and making this scientist's boyhood dream a reality.

A LANL Scientist's Dream Takes Off to Zap Rocks on Mars

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wiens, Roger

2012-07-12

Roger Wiens, with a team of 40 people at Los Alamos National Laboratory and the collaboration of the French space institute IRAP, created ChemCam, a laser spectrometer and telescope device aboard the Curiosity rover. ChemCam will blast rocks from as far as 7 meters, vaporize bits of their surfaces, and spectroscopically determine their chemical composition, aiding in the search for life on Mars, and making this scientist's boyhood dream a reality.

CT brush and CancerZap!: two video games for computed tomography dose minimization.

PubMed

Alvare, Graham; Gordon, Richard

2015-05-12

X-ray dose from computed tomography (CT) scanners has become a significant public health concern. All CT scanners spray x-ray photons across a patient, including those using compressive sensing algorithms. New technologies make it possible to aim x-ray beams where they are most needed to form a diagnostic or screening image. We have designed a computer game, CT Brush, that takes advantage of this new flexibility. It uses a standard MART algorithm (Multiplicative Algebraic Reconstruction Technique), but with a user defined dynamically selected subset of the rays. The image appears as the player moves the CT brush over an initially blank scene, with dose accumulating with every "mouse down" move. The goal is to find the "tumor" with as few moves (least dose) as possible. We have successfully implemented CT Brush in Java and made it available publicly, requesting crowdsourced feedback on improving the open source code. With this experience, we also outline a "shoot 'em up game" CancerZap! for photon limited CT. We anticipate that human computing games like these, analyzed by methods similar to those used to understand eye tracking, will lead to new object dependent CT algorithms that will require significantly less dose than object independent nonlinear and compressive sensing algorithms that depend on sprayed photons. Preliminary results suggest substantial dose reduction is achievable.

p56Lck and p59Fyn Regulate CD28 Binding to Phosphatidylinositol 3-Kinase, Growth Factor Receptor-Bound Protein GRB-2, and T Cell-Specific Protein-Tyrosine Kinase ITK: Implications for T-Cell Costimulation

NASA Astrophysics Data System (ADS)

Raab, Monika; Cai, Yun-Cai; Bunnell, Stephen C.; Heyeck, Stephanie D.; Berg, Leslie J.; Rudd, Christopher E.

1995-09-01

T-cell activation requires cooperative signals generated by the T-cell antigen receptor ξ-chain complex (TCRξ-CD3) and the costimulatory antigen CD28. CD28 interacts with three intracellular proteins-phosphatidylinositol 3-kinase (PI 3-kinase), T cell-specific protein-tyrosine kinase ITK (formerly TSK or EMT), and the complex between growth factor receptor-bound protein 2 and son of sevenless guanine nucleotide exchange protein (GRB-2-SOS). PI 3-kinase and GRB-2 bind to the CD28 phosphotyrosine-based Tyr-Met-Asn-Met motif by means of intrinsic Src-homology 2 (SH2) domains. The requirement for tyrosine phosphorylation of the Tyr-Met-Asn-Met motif for SH2 domain binding implicates an intervening protein-tyrosine kinase in the recruitment of PI 3-kinase and GRB-2 by CD28. Candidate kinases include p56Lck, p59Fyn, ξ-chain-associated 70-kDa protein (ZAP-70), and ITK. In this study, we demonstrate in coexpression studies that p56Lck and p59Fyn phosphorylate CD28 primarily at Tyr-191 of the Tyr-Met-Asn-Met motif, inducing a 3- to 8-fold increase in p85 (subunit of PI 3-kinase) and GRB-2 SH2 binding to CD28. Phosphatase digestion of CD28 eliminated binding. In contrast to Src kinases, ZAP-70 and ITK failed to induce these events. Further, ITK binding to CD28 was dependent on the presence of p56Lck and is thus likely to act downstream of p56Lck/p59Fyn in a signaling cascade. p56Lck is therefore likely to be a central switch in T-cell activation, with the dual function of regulating CD28-mediated costimulation as well as TCR-CD3-CD4 signaling.

Spectroscopic confirmation and photometry of the Fe II novae M31N 2017-11a (AT2017hvi) and M31N 2017-11c

NASA Astrophysics Data System (ADS)

Fabrika, S.; Sholukhova, O.; Vinokurov, A.; Valeev, A. F.; Solovyeva, Yu.; Hornoch, K.; Henze, M.; Shafter, A. W.

2017-11-01

We report optical spectroscopic confirmation of the two recent M31 nova candidates M31N 2017-11a (AT2017hvi) and M31N 2017-11c. The first nova was discovered on 2017-11-04.695 by PMO-Tsinghua Supernova Survey (PTSS-17zap); the second was discovered on 2017-11-12.465 by K. Nishiyama and F. Kabashima (=TCP J00414435+4108287).

Deconvolution of Stark broadened spectra for multi-point density measurements in a flow Z-pinch

DOE PAGES

Vogman, G. V.; Shumlak, U.

2011-10-13

Stark broadened emission spectra, once separated from other broadening effects, provide a convenient non-perturbing means of making plasma density measurements. A deconvolution technique has been developed to measure plasma densities in the ZaP flow Z-pinch experiment. The ZaP experiment uses sheared flow to mitigate MHD instabilities. The pinches exhibit Stark broadened emission spectra, which are captured at 20 locations using a multi-chord spectroscopic system. Spectra that are time- and chord-integrated are well approximated by a Voigt function. The proposed method simultaneously resolves plasma electron density and ion temperature by deconvolving the spectral Voigt profile into constituent functions: a Gaussian functionmore » associated with instrument effects and Doppler broadening by temperature; and a Lorentzian function associated with Stark broadening by electron density. The method uses analytic Fourier transforms of the constituent functions to fit the Voigt profile in the Fourier domain. The method is discussed and compared to a basic least-squares fit. The Fourier transform fitting routine requires fewer fitting parameters and shows promise in being less susceptible to instrumental noise and to contamination from neighboring spectral lines. The method is evaluated and tested using simulated lines and is applied to experimental data for the 229.69 nm C III line from multiple chords to determine plasma density and temperature across the diameter of the pinch. As a result, these measurements are used to gain a better understanding of Z-pinch equilibria.« less

Deconvolution of Stark broadened spectra for multi-point density measurements in a flow Z-pinch

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vogman, G. V.; Shumlak, U.

2011-10-15

Stark broadened emission spectra, once separated from other broadening effects, provide a convenient non-perturbing means of making plasma density measurements. A deconvolution technique has been developed to measure plasma densities in the ZaP flow Z-pinch experiment. The ZaP experiment uses sheared flow to mitigate MHD instabilities. The pinches exhibit Stark broadened emission spectra, which are captured at 20 locations using a multi-chord spectroscopic system. Spectra that are time- and chord-integrated are well approximated by a Voigt function. The proposed method simultaneously resolves plasma electron density and ion temperature by deconvolving the spectral Voigt profile into constituent functions: a Gaussian functionmore » associated with instrument effects and Doppler broadening by temperature; and a Lorentzian function associated with Stark broadening by electron density. The method uses analytic Fourier transforms of the constituent functions to fit the Voigt profile in the Fourier domain. The method is discussed and compared to a basic least-squares fit. The Fourier transform fitting routine requires fewer fitting parameters and shows promise in being less susceptible to instrumental noise and to contamination from neighboring spectral lines. The method is evaluated and tested using simulated lines and is applied to experimental data for the 229.69 nm C III line from multiple chords to determine plasma density and temperature across the diameter of the pinch. These measurements are used to gain a better understanding of Z-pinch equilibria.« less

Impaired NFAT and NFκB activation are involved in suppression of CD40 ligand expression by Δ{sup 9}-tetrahydrocannabinol in human CD4{sup +} T cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ngaotepprutaram, Thitirat; Center for Integrative Toxicology, Michigan State University; Kaplan, Barbara L.F.

We have previously reported that Δ{sup 9}-tetrahydrocannabinol (Δ{sup 9}-THC), the main psychoactive cannabinoid in marijuana, suppresses CD40 ligand (CD40L) expression by activated mouse CD4{sup +} T cells. CD40L is involved in pathogenesis of many autoimmune and inflammatory diseases. In the present study, we investigated the molecular mechanism of Δ{sup 9}-THC-mediated suppression of CD40L expression using peripheral blood human T cells. Pretreatment with Δ{sup 9}-THC attenuated CD40L expression in human CD4{sup +} T cells activated by anti-CD3/CD28 at both the protein and mRNA level, as determined by flow cytometry and quantitative real-time PCR, respectively. Electrophoretic mobility shift assays revealed that Δ{supmore » 9}-THC suppressed the DNA-binding activity of both NFAT and NFκB to their respective response elements within the CD40L promoter. An assessment of the effect of Δ{sup 9}-THC on proximal T cell-receptor (TCR) signaling induced by anti-CD3/CD28 showed significant impairment in the rise of intracellular calcium, but no significant effect on the phosphorylation of ZAP70, PLCγ1/2, Akt, and GSK3β. Collectively, these findings identify perturbation of the calcium-NFAT and NFκB signaling cascade as a key mechanistic event by which Δ{sup 9}-THC suppresses human T cell function. - Highlights: • Δ{sup 9}-THC attenuated CD40L expression in activated human CD4+ T cells. • Δ{sup 9}-THC suppressed DNA-binding activity of NFAT and NFκB. • Δ{sup 9}-THC impaired elevation of intracellular Ca2+. • Δ{sup 9}-THC did not affect phosphorylation of ZAP70, PLCγ1/2, Akt, and GSK3β.« less

Src-like adaptor protein down-regulates T cell receptor (TCR)-CD3 expression by targeting TCRzeta for degradation.

PubMed

Myers, Margaret D; Dragone, Leonard L; Weiss, Arthur

2005-07-18

Src-like adaptor protein (SLAP) down-regulates expression of the T cell receptor (TCR)-CD3 complex during a specific stage of thymocyte development when the TCR repertoire is selected. Consequently, SLAP-/- thymocytes display alterations in thymocyte development. Here, we have studied the mechanism of SLAP function. We demonstrate that SLAP-deficient thymocytes have increased TCRzeta chain expression as a result of a defect in TCRzeta degradation. Failure to degrade TCRzeta leads to an increased pool of fully assembled TCR-CD3 complexes that are capable of recycling back to the cell surface. We also provide evidence that SLAP functions in a pathway that requires the phosphorylated TCRzeta chain and the Src family kinase Lck, but not ZAP-70 (zeta-associated protein of 70 kD). These studies reveal a unique mechanism by which SLAP contributes to the regulation of TCR expression during a distinct stage of thymocyte development.

Science& Technology Review December 2002

DOE Office of Scientific and Technical Information (OSTI.GOV)

Budil, K S

2002-10-28

This issue has the following articles: (1) ''Doing It All: Sustaining Our Working Solutions, Rising to New Challenges''; (2) ''Emerging from the Cold War: Stockpile Stewardship and Beyond''--When the Cold War ended, Lawrence Livermore stepped up to a new national challenge--maintaining the safety and reliability of the U.S. nuclear stockpile without underground testing. (3) ''Machines from Interlocking Molecules''--Fundamental chemistry and physics research will enable scientists to control and use individual molecules. (4) ''Laser Zaps Communication Bottleneck''--Using laser communications, the U.S. military will be able to transmit data from advanced remote sensors in real time.

Apollo 14 rock samples

NASA Technical Reports Server (NTRS)

Carlson, I. C.

1978-01-01

Petrographic descriptions of all Apollo 14 samples larger than 1 cm in any dimension are presented. The sample description format consists of: (1) an introductory section which includes information on lunar sample location, orientation, and return containers, (2) a section on physical characteristics, which contains the sample mass, dimensions, and a brief description; (3) surface features, including zap pits, cavities, and fractures as seen in binocular view; (4) petrographic description, consisting of a binocular description and, if possible, a thin section description; and (5) a discussion of literature relevant to sample petrology is included for samples which have previously been examined by the scientific community.

Proteomic profiling of the human T-cell nucleolus.

PubMed

Jarboui, Mohamed Ali; Wynne, Kieran; Elia, Giuliano; Hall, William W; Gautier, Virginie W

2011-12-01

The nucleolus, site of ribosome biogenesis, is a dynamic subnuclear organelle involved in diverse cellular functions. The size, number and organisation of nucleoli are cell-specific and while it remains to be established, the nucleolar protein composition would be expected to reflect lineage-specific transcriptional regulation of rDNA genes and have cell-type functional components. Here, we describe the first characterisation of the human T-cell nucleolar proteome. Using the Jurkat T-cell line and a reproducible organellar proteomic approach, we identified 872 nucleolar proteins. In addition to ribosome biogenesis and RNA processing networks, network modeling and topological analysis of nucleolar proteome revealed distinct macromolecular complexes known to orchestrate chromatin structure and to contribute to the regulation of gene expression, replication, recombination and repair, and chromosome segregation. Furthermore, among our dataset, we identified proteins known to functionally participate in T-cell biology, including RUNX1, ILF3, ILF2, STAT3, LSH, TCF-1, SATB1, CTCF, HMGB3, BCLAF1, FX4L1, ZAP70, TIAM1, RAC2, THEMIS, LCP1, RPL22, TOPK, RETN, IFI-16, MCT-1, ISG15, and 14-3-3τ, which support cell-specific composition of the Jurkat nucleolus. Subsequently, the nucleolar localisation of RUNX1, ILF3, STAT3, ZAP70 and RAC2 was further validated by Western Blot analysis and immunofluorescence microscopy. Overall, our T-cell nucleolar proteome dataset not only further expands the existing repertoire of the human nucleolar proteome but support a cell type-specific composition of the nucleolus in T cell and highlights the potential roles of the nucleoli in lymphocyte biology. Copyright © 2011 Elsevier Ltd. All rights reserved.

Signaling of the ITK (interleukin 2-inducible T cell kinase)-SYK (spleen tyrosine kinase) fusion kinase is dependent on adapter SLP-76 and on the adapter function of the kinases SYK and ZAP70.

PubMed

Hussain, Alamdar; Mohammad, Dara K; Gustafsson, Manuela O; Uslu, Merve; Hamasy, Abdulrahman; Nore, Beston F; Mohamed, Abdalla J; Smith, C I Edvard

2013-03-08

The inducible T cell kinase-spleen tyrosine kinase (ITK-SYK) oncogene consists of the Tec homology-pleckstrin homology domain of ITK and the kinase domain of SYK, and it is believed to be the cause of peripheral T cell lymphoma. We and others have recently demonstrated that this fusion protein is constitutively tyrosine-phosphorylated and is transforming both in vitro and in vivo. To gain a deeper insight into the molecular mechanism(s) underlying its activation and signaling, we mutated a total of eight tyrosines located in the SYK portion of the chimera into either phenylalanine or to the negatively charged glutamic acid. Although mutations in the interdomain-B region affected ITK-SYK kinase activity, they only modestly altered downstream signaling events. In contrast, mutations that were introduced in the kinase domain triggered severe impairment of downstream signaling. Moreover, we show here that SLP-76 is critical for ITK-SYK activation and is particularly required for the ITK-SYK-dependent phosphorylation of SYK activation loop tyrosines. In Jurkat cell lines, we demonstrate that expression of ITK-SYK fusion requires an intact SLP-76 function and significantly induces IL-2 secretion and CD69 expression. Furthermore, the SLP-76-mediated induction of IL-2 and CD69 could be further enhanced by SYK or ZAP-70, but it was independent of their kinase activity. Notably, ITK-SYK expression in SYF cells phosphorylates SLP-76 in the absence of SRC family kinases. Altogether, our data suggest that ITK-SYK exists in the active conformation state and is therefore capable of signaling without SRC family kinases or stimulation of the T cell receptor.

HIV-1 Nef limits communication between linker of activated T cells and SLP-76 to reduce formation of SLP-76-signaling microclusters following TCR stimulation.

PubMed

Abraham, Libin; Bankhead, Peter; Pan, Xiaoyu; Engel, Ulrike; Fackler, Oliver T

2012-08-15

Signal initiation by engagement of the TCR triggers actin rearrangements, receptor clustering, and dynamic organization of signaling complexes to elicit and sustain downstream signaling. Nef, a pathogenicity factor of HIV, disrupts early TCR signaling in target T cells. To define the mechanism underlying this Nef-mediated signal disruption, we employed quantitative single-cell microscopy following surface-mediated TCR stimulation that allows for dynamic visualization of distinct signaling complexes as microclusters (MCs). Despite marked inhibition of actin remodeling and cell spreading, the induction of MCs containing TCR-CD3 or ZAP70 was not affected significantly by Nef. However, Nef potently inhibited the subsequent formation of MCs positive for the signaling adaptor Src homology-2 domain-containing leukocyte protein of 76 kDa (SLP-76) to reduce MC density in Nef-expressing and HIV-1-infected T cells. Further analyses suggested that Nef prevents formation of SLP-76 MCs at the level of the upstream adaptor protein, linker of activated T cells (LAT), that couples ZAP70 to SLP-76. Nef did not disrupt pre-existing MCs positive for LAT. However, the presence of the viral protein prevented de novo recruitment of active LAT into MCs due to retargeting of LAT to an intracellular compartment. These modulations in MC formation and composition depended on Nef's ability to simultaneously disrupt both actin remodeling and subcellular localization of TCR-proximal machinery. Nef thus employs a dual mechanism to disturb early TCR signaling by limiting the communication between LAT and SLP-76 and preventing the dynamic formation of SLP-76-signaling MCs.

Signaling of the ITK (Interleukin 2-inducible T Cell Kinase)-SYK (Spleen Tyrosine Kinase) Fusion Kinase Is Dependent on Adapter SLP-76 and on the Adapter Function of the Kinases SYK and ZAP70*

PubMed Central

Hussain, Alamdar; Mohammad, Dara K.; Gustafsson, Manuela O.; Uslu, Merve; Hamasy, Abdulrahman; Nore, Beston F.; Mohamed, Abdalla J.; Smith, C. I. Edvard

2013-01-01

The inducible T cell kinase-spleen tyrosine kinase (ITK-SYK) oncogene consists of the Tec homology-pleckstrin homology domain of ITK and the kinase domain of SYK, and it is believed to be the cause of peripheral T cell lymphoma. We and others have recently demonstrated that this fusion protein is constitutively tyrosine-phosphorylated and is transforming both in vitro and in vivo. To gain a deeper insight into the molecular mechanism(s) underlying its activation and signaling, we mutated a total of eight tyrosines located in the SYK portion of the chimera into either phenylalanine or to the negatively charged glutamic acid. Although mutations in the interdomain-B region affected ITK-SYK kinase activity, they only modestly altered downstream signaling events. In contrast, mutations that were introduced in the kinase domain triggered severe impairment of downstream signaling. Moreover, we show here that SLP-76 is critical for ITK-SYK activation and is particularly required for the ITK-SYK-dependent phosphorylation of SYK activation loop tyrosines. In Jurkat cell lines, we demonstrate that expression of ITK-SYK fusion requires an intact SLP-76 function and significantly induces IL-2 secretion and CD69 expression. Furthermore, the SLP-76-mediated induction of IL-2 and CD69 could be further enhanced by SYK or ZAP-70, but it was independent of their kinase activity. Notably, ITK-SYK expression in SYF cells phosphorylates SLP-76 in the absence of SRC family kinases. Altogether, our data suggest that ITK-SYK exists in the active conformation state and is therefore capable of signaling without SRC family kinases or stimulation of the T cell receptor. PMID:23293025

CD25 and CD69 induction by α4β1 outside-in signalling requires TCR early signalling complex proteins

PubMed Central

Cimo, Ann-Marie; Ahmed, Zamal; McIntyre, Bradley W.; Lewis, Dorothy E.; Ladbury, John E.

2013-01-01

Distinct signalling pathways producing diverse cellular outcomes can utilize similar subsets of proteins. For example, proteins from the TCR (T-cell receptor) ESC (early signalling complex) are also involved in interferon-α receptor signalling. Defining the mechanism for how these proteins function within a given pathway is important in understanding the integration and communication of signalling networks with one another. We investigated the contributions of the TCR ESC proteins Lck (lymphocyte-specific kinase), ZAP-70 (ζ-chain-associated protein of 70 kDa), Vav1, SLP-76 [SH2 (Src homology 2)-domain-containing leukocyte protein of 76 kDa] and LAT (linker for activation of T-cells) to integrin outside-in signalling in human T-cells. Lck, ZAP-70, SLP-76, Vav1 and LAT were activated by α4β1 outside-in signalling, but in a manner different from TCR signalling. TCR stimulation recruits ESC proteins to activate the mitogen-activated protein kinase ERK (extracellular-signal-regulated kinase). α4β1 outside-in-mediated ERK activation did not require TCR ESC proteins. However, α4β1 outside-in signalling induced CD25 and co-stimulated CD69 and this was dependent on TCR ESC proteins. TCR and α4β1 outside-in signalling are integrated through the common use of TCR ESC proteins; however, these proteins display functionally distinct roles in these pathways. These novel insights into the cross-talk between integrin outside-in and TCR signalling pathways are highly relevant to the development of therapeutic strategies to overcome disease associated with T-cell deregulation. PMID:23758320

LANL Researcher Roger Wiens Discusses ChemCam

ScienceCinema

Wiens, Roger

2018-01-16

Discussion of the ChemCam instrument on the Curiosity Rover that occurred during the NASA press conference prior to launch of the Mars Science Laboratory. The ChemCam instrument was developed by Los Alamos National Laboratory and the French Space Institute. Los Alamos National Laboratory researcher Roger Wiens discusses the instrument on this video. ChemCam uses a laser to "zap" features of the Martian landscape and then uses a spectrometer to gather information about the composition of the sample. ChemCam will help the Curiosity Rover determine whether Mars is or was habitable. The Rover is expected to touch down on the Red Planet on August 5, 2012.

ZAP! The X-Ray Laser is Born

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ratner, Daniel

2009-11-17

SLAC has converted its giant particle accelerator into the world's first X-ray laser. By a billion fold the world's brightest X-ray source, the laser packs a trillion photons into pulses as short as a millionth of a billionth of a second. The ultra-bright, ultra-short X-ray pulses will drive a wide range of new experiments, as scientists strip electrons from atoms, photograph single molecules and make movies of chemical reactions. How has SLAC accomplished such feats of X-ray wizardry? Attend this public lecture to learn about the basics of an X-ray laser, the technologies at SLAC that make it possible, andmore » the exciting new experiments now underway.« less

LANL Researcher Roger Wiens Discusses ChemCam

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wiens, Roger

2012-02-15

Discussion of the ChemCam instrument on the Curiosity Rover that occurred during the NASA press conference prior to launch of the Mars Science Laboratory. The ChemCam instrument was developed by Los Alamos National Laboratory and the French Space Institute. Los Alamos National Laboratory researcher Roger Wiens discusses the instrument on this video. ChemCam uses a laser to "zap" features of the Martian landscape and then uses a spectrometer to gather information about the composition of the sample. ChemCam will help the Curiosity Rover determine whether Mars is or was habitable. The Rover is expected to touch down on the Redmore » Planet on August 5, 2012.« less

Microgravity and Cellular Consequences in Lymphocyte Function

NASA Technical Reports Server (NTRS)

Pellis, Neal R.; Sundaresan, Alamelu

2004-01-01

Mammalian cells adapt to the environment of low gravity and express a series of responses, some possibly from direct effects on cells and others based on environmental conditions created by microgravity. Human lymphocytes in microgravity culture are functionally diminished in activation and locomotion. Both processes are integral to optimal immune response to fight pathogens. The NASA Rotating-wall vessel (RWV) is a well-accepted analog for microgravity culture on the ground. Gene array experiments and immunoblotting identified upstream events in human lymphocytes adapting to microgravity analog culture. Microgravity induces selective changes, many of which are cell membrane related. Results showed that upstream of PKC in the T cell activation cascade, PLC-gamma and LAT are significantly diminished. ZAP 70 which controls LAT activation is also down regulated in modeled microgravity. Thus events governing cell shape might warrant attention in microgravity conditions. The goal of this study is to delineate response suites that are consequential, direct or indirect effects of the microgravity environment and which of these are essential to lymphocytes

The Role of Positive Psychology in the Modern Medical Practice.

PubMed

Friedman, Scott E; Baum, Neil

2016-01-01

American healthcare has been shrouded in a cloak of negativity for too long. Many doctors are more pessimistic about the state of healthcare than they were even a few decades ago. In spite of extraordinary advances in the clinical aspects of healthcare, the business side of medicine has created a downward spiral in physician spirit, resulting in unnecessary stress, zapped energy, a rise in interpersonal conflict and, ultimately and as a byproduct of this negativity, reduced patient satisfaction. This downward spiral needs to be--and can be--stemmed. This article discuss how insights from the field of positive psychology and social neuroscience can help healthcare providers and their organizations flourish, in both their professional practices and personal lives.

Design and calibration of zero-additional-phase SPIDER

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baum, Peter; Riedle, Eberhard

2005-09-01

Zero-additional-phase spectral phase interferometry for direct electric field reconstruction (ZAP-SPIDER) is a novel technique for measuring the temporal shape and phase of ultrashort optical pulses directly at the interaction point of a spectroscopic experiment. The scheme is suitable for an extremely wide wavelength region from the ultraviolet to the near infrared. We present a comprehensive description of the experimental setup and design guidelines to effectively apply the technique to various wavelengths and pulse durations. The calibration of the setup and procedures to check the consistency of the measurement are discussed in detail. We show experimental data for various center wavelengthsmore » and pulse durations down to 7 fs to verify the applicability to a wide range of pulse parameters.« less

SH2 Domains Serve as Lipid-Binding Modules for pTyr-Signaling Proteins.

PubMed

Park, Mi-Jeong; Sheng, Ren; Silkov, Antonina; Jung, Da-Jung; Wang, Zhi-Gang; Xin, Yao; Kim, Hyunjin; Thiagarajan-Rosenkranz, Pallavi; Song, Seohyeon; Yoon, Youngdae; Nam, Wonhee; Kim, Ilshin; Kim, Eui; Lee, Dong-Gyu; Chen, Yong; Singaram, Indira; Wang, Li; Jang, Myoung Ho; Hwang, Cheol-Sang; Honig, Barry; Ryu, Sungho; Lorieau, Justin; Kim, You-Me; Cho, Wonhwa

2016-04-07

The Src-homology 2 (SH2) domain is a protein interaction domain that directs myriad phosphotyrosine (pY)-signaling pathways. Genome-wide screening of human SH2 domains reveals that ∼90% of SH2 domains bind plasma membrane lipids and many have high phosphoinositide specificity. They bind lipids using surface cationic patches separate from pY-binding pockets, thus binding lipids and the pY motif independently. The patches form grooves for specific lipid headgroup recognition or flat surfaces for non-specific membrane binding and both types of interaction are important for cellular function and regulation of SH2 domain-containing proteins. Cellular studies with ZAP70 showed that multiple lipids bind its C-terminal SH2 domain in a spatiotemporally specific manner and thereby exert exquisite spatiotemporal control over its protein binding and signaling activities in T cells. Collectively, this study reveals how lipids control SH2 domain-mediated cellular protein-protein interaction networks and suggest a new strategy for therapeutic modulation of pY-signaling pathways. Copyright © 2016 Elsevier Inc. All rights reserved.

PARP13 and RNA regulation in immunity and cancer

PubMed Central

Todorova, Tanya; Bock, Florian; Chang, Paul

2015-01-01

Posttranscriptional regulation of RNA is an important mechanism for activating and resolving cellular stress responses. Poly(ADP-ribose) Polymerase-13 (PARP13), also known as ZC3HAV1 and Zinc-finger Antiviral Protein (ZAP), is an RNA-binding protein that regulates the stability, and translation of specific mRNAs, and modulates the miRNA silencing pathway to globally impact miRNA targets. These functions of PARP13 are important components of the cellular response to stress. In addition, the ability of PARP13 to restrict oncogenic viruses and to repress the pro-survival cytokine receptor TRAILR4 suggests that it can be protective against malignant transformation and cancer development. The relevance of PARP13 to human health and disease make it a promising therapeutic target. PMID:25851173

The optical re-brightening of nova M31N 2017-11a

NASA Astrophysics Data System (ADS)

Xu, Zhijian; Gao, Xing; Li, Yanxi; Zhao, Jingyuan; Zhang, Mi

2017-12-01

We report the initial discovery of the optical re-brightening of the Fe II class nova M31N 2017-11a (AT2017hvi = PTSS-17zap) which was first reported by PMO-Tsinghua Supernova Survey (PTSS, http://www.cneost.org/ptss/ ), (2017, TNS Discovery Report https://wis-tns.weizmann.ac.il/object/2017hvi) at r-Sloan magnitude 18.5 on 2017-11-04 16:41:02 UT. Spectroscopy by Williams & Darnley using the 2-m Liverpool telescope (ATel #10990) on 2017 Nov 20.11 UT, and by Fabrika et al., (ATel #10998) taken two days later at the Russian BTA telescope, showed Balmer emission lines together with numerous strong Fe II lines, confirming its classification as a classical Fe II class nova.

Diffuse large B-cell lymphoma (Richter syndrome) in patients with chronic lymphocytic leukaemia (CLL): a cohort study of newly diagnosed patients.

PubMed

Parikh, Sameer A; Rabe, Kari G; Call, Timothy G; Zent, Clive S; Habermann, Thomas M; Ding, Wei; Leis, Jose F; Schwager, Susan M; Hanson, Curtis A; Macon, William R; Kay, Neil E; Slager, Susan L; Shanafelt, Tait D

2013-09-01

Nearly all information about patients with chronic lymphocytic leukaemia (CLL) who develop diffuse large B-cell lymphoma [Richter syndrome (RS)] is derived from retrospective case series or patients treated on clinical trials. We used the Mayo Clinic CLL Database to identify patients with newly diagnosed CLL between January 2000 and July 2011. Individuals who developed biopsy-proven RS during follow-up were identified. After a median follow-up of 4 years, 37/1641 (2·3%) CLL patients developed RS. The rate of RS was approximately 0·5%/year. Risk of RS was associated with advanced Rai stage at diagnosis (P < 0·001), high-risk genetic abnormalitites on fluorescence in situ hybridization (P < 0·0001), unmutated IGHV (P = 0·003), and expression of ZAP70 (P = 0·02) and CD38 (P = 0·001). The rate of RS doubled in patients after treatment for CLL (1%/year). Stereotyped B-cell receptors (odds-ratio = 4·2; P = 0·01) but not IGHV4-39 family usage was associated with increased risk of RS. Treatment with combination of purine analogues and alkylating agents increased the risk of RS three-fold (odds-ratio = 3·26, P = 0·0003). Median survival after RS diagnosis was 2·1 years. The RS prognosis score stratified patients into three risk groups with median survivals of 0·5 years, 2·1 years and not reached. Both underlying characteristics of the CLL clone and subsequent CLL therapy influence the risk of RS. Survival after RS remains poor and new therapies are needed. © 2013 John Wiley & Sons Ltd.

The effect of CD4 receptor downregulation and its downstream signaling molecules on HIV-1 latency

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, Kyung-Chang; School of Life Science and Biotechnology, Korea University, Seoul; Kim, Hyeon Guk

2011-01-14

Research highlights: {yields} CD4 receptors were downregulated on the surface of HIV-1 latently infected cells. {yields} CD4 downstream signaling molecules were suppressed in HIV-1 latently infected cells. {yields} HIV-1 progeny can be reactivated by induction of T-cell activation signal molecules. {yields} H3K4me3 and H3K9ac were highly enriched in CD4 downstream signaling molecules. {yields} HIV-1 latency can be maintained by the reduction of downstream signaling molecules. -- Abstract: HIV-1 can establish a latent infection in memory CD4 + T cells to evade the host immune response. CD4 molecules can act not only as the HIV-1 receptor for entry but also asmore » the trigger in an intracellular signaling cascade for T-cell activation and proliferation via protein tyrosine kinases. Novel chronic HIV-1-infected A3.01-derived (NCHA) cells were used to examine the involvement of CD4 downstream signaling in HIV-1 latency. CD4 receptors in NCHA cells were dramatically downregulated on its surface but were slightly decreased in whole-cell lysates. The expression levels of CD4 downstream signaling molecules, including P56{sup Lck}, ZAP-70, LAT, and c-Jun, were sharply decreased in NCHA cells. The lowered histone modifications of H3K4me3 and H3K9ac correlated with the downregulation of P56{sup Lck}, ZAP-70, and LAT in NCHA cells. AP-1 binding activity was also reduced in NCHA cells. LAT and c-Jun suppressed in NCHA cells were highly induced after PMA treatment. In epigenetic analysis, other signal transduction molecules which are associated with active and/or latent HIV-1 infection showed normal states in HIV-1 latently infected cells compared to A3.01 cells. In conclusion, we demonstrated that the HIV-1 latent state is sustained by the reduction of downstream signaling molecules via the downregulation of CD4 and the attenuated activity of transcription factor as AP-1. The HIV-1 latency model via T-cell deactivation may provide some clues for the development of the new antireservoir therapy.« less

Proteome-wide characterization of signalling interactions in the hippocampal CA4/DG subfield of patients with Alzheimer’s disease

PubMed Central

Ho Kim, Jae; Franck, Julien; Kang, Taewook; Heinsen, Helmut; Ravid, Rivka; Ferrer, Isidro; Hee Cheon, Mi; Lee, Joo-Yong; Shin Yoo, Jong; Steinbusch, Harry W; Salzet, Michel; Fournier, Isabelle; Mok Park, Young

2015-01-01

Alzheimer’s disease (AD) is the most common form of dementia; however, mechanisms and biomarkers remain unclear. Here, we examined hippocampal CA4 and dentate gyrus subfields, which are less studied in the context of AD pathology, in post-mortem AD and control tissue to identify possible biomarkers. We performed mass spectrometry-based proteomic analysis combined with label-free quantification for identification of differentially expressed proteins. We identified 4,328 proteins, of which 113 showed more than 2-fold higher or lower expression in AD hippocampi than in control tissues. Five proteins were identified as putative AD biomarkers (MDH2, PCLO, TRRAP, YWHAZ, and MUC19 isoform 5) and were cross-validated by immunoblotting, selected reaction monitoring, and MALDI imaging. We also used a bioinformatics approach to examine upstream signalling interactions of the 113 regulated proteins. Five upstream signalling (IGF1, BDNF, ZAP70, MYC, and cyclosporin A) factors showed novel interactions in AD hippocampi. Taken together, these results demonstrate a novel platform that may provide new strategies for the early detection of AD and thus its diagnosis. PMID:26059363

Achromobacter denitrificans SP1 produces pharmaceutically active 25C prodigiosin upon utilizing hazardous di(2-ethylhexyl)phthalate.

PubMed

Pradeep, S; Sarath Josh, M K; Balachandran, S; Sudha Devi, R; Sadasivam, R; Thirugnanam, P E; Doble, Mukesh; Anderson, Robin C; Benjamin, Sailas

2014-11-01

This first report describes the purification and identification of an orange-red pigment produced by Achromobacter denitrificans strain SP1 (isolated from sewage sludge heavily contaminated with plastics) during its growth in a simple basal salt medium supplemented with the hazardous di(2-ethylhexyl)phthalate (DEHP) blended in PVC blood bag (in situ) or free DEHP (ex situ) as carbon source. The cell-bound pigment was elucidated, characterized at molecular level, and described as an unusual 25C prodigiosin analog for the first time. At laboratory conditions (in flasks), the dry cell mass was 75.2mg/g blood bag, which upon extraction yielded 7.1mg prodigiosin; at this stage the pH of the medium was dropped from 7.2 to 3.5. Considering its pharmaceutical importance, taking 10 known prodigiosins as controls, this 25C prodigiosin was subjected to molecular docking studies, showed comparable and promising binding efficiencies with the crucial molecular human targets like cycloxygenase-2, ZAP-70 kinase and Jak-3 kinase. Copyright © 2014 Elsevier Ltd. All rights reserved.

FAA (Federal Aviation Administration) Air Traffic Activity FY 1983.

DTIC Science & Technology

1983-09-30

tI1148335 NEVA (A LAS VERAS FCCARRAN INTL ( LAT ) L PRIMARY OPERAT1ENS 2E3549 121tE* 5P ? c /619 1 4අ SECEUARY OPERATIONS 13P8 3( 113* 1 1I OVERFLIGHTS 8026A...34NLSCATINE IA N I 1L1 27E 14 204 0PILSCLE SHOALS AL S 1 HSV 1366 55p 26 769 15 MNSKEGCN MI N I MKG 2263 547 e34 911 111PlSKCGEE OK h ZF 17 ) 3 Ii...821756 2K10M EI-CUiK :KV PARK NY N4 ,NY 1i 0 1 8* R(OPUC OH N 7ZSf 4t. 16 1 26 ZAIONE 41. S 1 HSV 23! C VI i~e11 tbU1JC CALLS MN IK ZAP4 ’I-36i1 .(EC CITY

CpG location and methylation level are crucial factors for the early detection of oral squamous cell carcinoma in brushing samples using bisulfite sequencing of a 13-gene panel.

PubMed

Morandi, Luca; Gissi, Davide; Tarsitano, Achille; Asioli, Sofia; Gabusi, Andrea; Marchetti, Claudio; Montebugnoli, Lucio; Foschini, Maria Pia

2017-01-01

Oral squamous cell carcinoma (OSCC) is usually diagnosed at an advanced stage and is commonly preceded by oral premalignant lesions. The mortality rates have remained unchanged (50% within 5 years after diagnosis), and it is related to tobacco smoking and alcohol intake. Novel molecular markers for early diagnosis are urgently needed. The purpose of this study was to evaluate the diagnostic value of methylation level in a set of 18 genes by bisulfite next-generation sequencing. With minimally invasive oral brushing, 28 consecutive OSCC, one squamous cell carcinoma with sarcomatoid features, six high-grade squamous intraepithelial lesions (HGSIL), 30 normal contralateral mucosa from the same patients, and 65 healthy donors were evaluated for DNA methylation analyzing 18 target genes by quantitative bisulfite next-generation sequencing. We further evaluated an independent cohort (validation dataset) made of 20 normal donors, one oral fibroma, 14 oral lichen planus (OLP), three proliferative verrucous leukoplakia (PVL), and two OSCC. Comparing OSCC with normal healthy donors and contralateral mucosa in 355 CpGs, we identified the following epigenetically altered genes: ZAP70 , ITGA4 , KIF1A , PARP15 , EPHX3 , NTM , LRRTM1 , FLI1 , MIR193 , LINC00599 , PAX1 , and MIR137HG showing hypermethylation and MIR296 , TERT , and GP1BB showing hypomethylation . The behavior of ZAP70 , GP1BB , H19 , EPHX3 , and MIR193 fluctuated among different interrogated CpGs. The gap between normal and OSCC samples remained mostly the same (Kruskal-Wallis P values < 0.05), but the absolute values changed conspicuously. ROC curve analysis identified the most informative CpGs, and we correctly stratified OSCC and HGSIL from normal donors using a multiclass linear discriminant analysis in a 13-gene panel (AUC 0.981). Only the OSCC with sarcomatoid features was negative. Three contralateral mucosa were positive, a sign of a possible field cancerization. Among imprinted genes, only MIR296 showed loss of imprinting. DNMT1 , TERC , and H19 together with the global methylation of long interspersed element 1 were unchanged. In the validation dataset, values over the threshold were detected in 2/2 OSCC, in 3/3 PVL, and in 2/14 OLP. Our data highlight the importance of CpG location and correct estimation of DNA methylation level for highly accurate early diagnosis of OSCC.

Overview of the Fusion Z-Pinch Experiment FuZE

NASA Astrophysics Data System (ADS)

Weber, T. R.; Shumlak, U.; Nelson, B. A.; Golingo, R. P.; Claveau, E. L.; McLean, H. S.; Tummel, K. K.; Higginson, D. P.; Schmidt, A. E.; UW/LLNL Team

2016-10-01

Previously, the ZaP device, at the University of Washington, demonstrated sheared flow stabilized (SFS) Z-pinch plasmas. Instabilities that have historically plagued Z-pinch plasma confinement were mitigated using sheared flows generated from a coaxial plasma gun of the Marshall type. Based on these results, a new SFS Z-pinch experiment, the Fusion Z-pinch Experiment (FuZE), has been constructed. FuZE is designed to investigate the scaling of SFS Z-pinch plasmas towards fusion conditions. The experiment will be supported by high fidelity physics modeling using kinetic and fluid simulations. Initial plans are in place for a pulsed fusion reactor following the results of FuZE. Notably, the design relies on proven commercial technologies, including a modest discharge current (1.5 MA) and voltage (40 kV), and liquid metal electrodes. Supported by DoE FES, NNSA, and ARPA-E ALPHA.

A survey of noninteractive zero knowledge proof system and its applications.

PubMed

Wu, Huixin; Wang, Feng

2014-01-01

Zero knowledge proof system which has received extensive attention since it was proposed is an important branch of cryptography and computational complexity theory. Thereinto, noninteractive zero knowledge proof system contains only one message sent by the prover to the verifier. It is widely used in the construction of various types of cryptographic protocols and cryptographic algorithms because of its good privacy, authentication, and lower interactive complexity. This paper reviews and analyzes the basic principles of noninteractive zero knowledge proof system, and summarizes the research progress achieved by noninteractive zero knowledge proof system on the following aspects: the definition and related models of noninteractive zero knowledge proof system, noninteractive zero knowledge proof system of NP problems, noninteractive statistical and perfect zero knowledge, the connection between noninteractive zero knowledge proof system, interactive zero knowledge proof system, and zap, and the specific applications of noninteractive zero knowledge proof system. This paper also points out the future research directions.

Overview of the FuZE Fusion Z-Pinch Experiment

NASA Astrophysics Data System (ADS)

Shumlak, U.; Nelson, B. A.; Claveau, E. L.; Forbes, E. G.; Golingo, R. P.; Stepanov, A. D.; Weber, T. R.; Zhang, Y.; McLean, H. S.; Higginson, D. P.; Schmidt, A.; Tummel, K. K.

2017-10-01

Successful results of the sheared flow stabilized (SFS) Z-pinch from ZaP and ZaP-HD have motivated the new FuZE project to scale the plasma performance to fusion conditions. The SFS Z-pinch is immune to the instabilities that plague the conventional Z-pinch yet maintains the same favorable radial scaling. The plasma density and temperature increase rapidly with decreasing plasma radius, which naturally leads to a compact configuration at fusion conditions. The SFS Z-pinch is being investigated as a novel approach to a compact fusion device in a collaborative ARPA-E ALPHA project with the University of Washington and Lawrence Livermore National Laboratory. The project includes an experimental effort coupled with high-fidelity physics modeling using kinetic and fluid simulations. Along with scaling law analysis, computational and experimental results from the FuZE device are presented. This work is supported by an award from US ARPA-E.

Cell Division in genus Corynebacterium: protein-protein interaction and molecular docking of SepF and FtsZ in the understanding of cytokinesis in pathogenic species.

PubMed

Oliveira, Alberto F; Folador, Edson L; Gomide, Anne C P; Goes-Neto, Aristóteles; Azevedo, Vasco A C; Wattam, Alice R

2018-02-15

The genus Corynebacterium includes species of great importance in medical, veterinary and biotechnological fields. The genus-specific families (PLfams) from PATRIC have been used to observe conserved proteins associated to all species. Our results showed a large number of conserved proteins that are associated with the cellular division process. Was not observe in our results other proteins like FtsA and ZapA that interact with FtsZ. Our findings point that SepF overlaps the function of this proteins explored by molecular docking, protein-protein interaction and sequence analysis. Transcriptomic analysis showed that these two (Sepf and FtsZ) proteins can be expressed in different conditions together. The work presents novelties on molecules participating in the cell division event, from the interaction of FtsZ and SepF, as new therapeutic targets.

User-friendly chemistry takes center stage at ACS meeting

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pool, R.

1992-09-11

These days it seems that what chemistry needs more than anything else is a good p.r. agent. If you ask John or Joan Q. Public about the accomplishments of the chemical industry, chances are they'll mention Love Canal, CFCs destroying the ozone layer, or carcinogens in food. However, if the national meeting of the American Chemical Society in Washington, D.C., 2 weeks ago is any indication, chemists are working hard to fix the image problem. Nearly all of the two dozen press conferences held during the meeting focused on food, health topics, environment-friendly technology, or some other subject close tomore » consumers' hearts. And the scientific talks themselves reflected the same interests, with sessions such as Environmental Successes in the Chemical Industry', Food Phytochemicals for Cancer Prevention', Chemistry of Electrophilic Metal Complexes', New Advances in Polyolefin Polymers', Zapping acid rain with microwaves.'« less

Testing the potential of bacterial branched tetraether membrane lipids as temperature proxy in peat and immature coal deposits

NASA Astrophysics Data System (ADS)

Weijers, J. W. H.; Steinmann, P.; Hopmans, E. C.; Basiliko, N.; Finkelstein, S. A.; Johnson, K. R.; Schouten, S.; Sinninghe Damsté, J. S.

2012-04-01

Branched glycerol dialkyl glycerol tetraether (brGDGT) membrane lipids occur ubiquitously in peat and soil. In soil, the degree of methylation and cyclisation of branched tetraethers (MBT index and CBT ratio, respectively) has shown to relate to both soil pH and annual mean air temperature (MAT). Using this relation, past annual MATs can be reconstructed by analysing brGDGTs in marine sediment records near large river outflows. More recently, the potential of this MBT/CBT proxy is also being explored in lakes. Despite being more abundant in peat than soils, however, the utility of the proxy has not yet been fully explored in peat records. Present day peat records generally extent back to the early Holocene, but if the MBT/CBT proxy were shown to be applicable in peat deposits, there is also potential to apply it to immature coal deposits like lignites, which could provide valuable snapshots of continental climate back to the early Cenozoic. Here results are presented of analyses of different peats in south eastern Canada, showing that the pH of peat along a nutrient gradient is rather well reflected by the CBT. Annual MAT reconstructions based on the MBT/CBT soil calibration, however, tend to overestimate measured MAT. This is also the case for peat analysed from the surface of Etang de la Gruère peat bog in the Swiss Jura Mountains. Along the 6m depth profile of this bog (~13ka), CBT-reconstructed pH is compared with in-situ measured pore water pH showing that the brGDGT composition does not reflect present-day in-situ conditions. Instead, it reflects a stratigraphic boundary between Carex and Sphagnum dominated peat at 4 m depth that is not present in the pore water profile, testifying to a 'fossil' nature of the brGDGTs down the peat bog. Analyses of three immature coals of the Argonne Premium Coal Series reveal that branched GDGTs are present in the most immature coal, the Beulah Zap lignite (Ro = 0.25%), and only just above detection limit in the Wyodak Anderson coal (Ro = 0.32%), both of about the same age (Late Palaeocene). In the more mature Illinois #6 coal (Ro = 0.46%), brGDGTs are completely absent. In the Denver Basin, a comparison is made between outcrop and drilled core samples of Palaeocene lignites. BrGDGTs are preserved in the core samples, although in low quantities compared to peat. Outcrop samples are clearly overprinted by modern soil derived brGDGTs, despite digging a meters deep trench, which shows the need to obtain fresh non-weathered samples by coring. Reconstructed annual MAT for both the Beulah Zap and the Denver Basin lignites are several degrees higher than estimates based on leaf margin and oxygen isotope analyses from the same sites. Both reconstructions do testify, nevertheless, to the warm continental conditions during the early Cenozoic of the central U.S.A.. Although further validation is required, potentially in the form of a specific peat calibration, these results do show potential for application of the MBT/CBT temperature proxy in peat and lignite deposits.

Effects of ozone or sulfur dioxide on annual plants of the Mojave Desert. [Camissonia claviformis; Camissonia hirtella; Cryptantha nevadensis; Festuca octoflora; Lepidium lasiocarpum; Baileya pleniradiata; Perityle emoryi; Oenothera californica

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thompson, C.R.; Olszyk, D.M.; Kats, G.

1984-10-01

Forty-seven species of annual plants from the Mojave Desert were grown in pots and exposed in open-top field chambers located at Riverside, California to test their relative sensitivity to SO/sub 2/ and O/sub 3/. Exposures were 8 h/day for four or five days at 0, 0.05, 0.1, 0.2, 0.3 and 0.4 ppm O/sub 3/ or 0, 0.2, 0.5, 0.6, 0.8, 1.0 and 1.5 ppm SO/sub 2/. Species differed widely in their response to the pollutants. Three species, Camissonia claviformis, Camissonia hirtella and Cryptantha nevadensis, were quite sensitive to both pollutants, exhibiting leaf injury when exposed to 0.1 ppm O/sub 3/more » or 0.2 ppm SO/sub 2/. Two species: Festuca octoflora and Lepidium lasiocarpum were tolerant to both pollutants, exhibiting no leaf injury with concentrations of 0.3 ppm O/sub 3/ or 1.5 ppm SO/sub 2/. The other species were intermediate in sensitivity, with O/sub 3/ sensitivity not always corresponding to SO/sub 2/ sensitivity. Total sulfur concentration analysis indicated that for 8 of 11 species tested, plants exposed to 0.2 ppm SO/sub 2/ had a higher sulfur concentration than nonexposed plants. Baileya pleniradiata and Perityle emoryi had the greatest increases in sulfur concentration for exposed versus control plants. To test for effects of the environmental and exposure system on plant sensitivity, 29 of the species exposed in the open-top chambers were grown in the ground and exposed with a modified chamberless zonal air pollution (ZAP) system to SO/sub 2/ in the Mojave Desert area near Daggett, California. Exposures were for a total of 37 h over six days to 0, 0.4, 0.8 and 1.0 ppm SO/sub 2/ with exposures only during periods with due west winds. Most of the species were not injured with any of the SO/sub 2/ exposure concentrations used with the ZAP system; only Camissonia claviformis showed injury at 0.8 ppm SO/sub 2/ and Oenothera californica exhibited injury at 1.0 ppm SO/sub 2/.« less

Immunological Characterization of Intraocular Lymphoid Follicles in a Spontaneous Recurrent Uveitis Model.

PubMed

Kleinwort, Kristina J H; Amann, Barbara; Hauck, Stefanie M; Feederle, Regina; Sekundo, Walter; Deeg, Cornelia A

2016-08-01

Recently, formation of tertiary lymphoid structures was demonstrated and further characterized in the R161H mouse model of spontaneous autoimmune uveitis. In the horse model of spontaneous recurrent uveitis, intraocular lymphoid follicle formation is highly characteristic, and found in all stages and scores of disease, but in depth analyses of immunologic features of these structures are lacking to date. Paraffin-embedded eye sections of cases with equine spontaneous recurrent uveitis (ERU) were characterized with immunohistochemistry to gain insight into the distribution, localization, and signaling of immune cells in intraocular tertiary lymphoid tissues. Ectopic lymphoid tissues were located preferentially in the iris, ciliary body, and retina at the ora serrata of horses with naturally-occurring ERU. The majority of cells in the tertiary lymphoid follicles were T cells with a scattered distribution of B cells and PNA+ cells interspersed. A fraction of T cells was additionally positive for memory cell marker CD45RO. Almost all cells coexpressed CD166, a molecule associated with activation and transmigration of T cells into inflamed tissues. Several transcription factors that govern immune cell responses were detectable in the tertiary lymphoid follicles, among them Zap70, TFIIB, GATA3, and IRF4. A high expression of the phosphorylated signal transducers and activators of transcription (STAT) proteins 1 and 5 were found at the margin of the structures. Cellular composition and structural organization of these inflammation-associated tertiary lymphoid tissue structures and the expression of markers of matured T and B cells point to highly organized adaptive immune responses in these follicles in spontaneous recurrent uveitis.

Rapid evolution of PARP genes suggests a broad role for ADP-ribosylation in host-virus conflicts.

PubMed

Daugherty, Matthew D; Young, Janet M; Kerns, Julie A; Malik, Harmit S

2014-01-01

Post-translational protein modifications such as phosphorylation and ubiquitinylation are common molecular targets of conflict between viruses and their hosts. However, the role of other post-translational modifications, such as ADP-ribosylation, in host-virus interactions is less well characterized. ADP-ribosylation is carried out by proteins encoded by the PARP (also called ARTD) gene family. The majority of the 17 human PARP genes are poorly characterized. However, one PARP protein, PARP13/ZAP, has broad antiviral activity and has evolved under positive (diversifying) selection in primates. Such evolution is typical of domains that are locked in antagonistic 'arms races' with viral factors. To identify additional PARP genes that may be involved in host-virus interactions, we performed evolutionary analyses on all primate PARP genes to search for signatures of rapid evolution. Contrary to expectations that most PARP genes are involved in 'housekeeping' functions, we found that nearly one-third of PARP genes are evolving under strong recurrent positive selection. We identified a >300 amino acid disordered region of PARP4, a component of cytoplasmic vault structures, to be rapidly evolving in several mammalian lineages, suggesting this region serves as an important host-pathogen specificity interface. We also found positive selection of PARP9, 14 and 15, the only three human genes that contain both PARP domains and macrodomains. Macrodomains uniquely recognize, and in some cases can reverse, protein mono-ADP-ribosylation, and we observed strong signatures of recurrent positive selection throughout the macro-PARP macrodomains. Furthermore, PARP14 and PARP15 have undergone repeated rounds of gene birth and loss during vertebrate evolution, consistent with recurrent gene innovation. Together with previous studies that implicated several PARPs in immunity, as well as those that demonstrated a role for virally encoded macrodomains in host immune evasion, our evolutionary analyses suggest that addition, recognition and removal of ADP-ribosylation is a critical, underappreciated currency in host-virus conflicts.

Rapid Evolution of PARP Genes Suggests a Broad Role for ADP-Ribosylation in Host-Virus Conflicts

PubMed Central

Daugherty, Matthew D.; Young, Janet M.; Kerns, Julie A.; Malik, Harmit S.

2014-01-01

Post-translational protein modifications such as phosphorylation and ubiquitinylation are common molecular targets of conflict between viruses and their hosts. However, the role of other post-translational modifications, such as ADP-ribosylation, in host-virus interactions is less well characterized. ADP-ribosylation is carried out by proteins encoded by the PARP (also called ARTD) gene family. The majority of the 17 human PARP genes are poorly characterized. However, one PARP protein, PARP13/ZAP, has broad antiviral activity and has evolved under positive (diversifying) selection in primates. Such evolution is typical of domains that are locked in antagonistic ‘arms races’ with viral factors. To identify additional PARP genes that may be involved in host-virus interactions, we performed evolutionary analyses on all primate PARP genes to search for signatures of rapid evolution. Contrary to expectations that most PARP genes are involved in ‘housekeeping’ functions, we found that nearly one-third of PARP genes are evolving under strong recurrent positive selection. We identified a >300 amino acid disordered region of PARP4, a component of cytoplasmic vault structures, to be rapidly evolving in several mammalian lineages, suggesting this region serves as an important host-pathogen specificity interface. We also found positive selection of PARP9, 14 and 15, the only three human genes that contain both PARP domains and macrodomains. Macrodomains uniquely recognize, and in some cases can reverse, protein mono-ADP-ribosylation, and we observed strong signatures of recurrent positive selection throughout the macro-PARP macrodomains. Furthermore, PARP14 and PARP15 have undergone repeated rounds of gene birth and loss during vertebrate evolution, consistent with recurrent gene innovation. Together with previous studies that implicated several PARPs in immunity, as well as those that demonstrated a role for virally encoded macrodomains in host immune evasion, our evolutionary analyses suggest that addition, recognition and removal of ADP-ribosylation is a critical, underappreciated currency in host-virus conflicts. PMID:24875882

Kinome signaling through regulated protein-protein interactions in normal and cancer cells.

PubMed

Pawson, Tony; Kofler, Michael

2009-04-01

The flow of molecular information through normal and oncogenic signaling pathways frequently depends on protein phosphorylation, mediated by specific kinases, and the selective binding of the resulting phosphorylation sites to interaction domains present on downstream targets. This physical and functional interplay of catalytic and interaction domains can be clearly seen in cytoplasmic tyrosine kinases such as Src, Abl, Fes, and ZAP-70. Although the kinase and SH2 domains of these proteins possess similar intrinsic properties of phosphorylating tyrosine residues or binding phosphotyrosine sites, they also undergo intramolecular interactions when linked together, in a fashion that varies from protein to protein. These cooperative interactions can have diverse effects on substrate recognition and kinase activity, and provide a variety of mechanisms to link the stimulation of catalytic activity to substrate recognition. Taken together, these data have suggested how protein kinases, and the signaling pathways in which they are embedded, can evolve complex properties through the stepwise linkage of domains within single polypeptides or multi-protein assemblies.

Parameter scaling toward high-energy density in a quasi-steady flow Z-pinch

NASA Astrophysics Data System (ADS)

Hughes, M. C.; Shumlak, U.; Nelson, B. A.; Golingo, R. P.; Claveau, E. L.; Doty, S. A.; Forbes, E. G.; Kim, B.; Ross, M. P.

2016-10-01

Sheared axial flows are utilized by the ZaP Flow Z-Pinch Experiment to stabilize MHD instabilities. The pinches formed are 50 cm long with radii ranging from 0.3 to 1.0 cm. The plasma is generated in a coaxial acceleration region, similar to a Marshall gun, which provides a steady supply of plasma for approximately 100 us. The power to the plasma is partially decoupled between the acceleration and pinch assembly regions through the use of separate power supplies. Adiabatic scaling of the Bennett relation gives targets for future devices to reach high-energy density conditions or fusion reactors. The applicability of an adiabatic assumption is explored and work is done experimentally to clarify the plasma compression process, which may be more generally polytropic. The device is capable of a much larger parameter space than previous machine iterations, allowing flexibility in the initial conditions of the compression process to preserve stability. This work is supported by DoE FES and NNSA.

Conformational changes induced in the protein tyrosine kinase p72syk by tyrosine phosphorylation or by binding of phosphorylated immunoreceptor tyrosine-based activation motif peptides.

PubMed Central

Kimura, T; Sakamoto, H; Appella, E; Siraganian, R P

1996-01-01

A critical event in signaling in immune cells is the interaction of Syk or ZAP-70 protein tyrosine kinases with multisubunit receptors that contain an approximately 18-amino-acid domain called the immunoreceptor tyrosine-based activation motif (ITAM). Tyrosine-phosphorylated Syk from activated cells was in a conformation different from that in nonstimulated cells as demonstrated by changes in immunoreactivity. The addition of tyrosine-diphosphorylated ITAM peptides resulted in a similar conformational change in Syk from nonactivated cells. The peptides based on FcepsilonRIgamma were more active than those based on Fcepsilon RIbeta. In vitro autophosphorylation of Syk was dramatically enhanced by the addition of the diphosphorylated ITAM peptides. The conformational change and the enhanced autophosphorylation required the presence of both phosphorylated tyrosines on the same molecule. These conformational changes in Syk by tyrosine phosphorylation or binding to diphosphorylated ITAM could be critical for Syk activation and downstream propagation of intracellular signals. PMID:8657120

A Survey of Noninteractive Zero Knowledge Proof System and Its Applications

PubMed Central

Wu, Huixin; Wang, Feng

2014-01-01

Zero knowledge proof system which has received extensive attention since it was proposed is an important branch of cryptography and computational complexity theory. Thereinto, noninteractive zero knowledge proof system contains only one message sent by the prover to the verifier. It is widely used in the construction of various types of cryptographic protocols and cryptographic algorithms because of its good privacy, authentication, and lower interactive complexity. This paper reviews and analyzes the basic principles of noninteractive zero knowledge proof system, and summarizes the research progress achieved by noninteractive zero knowledge proof system on the following aspects: the definition and related models of noninteractive zero knowledge proof system, noninteractive zero knowledge proof system of NP problems, noninteractive statistical and perfect zero knowledge, the connection between noninteractive zero knowledge proof system, interactive zero knowledge proof system, and zap, and the specific applications of noninteractive zero knowledge proof system. This paper also points out the future research directions. PMID:24883407

A Mini HIP HOP Assay Uncovers a Central Role for Copper and Zinc in the Antifungal Mode of Action of Allicin.

PubMed

Prescott, Thomas A K; Panaretou, Barry

2017-05-10

Garlic contains the organosulfur compound allicin which exhibits potent antifungal activity. Here we demonstrate the use of a highly simplified yeast chemical genetic screen to characterize its mode of action. By screening 24 validated yeast gene deletion "signature" strains for which hypersensitivity is characteristic for common antifungal modes of action, yeast lacking the high affinity Cu 2+ transporter Ctr1 was found to be hypersensitive to allicin. Focusing on transition metal related genes identified two more hypersensitive strains lacking the Cu 2+ and Zn 2+ transcription factors Mac1 and Zap1. Hypersensitivity in these strains was reversed by the addition of Cu 2+ and Zn 2+ ions, respectively. The results suggest the antifungal activity of allicin is mediated through restricted Cu 2+ and Zn 2+ uptake or inhibition of Cu 2+ and Zn 2+ metalloproteins. As certain antimicrobial modes of action are much more common than others, the approach taken here provides a useful way to identify them early on.

A Phosphosite within the SH2 Domain of Lck Regulates Its Activation by CD45.

PubMed

Courtney, Adam H; Amacher, Jeanine F; Kadlecek, Theresa A; Mollenauer, Marianne N; Au-Yeung, Byron B; Kuriyan, John; Weiss, Arthur

2017-08-03

The Src Family kinase Lck sets a critical threshold for T cell activation because it phosphorylates the TCR complex and the Zap70 kinase. How a T cell controls the abundance of active Lck molecules remains poorly understood. We have identified an unappreciated role for a phosphosite, Y192, within the Lck SH2 domain that profoundly affects the amount of active Lck in cells. Notably, mutation of Y192 blocks critical TCR-proximal signaling events and impairs thymocyte development in retrogenic mice. We determined that these defects are caused by hyperphosphorylation of the inhibitory C-terminal tail of Lck. Our findings reveal that modification of Y192 inhibits the ability of CD45 to associate with Lck in cells and dephosphorylate the C-terminal tail of Lck, which prevents its adoption of an active open conformation. These results suggest a negative feedback loop that responds to signaling events that tune active Lck amounts and TCR sensitivity. Copyright © 2017 Elsevier Inc. All rights reserved.

ERK-dependent T cell receptor threshold calibration in rheumatoid arthritis.

PubMed

Singh, Karnail; Deshpande, Pratima; Pryshchep, Sergey; Colmegna, Inés; Liarski, Vladimir; Weyand, Cornelia M; Goronzy, Jörg J

2009-12-15

Immune responses to citrullinated neoantigens and clinical efficacy of costimulation blockade indicate a general defect in maintaining T cell tolerance in rheumatoid arthritis (RA). To examine whether TCR threshold calibration contributes to disease pathogenesis, signaling in RA T cells was quantified. RA patients had a selective increase in ERK phosphorylation compared with demographically matched controls due to a mechanism distal of Ras activation. Increased ERK responses included naive and memory CD4 and CD8 T cells and did not correlate with disease activity. The augmented ERK activity delayed SHP-1 recruitment to the TCR synapse and sustained TCR-induced Zap70 and NF-kappaB signaling, facilitating responses to suboptimal stimulation. Increased responsiveness of the ERK pathway was also a characteristic finding in the SKG mouse model of RA where it preceded clinical symptoms. Treatment with subtherapeutic doses of a MEK-1/2 inhibitor delayed arthritis onset and reduced severity, suggesting that increased ERK phosphorylation predisposes for autoimmunity and can be targeted to prevent disease.

[Etiological and molecular characteristics of diarrhea caused Proteus mirabilis].

PubMed

Shi, Xiaolu; Hu, Qinghua; Lin, Yiman; Qiu, Yaqun; Li, Yinghui; Jiang, Min; Chen, Qiongcheng

2014-06-01

To analyze the etiological characteristics, virulence genes and plasmids that carrying diarrhea-causing Proteus mirabilis and to assess their relationship with drug resistance and pathogenicity. Proteus mirabilis coming from six different sources (food poisoning, external environment and healthy people) were analyzed biochemically, on related susceptibility and pulsed-field gel electrophoresis (PFGE). Virulence genes were detected by PCR. Plasmids were extracted and sequenced after gel electrophoresis purification. The biochemical characteristics of Proteus mirabilis from different sources seemed basically the same, and each of them showed having common virulence genes, as ureC, rsmA, hpmA and zapA. However, the PFGE patterns and susceptibility of these strains were different, so as the plasmids that they carried. Plasmid that presented in the sequenced strain showed that the 2 683 bp length plasmid encodes qnrD gene was associated with the quinolone resistance. Etiological characteristics and molecular characteristics of Proteus mirabilis gathered from different sources, were analyzed. Results indicated that traditional biochemical analysis and common virulence gene identification might be able to distinguish the strains with different sources. However, PFGE and plasmids analysis could distinguish the sources of strains and to identify those plasmids that commonly carried by the drug-resistant strains. These findings also provided theoretical basis for further study on the nature of resistance and pathogenicity in Proteus mirabilis.

[Prognostic value of absolute monocyte count in chronic lymphocytic leukaemia].

PubMed

Szerafin, László; Jakó, János; Riskó, Ferenc

2015-04-01

The low peripheral absolute lymphocyte and high monocyte count have been reported to correlate with poor clinical outcome in various lymphomas and other cancers. However, a few data known about the prognostic value of absolute monocyte count in chronic lymphocytic leukaemia. The aim of the authors was to investigate the impact of absolute monocyte count measured at the time of diagnosis in patients with chronic lymphocytic leukaemia on the time to treatment and overal survival. Between January 1, 2005 and December 31, 2012, 223 patients with newly-diagnosed chronic lymphocytic leukaemia were included. The rate of patients needing treatment, time to treatment, overal survival and causes of mortality based on Rai stages, CD38, ZAP-70 positivity and absolute monocyte count were analyzed. Therapy was necessary in 21.1%, 57.4%, 88.9%, 88.9% and 100% of patients in Rai stage 0, I, II, III an IV, respectively; in 61.9% and 60.8% of patients exhibiting CD38 and ZAP-70 positivity, respectively; and in 76.9%, 21.2% and 66.2% of patients if the absolute monocyte count was <0.25 G/l, between 0.25-0.75 G/l and >0.75 G/l, respectively. The median time to treatment and the median overal survival were 19.5, 65, and 35.5 months; and 41.5, 65, and 49.5 months according to the three groups of monocyte counts. The relative risk of beginning the therapy was 1.62 (p<0.01) in patients with absolute monocyte count <0.25 G/l or >0.75 G/l, as compared to those with 0.25-0.75 G/l, and the risk of overal survival was 2.41 (p<0.01) in patients with absolute monocyte count lower than 0.25 G/l as compared to those with higher than 0.25 G/l. The relative risks remained significant in Rai 0 patients, too. The leading causes of mortality were infections (41.7%) and the chronic lymphocytic leukaemia (58.3%) in patients with low monocyte count, while tumours (25.9-35.3%) and other events (48.1 and 11.8%) occurred in patients with medium or high monocyte counts. Patients with low and high monocyte counts had a shorter time to treatment compared to patients who belonged to the intermediate monocyte count group. The low absolute monocyte count was associated with increased mortality caused by infectious complications and chronic lymphocytic leukaemia. The absolute monocyte count may give additional prognostic information in Rai stage 0, too.

Serum level of CD26 predicts time to first treatment in early B-chronic lymphocytic leukemia.

PubMed

Molica, Stefano; Digiesi, Giovanna; Mirabelli, Rosanna; Cutrona, Giovanna; Antenucci, Anna; Molica, Matteo; Giannarelli, Diana; Sperduti, Isabella; Morabito, Fortunato; Neri, Antonino; Baldini, Luca; Ferrarini, Manlio

2009-09-01

We analyzed the correlation between well-established biological parameters of prognostic relevance in B-cell chronic lymphocytic leukemia (CLL) [i.e. mutational status of the immunoglobulin heavy chain variable region (IgV(H)), ZAP-70- and CD38-expression] and serum levels of CD26 (dipeptidyl peptidase IV, DPP IV) by evaluating the impact of these variables on the time to first treatment (TFT) in a series of 69 previously untreated Binet stage A B-cell CLL patients. By using a commercial ELISA we found that with exception of a borderline significance for ZAP-70 (P = 0.07) and CD38 (P = 0.08), circulating levels of CD26 did not correlate with either Rai substages (P = 0.520) or other biomarker [beta2-microglobulin (P = 0.933), LDH (P = 0.101), mutational status of IgV(H) (P = 0.320)]. Maximally selected log-rank statistic plots identified a CD26 serum concentration of 371 ng/mL as the best cut-off. This threshold allowed the identification of two subsets of patients with CD26 serum levels higher and lower that 371 ng/mL respectively, whose clinical outcome was different with respect to TFT (i.e. 46% and 71% at 5 yr respectively; P = 0.005). Along with higher serum levels of CD26, the univariate Cox proportional hazard model identified absence of mutation in IgV(H) (P < 0.0001) as predictor of shorter TFT. As in multivariate analysis all these parameters maintained their discriminating power (mutational status of IgV(H,)P < 0.0001; soluble CD26, P = 0.02) their combined effect on clinical outcome was assessed. When three groups were considered: (1) Low-risk group (n = 31), patients with concordant IgVH(mut) and low level of soluble CD26; (2) intermediate risk group (n = 26), patients with discordant pattern; (3) high-risk group (n = 12), patients with concordant IgVH(unmut) and high level of soluble CD26, differences in the TFT were statistically significant, with a TFT at 5 yr of respectively 88%, 51% and 43% (P < 0.0001). Our results indicate that in early B-cell CLL biological profile including among other parameters soluble CD26 may provide a useful insight into the complex interrelationship of prognostic variables. Furthermore, CD26 along with mutational status of IgV(H) can be adequately used to predict clinical behavior of patients with low risk disease.

cDNA Cloning, expression and characterization of an allergenic 60s ribosomal protein of almond (prunus dulcis).

PubMed

Abolhassani, Mohsen; Roux, Kenneth H

2009-06-01

Tree nuts, including almond (prunus dulcis) are a source of food allergens often associated with life-threatening allergic reactions in susceptible individuals. Although the proteins in almonds have been biochemically characterized, relatively little has been reported regarding the identity of the allergens involved in almond sensitivity. The present study was undertaken to identify the allergens of the almond by cDNA library approach. cDNA library of almond seeds was constructed in Uni-Zap XR lamda vector and expressed in E. coli XL-1 blue. Plaques were immunoscreened with pooled sera of allergic patients. The cDNA clone reacting significantly with specific IgE antibodies was selected and subcloned and subsequently expressed in E. coli. The amino acids deducted from PCR product of clone showed homology to 60s acidic ribosomal protein of almond. The expressed protein was 11,450 Dalton without leader sequence. Immunoreactivity of the recombinant 60s ribosomal protein (r60sRP) was evaluated with dot blot analysis using pooled and individual sera of allergic patients. The data showed that r60sRP and almond extract (as positive control) possess the ability to bind the IgE antibodies. The results showed that expressed protein is an almond allergen.Whether this r60sRP represents a major allergen of almond needs to be further studied which requires a large number of sera from the almond atopic patients and also need to determine the IgE-reactive frequencies of each individual allergen.

Piperine from black pepper inhibits activation-induced proliferation and effector function of T lymphocytes.

PubMed

Doucette, Carolyn D; Rodgers, Gemma; Liwski, Robert S; Hoskin, David W

2015-11-01

Piperine is a major alkaloid component of black pepper (Piper nigrum Linn), which is a widely consumed spice. Here, we investigated the effect of piperine on mouse T lymphocyte activation. Piperine inhibited polyclonal and antigen-specific T lymphocyte proliferation without affecting cell viability. Piperine also suppressed T lymphocyte entry into the S and G2 /M phases of the cell cycle, and decreased expression of G1 -associated cyclin D3, CDK4, and CDK6. In addition, piperine inhibited CD25 expression, synthesis of interferon-γ, interleukin (IL)-2, IL-4, and IL-17A, and the generation of cytotoxic effector cells. The inhibitory effect of piperine on T lymphocytes was associated with hypophosphorylation of Akt, extracellular signal-regulated kinase, and inhibitor of κBα, but not ZAP-70. The ability of piperine to inhibit several key signaling pathways involved in T lymphocyte activation and the acquisition of effector function suggests that piperine might be useful in the management of T lymphocyte-mediated autoimmune and chronic inflammatory disorders. © 2015 Wiley Periodicals, Inc.

T cell costimulation via the integrin VLA-4 inhibits the actin-dependent centralization of signaling microclusters containing the adaptor SLP-76.

PubMed

Nguyen, Ken; Sylvain, Nicholas R; Bunnell, Stephen C

2008-06-01

Antigen-dependent T cell activation drives the formation of signaling microclusters containing the adaptor SLP-76. Costimulatory integrins regulate SLP-76 phosphorylation and could influence SLP-76 microclusters in the integrin-rich periphery of the immune synapse. We report that costimulation by the integrin VLA-4 (alpha4beta1) required SLP-76 domains implicated in microcluster assembly. Pro-adhesive ligands enlarged the contact and increased the number of SLP-76 microclusters regardless of their costimulatory potential. Costimulatory VLA-4 ligands also prevented the centralization of SLP-76, promoted microcluster persistence, prolonged lateral interactions between SLP-76 and its upstream kinase, ZAP-70, and retained SLP-76 in tyrosine-phosphorylated peripheral structures. SLP-76 centralization was driven by dynamic actin polymerization and was correlated with inward actin flows. VLA-4 ligation retarded these flows, even in the absence of SLP-76. These data suggest a widely applicable model of costimulation, in which integrins promote sustained signaling by attenuating cytoskeletal movements that drive the centralization and inactivation of SLP-76 microclusters.

A PLC-γ1-independent, RasGRP1-ERK dependent pathway drives lymphoproliferative disease in LAT-Y136F mutant mice

PubMed Central

Kortum, Robert L.; Rouquette-Jazdanian, Alexandre K.; Miyaji, Michihiko; Merrill, Robert K.; Markegard, Evan; Pinski, John M.; Wesselink, Amelia; Nath, Nandan N.; Alexander, Clayton P.; Li, Wenmei; Kedei, Noemi; Roose, Jeroen P.; Blumberg, Peter M.; Samelson, Lawrence E.; Sommers, Connie L.

2012-01-01

Mice expressing a germline mutation in the PLC-γ1 binding site of LAT (linker for activation of T cells) show progressive lymphoproliferation and ultimately die at 4–6 months of age. The hyper-activated T cells in these mice show defective TCR-induced calcium flux, but enhanced Ras/ERK activation that is critical for disease progression. Despite the loss of LAT-dependent PLC-γ1 binding and activation, genetic analysis revealed RasGRP1, and not Sos1 or Sos2, to be the major RasGEF responsible for ERK activation and the lymphoproliferative phenotype in these mice. Analysis of isolated CD4+ T cells from LAT-Y136F mice showed altered proximal TCR-dependent kinase signaling, which activated a Zap70- and LAT-independent pathway. Moreover, LAT-Y136F T cells showed ERK activation that was dependent on Lck and/or Fyn, PKCθ, and RasGRP1. These data demonstrate a novel route to Ras activation in vivo in a pathological setting. PMID:23209318

Cloning and expression of Bartonella henselae sucB gene encoding an immunogenic dihydrolipoamide succinyltransferase homologous protein.

PubMed

Kabeya, Hidenori; Maruyama, Soichi; Hirano, Kouji; Mikami, Takeshi

2003-01-01

Immunoscreening of a ZAP genomic library of Bartonella henselae strain Houston-1 expressed in Escherichia coli resulted in the isolation of a clone containing 3.5 kb BamHI genomic DNA fragment. This 3.5 kb DNA fragment was found to contain a sequence of a gene encoding a protein with significant homology to the dihydrolipoamide succinyltransferase of Brucella melitensis (sucB). Subsequent cloning and DNA sequence analysis revealed that the deduced amino acid sequence from the cloned gene showed 66.5% identity to SucB protein of B. melitensis, and 43.4 and 47.2% identities to those of Coxiella burnetii and E. coli, respectively. The gene was expressed as a His-Nus A-tagged fusion protein. The recombinant SucB protein (rSucB) was shown to be an immunoreactive protein of about 115 kDa by Western blot analysis with sera from B. henselae-immunized mice. Therefore the rSucB may be a candidate antigen for a specific serological diagnosis of B. henselae infection.

Physiological and Transcriptional Responses of Saccharomyces cerevisiae to Zinc Limitation in Chemostat Cultures †

PubMed Central

De Nicola, Raffaele; Hazelwood, Lucie A.; De Hulster, Erik A. F.; Walsh, Michael C.; Knijnenburg, Theo A.; Reinders, Marcel J. T.; Walker, Graeme M.; Pronk, Jack T.; Daran, Jean-Marc; Daran-Lapujade, Pascale

2007-01-01

Transcriptional responses of the yeast Saccharomyces cerevisiae to Zn availability were investigated at a fixed specific growth rate under limiting and abundant Zn concentrations in chemostat culture. To investigate the context dependency of this transcriptional response and eliminate growth rate-dependent variations in transcription, yeast was grown under several chemostat regimens, resulting in various carbon (glucose), nitrogen (ammonium), zinc, and oxygen supplies. A robust set of genes that responded consistently to Zn limitation was identified, and the set enabled the definition of the Zn-specific Zap1p regulon, comprised of 26 genes and characterized by a broader zinc-responsive element consensus (MHHAACCBYNMRGGT) than so far described. Most surprising was the Zn-dependent regulation of genes involved in storage carbohydrate metabolism. Their concerted down-regulation was physiologically relevant as revealed by a substantial decrease in glycogen and trehalose cellular content under Zn limitation. An unexpectedly large number of genes were synergistically or antagonistically regulated by oxygen and Zn availability. This combinatorial regulation suggested a more prominent involvement of Zn in mitochondrial biogenesis and function than hitherto identified. PMID:17933919

T cell receptor for antigen induces linker for activation of T cell–dependent activation of a negative signaling complex involving Dok-2, SHIP-1, and Grb-2

PubMed Central

Dong, Shen; Corre, Béatrice; Foulon, Eliane; Dufour, Evelyne; Veillette, André; Acuto, Oreste; Michel, Frédérique

2006-01-01

Adaptor proteins positively or negatively regulate the T cell receptor for antigen (TCR) signaling cascade. We report that after TCR stimulation, the inhibitory adaptor downstream of kinase (Dok)-2 and its homologue Dok-1 are involved in a multimolecular complex including the lipid phosphatase Src homology 2 domain–containing inositol polyphosphate 5′-phosphatase (SHIP)-1 and Grb-2 which interacts with the membrane signaling scaffold linker for activation of T cells (LAT). Knockdown of LAT and SHIP-1 expression indicated that SHIP-1 favored recruitment of Dok-2 to LAT. Knockdown of Dok-2 and Dok-1 revealed their negative control on Akt and, unexpectedly, on Zap-70 activation. Our findings support the view that Dok-1 and -2 are critical elements of a LAT-dependent negative feedback loop that attenuates early TCR signal. Dok-1 and -2 may therefore exert a critical role in shaping the immune response and as gatekeepers for T cell tolerance. PMID:17043143

Analysis of chronic lymphotic leukemia transcriptomic profile: differences between molecular subgroups.

PubMed

Jantus Lewintre, Eloisa; Reinoso Martín, Cristina; Montaner, David; Marín, Miguel; José Terol, María; Farrás, Rosa; Benet, Isabel; Calvete, Juan J; Dopazo, Joaquín; García-Conde, Javier

2009-01-01

B cell chronic lymphocytic leukemia (CLL) is a lymphoproliferative disorder with a variable clinical course. Patients with unmutated IgV(H) gene show a shorter progression-free and overall survival than patients with immunoglobulin heavy chain variable regions (IgV(H)) gene mutated. In addition, BCL6 mutations identify a subgroup of patients with high risk of progression. Gene expression was analysed in 36 early-stage patients using high-density microarrays. Around 150 genes differentially expressed were found according to IgV(H) mutations, whereas no difference was found according to BCL6 mutations. Functional profiling methods allowed us to distinguish KEGG and gene ontology terms showing coordinated gene expression changes across subgroups of CLL. We validated a set of differentially expressed genes according to IgV(H) status, scoring them as putative prognostic markers in CLL. Among them, CRY1, LPL, CD82 and DUSP22 are the ones with at least equal or superior performance to ZAP70 which is actually the most used surrogate marker of IgV(H) status.

PET imaging of T cells: Target identification and feasibility assessment.

PubMed

Auberson, Yves P; Briard, Emmanuelle; Rudolph, Bettina; Kaupmann, Klemen; Smith, Paul; Oberhauser, Berndt

2018-06-01

Imaging T cells using positron emission tomography (PET) would be highly useful for diagnosis and monitoring in immunology and oncology patients. There are however no obvious targets that can be used to develop imaging agents for this purpose. We evaluated several potential target proteins with selective expression in T cells, and for which lead molecules were available: PKC , Lck, ZAP70 and Itk. Ultimately, we focused on Itk (interleukin-2-inducible T cell kinase) and identified a tool molecule with properties suitable for in vivo imaging of T cells, (5aR)-5,5-difluoro-5a-methyl-N-(1-((S)-3-(methylsulfonyl)-phenyl)(tetrahydro-2H-pyran-4-yl)methyl)-1H-pyrazol-4-yl)-1,4,4a,5,5a,6-hexahydro-cyclopropa[f]-indazole-3-carboxamide (23). While not having the optimal profile for clinical use, this molecule indicates that it might be possible to develop Itk-selective PET ligands for imaging the distribution of T cells in patients. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Problems of deep drilling in abnormally pressured zones of the Kara Sea continental shelf

DOE Office of Scientific and Technical Information (OSTI.GOV)

Simonov, V.I.

1996-12-31

There are discussed results of drilling operations in shelf hydrocarbon areas of the Far North of Tyumen Region (Kharassavieskaya, Bovanenkovskaya and Krusenshternskaya ones) and on the Bely Island. The author describes equipment and technologies used, problems arising in the process of operations and possible ways of solving them. Application of the results discussed in the report seems rather attractive in connection with possible realization of joint projects on development of the mentioned areas. Thus, Amoco Eurasia plans to participate in development of Bovanenkovskoye and Novoportovskoye fields. Well planning for Amoco has been done of specialists of ZapSibBurNIPI. Experience of Russianmore » drilling companies in the Yamal area (Far North of Tyumen Region) has proved that well planning for shelf areas requires special attention as drilling-in both overpressured zones (Bovanenkovskoye field) and underpressured ones (Novoportovskoye field) is done actually in balance. Investigated are reasons for such drilling problems as kicks and lost circulation. Taking them into consideration will help to decrease considerably the cost of well drilling in shelf areas.« less

The convergence of complete active space self-consistent-field configuration interaction including all single and double excitation energies to the complete basis set limit

NASA Astrophysics Data System (ADS)

Petersson, George A.; Malick, David K.; Frisch, Michael J.; Braunstein, Matthew

2006-07-01

Examination of the convergence of full valence complete active space self-consistent-field configuration interaction including all single and double excitation (CASSCF-CISD) energies with expansion of the one-electron basis set reveals a pattern very similar to the convergence of single determinant energies. Calculations on the lowest four singlet states and the lowest four triplet states of N2 with the sequence of n-tuple-ζ augmented polarized (nZaP) basis sets (n =2, 3, 4, 5, and 6) are used to establish the complete basis set limits. Full configuration-interaction (CI) and core electron contributions must be included for very accurate potential energy surfaces. However, a simple extrapolation scheme that has no adjustable parameters and requires nothing more demanding than CAS(10e -,8orb)-CISD/3ZaP calculations gives the Re, ωe, ωeXe, Te, and De for these eight states with rms errors of 0.0006Å, 4.43cm-1, 0.35cm-1, 0.063eV, and 0.018eV, respectively.

FcγRIIb expression in early stage chronic lymphocytic leukemia.

PubMed

Bosch, Rosa; Mora, Alba; Vicente, Eva Puy; Ferrer, Gerardo; Jansà, Sonia; Damle, Rajendra; Gorlatov, Sergey; Rai, Kanti; Montserrat, Emili; Nomdedeu, Josep; Pratcorona, Marta; Blanco, Laura; Saavedra, Silvana; Garrido, Ana; Esquirol, Albert; Garcia, Irene; Granell, Miquel; Martino, Rodrigo; Delgado, Julio; Sierra, Jorge; Chiorazzi, Nicholas; Moreno, Carol

2017-11-01

In normal B-cells, B-cell antigen receptor (BCR) signaling can be negatively regulated by the low-affinity receptor FcγRIIb (CD32b). To better understand the role of FcγRIIb in chronic lymphocytic leukemia (CLL), we correlated its expression on 155 samples from newly-diagnosed Binet A patients with clinical characteristics and outcome. FcγRIIb expression was similar in normal B-cells and leukemic cells, this being heterogenous among patients and within CLL clones. FcγRIIb expression did not correlate with well known prognostic markers [disease stage, serum beta-2 microglobulin (B2M), IGHV mutational status, expression of ZAP-70 and CD38, and cytogenetics] except for a weak concordance with CD49d. Moreover, patients with low FcγRIIb expression (69/155, 44.5%) required therapy earlier than those with high FcγRIIb expression (86/155, 55.5%) (median 151.4 months vs. not reached; p=.071). These results encourage further investigation on the role of FcγRIIb in CLL biology and prognostic significance in larger series of patients.

Dipeptidyl peptidase III is a zinc metallo-exopeptidase. Molecular cloning and expression.

PubMed Central

Fukasawa, K; Fukasawa, K M; Kanai, M; Fujii, S; Hirose, J; Harada, M

1998-01-01

We have purified dipeptidyl peptidase III (EC 3.4.14.4) from human placenta. It had a pH optimum of 8.8 and readily hydrolysed Arg-Arg-beta-naphthylamide. Monoamino acid-, Gly-Phe-, Gly-Pro- and Bz-Arg-beta-naphthylamides were not hydrolysed at all. The enzyme was inhibited by p-chloromercuriphenylsulphonic acid, metal chelators and 3,4-dichloroisocoumarin and contained 1 mol of zinc per mol of enzyme. The zinc dissociation constant was 250 fM at pH 7. 4 as determined by the zinc binding study. We isolated, by immunological screening of a Uni-ZAP XR cDNA library constructed from rat liver mRNA species, a cDNA clone with 2633 bp encoding the rat enzyme. The longest open reading frame encodes a 827-residue protein with a theoretical molecular mass of 92790 Da. Escherichia coli SOLR cells were infected with the pBluescript phagemid containing the cloned cDNA and established the overexpression of a protein that hydrolysed Arg-Arg-beta-naphthylamide. The recombinant protein was purified and the amino acid sequence of the protein was confirmed. We presumed that the putative zinc-binding domain involved in catalysis was present in the recombinant enzyme. It was a novel zinc-binding motif in that one amino acid residue was inserted into the conserved HEXXH motif characteristic of the metalloproteinases. PMID:9425109

Atypical rearrangement involving 3′-IGH@ and a breakpoint at least 400 Kb upstream of an intact MYC in a CLL patient with an apparently balanced t(8;14)(q24.1;q32) and negative MYC expression

PubMed Central

2013-01-01

The t(8;14)(q24.1;q32), the cytogenetic hallmark of Burkitt’s lymphoma, is also found, but rarely, in cases of chronic lymphocytic leukemia (CLL). Such translocation typically results in a MYC-IGH@ fusion subsequently deregulating and overexpressing MYC on der 14q32. In CLL, atypical rearrangements resulting in its gain or loss, within or outside of IGH@ or MYC locus, have been reported, but their clinical significance remains uncertain. Herein, we report a 67 year-old male with complex cytogenetic findings of apparently balanced t(8;14) and unreported complex rearrangements of IGH@ and MYC loci. His clinical, morphological and immunophenotypic features were consistent with the diagnosis of CLL. Interphase FISH studies revealed deletions of 11q22.3 and 13q14.3, and an extra copy of IGH@, indicative of rearrangement. Karyotype analysis showed an apparently balanced t(8;14)(q24.1;q32). Sequential GPG-metaphase FISH studies revealed abnormal signal patterns: rearrangement of IGH break apart probe with the 5’-IGH@ on derivative 8q24.1 and the 3’-IGH@ retained on der 14q; absence of MYC break apart-specific signal on der 8q; and, the presence of unsplit 5’-MYC-3’ break apart probe signals on der 14q. The breakpoint on 8q24.1 was found to be at least 400 Kb upstream of 5’ of MYC. In addition, FISH studies revealed two abnormal clones; one with 13q14.3 deletion, and the other, with concurrent 11q deletion and atypical rearrangements. Chromosome microarray analysis (CMA) detected a 7.1 Mb deletion on 11q22.3-q23.3 including ATM, a finding consistent with FISH results. While no significant copy number gain or loss observed on chromosomes 8, 12 and 13, a 455 Kb microdeletion of uncertain clinical significance was detected on 14q32.33. Immunohistochemistry showed co-expression of CD19, CD5, and CD23, positive ZAP-70 expression and absence of MYC expression. Overall findings reveal an apparently balanced t(8;14) and atypical complex rearrangements involving 3’-IGH@ and a breakpoint at least 400 Kb upstream of MYC, resulting in the relocation of the intact 5’-MYC-3’ from der 8q, and apposition to 3’-IGH@ at der 14q. This case report provides unique and additional cytogenetic data that may be of clinical significance in such a rare finding in CLL. It also highlights the utility of conventional and sequential metaphase FISH in understanding complex chromosome anomalies and their association with other clinical findings in patients with CLL. To the best of our knowledge, this is the first CLL reported case with such an atypical rearrangement in a patient with a negative MYC expression. PMID:23369149

Curiosity's ChemCam Checks 'Christmas Cove' Colors

NASA Image and Video Library

2017-11-01

The Chemistry and Camera (ChemCam) instrument on NASA's Curiosity Mars rover examined a freshly brushed area on target rock "Christmas Cove" and found spectral evidence of hematite, an iron-oxide mineral. ChemCam sometimes zaps rocks with a laser, but can also be used, as in this case, in a "passive" mode. In this type of investigation, the instrument's telescope delivers to spectrometers the sunlight reflected from a small target point. The upper-left inset of this graphic is an image from ChemCam's Remote Micro-Imager with five labeled points that the instrument analyzed. The image covers an area about 2 inches (5 centimeters) wide, and the bright lines are fractures in the rock filled with calcium sulfate minerals. The five charted lines of the graphic correspond to those five points and show the spectrometer measurements of brightness at thousands of different wavelengths, from 400 nanometers (at the violet end of the visible-light spectrum) to 840 nanometers (in near-infrared). Sections of the spectrum measurements that are helpful for identifying hematite are annotated. These include a dip around 535 nanometers, the green-light portion of the spectrum at which fine-grained hematite tends to absorb more light and reflect less compared to other parts of the spectrum. That same green-absorbing characteristic of the hematite makes it appear purplish when imaged through special filters of Curiosity's Mast Camera and even in usual color images. The spectra also show maximum reflectance values near 750 nanometers, followed by a steep decrease in the spectral slope toward 840 nanometers, both of which are consistent with hematite. This ChemCam examination of Christmas Cove was part of an experiment to determine whether the rock had evidence of hematite under a tan coating of dust. The target area was brushed with Curiosity's Dust Removal Tool prior to these ChemCam passive observations on Sept. 17, 2017, during the 1,819th Martian day, or sol, of Curiosity's work on Mars. https://photojournal.jpl.nasa.gov/catalog/PIA22068

LPL is the strongest prognostic factor in a comparative analysis of RNA-based markers in early chronic lymphocytic leukemia.

PubMed

Kaderi, Mohd Arifin; Kanduri, Meena; Buhl, Anne Mette; Sevov, Marie; Cahill, Nicola; Gunnarsson, Rebeqa; Jansson, Mattias; Smedby, Karin Ekström; Hjalgrim, Henrik; Jurlander, Jesper; Juliusson, Gunnar; Mansouri, Larry; Rosenquist, Richard

2011-08-01

The expression levels of LPL, ZAP70, TCL1A, CLLU1 and MCL1 have recently been proposed as prognostic factors in chronic lymphocytic leukemia. However, few studies have systematically compared these different RNA-based markers. Using real-time quantitative PCR, we measured the mRNA expression levels of these genes in unsorted samples from 252 newly diagnosed chronic lymphocytic leukemia patients and correlated our data with established prognostic markers (for example Binet stage, CD38, IGHV gene mutational status and genomic aberrations) and clinical outcome. High expression levels of all RNA-based markers, except MCL1, predicted shorter overall survival and time to treatment, with LPL being the most significant. In multivariate analysis including the RNA-based markers, LPL expression was the only independent prognostic marker for overall survival and time to treatment. When studying LPL expression and the established markers, LPL expression retained its independent prognostic strength for overall survival. All of the RNA-based markers, albeit with varying ability, added prognostic information to established markers, with LPL expression giving the most significant results. Notably, high LPL expression predicted a worse outcome in good-prognosis subgroups, such as patients with mutated IGHV genes, Binet stage A, CD38 negativity or favorable cytogenetics. In particular, the combination of LPL expression and CD38 could further stratify Binet stage A patients. LPL expression is the strongest RNA-based prognostic marker in chronic lymphocytic leukemia that could potentially be applied to predict outcome in the clinical setting, particularly in the large group of patients with favorable prognosis.

Patient satisfaction with ambulatory care in Germany: effects of patient- and medical practice-related factors.

PubMed

Auras, Silke; Ostermann, Thomas; de Cruppé, Werner; Bitzer, Eva-Maria; Diel, Franziska; Geraedts, Max

2016-12-01

The study aimed to illustrate the effect of the patients' sex, age, self-rated health and medical practice specialization on patient satisfaction. Secondary analysis of patient survey data using multilevel analysis (generalized linear mixed model, medical practice as random effect) using a sequential modelling strategy. We examined the effects of the patients' sex, age, self-rated health and medical practice specialization on four patient satisfaction dimensions: medical practice organization, information, interaction, professional competence. The study was performed in 92 German medical practices providing ambulatory care in general medicine, internal medicine or gynaecology. In total, 9888 adult patients participated in a patient survey using the validated 'questionnaire on satisfaction with ambulatory care-quality from the patient perspective [ZAP]'. We calculated four models for each satisfaction dimension, revealing regression coefficients with 95% confidence intervals (CIs) for all independent variables, and using Wald Chi-Square statistic for each modelling step (model validity) and LR-Tests to compare the models of each step with the previous model. The patients' sex and age had a weak effect (maximum regression coefficient 1.09, CI 0.39; 1.80), and the patients' self-rated health had the strongest positive effect (maximum regression coefficient 7.66, CI 6.69; 8.63) on satisfaction ratings. The effect of medical practice specialization was heterogeneous. All factors studied, specifically the patients' self-rated health, affected patient satisfaction. Adjustment should always be considered because it improves the comparability of patient satisfaction in medical practices with atypically varying patient populations and increases the acceptance of comparisons. © The Author 2016. Published by Oxford University Press in association with the International Society for Quality in Health Care. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Cell-intrinsic determinants of ibrutinib-induced apoptosis in Chronic Lymphocytic Leukemia

PubMed Central

Amin, Nisar A.; Balasubramanian, Sriram; Saiya-Cork, Kamlai; Shedden, Kerby; Hu, Nan; Malek, Sami N.

2016-01-01

Purpose Ibrutinib, a Bruton’s tyrosine kinase (BTK) inhibitor, is approved for the treatment of relapsed CLL and CLL with del17p. Mechanistically, ibrutinib interferes with BCR signaling as well as multiple CLL cell to microenvironment interactions. Given the importance of ibrutinib in the management of CLL, a deeper understanding of factors governing sensitivity and resistance is warranted. Experimental Design We studied 48 longitudinally sampled paired CLL samples, 42 of which were procured before and after standard CLL chemotherapies, and characterized them for well-studied CLL molecular traits as well as by whole exome sequencing and SNP 6.0 array profiling. We exposed these samples to 0.25 μM – 5 μM of ibrutinib ex vivo and measured apoptosis fractions as well as BCR signaling by immunoblotting. We disrupted TP53 in HG3, PGA1 and PG-EBV cell lines and measured BCR signaling and ibrutinib responses. Results CLL samples demonstrated a surprisingly wide range of ex vivo sensitivities to ibrutinib with IC50 values ranging from 0.4 μM – 9.7 μM. Unmutated IGVH status, elevated ZAP70 expression and trisomy 12 were associated with heightened sensitivity to ibrutinib treatment. Five CLL samples were substantially more resistant to ibrutinib following relapse from chemotherapy; of these, three had acquired a del17p/TP53 mutated status. A validation sample of 15 CLL carrying TP53 mutations, of which 13 carried both del17p and a TP53 mutation confirmed substantially less sensitivity to ibrutinib-induced apoptosis. Conclusions This study identifies that CLL harboring del17p/TP53 mutated cells are substantially less sensitive to ibrutinib-induced apoptosis than del17p/TP53 wild type cells. PMID:27535981

Distinct regulatory functions of SLP-76 and MIST in NK cell cytotoxicity and IFN-gamma production.

PubMed

Hidano, Shinya; Sasanuma, Hiroki; Ohshima, Keiko; Seino, Ken-ichiro; Kumar, Lalit; Hayashi, Katsuhiko; Hikida, Masaki; Kurosaki, Tomohiro; Taniguchi, Masaru; Geha, Raif S; Kitamura, Daisuke; Goitsuka, Ryo

2008-03-01

Activation of NK cells is triggered by multiple receptors. We demonstrate here that SLP-76 is required for CD16- and NKG2D-mediated NK cell cytotoxicity, while MIST negatively regulates these responses in an SLP-76-dependent manner. Exceptionally, MIST acts as a positive regulator of cytotoxicity against YAC-1 cells, although SLP-76 plays a more key role. SLP-76 acts as a dominant positive regulator for both NKG2D-mediated and YAC-1 cell-triggered IFN-gamma production. Although NKG2D-mediated IFN-gamma production depends on phospholipase C (PLC) gamma 2, YAC-1 cell-triggered IFN-gamma production is PLC gamma 2- and Syk/ZAP-70 independent and nuclear factor-kappa B mediated. SLP-76 is required for this process in the presence of MIST but is dispensable in the absence of MIST. Thus, YAC-1 cell-triggered NKG2D-independent IFN-gamma production appears to be regulated by SLP-76-dependent and -independent pathways, in which the latter is negatively regulated by MIST. Taken together, these results suggest that SLP-76 and MIST distinctly but interactively regulate NK cell cytotoxicity and IFN-gamma production.

SLAP deficiency enhances number and function of regulatory T cells preventing chronic autoimmune arthritis in SKG mice.

PubMed

Peterson, Lisa K; Shaw, Laura A; Joetham, Anthony; Sakaguchi, Shimon; Gelfand, Erwin W; Dragone, Leonard L

2011-02-15

To test if manipulating TCR complex-mediated signaling (TCR signaling) could treat autoimmune disease, we generated the double SKG Src-like adapter protein (SLAP) knockout (DSSKO) mouse model. The SKG mutation in ZAP70 and SLAP have opposing functions on the regulation of TCR signaling. The combination of these two mutations alters TCR signaling in the context of a defined genetic background, uniform environmental conditions, and a well-characterized signaling disruption. In contrast to SKG mice, DSSKO mice do not develop zymosan-induced chronic autoimmune arthritis. This arthritis prevention is not due to significant alterations in thymocyte development or repertoire selection but instead enhanced numbers of regulatory T cells (Tregs) and decreased numbers of Th17 cells skewing the ratio of Tregs to autoreactive effector T cells. Treg depletion and/or functional blockade led to the development of arthritis in DSSKO mice. In vitro suppression of effector T cell proliferation was also enhanced, demonstrating that DSSKO mice have increased numbers of Tregs with increased function. Understanding how TCR signals influence development, expansion, and function of Tregs in DSSKO mice could advance our ability to manipulate Treg biology to treat ultimately autoimmune disease.

SLAP Deficiency Enhances Number and Function of Regulatory T Cells Preventing Chronic Autoimmune Arthritis in SKG Mice

PubMed Central

Peterson, Lisa K.; Shaw, Laura A.; Joetham, Anthony; Sakaguchi, Shimon; Gelfand, Erwin W.; Dragone, Leonard L.

2011-01-01

To test if manipulating TCR complex-mediated signaling (TCR signaling) could treat autoimmune disease, we generated the double SKG Src-like adapter protein (SLAP) knockout (DSSKO) mouse model. The SKG mutation in ZAP70 and SLAP have opposing functions on the regulation of TCR signaling. The combination of these two mutations alters TCR signaling in the context of a defined genetic background, uniform environmental conditions, and a well-characterized signaling disruption. In contrast to SKG mice, DSSKO mice do not develop zymosan-induced chronic autoimmune arthritis. This arthritis prevention is not due to significant alterations in thymocyte development or repertoire selection but instead enhanced numbers of regulatory T cells (Tregs) and decreased numbers of Th17 cells skewing the ratio of Tregs to autoreactive effector T cells. Treg depletion and/or functional blockade led to the development of arthritis in DSSKO mice. In vitro suppression of effector T cell proliferation was also enhanced, demonstrating that DSSKO mice have increased numbers of Tregs with increased function. Understanding how TCR signals influence development, expansion, and function of Tregs in DSSKO mice could advance our ability to manipulate Treg biology to treat ultimately autoimmune disease. PMID:21248251

MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias

PubMed Central

Calin, George Adrian; Liu, Chang-Gong; Sevignani, Cinzia; Ferracin, Manuela; Felli, Nadia; Dumitru, Calin Dan; Shimizu, Masayoshi; Cimmino, Amelia; Zupo, Simona; Dono, Mariella; Dell'Aquila, Marie L.; Alder, Hansjuerg; Rassenti, Laura; Kipps, Thomas J.; Bullrich, Florencia; Negrini, Massimo; Croce, Carlo M.

2004-01-01

Little is known about the expression levels or function of micro-RNAs (miRNAs) in normal and neoplastic cells, although it is becoming clear that miRNAs play important roles in the regulation of gene expression during development [Ambros, V. (2003) Cell 113, 673–676; McManus, M. T. (2003) Semin. Cancer Biol. 13, 253–258]. We now report the genomewide expression profiling of miRNAs in human B cell chronic lymphocytic leukemia (CLL) by using a microarray containing hundreds of human precursor and mature miRNA oligonucleotide probes. This approach allowed us to identify significant differences in miRNome expression between CLL samples and normal CD5+ B cells; data were confirmed by Northern blot analyses and real-time RT-PCR. At least two distinct clusters of CLL samples can be identified that were associated with the presence or absence of Zap-70 expression, a predictor of early disease progression. Two miRNA signatures were associated with the presence or absence of mutations in the expressed Ig variableregion genes or with deletions at 13q14, respectively. These data suggest that miRNA expression patterns have relevance to the biological and clinical behavior of this leukemia. PMID:15284443

SSRI and SNRI withdrawal symptoms reported on an internet forum.

PubMed

Stockmann, Tom; Odegbaro, Dolapo; Timimi, Sami; Moncrieff, Joanna

2018-05-09

Antidepressant withdrawal symptoms are well-recognised, but their potential duration remains uncertain. We aimed to describe the characteristics of withdrawal associated with two popular classes of antidepressants, including duration. We analysed the content of a sample of posts on an antidepressant withdrawal website. We compared the characteristics of withdrawal associated with SSRIs and SNRIs, including time of onset, duration and nature of symptoms. 110 posts about SSRI withdrawal, and 63 concerning SNRI withdrawal, were analysed. The mean duration of withdrawal symptoms was significantly longer with SSRIs than SNRIs: 90.5 weeks (standard deviation, SD, 150.0) and 50.8 weeks (SD 76.0) respectively; p = 0.043). Neurological symptoms, such as 'brain zaps,' were more common among SNRI users (p = 0.023). Psychosexual/genitourinary symptoms may be more common among SSRI users (p = 0.054). The website aims to help people with antidepressant withdrawal, and is therefore likely to attract people who have difficulties. Length of prior use of antidepressants was long, with a mean of 252.2 weeks (SD 250.8). People accessing antidepressant withdrawal websites report experiencing protracted withdrawal symptoms. There are some differences in the characteristics of withdrawal associated with different classes of antidepressants.

A Reactor Development Scenario for the FUZE Shear-flow Stabilized Z-pinch

NASA Astrophysics Data System (ADS)

McLean, H. S.; Higginson, D. P.; Schmidt, A.; Tummel, K. K.; Shumlak, U.; Nelson, B. A.; Claveau, E. L.; Golingo, R. P.; Weber, T. R.

2016-10-01

We present a conceptual design, scaling calculations, and a development path for a pulsed fusion reactor based on the shear-flow-stabilized Z-pinch device. Experiments performed on the ZaP device have demonstrated stable operation for 40 us at 150 kA total discharge current (with 100 kA in the pinch) for pinches that are 1cm in diameter and 100 cm long. Scaling calculations show that achieving stabilization for a pulse of 100 usec, for discharge current 1.5 MA, in a shortened pinch 50 cm, results in a pinch diameter of 200 um and a reactor plant Q 5 for reasonable assumptions of the various system efficiencies. We propose several key intermediate performance levels in order to justify further development. These include achieving operation at pinch currents of 300 kA, where Te and Ti are calculated to exceed 1 keV, 700 kA where fusion power exceeds pinch input power, and 1 MA where fusion energy per pulse exceeds input energy per pulse. This work funded by USDOE ARPAe ALPHA Program and performed under the auspices of Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. LLNL-ABS-697801.

Antimatter Driven P-B11 Fusion Propulsion System

NASA Technical Reports Server (NTRS)

Kammash, Terry; Martin, James; Godfroy, Thomas

2002-01-01

One of the major advantages of using P-B11 fusion fuel is that the reaction produces only charged particles in the form of three alpha particles and no neutrons. A fusion concept that lends itself to this fuel cycle is the Magnetically Insulated Inertial Confinement Fusion (MICF) reactor whose distinct advantage lies in the very strong magnetic field that is created when an incident particle (or laser) beam strikes the inner wall of the target pellet. This field serves to thermally insulate the hot plasma from the metal wall thereby allowing thc plasma to burn for a long time and produce a large energy magnification. If used as a propulsion device, we propose using antiprotons to drive the system which we show to be capable of producing very large specific impulse and thrust. By way of validating the confinement propenies of MICF we will address a proposed experiment in which pellets coated with P-B11 fuel at the appropriate ratio will be zapped by a beam of antiprotons that enter the target through a hole. Calculations showing the density and temperature of the generated plasma along with the strength of the magnetic field and other properties of the system will be presented and discussed.

Proteus mirabilis interkingdom swarming signals attract blow flies

PubMed Central

Ma, Qun; Fonseca, Alicia; Liu, Wenqi; Fields, Andrew T; Pimsler, Meaghan L; Spindola, Aline F; Tarone, Aaron M; Crippen, Tawni L; Tomberlin, Jeffery K; Wood, Thomas K

2012-01-01

Flies transport specific bacteria with their larvae that provide a wider range of nutrients for those bacteria. Our hypothesis was that this symbiotic interaction may depend on interkingdom signaling. We obtained Proteus mirabilis from the salivary glands of the blow fly Lucilia sericata; this strain swarmed significantly and produced a strong odor that attracts blow flies. To identify the putative interkingdom signals for the bacterium and flies, we reasoned that as swarming is used by this bacterium to cover the food resource and requires bacterial signaling, the same bacterial signals used for swarming may be used to communicate with blow flies. Using transposon mutagenesis, we identified six novel genes for swarming (ureR, fis, hybG, zapB, fadE and PROSTU_03490), then, confirming our hypothesis, we discovered that fly attractants, lactic acid, phenol, NaOH, KOH and ammonia, restore swarming for cells with the swarming mutations. Hence, compounds produced by the bacterium that attract flies also are utilized for swarming. In addition, bacteria with the swarming mutation rfaL attracted fewer blow flies and reduced the number of eggs laid by the flies. Therefore, we have identified several interkingdom signals between P. mirabilis and blow flies. PMID:22237540

Enabling Broadband as Commodity within Access Networks: A QoS Recipe

NASA Astrophysics Data System (ADS)

Areizaga, Enrique; Foglar, Andreas; Elizondo, Antonio J.; Geilhardt, Frank

This paper describes the QoS features that will transform the access networks landscape in order to bring “Broadband” as a commodity while setting up the pillars of the “Future Media Internet”. Quality of Experience is obviously key for emerging and future services. Broadcasting services will first need to equal the QoE of their counterparts in the Open-air market (for IP-TV examples would be artifact-free, no picture freezing, fast zapping times) and offer new features often using interactivity (Time-shifted TV, access to more content, 3DTV with feeling of presence). The huge variety of communications alternatives will lead to different requirements per customer, whose needs will also be dependent on parameters like where the connection is made, the time of the day/day of the week/period of the year or even his/her mood. Today’s networks, designed for providing just Broadband connectivity, will not be enough to satisfy customer’s needs and will necessarily support the introduction of new and innovative services. The Networks of the future should learn from the way the users are communicating, what services they are using, where, when, and how, and adapt accordingly.

Iron-Nickel Meteorite Zapped by Mars Rover Laser

NASA Image and Video Library

2016-11-02

The dark, golf-ball-size object in this composite, colorized view from the Chemistry and Camera (ChemCam) instrument on NASA's Curiosity Mars rover shows a grid of shiny dots where ChemCam had fired laser pulses used for determining the chemical elements in the target's composition. The analysis confirmed that this object, informally named "Egg Rock," is an iron-nickel meteorite. Iron-nickel meteorites are a common class of space rocks found on Earth, and previous examples have been found on Mars, but Egg Rock is the first on Mars to be examined with a laser-firing spectrometer. The laser pulses on Oct. 30, 2016, induced bursts of glowing gas at the target, and ChemCam's spectrometer read the wavelengths of light from those bursts to gain information about the target's composition. The laser pulses also burned through the dark outer surface, exposing bright interior material. This view combines two images taken later the same day by ChemCam's remote micro-imager (RMI) camera, with color added from an image taken by Curiosity's Mast Camera (Mastcam). A Mastcam image of Egg Rock is at PIA21134. http://photojournal.jpl.nasa.gov/catalog/PIA21133

A PLC-γ1 Feedback Pathway Regulates Lck Substrate Phosphorylation at the T-Cell Receptor and SLP-76 Complex.

PubMed

Belmont, Judson; Gu, Tao; Mudd, Ashley; Salomon, Arthur R

2017-08-04

Phospholipase C gamma 1 (PLC-γ1) occupies a critically important position in the T-cell signaling pathway. While its functions as a regulator of both Ca 2+ signaling and PKC-family kinases are well characterized, PLC-γ1's role in the regulation of early T-cell receptor signaling events is incompletely understood. Activation of the T-cell receptor leads to the formation of a signalosome complex between SLP-76, LAT, PLC-γ1, Itk, and Vav1. Recent studies have revealed the existence of both positive and negative feedback pathways from SLP-76 to the apical kinase in the pathway, Lck. To determine if PLC-γ1 contributes to the regulation of these feedback networks, we performed a quantitative phosphoproteomic analysis of PLC-γ1-deficient T cells. These data revealed a previously unappreciated role for PLC-γ1 in the positive regulation of Zap-70 and T-cell receptor tyrosine phosphorylation. Conversely, PLC-γ1 negatively regulated the phosphorylation of SLP-76-associated proteins, including previously established Lck substrate phosphorylation sites within this complex. While the positive and negative regulatory phosphorylation sites on Lck were largely unchanged, Tyr 192 phosphorylation was elevated in Jgamma1. The data supports a model wherein Lck's targeting, but not its kinase activity, is altered by PLC-γ1, possibly through Lck Tyr 192 phosphorylation and increased association of the kinase with protein scaffolds SLP-76 and TSAd.

Initiating Molecular Growth in the Interstellar Medium via Dimeric Complexes of Observed Ions and Molecules

NASA Technical Reports Server (NTRS)

Bera, Partha P.; Head-Gordon, Martin; Lee, Timothy J.

2011-01-01

A feasible initiation step for particle growth in the interstellar medium (ISM) is simulated by means of ab quantum chemistry methods. The systems studied are dimer ions formed by pairing nitrogen containing small molecules known to exist in the ISM with ions of unsaturated hydrocarbons or vice versa. Complexation energies, structures of ensuing complexes and electronic excitation spectra of the encounter complexes are estimated using various quantum chemistry methods. Moller-Plesset perturbation theory (MP2, Z-averaged perturbation theory (ZAP2), coupled cluster singles and doubles with perturbative triples corrections (CCSD(T)), and density functional theory (DFT) methods (B3LYP) were employed along with the correlation consistent cc-pVTZ and aug-cc-pVTZ basis sets. Two types of complexes are predicted. One type of complex has electrostatic binding with moderate (7-20 kcal per mol) binding energies, that are nonetheless significantly stronger than typical van der Waals interactions between molecules of this size. The other type of complex develops strong covalent bonds between the fragments. Cyclic isomers of the nitrogen containing complexes are produced very easily by ion-molecule reactions. Some of these complexes show intense ultraviolet visible spectra for electronic transitions with large oscillator strengths at the B3LYP, omegaB97, and equations of motion coupled cluster (EOM-CCSD) levels. The open shell nitrogen containing carbonaceous complexes especially exhibit a large oscillator strength electronic transition in the visible region of the electromagnetic spectrum.

Impaired NFAT and NFκB activation are involved in suppression of CD40 ligand expression by Δ(9)-tetrahydrocannabinol in human CD4(+) T cells.

PubMed

Ngaotepprutaram, Thitirat; Kaplan, Barbara L F; Kaminski, Norbert E

2013-11-15

We have previously reported that Δ(9)-tetrahydrocannabinol (Δ(9)-THC), the main psychoactive cannabinoid in marijuana, suppresses CD40 ligand (CD40L) expression by activated mouse CD4(+) T cells. CD40L is involved in pathogenesis of many autoimmune and inflammatory diseases. In the present study, we investigated the molecular mechanism of Δ(9)-THC-mediated suppression of CD40L expression using peripheral blood human T cells. Pretreatment with Δ(9)-THC attenuated CD40L expression in human CD4(+) T cells activated by anti-CD3/CD28 at both the protein and mRNA level, as determined by flow cytometry and quantitative real-time PCR, respectively. Electrophoretic mobility shift assays revealed that Δ(9)-THC suppressed the DNA-binding activity of both NFAT and NFκB to their respective response elements within the CD40L promoter. An assessment of the effect of Δ(9)-THC on proximal T cell-receptor (TCR) signaling induced by anti-CD3/CD28 showed significant impairment in the rise of intracellular calcium, but no significant effect on the phosphorylation of ZAP70, PLCγ1/2, Akt, and GSK3β. Collectively, these findings identify perturbation of the calcium-NFAT and NFκB signaling cascade as a key mechanistic event by which Δ(9)-THC suppresses human T cell function. © 2013.

A Candida Biofilm-Induced Pathway for Matrix Glucan Delivery: Implications for Drug Resistance

PubMed Central

Taff, Heather T.; Nett, Jeniel E.; Zarnowski, Robert; Ross, Kelly M.; Sanchez, Hiram; Cain, Mike T.; Hamaker, Jessica; Mitchell, Aaron P.; Andes, David R.

2012-01-01

Extracellular polysaccharides are key constituents of the biofilm matrix of many microorganisms. One critical carbohydrate component of Candida albicans biofilms, β-1,3 glucan, has been linked to biofilm protection from antifungal agents. In this study, we identify three glucan modification enzymes that function to deliver glucan from the cell to the extracellular matrix. These enzymes include two predicted glucan transferases and an exo-glucanase, encoded by BGL2, PHR1, and XOG1, respectively. We show that the enzymes are crucial for both delivery of β-1,3 glucan to the biofilm matrix and for accumulation of mature matrix biomass. The enzymes do not appear to impact cell wall glucan content of biofilm cells, nor are they necessary for filamentation or biofilm formation. We demonstrate that mutants lacking these genes exhibit enhanced susceptibility to the commonly used antifungal, fluconazole, during biofilm growth only. Transcriptional analysis and biofilm phenotypes of strains with multiple mutations suggest that these enzymes act in a complementary fashion to distribute matrix downstream of the primary β-1,3 glucan synthase encoded by FKS1. Furthermore, our observations suggest that this matrix delivery pathway works independently from the C. albicans ZAP1 matrix formation regulatory pathway. These glucan modification enzymes appear to play a biofilm-specific role in mediating the delivery and organization of mature biofilm matrix. We propose that the discovery of inhibitors for these enzymes would provide promising anti-biofilm therapeutics. PMID:22876186

Multisite Phosphorylation Modulates the T Cell Receptor ζ-Chain Potency but not the Switchlike Response.

PubMed

Mukhopadhyay, Himadri; de Wet, Ben; Clemens, Lara; Maini, Philip K; Allard, Jun; van der Merwe, P Anton; Dushek, Omer

2016-04-26

Multisite phosphorylation is ubiquitous in cellular signaling and is thought to provide signaling proteins with additional regulatory mechanisms. Indeed, mathematical models have revealed a large number of mechanisms by which multisite phosphorylation can produce switchlike responses. The T cell antigen receptor (TCR) is a multisubunit receptor on the surface of T cells that is a prototypical multisite substrate as it contains 20 sites that are distributed on 10 conserved immunoreceptor tyrosine-based activation motifs (ITAMs). The TCR ζ-chain is a homodimer subunit that contains six ITAMs (12 sites) and exhibits a number of properties that are predicted to be sufficient for a switchlike response. We have used cellular reconstitution to systematically study multisite phosphorylation of the TCR ζ-chain. We find that multisite phosphorylation proceeds by a nonsequential random mechanism, and find no evidence that multiple ITAMs modulate a switchlike response but do find that they alter receptor potency and maximum phosphorylation. Modulation of receptor potency can be explained by a reduction in molecular entropy of the disordered ζ-chain upon phosphorylation. We further find that the tyrosine kinase ZAP-70 increases receptor potency but does not modulate the switchlike response. In contrast to other multisite proteins, where phosphorylations act in strong concert to modulate protein function, we suggest that the multiple ITAMs on the TCR function mainly to amplify subsequent signaling. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

The TGA codons are present in the open reading frame of selenoprotein P cDNA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hill, K.E.; Lloyd, R.S.; Read, R.

1991-03-11

The TGA codon in DNA has been shown to direct incorporation of selenocysteine into protein. Several proteins from bacteria and animals contain selenocysteine in their primary structures. Each of the cDNA clones of these selenoproteins contains one TGA codon in the open reading frame which corresponds to the selenocysteine in the protein. A cDNA clone for selenoprotein P (SeP), obtained from a {gamma}ZAP rat liver library, was sequenced by the dideoxy termination method. The correct reading frame was determined by comparison of the deduced amino acid sequence with the amino acid sequence of several peptides from SeP. Using SeP labelledmore » with {sup 75}Se in vivo, the selenocysteine content of the peptides was verified by the collection of carboxymethylated {sup 77}Se-selenocysteine as it eluted from the amino acid analyzer and determination of the radioactivity contained in the collected samples. Ten TGA codons are present in the open reading frame of the cDNA. Peptide fragmentation studies and the deduced sequence indicate that selenium-rich regions are located close to the carboxy terminus. Nine of the 10 selenocysteines are located in the terminal 26% of the sequence with four in the terminal 15 amino acids. The deduced sequence codes for a protein of 385 amino acids. Cleavage of the signal peptide gives the mature protein with 366 amino acids and a calculated mol wt of 41,052 Da. Searches of PIR and SWISSPROT protein databases revealed no similarity with glutathione peroxidase or other selenoproteins.« less

CD81 Controls Sustained T Cell Activation Signaling and Defines the Maturation Stages of Cognate Immunological Synapses

PubMed Central

Rocha-Perugini, V.; Zamai, M.; González-Granado, J. M.; Barreiro, O.; Tejera, E.; Yañez-Mó, M.; Caiolfa, V. R.

2013-01-01

In this study, we investigated the dynamics of the molecular interactions of tetraspanin CD81 in T lymphocytes, and we show that CD81 controls the organization of the immune synapse (IS) and T cell activation. Using quantitative microscopy, including fluorescence recovery after photobleaching (FRAP), phasor fluorescence lifetime imaging microscopy-Föster resonance energy transfer (phasorFLIM-FRET), and total internal reflection fluorescence microscopy (TIRFM), we demonstrate that CD81 interacts with ICAM-1 and CD3 during conjugation between T cells and antigen-presenting cells (APCs). CD81 and ICAM-1 exhibit distinct mobilities in central and peripheral areas of early and late T cell-APC contacts. Moreover, CD81–ICAM-1 and CD81-CD3 dynamic interactions increase over the time course of IS formation, as these molecules redistribute throughout the contact area. Therefore, CD81 associations unexpectedly define novel sequential steps of IS maturation. Our results indicate that CD81 controls the temporal progression of the IS and the permanence of CD3 in the membrane contact area, contributing to sustained T cell receptor (TCR)-CD3-mediated signaling. Accordingly, we find that CD81 is required for proper T cell activation, regulating CD3ζ, ZAP-70, LAT, and extracellular signal-regulated kinase (ERK) phosphorylation; CD69 surface expression; and interleukin-2 (IL-2) secretion. Our data demonstrate the important role of CD81 in the molecular organization and dynamics of the IS architecture that sets the signaling threshold in T cell activation. PMID:23858057

WASp family verprolin-homologous protein-2 (WAVE2) and Wiskott-Aldrich syndrome protein (WASp) engage in distinct downstream signaling interactions at the T cell antigen receptor site.

PubMed

Pauker, Maor H; Reicher, Barak; Joseph, Noah; Wortzel, Inbal; Jakubowicz, Shlomi; Noy, Elad; Perl, Orly; Barda-Saad, Mira

2014-12-12

T cell antigen receptor (TCR) engagement has been shown to activate pathways leading to actin cytoskeletal polymerization and reorganization, which are essential for lymphocyte activation and function. Several actin regulatory proteins were implicated in regulating the actin machinery, such as members of the Wiskott-Aldrich syndrome protein (WASp) family. These include WASp and the WASp family verprolin-homologous protein-2 (WAVE2). Although WASp and WAVE2 share several structural features, the precise regulatory mechanisms and potential redundancy between them have not been fully characterized. Specifically, unlike WASp, the dynamic molecular interactions that regulate WAVE2 recruitment to the cell membrane and specifically to the TCR signaling complex are largely unknown. Here, we identify the molecular mechanism that controls the recruitment of WAVE2 in comparison with WASp. Using fluorescence resonance energy transfer (FRET) and novel triple-color FRET (3FRET) technology, we demonstrate how WAVE2 signaling complexes are dynamically regulated during lymphocyte activation in vivo. We show that, similar to WASp, WAVE2 recruitment to the TCR site depends on protein-tyrosine kinase, ZAP-70, and the adaptors LAT, SLP-76, and Nck. However, in contrast to WASp, WAVE2 leaves this signaling complex and migrates peripherally together with vinculin to the membrane leading edge. Our experiments demonstrate that WASp and WAVE2 differ in their dynamics and their associated proteins. Thus, this study reveals the differential mechanisms regulating the function of these cytoskeletal proteins. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

WASp Family Verprolin-homologous Protein-2 (WAVE2) and Wiskott-Aldrich Syndrome Protein (WASp) Engage in Distinct Downstream Signaling Interactions at the T Cell Antigen Receptor Site*

PubMed Central

Pauker, Maor H.; Reicher, Barak; Joseph, Noah; Wortzel, Inbal; Jakubowicz, Shlomi; Noy, Elad; Perl, Orly; Barda-Saad, Mira

2014-01-01

T cell antigen receptor (TCR) engagement has been shown to activate pathways leading to actin cytoskeletal polymerization and reorganization, which are essential for lymphocyte activation and function. Several actin regulatory proteins were implicated in regulating the actin machinery, such as members of the Wiskott-Aldrich syndrome protein (WASp) family. These include WASp and the WASp family verprolin-homologous protein-2 (WAVE2). Although WASp and WAVE2 share several structural features, the precise regulatory mechanisms and potential redundancy between them have not been fully characterized. Specifically, unlike WASp, the dynamic molecular interactions that regulate WAVE2 recruitment to the cell membrane and specifically to the TCR signaling complex are largely unknown. Here, we identify the molecular mechanism that controls the recruitment of WAVE2 in comparison with WASp. Using fluorescence resonance energy transfer (FRET) and novel triple-color FRET (3FRET) technology, we demonstrate how WAVE2 signaling complexes are dynamically regulated during lymphocyte activation in vivo. We show that, similar to WASp, WAVE2 recruitment to the TCR site depends on protein-tyrosine kinase, ZAP-70, and the adaptors LAT, SLP-76, and Nck. However, in contrast to WASp, WAVE2 leaves this signaling complex and migrates peripherally together with vinculin to the membrane leading edge. Our experiments demonstrate that WASp and WAVE2 differ in their dynamics and their associated proteins. Thus, this study reveals the differential mechanisms regulating the function of these cytoskeletal proteins. PMID:25342748

Metabolism and Regulation of Glycerolipids in the Yeast Saccharomyces cerevisiae

PubMed Central

Henry, Susan A.; Kohlwein, Sepp D.; Carman, George M.

2012-01-01

Due to its genetic tractability and increasing wealth of accessible data, the yeast Saccharomyces cerevisiae is a model system of choice for the study of the genetics, biochemistry, and cell biology of eukaryotic lipid metabolism. Glycerolipids (e.g., phospholipids and triacylglycerol) and their precursors are synthesized and metabolized by enzymes associated with the cytosol and membranous organelles, including endoplasmic reticulum, mitochondria, and lipid droplets. Genetic and biochemical analyses have revealed that glycerolipids play important roles in cell signaling, membrane trafficking, and anchoring of membrane proteins in addition to membrane structure. The expression of glycerolipid enzymes is controlled by a variety of conditions including growth stage and nutrient availability. Much of this regulation occurs at the transcriptional level and involves the Ino2–Ino4 activation complex and the Opi1 repressor, which interacts with Ino2 to attenuate transcriptional activation of UASINO-containing glycerolipid biosynthetic genes. Cellular levels of phosphatidic acid, precursor to all membrane phospholipids and the storage lipid triacylglycerol, regulates transcription of UASINO-containing genes by tethering Opi1 to the nuclear/endoplasmic reticulum membrane and controlling its translocation into the nucleus, a mechanism largely controlled by inositol availability. The transcriptional activator Zap1 controls the expression of some phospholipid synthesis genes in response to zinc availability. Regulatory mechanisms also include control of catalytic activity of glycerolipid enzymes by water-soluble precursors, products and lipids, and covalent modification of phosphorylation, while in vivo function of some enzymes is governed by their subcellular location. Genome-wide genetic analysis indicates coordinate regulation between glycerolipid metabolism and a broad spectrum of metabolic pathways. PMID:22345606

Scaling the Shear-flow Stabilized Z-pinch to Reactor Conditions

NASA Astrophysics Data System (ADS)

McLean, H. S.; Schmidt, A.; Shumlak, U.; Nelson, B. A.; Golingo, R. P.; Cleveau, E.

2015-11-01

We present a conceptual design along with scaling calculations for a pulsed fusion reactor based on the shear-flow-stabilized Z-pinch device. Experiments performed on the ZaP device, at the University of Washington, have demonstrated stable operation for durations of 20 usec at ~100kA discharge current for pinches that are ~1 cm in diameter and 100 cm long. The inverse of the pinch diameter and plasma energy density scale strongly with pinch current and calculations show that maintaining stabilization durations of ~7 usec for increased discharge current (~15x) in a shortened pinch (10 cm) results in a pinch diameter of ~200 um and plasma conditions that approach those needed to support significant fusion burn and energy gain (Ti ~ 30keV, density ~ 3e26/m3, ntau ~1.4e20 sec/m3). Compelling features of the concept include operation at modest discharge current (1.5 MA) and voltage (40kV) along with direct adoption of liquid metals for at least one electrode--technological capabilities that have been proven in existing, commercial, pulse power devices such as large ignitrons. LLNL-ABS-674920. This work performed under the auspices of the U.S. Department of Energy ARPAe ALPHA Program by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

Polytropic scaling of a flow Z-pinch

NASA Astrophysics Data System (ADS)

Hughes, M. C.; Shumlak, U.; Nelson, B. A.; Golingo, R. P.; Claveau, E. L.; Doty, S. A.; Forbes, E. G.; Kim, B.; Ross, M. P.; Weed, J. R.

2015-11-01

The ZaP Flow Z-Pinch project investigates the use of velocity shear to mitigate MHD instabilities. The ZaP-HD experiment produces 50 cm long pinches of varying radii. The power to the experiment is split between the plasma formation and acceleration process and the pinch assembly and compression process. Once the pinch is formed, low magnetic fluctuations indicate a quiescent, long-lived pinch. The split power supply allows more control of the pinch current than previous machine iterations, with a designed range from 50 to 150 kA. Radial force balance leads to the Bennett relation which indicates that as the pinch compresses due to increasing currents, the plasma pressure and/or linear density must change. Through ion spectroscopy and digital holographic interferometry coupled with magnetic measurements of the pinch current, the components of the Bennett relation can be fully measured. A scaling relation is then assumed to follow a polytrope as the pinch pressure, initially approximately 250 kPa, increases from an initially formed state to much higher values, approaching 100 MPa. A preliminary analysis of pinch scaling is shown corroborating with other diagnostics on the machine along with extrapolations to required currents for an HEDLP machine. This work is supported by grants from the U.S. Department of Energy and the U.S. National Nuclear Security Administration.

Cytotoxic-T-lymphocyte antigen 4 receptor signaling for lymphocyte adhesion is mediated by C3G and Rap1.

PubMed

Kloog, Yoel; Mor, Adam

2014-03-01

T-lymphocyte adhesion plays a critical role in both inflammatory and autoimmune responses. The small GTPase Rap1 is the key coordinator mediating T-cell adhesion to endothelial cells, antigen-presenting cells, and virus-infected cells. We describe a signaling pathway, downstream of the cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor, leading to Rap1-mediated adhesion. We identified a role for the Rap1 guanine nucleotide exchange factor C3G in the regulation of T-cell adhesion and showed that this factor is required for both T-cell receptor (TCR)-mediated and CTLA-4-mediated T-cell adhesion. Our data indicated that C3G translocates to the plasma membrane downstream of TCR signaling, where it regulates activation of Rap1. We also showed that CTLA-4 receptor signaling mediates tyrosine phosphorylation in the C3G protein, and that this is required for augmented activation of Rap1 and increased adhesion mediated by leukocyte function-associated antigen type 1 (LFA-1). Zap70 is required for C3G translocation to the plasma membrane, whereas the Src family member Hck facilitates C3G phosphorylation. These findings point to C3G and Hck as promising potential therapeutic targets for the treatment of T-cell-dependent autoimmune disorders.

Overexpressed BAG3 is a potential therapeutic target in chronic lymphocytic leukemia.

PubMed

Zhu, Huayuan; Wu, Wei; Fu, Yuan; Shen, Wenyi; Miao, Kourong; Hong, Min; Xu, Wei; Young, Ken H; Liu, Peng; Li, Jianyong

2014-03-01

Bcl-2-associated athanogene 3 (BAG3), a member of BAG family, is shown to sustain cell survival and underlie resistance to chemotherapy in human neoplastic cells. We aimed to determine the exact role and underlying mechanisms of BAG3 in human chronic lymphocytic leukemia (CLL). One hundred human CLL samples and 20 normal B-cell samples from healthy controls were collected. We measured the BAG3 expression in these cells and explored its relationship with known prognostic factors for CLL. The roles of BAG3 in cell apoptosis and migration were evaluated by small interfering RNA-mediated knockdown of BAG3 in primary CLL cells. We showed that BAG3 expression level was increased in CLL cells compared with normal B cells. Moreover, BAG3 expression was particularly upregulated in CD38 positive, unmutated immunoglobulin heavy-chain patients and those with lymphadenopathy and/or splenomegaly. Importantly, patients with increased BAG3 expression level have poor overall survival in subgroups with positive ZAP-70 or those without any "p53 abnormality". In addition, knocking down of BAG3 expression resulted in increased apoptotic ratio and decreased migration in primary CLL cells. Our data indicate that BAG3 is a marker of poor prognostic in specific subgroups of CLL patients and may be a potential therapeutic target for this disease.

Dark, Thin Fracture-Filling Material

NASA Image and Video Library

2015-11-11

These images and overlay bar charts from the Chemistry and Camera (ChemCam) instrument on NASA's Curiosity Mars rover indicate where some high-potassium material is localized within mineral veins at "Garden City." The two images are from ChemCam's Remote Micro-Imager. Each covers an area just over an inch wide (scale bars are in millimeters) in veins at the Garden City site on lower Mount Sharp. The overlay charts show comparisons of potassium (blue) and iron (red) in the mineral veins' compositions determined by reading the spectra of light induced by zapping points in each area with ChemCam's laser. Mineral veins such as these form where fluids move through fractured rocks, depositing minerals in the fractures and affecting chemistry of the surrounding rock. The thin layer of dark fracture-filling material in the image on the right contains much more potassium than the other local material on the left, indicating either different fluid compositions or local variations in the rock. The image on the left was taken on April 4, 2015, during the 946th Martian day, or sol, of Curiosity's work on Mars. The image on the right was taken on Sol 936, on March 25, 2015. A broader view of the prominent mineral veins at Garden City is at PIA19161. http://photojournal.jpl.nasa.gov/catalog/PIA19923

Stimulus specific effect of ibuprofen on chemiluminescence of sheep neutrophils

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tahamont, M.V.; Margiotta, M.; Gee, M.H.

1986-03-05

The authors have shown that pretreatment with ibuprofen inhibits free radical release from complement stimulated neutrophils. To further examine the effect of ibuprofen on neutrophil free radical release, they stimulated neutrophils with the synthetic peptide, FMLP, phorbol myristate acetate (PMA), or zymosan-activated plasma (ZAP). Pure (>95%), viable (>95%) sheep neutrophils (2 x 10/sup 6/) were placed in HEPES buffer, luminol, drug or vehicle and stimulated in the luminometer with one of the stimuli. The chemiluminescence (CL) response was recorded and the drug treated samples were compared to vehicle treated controls. Ibuprofen had a dose dependent effect on CL in ZAPmore » stimulated neutrophils. At the highest dose (10/sup -2/M) these cells produced only 37 +/- 7% of the CL response observed in the control cells. In contrast, at the same dose, ibuprofen did not significantly attenuate CL seen in FMLP stimulated cells, with these cells producing 79 +/- 7% of the control cells; nor did ibuprofen effect PMA stimulated CL, as these cells produced a CL response that was 85 +/- 8% of the control cells. Ibuprofen appears to have a stimulus specific effect on free radical release in activated neutrophils. It is also apparent that ibuprofen inhibits complement stimulated free radical release by some mechanism independent of its cyclooxygenase inhibitory effect.« less

The impact of ex vivo clinical grade activation protocols on human T cell phenotype and function for the generation of genetically modified cells for adoptive cell transfer therapy

PubMed Central

Tumeh, Paul C.; Koya, Richard C.; Chodon, Thinle; Graham, Nicholas A.; Graeber, Thomas G.; Comin-Anduix, Begoña; Ribas, Antoni

2011-01-01

Optimized conditions for the ex vivo activation, genetic manipulation, and expansion of human lymphocytes for adoptive cell therapy (ACT) may lead to protocols that maximize their in vivo function. We analyzed the effects of four clinical grade activation and expansion protocols over three weeks on cell proliferative rate, immunophenotype, cell metabolism, and transduction efficiency of human peripheral blood mononuclear cells (PBMCs). Peak lentiviral transduction efficiency was early (days 2 to 4), at a time when cells demonstrated a larger size, maximal uptake of metabolic substrates, and the highest level of proximal TCR signaling engagement. Anti-CD2/3/28 activation beads induced greater proliferation rate and skewed PBMCs early on to a CD4 phenotype when compared to the cells cultured in OKT3. Multicolor surface phenotyping demonstrated that changes in T cell surface markers that define T cell functional phenotypes were dependent on the time spent in culture as opposed to the particular activation protocol. In conclusion, ex vivo activation of human PBMCs for ACT demonstrate defined immunophenotypic and functional signatures over time, with cells early on showing larger sizes, higher transduction efficiency, maximal metabolic activity and ZAP-70 activation. PMID:20842061

Prolonged lymphocytosis during ibrutinib therapy is associated with distinct molecular characteristics and does not indicate a suboptimal response to therapy

PubMed Central

Smucker, Kelly; Smith, Lisa L.; Lozanski, Arletta; Zhong, Yiming; Ruppert, Amy S.; Lucas, David; Williams, Katie; Zhao, Weiqiang; Rassenti, Laura; Ghia, Emanuela; Kipps, Thomas J.; Mantel, Rose; Jones, Jeffrey; Flynn, Joseph; Maddocks, Kami; O’Brien, Susan; Furman, Richard R.; James, Danelle F.; Clow, Fong; Lozanski, Gerard; Johnson, Amy J.; Byrd, John C.

2014-01-01

The Bruton’s tyrosine kinase (BTK) inhibitor ibrutinib has outstanding activity in patients with chronic lymphocytic leukemia. Most patients experience lymphocytosis, representing lymphocyte egress from nodal compartments. This resolves within 8 months in the majority of patients, but a subgroup has lymphocytosis lasting >12 months. Here we report a detailed characterization of patients with persistent lymphocytosis during ibrutinib therapy. Signaling evaluation showed that while BTK is inhibited, downstream mediators of B-cell receptor (BCR) signaling are activated in persistent lymphocytes. These cells cannot be stimulated through the BCR and do not show evidence of target gene activation. Flow cytometry for κ and λ expression, IGHV sequencing, Zap-70 methylation, and targeted gene sequencing in these patients are identical at baseline and later time points, suggesting that persistent lymphocytes do not represent clonal evolution. In vitro treatment with targeted kinase inhibitors shows that they are not addicted to a single survival pathway. Finally, progression-free survival is not inferior for patients with prolonged lymphocytosis vs those with traditional responses. Thus, prolonged lymphocytosis is common following ibrutinib treatment, likely represents the persistence of a quiescent clone, and does not predict a subgroup of patients likely to relapse early. PMID:24415539

Cell-Intrinsic Determinants of Ibrutinib-Induced Apoptosis in Chronic Lymphocytic Leukemia.

PubMed

Amin, Nisar A; Balasubramanian, Sriram; Saiya-Cork, Kamlai; Shedden, Kerby; Hu, Nan; Malek, Sami N

2017-02-15

Purpose: Ibrutinib, a Bruton tyrosine kinase (BTK) inhibitor, is approved for the treatment of relapsed chronic lymphocytic leukemia (CLL) and CLL with del17p. Mechanistically, ibrutinib interferes with B-cell receptor (BCR) signaling as well as multiple CLL cell-to-microenvironment interactions. Given the importance of ibrutinib in the management of CLL, a deeper understanding of factors governing sensitivity and resistance is warranted. Experimental Design: We studied 48 longitudinally sampled paired CLL samples, 42 of which were procured before and after standard CLL chemotherapies, and characterized them for well-studied CLL molecular traits as well as by whole-exome sequencing and SNP 6.0 array profiling. We exposed these samples to 0.25 to 5 μmol/L of ibrutinib ex vivo and measured apoptosis fractions as well as BCR signaling by immunoblotting. We disrupted TP53 in HG3, PGA1, and PG-EBV cell lines and measured BCR signaling and ibrutinib responses. Results: CLL samples demonstrated a surprisingly wide range of ex vivo sensitivities to ibrutinib, with IC 50 values ranging from 0.4 to 9.7 μmol/L. Unmutated IGVH status, elevated ZAP70 expression, and trisomy 12 were associated with heightened sensitivity to ibrutinib treatment. Five CLL samples were substantially more resistant to ibrutinib following relapse from chemotherapy; of these, three had acquired a del17p/ TP53 -mutated status. A validation sample of 15 CLL carrying TP53 mutations, of which 13 carried both del17p and a TP53 mutation, confirmed substantially less sensitivity to ibrutinib-induced apoptosis. Conclusions: This study identifies that CLL harboring del17p/ TP53 -mutated cells are substantially less sensitive to ibrutinib-induced apoptosis than del17p/ TP53 wild-type cells. Clin Cancer Res; 23(4); 1049-59. ©2016 AACR . ©2016 American Association for Cancer Research.

Augmented reality on poster presentations, in the field and in the classroom

NASA Astrophysics Data System (ADS)

Hawemann, Friedrich; Kolawole, Folarin

2017-04-01

Augmented reality (AR) is the direct addition of virtual information through an interface to a real-world environment. In practice, through a mobile device such as a tablet or smartphone, information can be projected onto a target- for example, an image on a poster. Mobile devices are widely distributed today such that augmented reality is easily accessible to almost everyone. Numerous studies have shown that multi-dimensional visualization is essential for efficient perception of the spatial, temporal and geometrical configuration of geological structures and processes. Print media, such as posters and handouts lack the ability to display content in the third and fourth dimensions, which might be in space-domain as seen in three-dimensional (3-D) objects, or time-domain (four-dimensional, 4-D) expressible in the form of videos. Here, we show that augmented reality content can be complimentary to geoscience poster presentations, hands-on material and in the field. In the latter example, location based data is loaded and for example, a virtual geological profile can be draped over a real-world landscape. In object based AR, the application is trained to recognize an image or object through the camera of the user's mobile device, such that specific content is automatically downloaded and displayed on the screen of the device, and positioned relative to the trained image or object. We used ZapWorks, a commercially-available software application to create and present examples of content that is poster-based, in which important supplementary information is presented as interactive virtual images, videos and 3-D models. We suggest that the flexibility and real-time interactivity offered by AR makes it an invaluable tool for effective geoscience poster presentation, class-room and field geoscience learning.

Novel host restriction factors implicated in HIV-1 replication.

PubMed

Ghimire, Dibya; Rai, Madhu; Gaur, Ritu

2018-04-01

Human immunodeficiency virus-1 (HIV-1) is known to interact with multiple host cellular proteins during its replication in the target cell. While many of these host cellular proteins facilitate viral replication, a number of them are reported to inhibit HIV-1 replication at various stages of its life cycle. These host cellular proteins, which are known as restriction factors, constitute an integral part of the host's first line of defence against the viral pathogen. Since the discovery of apolipoprotein B mRNA-editing enzyme 3G (APOBEC3G) as an HIV-1 restriction factor, several human proteins have been identified that exhibit anti-HIV-1 restriction. While each restriction factor employs a distinct mechanism of inhibition, the HIV-1 virus has equally evolved complex counter strategies to neutralize their inhibitory effect. APOBEC3G, tetherin, sterile alpha motif and histidine-aspartate domain 1 (SAMHD1), and trim-5α are some of the best known HIV-1 restriction factors that have been studied in great detail. Recently, six novel restriction factors were discovered that exhibit significant antiviral activity: endoplasmic reticulum α1,2-mannosidase I (ERManI), translocator protein (TSPO), guanylate-binding protein 5 (GBP5), serine incorporator (SERINC3/5) and zinc-finger antiviral protein (ZAP). The focus of this review is to discuss the antiviral mechanism of action of these six restriction factors and provide insights into the probable counter-evasion strategies employed by the HIV-1 virus. The recent discovery of new restriction factors substantiates the complex host-pathogen interactions occurring during HIV-1 pathogenesis and makes it imperative that further investigations are conducted to elucidate the molecular basis of HIV-1 replication.

The pan-ErbB tyrosine kinase inhibitor canertinib induces caspase-mediated cell death in human T-cell leukemia (Jurkat) cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Trinks, Cecilia, E-mail: Cecilia.trinks@liu.se; Severinsson, Emelie A., E-mail: Emelie.severinsson@liu.se; Holmlund, Birgitta, E-mail: Birgitta.holmlund@lio.se

2011-07-08

Highlights: {yields} Canertinib induces caspase-mediated apoptosis in T-cell leukemia cells in vitro. {yields} Canertinib mediates activation of the intrinsic apoptotic pathway. {yields} Canertinib induces apoptosis in an ErbB receptor independent manner. {yields} Lymphocyte specific proteins as well as survival kinases are inhibited. {yields} Canertinib may act as a multi-kinase inhibiting drug in human T-cell malignancies. -- Abstract: Canertinib is a novel ErbB-receptor inhibitor currently in clinical development for the treatment of solid tumors overexpressing ErbB-receptors. We have recently demonstrated that canertinib displays anti-proliferative and pro-apoptotic effects in human myeloid leukemia cells devoid of ErbB-receptors. The mechanism mediating these effects aremore » however unknown. In this study, we show that canertinib is able to act as a multi-kinase inhibitor by inhibition of several intracellular kinases involved in T-cell signaling such as Akt, Erk1/2 and Zap-70, and reduced Lck protein expression in the human T-cell leukemia cell line Jurkat. Treatment with canertinib at a concentration of 2 {mu}M caused accumulation of Jurkat cells in the G{sub 1} cell cycle phase and increased doses induced apoptosis in a time-dependent manner. Apoptotic signs of treated cells were detected by Annexin V staining and cleavage of PARP, caspase-3, -8, -9, -10 and Bid. A subset of the pro-apoptotic signals mediated by canertinib could be significantly reduced by specific caspase inhibitors. Taken together, these results demonstrate the dual ability of canertinib to downregulate important signaling pathways and to activate caspase-mediated intrinsic apoptosis pathway in human T-cell leukemia cells.« less

Multi-gene phylogenetic analysis reveals that shochu-fermenting Saccharomyces cerevisiae strains form a distinct sub-clade of the Japanese sake cluster.

PubMed

Futagami, Taiki; Kadooka, Chihiro; Ando, Yoshinori; Okutsu, Kayu; Yoshizaki, Yumiko; Setoguchi, Shinji; Takamine, Kazunori; Kawai, Mikihiko; Tamaki, Hisanori

2017-10-01

Shochu is a traditional Japanese distilled spirit. The formation of the distinguishing flavour of shochu produced in individual distilleries is attributed to putative indigenous yeast strains. In this study, we performed the first (to our knowledge) phylogenetic classification of shochu strains based on nucleotide gene sequences. We performed phylogenetic classification of 21 putative indigenous shochu yeast strains isolated from 11 distilleries. All of these strains were shown or confirmed to be Saccharomyces cerevisiae, sharing species identification with 34 known S. cerevisiae strains (including commonly used shochu, sake, ale, whisky, bakery, bioethanol and laboratory yeast strains and clinical isolate) that were tested in parallel. Our analysis used five genes that reflect genome-level phylogeny for the strain-level classification. In a first step, we demonstrated that partial regions of the ZAP1, THI7, PXL1, YRR1 and GLG1 genes were sufficient to reproduce previous sub-species classifications. In a second step, these five analysed regions from each of 25 strains (four commonly used shochu strains and the 21 putative indigenous shochu strains) were concatenated and used to generate a phylogenetic tree. Further analysis revealed that the putative indigenous shochu yeast strains form a monophyletic group that includes both the shochu yeasts and a subset of the sake group strains; this cluster is a sister group to other sake yeast strains, together comprising a sake-shochu group. Differences among shochu strains were small, suggesting that it may be possible to correlate subtle phenotypic differences among shochu flavours with specific differences in genome sequences. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

The Structural Basis for Activation and Inhibition of ZAP-70 Kinase Domain

PubMed Central

Huber, Roland G.; Fan, Hao; Bond, Peter J.

2015-01-01

ZAP–70 (Zeta-chain-associated protein kinase 70) is a tyrosine kinase that interacts directly with the activated T-cell receptor to transduce downstream signals, and is hence a major player in the regulation of the adaptive immune response. Dysfunction of ZAP–70 causes selective T cell deficiency that in turn results in persistent infections. ZAP–70 is activated by a variety of signals including phosphorylation of the kinase domain (KD), and binding of its regulatory tandem Src homology 2 (SH2) domains to the T cell receptor. The present study investigates molecular mechanisms of activation and inhibition of ZAP–70 via atomically detailed molecular dynamics simulation approaches. We report microsecond timescale simulations of five distinct states of the ZAP–70 KD, comprising apo, inhibited and three phosphorylated variants. Extensive analysis of local flexibility and correlated motions reveal crucial transitions between the states, thus elucidating crucial steps in the activation mechanism of the ZAP–70 KD. Furthermore, we rationalize previously observed staurosporine-bound crystal structures, suggesting that whilst the KD superficially resembles an “active-like” conformation, the inhibitor modulates the underlying protein dynamics and restricts it in a compact, rigid state inaccessible to ligands or cofactors. Finally, our analysis reveals a novel, potentially druggable pocket in close proximity to the activation loop of the kinase, and we subsequently use its structure in fragment-based virtual screening to develop a pharmacophore model. The pocket is distinct from classical type I or type II kinase pockets, and its discovery offers promise in future design of specific kinase inhibitors, whilst mutations in residues associated with this pocket are implicated in immunodeficiency in humans. PMID:26473606

Hydrologic characteristics and possible effects of surface mining in the northwestern part of West Branch Antelope Creek basin, Mercer County, North Dakota

USGS Publications Warehouse

Crawley, Mark E.; Emerson, Douglas G.

1981-01-01

Lignite beds and abundant discontinuous sandstone beds of the Paleocene Sentinel Butte Member of the Fort Union Formation and sand and gravel beds in the Quarternary glaciofluvial deposits (Antelope Creek aquifer) are the most important aquifers for domestic and livestock water supplies in the West Branch Antelope Creek basin. In the Beulah-Zap lignite, ground water moves from highland area in the west toward the Antelope Creek aquifer. Water levels in the basal Sentinel Butte sandstone appear to be controlled by the level of Lake Sakakawea. In the glaciofluvial deposits of the Antelope Creek aquifer water moves from a ground-water divide northwestward to Lake Sakakawea and southeastward toward the Knife River. Large water-level declines in wells completed in the lignite and shallower aquifers could be expected with mining. The effects probably would be limited to within 1 to 2 miles of an active mine. Surface-runoff duration could be altered by increased infiltration and retention in the reclaimed are and possible temporal extension of base flow could occur. Shallow ground water beneath mine sites would be expected to increase in dissolved solids and locally to contain large sodium and sulfate concentrations. In some locations movement of poor quality water toward the Antelope Creek aquifer would be expected. (USGS)

Producing High-Performance, Stable, Sheared-Flow Z-Pinches in the FuZE project

NASA Astrophysics Data System (ADS)

Golingo, R. P.; Shumlak, U.,; Nelson, B. A.; Claveau, E. L.; Forbes, E. G.; Stepanov, A. D.; Weber, T. R.; Zhang, Y.; McLean, H. S.; Tummel, K. K.; Higginson, D. P.; Schmidt, A. E.; University of Washington (UW) Collaboration; Lawrence Livermore National Laboratory (LLNL) Collaboration

2017-10-01

The Fusion Z-Pinch Experiment (FuZE) has made significant strides towards generating high-performance, stable Z-pinch plasmas with goals of ne = 1018 cm-3 and T =1 keV. The Z-pinch plasmas are stabilized with a sheared axial flow that is driven by a coaxial accelerator. The new FuZE device has been constructed and reproduces the major scientific achievements the ZaP project at the University of Washington; ne = 1016 cm-3,T = 100 eV, r<1 cm, and tstable >20 μs. These parameters are measured with an array of magnetic field probes, spectroscopy, and fast framing cameras. The plasma parameters are achieved using a small fraction of the maximum energy storage and gas injection capability of the FuZE device. Higher density, ne = 5×1017 cm-3, and temperature, T = 500 eV, Z-pinch plasmas are formed by increasing the pinch current. At the higher voltages and currents, the ionization rates in the accelerator increase. By modifying the neutral gas profile in the accelerator, the plasma flow from the accelerator is maintained, driving the flow shear. Formation and sustainment of the sheared-flow Z-pinch plasma will be discussed. Experimental data demonstrating high performance plasmas in a stable Z-pinches will be shown. This work is supported by an award from US ARPA-E.

Rab11-FIP3 Regulation of Lck Endosomal Traffic Controls TCR Signal Transduction.

PubMed

Bouchet, Jérôme; Del Río-Iñiguez, Iratxe; Vázquez-Chávez, Elena; Lasserre, Rémi; Agüera-González, Sonia; Cuche, Céline; McCaffrey, Mary W; Di Bartolo, Vincenzo; Alcover, Andrés

2017-04-01

The role of endosomes in receptor signal transduction is a long-standing question, which remains largely unanswered. The T cell Ag receptor and various components of its proximal signaling machinery are associated with distinct endosomal compartments, but how endosomal traffic affects T cell signaling remains ill-defined. In this article, we demonstrate in human T cells that the subcellular localization and function of the protein tyrosine kinase Lck depends on the Rab11 effector FIP3 (Rab11 family interacting protein-3). FIP3 overexpression or silencing and its ability to interact with Rab11 modify Lck subcellular localization and its delivery to the immunological synapse. Importantly, FIP3-dependent Lck localization controls early TCR signaling events, such as tyrosine phosphorylation of TCRζ, ZAP70, and LAT and intracellular calcium concentration, as well as IL-2 gene expression. Interestingly, FIP3 controls both steady-state and poststimulation phosphotyrosine and calcium levels. Finally, our findings indicate that FIP3 modulates TCR-CD3 cell surface expression via the regulation of steady-state Lck-mediated TCRζ phosphorylation, which in turn controls TCRζ protein levels. This may influence long-term T cell activation in response to TCR-CD3 stimulation. Therefore, our data underscore the importance of finely regulated endosomal traffic in TCR signal transduction and T cell activation leading to IL-2 production. Copyright © 2017 by The American Association of Immunologists, Inc.

Construction of a complementary DNA library for Parelaphostrongylus tenuis and identification of a potentially sero-diagnostic recombinant antigen.

PubMed

Ogunremi, Oladele; Benjamin, Jane; MacDonald, Lily; Schimpf, Robert

2008-12-01

Newly developed serological tests for diagnosing parelaphostrongylosis in cervids, using the excretory-secretory products (ES) of the infective larvae of Parelaphostrongylus tenuis in enzyme-linked immunosorbent assays (ELISAs), have demonstrable superiority over the traditional method of larval recovery and microscopic identification. To generate a source of ELISA antigen by genetic engineering, we created a complementary DNA (cDNA) expression library by the reverse transcription of mRNA of P. tenuis adult worms, and ligation with the vector lambda-ZAP II. The library was screened using antisera produced in mice by immunization with a somatic antigen preparation of adult worms. Seventeen clones were isolated, sequenced, and checked for similarity to other DNA sequences in GenBank. A previously identified parasite gene encoding an aspartyl protease inhibitor (API) was isolated from the cDNA library, subcloned and expressed using the pET expression vector to produce a glutathione S transferase (GST)-His-S.Tag-P. tenuis API fusion protein (molecular weight = 63 kDa). An enzyme-linked immunosorbent assay utilizing the API fusion protein as the coating antigen was used to serologically diagnose all white-tailed deer (WTD, 10 out of 10) that had been inoculated with 6 - 150 L3 P. tenuis, indicating that the antigen may be a useful serodiagnostic antigen for P. tenuis infection in this cervid species.

Cytotoxic-T-Lymphocyte Antigen 4 Receptor Signaling for Lymphocyte Adhesion Is Mediated by C3G and Rap1

PubMed Central

Kloog, Yoel

2014-01-01

T-lymphocyte adhesion plays a critical role in both inflammatory and autoimmune responses. The small GTPase Rap1 is the key coordinator mediating T-cell adhesion to endothelial cells, antigen-presenting cells, and virus-infected cells. We describe a signaling pathway, downstream of the cytotoxic T-lymphocyte antigen 4 (CTLA-4) receptor, leading to Rap1-mediated adhesion. We identified a role for the Rap1 guanine nucleotide exchange factor C3G in the regulation of T-cell adhesion and showed that this factor is required for both T-cell receptor (TCR)-mediated and CTLA-4-mediated T-cell adhesion. Our data indicated that C3G translocates to the plasma membrane downstream of TCR signaling, where it regulates activation of Rap1. We also showed that CTLA-4 receptor signaling mediates tyrosine phosphorylation in the C3G protein, and that this is required for augmented activation of Rap1 and increased adhesion mediated by leukocyte function-associated antigen type 1 (LFA-1). Zap70 is required for C3G translocation to the plasma membrane, whereas the Src family member Hck facilitates C3G phosphorylation. These findings point to C3G and Hck as promising potential therapeutic targets for the treatment of T-cell-dependent autoimmune disorders. PMID:24396067

Genome-wide analysis of signal transducers and regulators of mitochondrial dysfunction in Saccharomyces cerevisiae.

PubMed

Singh, Keshav K; Rasmussen, Anne Karin; Rasmussen, Lene Juel

2004-04-01

Mitochondrial dysfunction is a hallmark of cancer cells. However, genetic response to mitochondrial dysfunction during carcinogenesis is unknown. To elucidate genetic response to mitochondrial dysfunction we used Saccharomyces cerevisiae as a model system. We analyzed genome-wide expression of nuclear genes involved in signal transduction and transcriptional regulation in a wild-type yeast and a yeast strain lacking the mitochondrial genome (rho(0)). Our analysis revealed that the gene encoding cAMP-dependent protein kinase subunit 3 (PKA3) was upregulated. However, the gene encoding cAMP-dependent protein kinase subunit 2 (PKA2) and the VTC1, PTK2, TFS1, CMK1, and CMK2 genes, involved in signal transduction, were downregulated. Among the known transcriptional factors, OPI1, MIG2, INO2, and ROX1 belonged to the upregulated genes, whereas MSN4, MBR1, ZMS1, ZAP1, TFC3, GAT1, ADR1, CAT8, and YAP4 including RFA1 were downregulated. RFA1 regulates DNA repair genes at the transcriptional level. RFA is also involved directly in DNA recombination, DNA replication, and DNA base excision repair. Downregulation of RFA1 in rho(0) cells is consistent with our finding that mitochondrial dysfunction leads to instability of the nuclear genome. Together, our data suggest that gene(s) involved in mitochondria-to-nucleus communication play a role in mutagenesis and may be implicated in carcinogenesis.

Pharmacogenetics of steroid-responsive acute graft-versus-host disease.

PubMed

Arora, Mukta; Weisdorf, Daniel J; Shanley, Ryan M; Thyagarajan, Bharat

2017-05-01

Glucocorticoids are central to effective therapy of acute graft-versus-host disease (GVHD). However, only about half of the patients respond to steroids in initial therapy. Based on postulated mechanisms for anti-inflammatory effectiveness, we explored genetic variations in glucocorticoid receptor, co-chaperone proteins, membrane transporters, inflammatory mediators, and variants in the T-cell receptor complex in hematopoietic cell transplant recipients with acute GVHD requiring treatment with steroids and their donors toward response at day 28 after initiation of therapy. A total of 300 recipient and donor samples were analyzed. Twenty-three SNPs in 17 genes affecting glucocorticoid pathways were included in the analysis. In multiple regression analysis, donor SNP rs3192177 in the ZAP70 gene (O.R. 2.8, 95% CI: 1.3-6.0, P=.008) and donor SNP rs34471628 in the DUSPI gene (O.R. 0.3, 95% CI: 0.1-1.0, P=.048) were significantly associated with complete or partial response. However, after adjustment for multiple testing, these SNPs did not remain statistically significant. Our results, on this small, exploratory, hypothesis generating analysis suggest that common genetic variation in glucocorticoid pathways may help identify subjects with differential response to glucocorticoids. This needs further assessment in larger datasets and if validated could help identify subjects for alternative treatments and design targeted treatments to overcome steroid resistance. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Digital holographic interferometry employing Fresnel transform reconstruction for the study of flow shear stabilized Z-pinch plasmas.

PubMed

Ross, M P; Shumlak, U

2016-10-01

The ZaP-HD flow Z-pinch project provides a platform to explore how shear flow stabilized Z-pinches could scale to high-energy-density plasma (plasma with pressures exceeding 1 Mbar) and fusion reactor conditions. The Z-pinch is a linear plasma confinement geometry in which the plasma carries axial electric current and is confined by its self-induced magnetic field. ZaP-HD generates shear stabilized, axisymmetric Z-pinches with stable lifetimes approaching 60 μs. The goal of the project is to increase the plasma density and temperature compared to the previous ZaP project by compressing the plasma to smaller radii (≈1 mm). Radial and axial plasma electron density structure is measured using digital holographic interferometry (DHI), which provides the necessary fine spatial resolution. ZaP-HD's DHI system uses a 2 ns Nd:YAG laser pulse with a second harmonic generator (λ = 532 nm) to produce holograms recorded by a Nikon D3200 digital camera. The holograms are numerically reconstructed with the Fresnel transform reconstruction method to obtain the phase shift caused by the interaction of the laser beam with the plasma. This provides a two-dimensional map of line-integrated electron density, which can be Abel inverted to determine the local number density. The DHI resolves line-integrated densities down to 3 × 10 20 m -2 with spatial resolution near 10 μm. This paper presents the first application of Fresnel transform reconstruction as an analysis technique for a plasma diagnostic, and it analyzes the method's accuracy through study of synthetic data. It then presents an Abel inversion procedure that utilizes data on both sides of a Z-pinch local number density profile to maximize profile symmetry. Error estimation and Abel inversion are applied to the measured data.

Elimination of textile dyes using activated carbons prepared from vegetable residues and their characterization.

PubMed

Peláez-Cid, Alejandra-Alicia; Herrera-González, Ana-María; Salazar-Villanueva, Martín; Bautista-Hernández, Alejandro

2016-10-01

In this study, three mesoporous activated carbons prepared from vegetable residues were used to remove acid, basic, and direct dyes from aqueous solutions, and reactive and vat dyes from textile wastewater. Granular carbons obtained by chemical activation at 673 K with phosphoric acid from prickly pear peels (CarTunaQ), broccoli stems (CarBrocQ), and white sapote seeds (CarZapQ) were highly efficient for the removal of dyes. Adsorption equilibrium studies were carried out in batch systems and treated with Langmuir and Freundlich isotherms. The maximum adsorption capacities calculated from the Langmuir isotherms ranged between 131.6 and 312.5 mg/g for acid dyes, and between 277.8 and 500.0 mg/g for basic dyes at 303 K. Our objective in this paper was to show that vegetable wastes can serve as precursors for activated carbons that can be used for the adsorption of dyes. Specifically CarBrocQ was the best carbon produced for the removal of textile dyes. The color removal of dyes present in textile wastewaters was compared with that of a commercial powdered carbon, and it was found that the carbons produced using waste material reached similar efficiency levels. Carbon samples were characterized by bulk density, point of zero charge, thermogravimetric analysis, elemental analysis, Fourier transform infrared spectroscopy, scanning electron microscopy, methylene blue adsorption isotherms at 303 K, and nitrogen adsorption isotherms at 77 K (SBET). The results show that the activated carbons possess a large specific surface area (1025-1177 m(2)/g) and high total pore volume (1.06-2.16 cm(3)/g) with average pore size diameters between 4.1 and 8.4 nm. Desorption and regeneration tests were made to test the viability of reusing the activated carbons. Copyright © 2016 Elsevier Ltd. All rights reserved.

Combination chemoimmunotherapy with pentostatin, cyclophosphamide, and rituximab shows significant clinical activity with low accompanying toxicity in previously untreated B chronic lymphocytic leukemia

PubMed Central

Kay, Neil E.; Geyer, Susan M.; Call, Timothy G.; Shanafelt, Tait D.; Zent, Clive S.; Jelinek, Diane F.; Tschumper, Renee; Bone, Nancy D.; Dewald, Gordon W.; Lin, Thomas S.; Heerema, Nyla A.; Smith, Lisa; Grever, Michael R.; Byrd, John C.

2007-01-01

Building on the prior work of use of pentostatin in chronic lymphocytic leukemia (CLL), we initiated a trial of combined pentostatin (2 mg/m2), cyclophosphamide (600 mg/m2), and rituximab (375 mg/m2) for 65 symptomatic, previously untreated patients. Of 64 evaluable patients, 34 (53%) were high Rai risk, 71% were nonmutated for the immunoglobulin heavy-chain variable region gene, 34% were CD38+, and 34% were ZAP-70+. Thirty patients (52%) had one anomaly detected by fluorescence in situ (FISH) hybridization, and 21 (36%) had complex FISH defects. Thirty-eight patients (58%) had grade 3+ hematologic toxicity but minimal transfusion needs and no major infections. Responses occurred in 58 patients (91%), with 26 (41%) complete responses (CRs), 14 (22%) nodular partial responses (nodular PRs), and 18 (28%) partial responses (PRs). Many patients with a CR also lacked evidence of minimal residual disease by 2-color flow cytometry. Examination of prognostic factors demonstrated poor response in the 3 patients with del(17p). In contrast, we found this regimen was equally effective in young versus older (> 70 years) patients and in del(11q22.3) versus other favorable prognostic factors. Thus, this novel regimen of pentostatin, cyclophosphamide, and rituximab for previously untreated patients with CLL demonstrated significant clinical activity despite poor risk-based prognoses, achievement of minimal residual disease in some, and modest toxicity. PMID:17008537

Phosphatidate Phosphatase Plays Role in Zinc-mediated Regulation of Phospholipid Synthesis in Yeast*

PubMed Central

Soto-Cardalda, Aníbal; Fakas, Stylianos; Pascual, Florencia; Choi, Hyeon-Son; Carman, George M.

2012-01-01

In the yeast Saccharomyces cerevisiae, the synthesis of phospholipids is coordinately regulated by mechanisms that control the homeostasis of the essential mineral zinc (Carman, G.M., and Han, G. S. (2007) Regulation of phospholipid synthesis in Saccharomyces cerevisiae by zinc depletion. Biochim. Biophys. Acta 1771, 322–330; Eide, D. J. (2009) Homeostatic and adaptive responses to zinc deficiency in Saccharomyces cerevisiae. J. Biol. Chem. 284, 18565–18569). The synthesis of phosphatidylcholine is balanced by the repression of CDP-diacylglycerol pathway enzymes and the induction of Kennedy pathway enzymes. PAH1-encoded phosphatidate phosphatase catalyzes the penultimate step in triacylglycerol synthesis, and the diacylglycerol generated in the reaction may also be used for phosphatidylcholine synthesis via the Kennedy pathway. In this work, we showed that the expression of PAH1-encoded phosphatidate phosphatase was induced by zinc deficiency through a mechanism that involved interaction of the Zap1p zinc-responsive transcription factor with putative upstream activating sequence zinc-responsive elements in the PAH1 promoter. The pah1Δ mutation resulted in the derepression of the CHO1-encoded phosphatidylserine synthase (CDP-diacylglycerol pathway enzyme) and loss of the zinc-mediated regulation of the enzyme. Loss of phosphatidate phosphatase also resulted in the derepression of the CKI1-encoded choline kinase (Kennedy pathway enzyme) but decreased the synthesis of phosphatidylcholine when cells were deficient of zinc. This result confirmed the role phosphatidate phosphatase plays in phosphatidylcholine synthesis via the Kennedy pathway. PMID:22128164

Differential control of the tyrosine kinases Lyn and Syk by the two signaling chains of the high affinity immunoglobulin E receptor.

PubMed

Jouvin, M H; Adamczewski, M; Numerof, R; Letourneur, O; Vallé, A; Kinet, J P

1994-02-25

Nonreceptor tyrosine kinases such as the newly described 70-kDa (ZAP-70/Syk) and Src-related tyrosine kinases are coupled to a variety of receptors, including the antigen receptors on B- and T-cells and the Fc receptors for IgE (Fc epsilon RI) and IgG (Fc gamma RI, Fc gamma RIII/CD16). Various subunits of these receptors contain homologous activation motifs which appear capable of autonomously triggering cell activation. Two forms of this motif are present in the Fc epsilon RI multimeric complex: one in the beta chain and one in the gamma chain. Here we show that each of the two tyrosine kinases known to be involved in Fc epsilon RI signaling is controlled by a distinct motif-containing chain. Lyn associates with the nonactivated beta chain, whereas gamma promotes the activation of Syk. We also show that neither the beta nor the gamma motif alone can account for the full signaling capacity of the entire receptor. We propose that, upon triggering of the tetrameric receptor, Lyn already bound to beta becomes activated and phosphorylates beta and gamma; the phosphorylation of gamma induces the association of Syk with gamma and also the activation of Syk, resulting in the phosphorylation and activation of phospholipase C gamma 1. Cooperative recruitment of specific kinases by the various signaling chains found in this family of antigen receptors could represent a way to achieve the full signaling capacity of the multimeric complexes.

Ci8 short, a novel LPS-induced peptide from the ascidian Ciona intestinalis, modulates responses of the human immune system.

PubMed

Bonura, Angela; Vizzini, Aiti; Vlah, Sara; Gervasi, Francesco; Longo, Alessandra; Melis, Mario R; Schildberg, Frank A; Colombo, Paolo

2018-02-01

The selective modulation of immunity is an emerging concept driven by the vast advances in our understanding of this crucial host defense system. Invertebrates have raised researchers' interest as potential sources of new bioactive molecules owing to their antibacterial, anticancer and immunomodulatory activities. A LipoPolySaccharide (LPS) challenge in the ascidian Ciona intestinalis generates the transcript, Ci8 short, with cis-regulatory elements in the 3' UTR region that are essential for shaping innate immune responses. The derived amino acidic sequence in silico analysis showed specific binding to human Major Histocompatibility Complex (MHC) Class I and Class II alleles. The role of Ci8 short peptide was investigated in a more evolved immune system using human Peripheral Blood Mononuclear Cells (PBMCs) as in vitro model. The biological activities of this molecule include the activation of 70kDa TCR ζ chain Associated Protein kinase (ZAP-70) and T Cell Receptor (TCR) Vβ oligo clonal selection on CD4 + T lymphocytes as well as increased proliferation and IFN-γ secretion. Furthermore Ci8 short affects CD4 + /CD25 high induced regulatory T cells (iTreg) subset selection which co-expressed the functional markers TGF-β1/Latency Associated Protein (LAP) and CD39/CD73. This paper describes a new molecule that modulates important responses of the human adaptive immune system. Copyright © 2017 Elsevier GmbH. All rights reserved.

Thinking Cap Plus Thinking Zap: tDCS of Frontopolar Cortex Improves Creative Analogical Reasoning and Facilitates Conscious Augmentation of State Creativity in Verb Generation.

PubMed

Green, Adam E; Spiegel, Katherine A; Giangrande, Evan J; Weinberger, Adam B; Gallagher, Natalie M; Turkeltaub, Peter E

2017-04-01

Recent neuroimaging evidence indicates neural mechanisms that support transient improvements in creative performance (augmented state creativity) in response to cognitive interventions (creativity cueing). Separately, neural interventions via tDCS show encouraging potential for modulating neuronal function during creative performance. If cognitive and neural interventions are separately effective, can they be combined? Does state creativity augmentation represent "real" creativity, or do interventions simply yield divergence by diminishing meaningfulness/appropriateness? Can augmenting state creativity bolster creative reasoning that supports innovation, particularly analogical reasoning? To address these questions, we combined tDCS with creativity cueing. Testing a regionally specific hypothesis from neuroimaging, high-definition tDCS-targeted frontopolar cortex activity recently shown to predict state creativity augmentation. In a novel analogy finding task, participants under tDCS formulated substantially more creative analogical connections in a large matrix search space (creativity indexed via latent semantic analysis). Critically, increased analogical creativity was not due to diminished accuracy in discerning valid analogies, indicating "real" creativity rather than inappropriate divergence. A simpler relational creativity paradigm (modified verb generation) revealed a tDCS-by-cue interaction; tDCS further enhanced creativity cue-related increases in semantic distance. Findings point to the potential of noninvasive neuromodulation to enhance creative relational cognition, including augmentation of the deliberate effort to formulate connections between distant concepts. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Rebamipide prevents peripheral arthritis and intestinal inflammation by reciprocally regulating Th17/Treg cell imbalance in mice with curdlan-induced spondyloarthritis.

PubMed

Min, Hong-Ki; Kim, Jae-Kyung; Lee, Seon-Yeong; Kim, Eun-Kyung; Lee, Seung Hoon; Lee, Jennifer; Kwok, Seung-Ki; Cho, Mi-La; Park, Sung-Hwan

2016-06-27

Spondyloarthritis (SpA) usually manifests as arthritis of the axial and peripheral joints but can also result in extra-articular manifestations such as inflammatory bowel disease. Proinflammatory cytokine interleukin-17 (IL-17) plays a crucial role in the pathogenesis of SpA. Rebamipide inhibits signal transducer and activator of transcription 3 that controls IL-17 production and Th17 cell differentiation. This study examined the effect of rebamipide on SpA development. SKG ZAP-70(W163C) mice were immunized with curdlan to induce SpA features. The mice were then intraperitoneally injected with rebamipide or vehicle 3 times a week for 14 weeks and their clinical scores were evaluated. Histological scores of the paw and spine and the length of the gut were measured at sacrifice. Immunohistochemical staining of IL-17 and tumor necrosis factor-α (TNF-α) was performed using tissue samples isolated from the axial joints, peripheral joints, and gut. Spleen tissue samples were isolated from both rebamipide- or vehicle-treated mice with SpA at 14 weeks after curdlan injection to determine the effect of rebamipide on Th17 and regulatory T (Treg) cell differentiation. Rebamipide decreased the clinical and histological scores of the peripheral joints. The total length of the gut was preserved in rebamipide-treated mice. IL-17 and TNF-α expression in the spine, peripheral joints, and gut was lower in rebamipide-treated mice than in control mice. Th17 cell differentiation was suppressed whereas Treg cell differentiation was upregulated in the spleen of rebamipide-treated mice. Rebamipide exerted beneficial effects in mice with SpA by preventing peripheral arthritis and intestinal inflammation and by regulating Th17/Treg cell imbalance, suggesting that it can be used as a potential therapeutic agent for treating arthritis to SpA patients.

SRC homology 2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) N-terminal tyrosine residues regulate a dynamic signaling equilibrium involving feedback of proximal T-cell receptor (TCR) signaling.

PubMed

Ji, Qinqin; Ding, Yiyuan; Salomon, Arthur R

2015-01-01

SRC homology 2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) is a cytosolic adaptor protein that plays an important role in the T-cell receptor-mediated T-cell signaling pathway. SLP-76 links proximal receptor stimulation to downstream effectors through interaction with many signaling proteins. Previous studies showed that mutation of three tyrosine residues, Tyr(112), Tyr(128), and Tyr(145), in the N terminus of SLP-76 results in severely impaired phosphorylation and activation of Itk and PLCγ1, which leads to defective calcium mobilization, Erk activation, and NFAT activation. To expand our knowledge of the role of N-terminal phosphorylation of SLP-76 from these three tyrosine sites, we characterized nearly 1000 tyrosine phosphorylation sites via mass spectrometry in SLP-76 reconstituted wild-type cells and SLP-76 mutant cells in which three tyrosine residues were replaced with phenylalanines (Y3F mutant). Mutation of the three N-terminal tyrosine residues of SLP-76 phenocopied SLP-76-deficient cells for the majority of tyrosine phosphorylation sites observed, including feedback on proximal T-cell receptor signaling proteins. Meanwhile, reversed phosphorylation changes were observed on Tyr(192) of Lck when we compared mutants to the complete removal of SLP-76. In addition, N-terminal tyrosine sites of SLP-76 also perturbed phosphorylation of Tyr(440) of Fyn, Tyr(702) of PLCγ1, Tyr(204), Tyr(397), and Tyr(69) of ZAP-70, revealing new modes of regulation on these sites. All these findings confirmed the central role of N-terminal tyrosine sites of SLP-76 in the pathway and also shed light on novel signaling events that are uniquely regulated by SLP-76 N-terminal tyrosine residues. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

SRC Homology 2 Domain-containing Leukocyte Phosphoprotein of 76 kDa (SLP-76) N-terminal Tyrosine Residues Regulate a Dynamic Signaling Equilibrium Involving Feedback of Proximal T-cell Receptor (TCR) Signaling*

PubMed Central

Ji, Qinqin; Ding, Yiyuan; Salomon, Arthur R.

2015-01-01

SRC homology 2 domain-containing leukocyte phosphoprotein of 76 kDa (SLP-76) is a cytosolic adaptor protein that plays an important role in the T-cell receptor–mediated T-cell signaling pathway. SLP-76 links proximal receptor stimulation to downstream effectors through interaction with many signaling proteins. Previous studies showed that mutation of three tyrosine residues, Tyr112, Tyr128, and Tyr145, in the N terminus of SLP-76 results in severely impaired phosphorylation and activation of Itk and PLCγ1, which leads to defective calcium mobilization, Erk activation, and NFAT activation. To expand our knowledge of the role of N-terminal phosphorylation of SLP-76 from these three tyrosine sites, we characterized nearly 1000 tyrosine phosphorylation sites via mass spectrometry in SLP-76 reconstituted wild-type cells and SLP-76 mutant cells in which three tyrosine residues were replaced with phenylalanines (Y3F mutant). Mutation of the three N-terminal tyrosine residues of SLP-76 phenocopied SLP-76-deficient cells for the majority of tyrosine phosphorylation sites observed, including feedback on proximal T-cell receptor signaling proteins. Meanwhile, reversed phosphorylation changes were observed on Tyr192 of Lck when we compared mutants to the complete removal of SLP-76. In addition, N-terminal tyrosine sites of SLP-76 also perturbed phosphorylation of Tyr440 of Fyn, Tyr702 of PLCγ1, Tyr204, Tyr397, and Tyr69 of ZAP-70, revealing new modes of regulation on these sites. All these findings confirmed the central role of N-terminal tyrosine sites of SLP-76 in the pathway and also shed light on novel signaling events that are uniquely regulated by SLP-76 N-terminal tyrosine residues. PMID:25316710

Inhibition of maternal embryonic leucine zipper kinase with OTSSP167 displays potent anti-leukemic effects in chronic lymphocytic leukemia.

PubMed

Zhang, Ya; Zhou, Xiangxiang; Li, Ying; Xu, Yangyang; Lu, Kang; Li, Peipei; Wang, Xin

2018-06-12

TP53 pathway defects contributed to therapy resistance and adverse clinical outcome in chronic lymphocytic leukemia (CLL), which represents an unmet clinical need with few therapeutic options. Maternal embryonic leucine zipper kinase (MELK) is a novel oncogene, which plays crucial roles in mitotic progression and stem cell maintenance. OTSSP167, an orally administrated inhibitor targeting MELK, is currently in a phase I/II clinical trial in patients with advanced breast cancer and acute myeloid leukemia. Yet, no investigation has been elucidated to date regarding the oncogenic role of MELK and effects of OTSSP167 in chronic lymphocytic leukemia (CLL). Previous studies confirmed MELK inhibition abrogated cancer cell survival via p53 signaling pathway. Thus, we aimed to determine the biological function of MELK and therapeutic potential of OTSSP167 in CLL. Herein, MELK over-expression was observed in CLL cells, and correlated with higher WBC count, advanced stage, elevated LDH, increased β2-MG level, unmutated IGHV, positive ZAP-70, deletion of 17p13 and inferior prognosis of CLL patients. In accordance with functional enrichment analyses in gene expression profiling, CLL cells with depletion or inhibition of MELK exhibited impaired cell proliferation, enhanced fast-onset apoptosis, induced G2/M arrest, attenuated cell chemotaxis and promoted sensitivity to fludarabine and ibrutinib. However, gain-of-function assay showed increased cell proliferation and cell chemotaxis. In addition, OTSSP167 treatment reduced phosphorylation of AKT and ERK1/2. It decreased FoxM1 phosphorylation, expression of FoxM1, cyclin B1 and CDK1, while up-regulating p53 and p21 expression. Taken together, MELK served as a candidate of therapeutic target in CLL. OTSSP167 exhibits potent anti-tumor activities in CLL cells, highlighting a novel molecule-based strategy for leukemic interventions.

Protein pathway activation associated with sustained virologic response in patients with chronic hepatitis C treated with pegylated interferon (PEG-IFN) and ribavirin (RBV).

PubMed

Younossi, Zobair M; Limongi, Dolores; Stepanova, Maria; Pierobon, Mariaelena; Afendy, Arian; Mehta, Rohini; Baranova, Ancha; Liotta, Lance; Petricoin, Emanuel

2011-02-04

Only half of chronic hepatitis C (CH-C) patients treated with pegylated interferon and ribavirin (PEG-IFN+RBV) achieve sustained virologic response) SVR. In addition to known factors, we postulated that activation of key protein signaling networks in the peripheral blood mononuclear cells (PBMCs) may contribute to SVR due to inherent patient-specific basal immune cell signaling architecture. In this study, we included 92 patients with CH-C. PBMCs were collected while patients were not receiving treatment and used for phosphoprotein-based network profiling. Patients received a full course of PEG-IFN+RBV with overall SVR of 55%. From PBMC, protein lysates were extracted and then used for Reverse Phase Protein Microarray (RPMA) analysis, which quantitatively measured the levels of cytokines and activation levels of 25 key protein signaling molecules involved in immune cell regulation and interferon alpha signaling. Regression models for predicting SVR were generated by stepwise bidirectional selection. Both clinical-laboratory and RPMA parameters were used as predictor variables. Model accuracies were estimated using 10-fold cross-validation. Our results show that by comparing patients who achieved SVR to those who did not, phosphorylation levels of 6 proteins [AKT(T308), JAK1(Y1022/1023), p70 S6 Kinase (S371), PKC zeta/lambda(T410/403), TYK2(Y1054/1055), ZAP-70(Y319)/Syk(Y352)] and overall levels of 6 unmodified proteins [IL2, IL10, IL4, IL5, TNF-alpha, CD5L] were significantly different (P < 0.05). For SVR, the model based on a combination of clinical and proteome parameters was developed, with an AUC = 0.914, sensitivity of 92.16%, and specificity of 85.0%. This model included the following parameters: viral genotype, previous treatment status, BMI, phosphorylated states of STAT2, AKT, LCK, and TYK2 kinases as well as steady state levels of IL4, IL5, and TNF-alpha. In conclusion, SVR could be predicted by a combination of clinical, cytokine, and protein signaling activation profiles. Signaling events elucidated in the study may shed some light into molecular mechanisms of response to anti-HCV treatment.

Whi3, an S. cerevisiae RNA-binding protein, is a component of stress granules that regulates levels of its target mRNAs.

PubMed

Holmes, Kristen J; Klass, Daniel M; Guiney, Evan L; Cyert, Martha S

2013-01-01

RNA binding proteins (RBPs) are vital to the regulation of mRNA transcripts, and can alter mRNA localization, degradation, translation, and storage. Whi3 was originally identified in a screen for small cell size mutants, and has since been characterized as an RBP. The identification of Whi3-interacting mRNAs involved in mediating cellular responses to stress suggested that Whi3 might be involved in stress-responsive RNA processing. We show that Whi3 localizes to stress granules in response to glucose deprivation or heat shock. The kinetics and pattern of Whi3 localization in response to a range of temperatures were subtly but distinctly different from those of known components of RNA processing granules. Deletion of Whi3 resulted in an increase in the relative abundance of Whi3 target RNAs, either in the presence or absence of heat shock. Increased levels of the CLN3 mRNA in whi3Δ cells may explain their decreased cell size. Another mRNA target of Whi3 encodes the zinc-responsive transcription factor Zap1, suggesting a role for Whi3 in response to zinc stress. Indeed, we found that whi3Δ cells have enhanced sensitivity to zinc toxicity. Together our results suggest an expanded model for Whi3 function: in addition to its role as a regulator of the cell cycle, Whi3 may have a role in stress-dependent RNA processing and responses to a variety of stress conditions.

The plant vacuolar Na+/H+ antiport.

PubMed

Barkla, B J; Apse, M P; Manolson, M F; Blumwald, E

1994-01-01

Salt stress imposes severe limitations on plant growth, however, the extent of growth reduction depends upon the soil salinity level and the plant species. One of the mechanisms employed by salt tolerant plants is the effective vacuolar compartmentalization of sodium. The sequestration of sodium into the vacuole occurs by the operation of a Na+/H+ antiport located at the tonoplast. Evidence for a plant vacuolar Na+/H+ antiport has been demonstrated in tissues, intact vacuoles and isolated tonoplast vesicles. In sugar beet cell suspensions, the activity of the vacuolar Na+/H+ antiport increased with increasing NaCl concentrations in the growth medium. This increased activity was correlated with the increased synthesis of a 170 kDa tonoplast polypeptide. In vivo labelling of tonoplast proteins showed the enhanced synthesis of the 170 kDa polypeptide not only upon exposure of the cells to salt, but also when the cells were grown in the presence of amiloride. Exposure of the cells to amiloride also resulted in increased vacuolar Na+/H+ antiport activity. Polyclonal antibodies raised against the 170 kDa polypeptide almost completely inhibited the antiport activity, suggesting the association of this protein with the plant vacuolar Na+/H+ antiport. Antibodies against the Na+/H+ antiport-associated polypeptide were used to screen a Beta lambda ZAP expression library. A partial clone of 1.65 kb was sequenced and found to encode a polypeptide with a putative transmembrane domain and a large hydrophilic C terminus. This clone showed no homology to any previously cloned gene at either the nucleic acid or the amino acid level.

Association of a novel regulatory polymorphism (-938C>A) in the BCL2 gene promoter with disease progression and survival in chronic lymphocytic leukemia.

PubMed

Nückel, Holger; Frey, Ulrich H; Bau, Maja; Sellmann, Ludger; Stanelle, Jens; Dürig, Jan; Jöckel, Karl-Heinz; Dührsen, Ulrich; Siffert, Winfried

2007-01-01

Bcl-2 plays a key role in the regulation of apoptosis. We investigated the role of a novel regulatory single-nucleotide polymorphism (-938C>A) in the inhibitory P2 BCL2 promoter in B-cell chronic lymphocytic leukemia (B-CLL). The -938C allele displayed significantly increased BCL2 promoter activity and binding of nuclear proteins compared with the A allele. Concomitantly, Bcl-2 protein expression in B cells from CLL patients carrying the -938 AA genotype was significantly increased compared with CC genotypes. Genotype distribution between 123 CLL patients (42 AA, 55 AC, 26 CC) and 120 genotyped healthy controls (36 AA, 63 AC, 21 CC) was not significantly different, suggesting that genotypes of this polymorphism do not increase the susceptibility for B-CLL. However, median time from first diagnosis to initiation of chemotherapy and median overall survival were significantly shorter in patients with -938AA genotype (38 and 199 months, respectively) compared with AC/CC genotypes (120 and 321 months, respectively; P = .008 and P = .003, respectively). Multivariable Cox regression identified the BCL2-938AA genotype as an independent prognostic factor for the time to first treatment (hazard ratio [HR] 1.9; P = .034) together with disease stage at diagnosis (HR 2.5; P = .004) and ZAP-70 status (HR 3.0; P = .001). The BCL2-938AA genotype is associated with increased Bcl-2 expression and a novel unfavorable genetic marker in patients with B-CLL.

Development of an optical Zn 2+ probe based on a single fluorescent protein

DOE PAGES

Qin, Yan; Sammond, Deanne W.; Braselmann, Esther; ...

2016-07-28

Various fluorescent probes have been developed to reveal the biological functions of intracellular labile Zn 2+. Here we present Green Zinc Probe (GZnP), a novel genetically encoded Zn 2+ sensor design based on a single fluorescent protein (single-FP). The GZnP sensor is generated by attaching two zinc fingers (ZF) of the transcription factor Zap1 (ZF1 and ZF2) to the two ends of a circularly permuted green fluorescent protein (cpGFP). Formation of ZF folds induces interaction between the two ZFs, which induces a change in the cpGFP conformation, leading to an increase in fluorescence. A small sensor library is created tomore » include mutations in the ZFs, cpGFP and linkers between ZF and cpGFP to improve signal stability, sensor brightness and dynamic range based on rational protein engineering and computational design by Rosetta. Using a cell-based library screen, we identify sensor GZnP1 which demonstrates a stable maximum signal, decent brightness (QY = 0.42 at apo state), as well as specific and sensitive response to Zn 2+ in HeLa cells (F max/F min = 2.6, K d = 58 pM, pH 7.4). The subcellular localizing sensors mito-GZnP1 (in mitochondria matrix) and Lck-GZnP1 (on plasma membrane) display sensitivity to Zn 2+ (F max/F min = 2.2). In conclusion, this sensor design provides freedom to be used in combination with other optical indicators and optogenetic tools for simultaneous imaging and advancing our understanding of cellular Zn 2+ function.« less

Genome Resequencing Identifies Unique Adaptations of Tibetan Chickens to Hypoxia and High-Dose Ultraviolet Radiation in High-Altitude Environments

PubMed Central

Zhang, Qian; Gou, Wenyu; Wang, Xiaotong; Zhang, Yawen; Ma, Jun; Zhang, Hongliang; Zhang, Ying; Zhang, Hao

2016-01-01

Tibetan chicken, unlike their lowland counterparts, exhibit specific adaptations to high-altitude conditions. The genetic mechanisms of such adaptations in highland chickens were determined by resequencing the genomes of four highland (Tibetan and Lhasa White) and four lowland (White Leghorn, Lindian, and Chahua) chicken populations. Our results showed an evident genetic admixture in Tibetan chickens, suggesting a history of introgression from lowland gene pools. Genes showing positive selection in highland populations were related to cardiovascular and respiratory system development, DNA repair, response to radiation, inflammation, and immune responses, indicating a strong adaptation to oxygen scarcity and high-intensity solar radiation. The distribution of allele frequencies of nonsynonymous single nucleotide polymorphisms between highland and lowland populations was analyzed using chi-square test, which showed that several differentially distributed genes with missense mutations were enriched in several functional categories, especially in blood vessel development and adaptations to hypoxia and intense radiation. RNA sequencing revealed that several differentially expressed genes were enriched in gene ontology terms related to blood vessel and respiratory system development. Several candidate genes involved in the development of cardiorespiratory system (FGFR1, CTGF, ADAM9, JPH2, SATB1, BMP4, LOX, LPR, ANGPTL4, and HYAL1), inflammation and immune responses (AIRE, MYO1F, ZAP70, DDX60, CCL19, CD47, JSC, and FAS), DNA repair, and responses to radiation (VCP, ASH2L, and FANCG) were identified to play key roles in the adaptation to high-altitude conditions. Our data provide new insights into the unique adaptations of highland animals to extreme environments. PMID:26907498

Orientation relationship between diamond and magnesiochromite inclusions

NASA Astrophysics Data System (ADS)

Alvaro, Matteo; Angel, Ross; Nimis, Paolo; Milani, Sula; Harris, Jeff; Nestola, Fabrizio

2017-04-01

The correct determination of the relative crystallographic orientations of single crystals has many applications. When single crystals undergo phase transitions, especially at high pressures, the relative orientations of the two phases yields insights into transition mechanisms (Dobson et al 2013). On the other hand, determination of the crystallographic orientations of minerals included in diamonds can provide insights into the mechanisms of their entrapment and the timing of their formation relative to the host diamond (e.g. Nestola et al. 2014, Milani et al. 2016). The reported occurrence of non-trivial orientations for some minerals in diamonds, suggesting an epitaxial relationship, has long been considered to reflect contemporaneous growth of the diamond and the inclusion (e.g. syngenesis). Correct interpretation of such orientations requires (i) a statistically significant crystallographic data set for single and multiple inclusions in a large number of diamonds, and (ii) a robust data-processing method, capable of removing ambiguities derived from the high symmetry of the diamond and the inclusion. We have developed a software to perform such processing (OrientXplot, Angel et al. 2015), starting from crystallographic orientation matrixes obtained by X-ray diffractometry or EBSD data. Previous studies of inclusions in lithospheric diamonds, by single-crystal X-ray diffraction and EBSD, indicate a wide variety in the orientations of different inclusion phases with respect to their diamond host (Futergendler & Frank-Kamenetsky 1961; Frank-Kamenetsky 1964; Wiggers de Vries et al. 2011; Nestola et al. 2014, Milani et al. 2016). For example, olivine inclusions in lithospheric diamonds from Udachnaya do not show any preferred orientations with respect to their diamond hosts, but multiple inclusions in a single diamond often show very similar orientations within few degrees. In the present work on magnesiochromite inclusions in diamonds from Udachnaya, there is a partial orientation between inclusion and host. A (111) plane of each inclusion is sub-parallel to a {111} plane of their diamond host, but with random orientations of the magnesiochromite [100], [010] and [001] relative to the diamond. In one case, where a single inclusion comprised a magnesiochromite-olivine touching pair, the magnesiochromite was oriented as noted above and the olivine showed a random orientation. The implications of these observations for the mechanisms of diamond growth will be explored and the results will be compared and combined with previous work. This work was supported by ERC starting grant "INDIMEDEA" (307322) to F. Nestola and by the MIUR-SIR grant "MILE DEEp" (RBSI140351) to M. Alvaro. References Angel R. J. et al., J. Appl. Crystallogr. 48, 1330-1334 (2015). Wiggers de Vries D.F. et al., Contrib Mineral Petr 161, 565-579 (2011). Dobson D. P. et al., Nat. Geosci. 6, 575-578 (2013). Frank-Kamenetsky V.A. The nature of structural impurities and inclusions in minerals. Leningrad, Gos. Univ. (1964) Futergendler S.I. and Frank-Kamenetsky V.A. Zap Vsesoyuzn Mineral Obs 90, 230 (1961). Nestola F. et al., Int. Geol. Rev. 56(13), 1658-1667 (2014).

DOE Office of Scientific and Technical Information (OSTI.GOV)

Culbert, A.A.; Wallis, G.A.; Kadler, K.E.

The brittleness of bone in people with lethal (type II) osteogenesis imperfecta, a heritable disorder caused by mutations in the type I collagen genes, arises from the deposition of abnormal collagen in the bone matrix. The inability of the abnormal collagen to participate in mineralization may be caused by its failure to interact with other bone proteins. Here, we have designed a strategy to isolate the genes important for mineralization of collagen during bone formation. Cells isolated from 16-day embryonic chick calvaria and seeded post-confluence in culture deposited a mineralized matrix over a period of 2 weeks. Chick skin fibroblastsmore » seeded and cultured under the same conditions did not mineralize. Using RT-PCR, we prepared short cDNAs ({approximately}300 bp) corresponding to the 3{prime} ends of mRNA from fibroblasts and separately from the mineralizing calvarial cells. Subtractive cDNA hybridization generated a pool of cDNAs that were specific to mineralizing calvarial cells but not to fibroblasts. Screening of 100,000 plaques of a chick bone ZAP Express cDNA library with this pool of mineralizing-specific cDNAs identified ten clones which comprised full-length cDNAs for the bone proteins osteopontin (eight of the ten positives), bone sialoprotein II (one of the ten positives), and cystatin (one of the ten positives). cDNAs for type I collagen, fibronectin, alkaline phosphatase, house-keeping genes, and other genes expressed in fibroblasts were not identified in this preliminary screen. The pool of short cDNAs is likely to comprise cDNAs for further bone-specific genes and will be used to screen the entire bone cDNA library of 4.2 million clones. 30 refs., 4 figs.« less

SDF-1 signaling via the CXCR4-TCR heterodimer requires PLC-β3 and PLC-γ1 for distinct cellular responses 1

PubMed Central

Kremer, Kimberly N.; Clift, Ian C.; Miamen, Alexander G.; Bamidele, Adebowale O.; Qian, Nan-Xin; Humphreys, Troy D.; Hedin, Karen E.

2011-01-01

The CXCR4 chemokine receptor is a G protein-coupled receptor (GPCR) that signals in T lymphocytes by forming a heterodimer with the T cell antigen receptor (TCR). CXCR4 and TCR functions are consequently highly cross-regulated, affecting T cell immune activation, cytokine secretion, and T cell migration. The CXCR4-TCR heterodimer stimulates T cell migration and activation of the ERK MAP kinase and downstream AP-1-dependent cytokine transcription in response to SDF-1, the sole chemokine ligand of CXCR4. These responses require Gi-type G proteins as well as TCR ITAM domains and the ZAP-70 tyrosine kinase, thus indicating that the CXCR4-TCR heterodimer signals to integrate GPCR-associated and TCR-associated signaling molecules in response to SDF-1. Yet, the phospholipase C (PLC) isozymes responsible for coupling the CXCR4-TCR heterodimer to distinct downstream cellular responses are incompletely characterized. Here, we demonstrate that PLC activity is required for SDF-1 to induce ERK activation, migration, and CXCR4 endocytosis in human T cells. SDF-1 signaling via the CXCR4-TCR heterodimer uses PLC-β3 to activate the Ras-ERK pathway and increase intracellular Ca2+ concentrations, while PLC-γ1 is dispensable for these outcomes. In contrast, PLC-γ1, but not PLC-β3, is required for SDF-1-mediated migration, via a mechanism independent of LAT. These results increase understanding of the signaling mechanisms employed by the CXCR4-TCR heterodimer, characterize new roles for PLC-β3 and PLC-γ1 in T cells, and suggest that multiple PLCs may also be activated downstream of other chemokine receptors in order to distinctly regulate migration versus other signaling functions. PMID:21705626

Mitochondria-derived hydrogen peroxide selectively enhances T cell receptor-initiated signal transduction.

PubMed

Gill, Tejpal; Levine, Alan D

2013-09-06

T cell receptor (TCR)-initiated signal transduction is reported to increase production of intracellular reactive oxygen species, such as superoxide (O2˙(-)) and hydrogen peroxide (H2O2), as second messengers. Although H2O2 can modulate signal transduction by inactivating protein phosphatases, the mechanism and the subcellular localization of intracellular H2O2 as a second messenger of the TCR are not known. The antioxidant enzyme superoxide dismutase (SOD) catalyzes the dismutation of highly reactive O2˙(-) into H2O2 and thus acts as an intracellular generator of H2O2. As charged O2˙(-) is unable to diffuse through intracellular membranes, cells express distinct SOD isoforms in the cytosol (Cu,Zn-SOD) and mitochondria (Mn-SOD), where they locally scavenge O2˙(-) leading to production of H2O2. A 2-fold organelle-specific overexpression of either SOD in Jurkat T cell lines increases intracellular production of H2O2 but does not alter the levels of intracellular H2O2 scavenging enzymes such as catalase, membrane-bound peroxiredoxin1 (Prx1), and cytosolic Prx2. We report that overexpression of Mn-SOD enhances tyrosine phosphorylation of TCR-associated membrane proximal signal transduction molecules Lck, LAT, ZAP70, PLCγ1, and SLP76 within 1 min of TCR cross-linking. This increase in mitochondrial H2O2 specifically modulates MAPK signaling through the JNK/cJun pathway, whereas overexpressing Cu,Zn-SOD had no effect on any of these TCR-mediated signaling molecules. As mitochondria translocate to the immunological synapse during TCR activation, we hypothesize this translocation provides the effective concentration of H2O2 required to selectively modulate downstream signal transduction pathways.

The dual Syk/JAK inhibitor cerdulatinib antagonises B-cell receptor and microenvironmental signaling in chronic lymphocytic leukemia

PubMed Central

Blunt, Matthew D; Koehrer, Stefan; Dobson, Rachel; Larrayoz, Marta; Wilmore, Sarah; Hayman, Alice; Parnell, Jack; Smith, Lindsay; Davies, Andrew; Johnson, Peter W; Conley, Pamela B; Pandey, Anjali; Strefford, Jon C; Stevenson, Freda K; Packham, Graham; Forconi, Francesco; Coffey, Greg; Burger, Jan A; Steele, Andrew J

2017-01-01

Purpose B-cell receptor (BCR)-associated kinase inhibitors such as ibrutinib have revolutionised the treatment of chronic lymphocytic leukemia (CLL). However, these agents are not curative and resistance is already emerging in a proportion of patients. Interleukin-4 (IL-4), expressed in CLL lymph nodes, can augment BCR-signalling and reduce the effectiveness of BCR-kinase inhibitors. Therefore simultaneous targeting of the IL-4- and BCR-signalling pathways by cerdulatinib, a novel dual Syk/JAK inhibitor currently in clinical trials (NCT01994382), may improve treatment responses in patients. Experimental Design PBMCs from CLL patients were treated with cerdulatinib alone or in combination with venetoclax. Cell death, chemokine and cell signalling assay were performed and analysed by flow cytometry, immunoblotting, Q-PCR and ELISA as indicated. Results At concentrations achievable in patients, cerdulatinib inhibited BCR- and IL-4-induced downstream signalling in CLL cells using multiple read-outs and prevented anti-IgM- and nurse-like cell (NLC)-mediated CCL3/CCL4 production. Cerdulatinib induced apoptosis of CLL cells, in a time- and concentration-dependent manner, and particularly in IGHV unmutated samples with greater BCR-signalling capacity and response to IL-4, or samples expressing higher levels of sIgM, CD49d+ or ZAP70+. Cerdulatinib overcame anti-IgM, IL-4/CD40L or NLC-mediated protection by preventing upregulation of MCL-1- and BCL-XL, however BCL-2 expression was unaffected. Furthermore in samples treated with IL-4/CD40L, cerdulatinib synergised with venetoclax in vitro to induce greater apoptosis than either drug alone. Conclusion Cerdulatinib is a promising therapeutic for the treatment of CLL either alone or in combination with venetoclax, with the potential to target critical survival pathways in this currently incurable disease. PMID:27697994

Modulation of the Fcepsilon receptor I signaling by tyrosine kinase inhibitors: search for therapeutic targets of inflammatory and allergy diseases.

PubMed

Lusková, Petra; Dráber, Petr

2004-01-01

Mast cells and basophils are major effector cells in the immunoglobulin E (IgE)-dependent allergic reactions as well as in the innate immunity. They are distributed throughout the body and, upon allergen exposure, are stimulated via the high affinity IgE receptor (FcepsilonRI) to release several pro-inflammatory mediators such as leukotrienes, immunoregulatory cytokines and histamine. FcepsilonRI-mediated signaling is initiated by tyrosine phosphorylation of FcepsilonRI subunits by Src family kinase Lyn, which is followed by an activation of Syk/Zap family kinase Syk. The activated kinases then in turn phosphorylate and activate other enzymes [phospholipase Cgamma (PLCgamma) isoforms, phosphatidylinositol-3 kinase (PI3K) isoforms, protein kinase C (PKC) isoforms, Bruton's tyrosine kinase (Btk) and others], adaptors [linker for activation of T cells (LAT), Cbl, Grb2 and others] and GTP exchange factors/GTPases (Vav, Ras, Rho, and others), and subsequently induce the mobilization of stored and extracellular Ca(2+). These and other biochemical events lead within seconds and minutes to the secretory response and later to the production of chemokines. This review is focused on the use of tyrosine kinase inhibitors specific for Src family kinases (PP1/PP2, SU6656 and CT5269), Syk kinase (piceatannol, ER-27319 and BAY 61-3606) and Btk (terreic acid and LFM-A13) for a modulation of FcepsilonRI-mediated signaling in mast cells. Potential use of the inhibitors in the treatment of inflammatory and allergy diseases as well as future directions in the development of highly specific tyrosine kinases inhibitors of new generations and their use in an intended modulation of mast cell signaling are discussed.

BT-benzo-29 inhibits bacterial cell proliferation by perturbing FtsZ assembly.

PubMed

Ray, Shashikant; Jindal, Bhavya; Kunal, Kishore; Surolia, Avadhesha; Panda, Dulal

2015-10-01

We have identified a potent antibacterial agent N-(4-sec-butylphenyl)-2-(thiophen-2-yl)-1H-benzo[d]imidazole-4-carboxamide (BT-benzo-29) from a library of benzimidazole derivatives that stalled bacterial division by inhibiting FtsZ assembly. A short (5 min) exposure of BT-benzo-29 disassembled the cytokinetic Z-ring in Bacillus subtilis cells without affecting the cell length and nucleoids. BT-benzo-29 also perturbed the localization of early and late division proteins such as FtsA, ZapA and SepF at the mid-cell. Further, BT-benzo-29 bound to FtsZ with a dissociation constant of 24 ± 3 μm and inhibited the assembly and GTPase activity of purified FtsZ. A docking analysis suggested that BT-benzo-29 may bind to FtsZ at the C-terminal domain near the T7 loop. BT-benzo-29 displayed significantly weaker inhibitory effects on the assembly and GTPase activity of two mutants (L272A and V275A) of FtsZ supporting the prediction of the docking analysis. Further, BT-benzo-29 did not appear to inhibit DNA duplication and nucleoid segregation and it did not perturb the membrane potential of B. subtilis cells. The results suggested that BT-benzo-29 exerts its potent antibacterial activity by inhibiting FtsZ assembly. Interestingly, BT-benzo-29 did not affect the membrane integrity of mammalian red blood cells. BT-benzo-29 bound to tubulin with a much weaker affinity than FtsZ and exerted significantly weaker effects on mammalian cells than on the bacterial cells indicating that the compound may have a strong antibacterial potential. © 2015 FEBS.

Adenosine regulates CD8 T-cell priming by inhibition of membrane-proximal T-cell receptor signalling

PubMed Central

Linnemann, Carsten; Schildberg, Frank A; Schurich, Anna; Diehl, Linda; Hegenbarth, Silke I; Endl, Elmar; Lacher, Svenja; Müller, Christa E; Frey, Jürgen; Simeoni, Luca; Schraven, Burkhart; Stabenow, Dirk; Knolle, Percy A

2009-01-01

Adenosine is a well-described anti-inflammatory modulator of immune responses within peripheral tissues. Extracellular adenosine accumulates in inflamed and damaged tissues and inhibits the effector functions of various immune cell populations, including CD8 T cells. However, it remains unclear whether extracellular adenosine also regulates the initial activation of naïve CD8 T cells by professional and semi-professional antigen-presenting cells, which determines their differentiation into effector or tolerant CD8 T cells, respectively. We show that adenosine inhibited the initial activation of murine naïve CD8 T cells after αCD3/CD28-mediated stimulation. Adenosine caused inhibition of activation, cytokine production, metabolic activity, proliferation and ultimately effector differentiation of naïve CD8 T cells. Remarkably, adenosine interfered efficiently with CD8 T-cell priming by professional antigen-presenting cells (dendritic cells) and semi-professional antigen-presenting cells (liver sinusoidal endothelial cells). Further analysis of the underlying mechanisms demonstrated that adenosine prevented rapid tyrosine phosphorylation of the key kinase ZAP-70 as well as Akt and ERK1/2 in naïve αCD3/CD28-stimulated CD8 cells. Consequently, αCD3/CD28-induced calcium-influx into CD8 cells was reduced by exposure to adenosine. Our results support the notion that extracellular adenosine controls membrane-proximal T-cell receptor signalling and thereby also differentiation of naïve CD8 T cells. These data raise the possibility that extracellular adenosine has a physiological role in the regulation of CD8 T-cell priming and differentiation in peripheral organs. PMID:19740334

Adenosine regulates CD8 T-cell priming by inhibition of membrane-proximal T-cell receptor signalling.

PubMed

Linnemann, Carsten; Schildberg, Frank A; Schurich, Anna; Diehl, Linda; Hegenbarth, Silke I; Endl, Elmar; Lacher, Svenja; Müller, Christa E; Frey, Jürgen; Simeoni, Luca; Schraven, Burkhart; Stabenow, Dirk; Knolle, Percy A

2009-09-01

Adenosine is a well-described anti-inflammatory modulator of immune responses within peripheral tissues. Extracellular adenosine accumulates in inflamed and damaged tissues and inhibits the effector functions of various immune cell populations, including CD8 T cells. However, it remains unclear whether extracellular adenosine also regulates the initial activation of naïve CD8 T cells by professional and semi-professional antigen-presenting cells, which determines their differentiation into effector or tolerant CD8 T cells, respectively. We show that adenosine inhibited the initial activation of murine naïve CD8 T cells after alphaCD3/CD28-mediated stimulation. Adenosine caused inhibition of activation, cytokine production, metabolic activity, proliferation and ultimately effector differentiation of naïve CD8 T cells. Remarkably, adenosine interfered efficiently with CD8 T-cell priming by professional antigen-presenting cells (dendritic cells) and semi-professional antigen-presenting cells (liver sinusoidal endothelial cells). Further analysis of the underlying mechanisms demonstrated that adenosine prevented rapid tyrosine phosphorylation of the key kinase ZAP-70 as well as Akt and ERK1/2 in naïve alphaCD3/CD28-stimulated CD8 cells. Consequently, alphaCD3/CD28-induced calcium-influx into CD8 cells was reduced by exposure to adenosine. Our results support the notion that extracellular adenosine controls membrane-proximal T-cell receptor signalling and thereby also differentiation of naïve CD8 T cells. These data raise the possibility that extracellular adenosine has a physiological role in the regulation of CD8 T-cell priming and differentiation in peripheral organs.

Quantal concept of T-cell activation: adhesion domains as immunological synapses

NASA Astrophysics Data System (ADS)

Sackmann, Erich

2011-06-01

Adhesion micro-domains (ADs) formed during encounters of lymphocytes with antigen-presenting cells (APC) mediate the genetic expression of quanta of cytokines interleukin-2 (IL-2). The IL-2-induced activation of IL-2 receptors promotes the stepwise progression of the T-cells through the cell cycle, hence their name, immunological synapses. The ADs form short-lived reaction centres controlling the recruitment of activators of the biochemical pathway (the kinases Lck and ZAP) while preventing the access of inhibitors (phosphatase CD45) through steric repulsion forces. CD45 acts as the generator of adhesion domains and, through its role as a spacer protein, also as the promoter of the reaction. In a second phase of T-cell-APC encounters, long-lived global reaction spaces (called supramolecular activation complexes (SMAC)) form by talin-mediated binding of the T-cell integrin (LFA-1) to the counter-receptor ICAM-1, resulting in the formation of ring-like tight adhesion zones (peripheral SMAC). The ADs move to the centre of the intercellular adhesion zone forming the central SMAC, which serve in the recycling of the AD. We propose that cell stimulation is triggered by integrating the effect evoked by the short-lived adhesion domains. Similar global reaction platforms are formed by killer cells to destruct APC. We present a testable mechanical model showing that global reaction spaces (SMAC or dome-like contacts between cytotoxic cells and APC) form by self-organization through delayed activation of the integrin-binding affinity and stabilization of the adhesion zones by F-actin recruitment. The mechanical stability and the polarization of the adhering T-cells are mediated by microtubule-actin cross-talk.

Signal transduction in primary human T lymphocytes in altered gravity during parabolic flight and clinostat experiments.

PubMed

Tauber, Svantje; Hauschild, Swantje; Paulsen, Katrin; Gutewort, Annett; Raig, Christiane; Hürlimann, Eva; Biskup, Josefine; Philpot, Claudia; Lier, Hartwin; Engelmann, Frank; Pantaleo, Antonella; Cogoli, Augusto; Pippia, Proto; Layer, Liliana E; Thiel, Cora S; Ullrich, Oliver

2015-01-01

Several limiting factors for human health and performance in microgravity have been clearly identified arising from the immune system, and substantial research activities are required in order to provide the basic information for appropriate integrated risk management. The gravity-sensitive nature of cells of the immune system renders them an ideal biological model in search for general gravity-sensitive mechanisms and to understand how the architecture and function of human cells is related to the gravitational force and therefore adapted to life on Earth. We investigated the influence of altered gravity in parabolic flight and 2D clinostat experiments on key proteins of activation and signaling in primary T lymphocytes. We quantified components of the signaling cascade 1.) in non-activated T lymphocytes to assess the "basal status" of the cascade and 2.) in the process of activation to assess the signal transduction. We found a rapid decrease of CD3 and IL-2R surface expression and reduced p-LAT after 20 seconds of altered gravity in non-activated primary T lymphocytes during parabolic flight. Furthermore, we observed decreased CD3 surface expression, reduced ZAP-70 abundance and increased histone H3-acetylation in activated T lymphocytes after 5 minutes of clinorotation and a transient downregulation of CD3 and stable downregulation of IL-2R during 60 minutes of clinorotation. CD3 and IL-2R are downregulated in primary T lymphocytes in altered gravity. We assume that a gravity condition around 1g is required for the expression of key surface receptors and appropriate regulation of signal molecules in T lymphocytes. © 2015 S. Karger AG, Basel.

Increasing Plasma Parameters using Sheared Flow Stabilization of a Z-Pinch

NASA Astrophysics Data System (ADS)

Shumlak, Uri

2016-10-01

Recent experiments on the ZaP Flow Z-Pinch at the University of Washington have been successful in compressing the plasma column to smaller radii, producing the predicted increases in plasma density (1018 cm-3), temperature (200 eV), and magnetic fields (4 T), while maintaining plasma stability for many Alfven times (over 40 μs) using sheared plasma flows. These results indicate the suitability of the device as a discovery science platform for astrophysical and high energy density plasma research, and keeps open a possible path to achieving burning plasma conditions in a compact fusion device. Long-lived Z-pinch plasmas have been produced with dimensions of 1 cm radius and 100 cm long that are stabilized by sheared axial flows for over 1000 Alfven radial transit times. The observed plasma stability is coincident with the presence of a sheared flow as measured by time-resolved multi-chord ion Doppler spectroscopy applied to impurity ion radiation. These measurements yield insights into the evolution of the velocity profile and show that the stabilizing behavior of flow shear agrees with theoretical calculations and 2-D MHD computational simulations. The flow shear value, extent, and duration are shown to be consistent with theoretical models of the plasma viscosity, which places a design constraint on the maximum axial length of a sheared flow stabilized Z-pinch. Measurements of the magnetic field topology indicate simultaneous azimuthal symmetry and axial uniformity along the entire 100 cm length of the Z-pinch plasma. Separate control of plasma acceleration and compression have increased the accessible plasma parameters and have generated stable plasmas with radii below 0.5 cm, as measured with a high resolution digital holographic interferometer. This work was supported by Grants from U.S. DOE, NNSA, and ARPA-E.

Molecular characterization, genomic arrangement, and expression of bmpD, a new member of the bmp class of genes encoding membrane proteins of Borrelia burgdorferi.

PubMed Central

Ramamoorthy, R; Povinelli, L; Philipp M, T

1996-01-01

An expression library made with Borrelia burgdorferi DNA in the vector lambda ZapII was screened with serum from a monkey infected with the Lyme disease agent. This serum killed B. burgdorferi in vitro by an antibody-dependent, complement-mediated mechanism and contained antibodies to at least seven spirochetal antigens, none of which were the major outer surface proteins OspA or OspB. Among several positive clones, a clone containing the B. burgdorferi bmpA gene encoding the immunodominant antigen P39 was obtained. Chromosome walking and DNA sequence analysis permitted the identification of two additional upstream genes homologous to the bmpA gene and its related companion, bmpB. The first of these was the recently characterized bmpC gene, and adjacent to it was the fourth and new member of this class, which has been designated bmpD. The gene product encoded by bmpD is 34l residues long, contains a signal sequence with a potential signal peptidase II cleavage site, and has 26% identity with TmpC of Treponema pallidum. Southern blotting confirmed the tandem arrangement of all four bmp genes in the chromosome of B. burgdorferi JD1. However, Northern (RNA) blotting revealed that bmpD is expressed as a monocistronic transcript, which indicates that it is not part of an operon at the bmp locus. The bmpD gene was found to be conserved in representative members of the three species of the B. burgdorferi sensu lato complex, suggesting that it serves an important biological function in the spirochete. PMID:8606088

Renal Integrin-Linked Kinase Depletion Induces Kidney cGMP-Axis Upregulation: Consequences on Basal and Acutely Damaged Renal Function

PubMed Central

Cano-Peñalver, José Luis; Griera, Mercedes; García-Jerez, Andrea; Hatem-Vaquero, Marco; Ruiz-Torres, María Piedad; Rodríguez-Puyol, Diego; de Frutos, Sergio; Rodríguez-Puyol, Manuel

2015-01-01

Soluble guanylyl cyclase (sGC) is activated by nitric oxide (NO) and produces cGMP, which activates cGMP-dependent protein kinases (PKG) and is hydrolyzed by specific phosphodiesterases (PDE). The vasodilatory and cytoprotective capacity of cGMP-axis activation results in a therapeutic strategy for several pathologies. Integrin-linked kinase (ILK), a major scaffold protein between the extracellular matrix and intracellular signaling pathways, may modulate the expression and functionality of the cGMP-axis–related proteins. We introduce ILK as a novel modulator in renal homeostasis as well as a potential target for cisplatin (CIS)-induced acute kidney injury (AKI) improvement. We used an adult mice model of depletion of ILK (cKD-ILK), which showed basal increase of sGC and PKG expressions and activities in renal cortex when compared with wildtype (WT) littermates. Twenty-four h activation of sGC activation with NO enhanced the filtration rate in cKD-ILK. During AKI, cKD-ILK maintained the cGMP-axis upregulation with consequent filtration rates enhancement and ameliorated CIS-dependent tubular epithelial-to-mesenchymal transition and inflammation and markers. To emphasize the role of cGMP-axis upregulation due to ILK depletion, we modulated the cGMP axis under AKI in vivo and in renal cultured cells. A suboptimal dose of the PDE inhibitor ZAP enhanced the beneficial effects of the ILK depletion in AKI mice. On the other hand, CIS increased contractility-related events in cultured glomerular mesangial cells and necrosis rates in cultured tubular cells; ILK depletion protected the cells while sGC blockade with ODQ fully recovered the damage. PMID:26562149

Constitutive Lck Activity Drives Sensitivity Differences between CD8+ Memory T Cell Subsets.

PubMed

Moogk, Duane; Zhong, Shi; Yu, Zhiya; Liadi, Ivan; Rittase, William; Fang, Victoria; Dougherty, Janna; Perez-Garcia, Arianne; Osman, Iman; Zhu, Cheng; Varadarajan, Navin; Restifo, Nicholas P; Frey, Alan B; Krogsgaard, Michelle

2016-07-15

CD8(+) T cells develop increased sensitivity following Ag experience, and differences in sensitivity exist between T cell memory subsets. How differential TCR signaling between memory subsets contributes to sensitivity differences is unclear. We show in mouse effector memory T cells (TEM) that >50% of lymphocyte-specific protein tyrosine kinase (Lck) exists in a constitutively active conformation, compared with <20% in central memory T cells (TCM). Immediately proximal to Lck signaling, we observed enhanced Zap-70 phosphorylation in TEM following TCR ligation compared with TCM Furthermore, we observed superior cytotoxic effector function in TEM compared with TCM, and we provide evidence that this results from a lower probability of TCM reaching threshold signaling owing to the decreased magnitude of TCR-proximal signaling. We provide evidence that the differences in Lck constitutive activity between CD8(+) TCM and TEM are due to differential regulation by SH2 domain-containing phosphatase-1 (Shp-1) and C-terminal Src kinase, and we use modeling of early TCR signaling to reveal the significance of these differences. We show that inhibition of Shp-1 results in increased constitutive Lck activity in TCM to levels similar to TEM, as well as increased cytotoxic effector function in TCM Collectively, this work demonstrates a role for constitutive Lck activity in controlling Ag sensitivity, and it suggests that differential activities of TCR-proximal signaling components may contribute to establishing the divergent effector properties of TCM and TEM. This work also identifies Shp-1 as a potential target to improve the cytotoxic effector functions of TCM for adoptive cell therapy applications. Copyright © 2016 by The American Association of Immunologists, Inc.

APT: Aperture Photometry Tool

NASA Astrophysics Data System (ADS)

Laher, Russ

2012-08-01

Aperture Photometry Tool (APT) is software for astronomers and students interested in manually exploring the photometric qualities of astronomical images. It has a graphical user interface (GUI) which allows the image data associated with aperture photometry calculations for point and extended sources to be visualized and, therefore, more effectively analyzed. Mouse-clicking on a source in the displayed image draws a circular or elliptical aperture and sky annulus around the source and computes the source intensity and its uncertainty, along with several commonly used measures of the local sky background and its variability. The results are displayed and can be optionally saved to an aperture-photometry-table file and plotted on graphs in various ways using functions available in the software. APT is geared toward processing sources in a small number of images and is not suitable for bulk processing a large number of images, unlike other aperture photometry packages (e.g., SExtractor). However, APT does have a convenient source-list tool that enables calculations for a large number of detections in a given image. The source-list tool can be run either in automatic mode to generate an aperture photometry table quickly or in manual mode to permit inspection and adjustment of the calculation for each individual detection. APT displays a variety of useful graphs, including image histogram, and aperture slices, source scatter plot, sky scatter plot, sky histogram, radial profile, curve of growth, and aperture-photometry-table scatter plots and histograms. APT has functions for customizing calculations, including outlier rejection, pixel “picking” and “zapping,” and a selection of source and sky models. The radial-profile-interpolation source model, accessed via the radial-profile-plot panel, allows recovery of source intensity from pixels with missing data and can be especially beneficial in crowded fields.

Genome Resequencing Identifies Unique Adaptations of Tibetan Chickens to Hypoxia and High-Dose Ultraviolet Radiation in High-Altitude Environments.

PubMed

Zhang, Qian; Gou, Wenyu; Wang, Xiaotong; Zhang, Yawen; Ma, Jun; Zhang, Hongliang; Zhang, Ying; Zhang, Hao

2016-02-23

Tibetan chicken, unlike their lowland counterparts, exhibit specific adaptations to high-altitude conditions. The genetic mechanisms of such adaptations in highland chickens were determined by resequencing the genomes of four highland (Tibetan and Lhasa White) and four lowland (White Leghorn, Lindian, and Chahua) chicken populations. Our results showed an evident genetic admixture in Tibetan chickens, suggesting a history of introgression from lowland gene pools. Genes showing positive selection in highland populations were related to cardiovascular and respiratory system development, DNA repair, response to radiation, inflammation, and immune responses, indicating a strong adaptation to oxygen scarcity and high-intensity solar radiation. The distribution of allele frequencies of nonsynonymous single nucleotide polymorphisms between highland and lowland populations was analyzed using chi-square test, which showed that several differentially distributed genes with missense mutations were enriched in several functional categories, especially in blood vessel development and adaptations to hypoxia and intense radiation. RNA sequencing revealed that several differentially expressed genes were enriched in gene ontology terms related to blood vessel and respiratory system development. Several candidate genes involved in the development of cardiorespiratory system (FGFR1, CTGF, ADAM9, JPH2, SATB1, BMP4, LOX, LPR, ANGPTL4, and HYAL1), inflammation and immune responses (AIRE, MYO1F, ZAP70, DDX60, CCL19, CD47, JSC, and FAS), DNA repair, and responses to radiation (VCP, ASH2L, and FANCG) were identified to play key roles in the adaptation to high-altitude conditions. Our data provide new insights into the unique adaptations of highland animals to extreme environments. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

High-pressure sorption of nitrogen, carbon dioxide, and their mixtures on Argonne Premium Coals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Andreas Busch; Yves Gensterblum; Bernhard M. Krooss

2007-06-15

Gas sorption isotherms have been measured for carbon dioxide and nitrogen and their binary mixture (N{sub 2}/CO{sub 2} {approximately} 80/20) on three different moisture-equilibrated coals from the Argonne Premium Coal Sample Program by the U.S. Department of Energy, varying in rank from 0.25 to 1.68% vitrinite reflectance (VR{sub r}). The measurements were conducted at 55 C and at pressures up to 27 MPa for the pure gases and up to 10 MPa for the gas mixture. The effects of the large differences in equilibrium moisture contents (0.8 to 32.2%) on sorption capacity were estimated on the basis of the aqueousmore » solubility of CO{sub 2} and N{sub 2} at experimental conditions. Especially for the Beulah-Zap coal with an equilibrium moisture content of {approximately} 32%, the amount of dissolved CO{sub 2} contributes significantly to the overall storage capacity, whereas the amounts of N{sub 2} dissolved in the moisture water are low and can be neglected. Sorption measurements with nitrogen/carbon dioxide mixtures showed very low capacities for N{sub 2}. For Illinois coal, these excess sorption values were even slightly negative, probably due to small volumetric effects (changes in condensed phase volume). The evolution of the composition of the free gas phase in contact with the coal sample has been monitored continuously during each pressure step of the sorption tests. This composition changed strongly over time. Apparently, CO{sub 2} reaches sorption sites very quickly initially and is subsequently partly replaced by N{sub 2} molecules until concentration equilibration is reached. 18 refs., 10 figs., 2 tabs.« less

The role of ZAP70 kinase in acute lymphoblastic leukemia infiltration into the central nervous system.

PubMed

Alsadeq, Ameera; Fedders, Henning; Vokuhl, Christian; Belau, Nele M; Zimmermann, Martin; Wirbelauer, Tim; Spielberg, Steffi; Vossen-Gajcy, Michaela; Cario, Gunnar; Schrappe, Martin; Schewe, Denis M

2017-02-01

Central nervous system infiltration and relapse are poorly understood in childhood acute lymphoblastic leukemia. We examined the role of zeta-chain-associated protein kinase 70 in preclinical models of central nervous system leukemia and performed correlative studies in patients. Zeta-chain-associated protein kinase 70 expression in acute lymphoblastic leukemia cells was modulated using short hairpin ribonucleic acid-mediated knockdown or ectopic expression. We show that zeta-chain-associated protein kinase 70 regulates CCR7/CXCR4 via activation of extracellular signal-regulated kinases. High expression of zeta-chain-associated protein kinase 70 in acute lymphoblastic leukemia cells resulted in a higher proportion of central nervous system leukemia in xenografts as compared to zeta-chain-associated protein kinase 70 low expressing counterparts. High zeta-chain-associated protein kinase 70 also enhanced the migration potential towards CCL19/CXCL12 gradients in vitro CCR7 blockade almost abrogated homing of acute lymphoblastic leukemia cells to the central nervous system in xenografts. In 130 B-cell precursor acute lymphoblastic leukemia and 117 T-cell acute lymphoblastic leukemia patients, zeta-chain-associated protein kinase 70 and CCR7/CXCR4 expression levels were significantly correlated. Zeta-chain-associated protein kinase 70 expression correlated with central nervous system disease in B-cell precursor acute lymphoblastic leukemia, and CCR7/CXCR4 correlated with central nervous system involvement in T-cell acute lymphoblastic leukemia patients. In multivariate analysis, zeta-chain-associated protein kinase 70 expression levels in the upper third and fourth quartiles were associated with central nervous system involvement in B-cell precursor acute lymphoblastic leukemia (odds ratio=7.48, 95% confidence interval, 2.06-27.17; odds ratio=6.86, 95% confidence interval, 1.86-25.26, respectively). CCR7 expression in the upper fourth quartile correlated with central nervous system positivity in T-cell acute lymphoblastic leukemia (odds ratio=11.00, 95% confidence interval, 2.00-60.62). We propose zeta-chain-associated protein kinase 70, CCR7 and CXCR4 as markers of central nervous system infiltration in acute lymphoblastic leukemia warranting prospective investigation. Copyright© Ferrata Storti Foundation.

CD147 and CD98 complex-mediated homotypic aggregation attenuates the CypA-induced chemotactic effect on Jurkat T cells.

PubMed

Guo, Na; Zhang, Kui; Lv, Minghua; Miao, Jinlin; Chen, Zhinan; Zhu, Ping

2015-02-01

Homotypic cell aggregation plays important roles in physiological and pathological processes, including embryogenesis, immune responses, angiogenesis, tumor cell invasion and metastasis. CD147 has been implicated in most of these phenomena, and it was identified as a T cell activation-associated antigen due to its obvious up-regulation in activated T cells. However, the explicit function and mechanism of CD147 in T cells have not been fully elucidated. In this study, large and compact aggregates were observed in Jurkat T cells after treatment with the specific CD147 monoclonal antibody HAb18 or after the expression of CD147 was silenced by RNA interference, which indicated an inhibitory effect of CD147 in T cell homotypic aggregation. Knocking down CD147 expression resulted in a significant decrease in CD98, along with prominent cell aggregation, similar to that treated by CD98 and CD147 monoclonal antibodies. Furthermore, decreased cell chemotactic activity was observed following CD147- and CD98-mediated cell aggregation, and increased aggregation was correlated with a decrease in the chemotactic ability of the Jurkat T cells, suggesting that CD147- and CD98-mediated homotypic cell aggregation plays a negative role in T cell chemotaxis. Our data also showed that p-ERK, p-ZAP70, p-CD3ζ and p-LCK were significantly decreased in the CD147- and CD98-knocked down Jurkat T cells, which suggested that decreased CD147- and/or CD98-induced homotypic T cell aggregation and aggregation-inhibited chemotaxis might be associated with these signaling pathways. A role for CD147 in cell aggregation and chemotaxis was further indicated in primary CD4(+) T cells. Similarly, low expression of CD147 in primary T cells induced prominent cell aggregation and this aggregation attenuated primary T cell chemotactic ability in response to CypA. Our results have demonstrated the correlation between homotypic cell aggregation and the chemotactic response of T cells to CypA, and these data indicate that CD147 and CD98 might play important roles in cyclophilin-induced cell migration. Copyright © 2014 Elsevier Ltd. All rights reserved.

Two forms of electrical resonance at theta frequencies, generated by M-current, h-current and persistent Na+ current in rat hippocampal pyramidal cells

PubMed Central

Hu, Hua; Vervaeke, Koen; Storm, Johan F

2002-01-01

Coherent network oscillations in the brain are correlated with different behavioural states. Intrinsic resonance properties of neurons provide a basis for such oscillations. In the hippocampus, CA1 pyramidal neurons show resonance at theta (θ) frequencies (2-7 Hz). To study the mechanisms underlying θ-resonance, we performed whole-cell recordings from CA1 pyramidal cells (n = 73) in rat hippocampal slices. Oscillating current injections at different frequencies (ZAP protocol), revealed clear resonance with peak impedance at 2-5 Hz at ≈33 °C (increasing to ≈7 Hz at ≈38 °C). The θ-resonance showed a U-shaped voltage dependence, being strong at subthreshold, depolarized (≈-60 mV) and hyperpolarized (≈-80 mV) potentials, but weaker near the resting potential (-72 mV). Voltage clamp experiments revealed three non-inactivating currents operating in the subthresold voltage range: (1) M-current (IM), which activated positive to -65 mV and was blocked by the M/KCNQ channel blocker XE991 (10 μm); (2) h-current (Ih), which activated negative to -65 mV and was blocked by the h/HCN channel blocker ZD7288 (10 μm); and (3) a persistent Na+ current (INaP), which activated positive to -65 mV and was blocked by tetrodotoxin (TTX, 1 μm). In current clamp, XE991 or TTX suppressed the resonance at depolarized, but not hyperpolarized membrane potentials, whereas ZD7288 abolished the resonance only at hyperpolarized potentials. We conclude that these cells show two forms of θ-resonance: ‘M-resonance’ generated by the M-current and persistent Na+ current in depolarized cells, and ‘H-resonance’ generated by the h-current in hyperpolarized cells. Computer simulations supported this interpretation. These results suggest a novel function for M/KCNQ channels in the brain: to facilitate neuronal resonance and network oscillations in cortical neurons, thus providing a basis for an oscillation-based neural code. PMID:12482886

The chemokine CXCL12 generates costimulatory signals in T cells to enhance phosphorylation and clustering of the adaptor protein SLP-76.

PubMed

Smith, Xin; Schneider, Helga; Köhler, Karsten; Liu, Hebin; Lu, Yuning; Rudd, Christopher E

2013-07-30

The CXC chemokine CXCL12 mediates the chemoattraction of T cells and enhances the stimulation of T cells through the T cell receptor (TCR). The adaptor SLP-76 [Src homology 2 (SH2) domain-containing leukocyte protein of 76 kD] has two key tyrosine residues, Tyr(113) and Tyr(128), that mediate signaling downstream of the TCR. We investigated the effect of CXCL12 on SLP-76 phosphorylation and the TCR-dependent formation of SLP-76 microclusters. Although CXCL12 alone failed to induce SLP-76 cluster formation, it enhanced the number, stability, and phosphorylation of SLP-76 microclusters formed in response to stimulation of the TCR by an activating antibody against CD3, a component of the TCR complex. Addition of CXCL12 to anti-CD3-stimulated cells resulted in F-actin polymerization that stabilized SLP-76 microclusters in the cells' periphery at the interface with antibody-coated coverslips and increased the interaction between SLP-76 clusters and those containing ZAP-70, the TCR-associated kinase that phosphorylates SLP-76, as well as increased TCR-dependent gene expression. Costimulation with CXCL12 and anti-CD3 increased the extent of phosphorylation of SLP-76 at Tyr(113) and Tyr(128), but not that of other TCR-proximal components, and mutation of either one of these residues impaired the CXCL12-dependent effect on SLP-76 microcluster formation, F-actin polymerization, and TCR-dependent gene expression. The effects of CXCL12 on SLP-76 microcluster formation were dependent on the coupling of its receptor CXCR4 to G(i)-family G proteins (heterotrimeric guanine nucleotide-binding proteins). Thus, we identified a costimulatory mechanism by which CXCL12 and antigen converge at SLP-76 microcluster formation to enhance T cell responses.

An MSC2 Promoter-lacZ Fusion Gene Reveals Zinc-Responsive Changes in Sites of Transcription Initiation That Occur across the Yeast Genome

PubMed Central

Wu, Yi-Hsuan; Taggart, Janet; Song, Pamela Xiyao; MacDiarmid, Colin; Eide, David J.

2016-01-01

The Msc2 and Zrg17 proteins of Saccharomyces cerevisiae form a complex to transport zinc into the endoplasmic reticulum. ZRG17 is transcriptionally induced in zinc-limited cells by the Zap1 transcription factor. In this report, we show that MSC2 mRNA also increases (~1.5 fold) in zinc-limited cells. The MSC2 gene has two in-frame ATG codons at its 5’ end, ATG1 and ATG2; ATG2 is the predicted initiation codon. When the MSC2 promoter was fused at ATG2 to the lacZ gene, we found that unlike the chromosomal gene this reporter showed a 4-fold decrease in lacZ mRNA in zinc-limited cells. Surprisingly, β-galactosidase activity generated by this fusion gene increased ~7 fold during zinc deficiency suggesting the influence of post-transcriptional factors. Transcription of MSC2ATG2-lacZ was found to start upstream of ATG1 in zinc-replete cells. In zinc-limited cells, transcription initiation shifted to sites just upstream of ATG2. From the results of mutational and polysome profile analyses, we propose the following explanation for these effects. In zinc-replete cells, MSC2ATG2-lacZ mRNA with long 5’ UTRs fold into secondary structures that inhibit translation. In zinc-limited cells, transcripts with shorter unstructured 5’ UTRs are generated that are more efficiently translated. Surprisingly, chromosomal MSC2 did not show start site shifts in response to zinc status and only shorter 5’ UTRs were observed. However, the shifts that occur in the MSC2ATG2-lacZ construct led us to identify significant transcription start site changes affecting the expression of ~3% of all genes. Therefore, zinc status can profoundly alter transcription initiation across the yeast genome. PMID:27657924

MicroRNAs in B-cell lymphomas: how a complex biology gets more complex.

PubMed

Musilova, K; Mraz, M

2015-05-01

MicroRNAs (miRNAs) represent important regulators of gene expression besides transcriptional control. miRNA regulation can be involved in the cell developmental fate decisions, but can also have more subtle roles in buffering stochastic fluctuations in gene expression. They participate in pathways fundamental to B-cell development like B-cell receptor (BCR) signalling, B-cell migration/adhesion, cell-cell interactions in immune niches, and the production and class-switching of immunoglobulins. miRNAs influence B-cell maturation, generation of pre-, marginal zone, follicular, B1, plasma and memory B cells. In this review, we discuss miRNAs with essential functions in malignant B-cell development (such as miR-150, miR-155, miR-21, miR-34a, miR-17-92 and miR-15-16). We also put these miRNAs in the context of normal B-cell differentiation, as this is intimately connected to neoplastic B-cell development. We review miRNAs' role in the most common B-cell malignancies, including chronic lymphocytic leukaemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL) and mantle cell lymphoma (MCL). We focus on miR-contribution to the regulation of important signalling pathways (such as NF-κB, PI3K/AKT and TGF-β), BCR signalling and its modulators (such as PTEN, SHIP-1, ZAP-70, GAB1 and BTK), anti- and pro-apoptotic proteins (such as BCL2, MCL1, TCL1, BIM, p53 and SIRT1) and transcription factors (such as MYC, MYB, PU.1, FOXP1 and BCL6). We also discuss the association of miRNAs' expression levels with the patients' survival and response to therapy, summarizing their potential use as predictive and prognostic markers. Importantly, the targeting of miRNAs (like use of anti-miR-155 or miR-34a mimic) could provide a novel therapeutic approach as evidenced by tumour regression in xenograft mouse models and initial promising data from clinical trials.

[Cloning, Expression and Immunodiagnostic Evaluation of the Fasciola gigantica Thioredoxin Peroxidase].

PubMed

Wang, Yue-qi; Zhou, Yan; Cheng, Na; Chen, Mu-xin; Ai, Lin; Liu, Yu-hua; Zhang, Jian-guo; Luo, Jia-jun; Xu, Xue-nian

2015-04-01

To immunoscreen the gene encoding thioredoxin peroxidase (TPx) from a cDNA library made from adult Fasciola gigantica worms, clone and express the gene, and evaluate the immunodiagnostic value of TPx recombinant protein. The A ZAP cDNA library was immunoscreened with pooled serum of fascioliasis gigantica patients. The obtained positive clones were sequenced and analyzed by multiple sequence alignment. The full-length (rFgTPx) and N-termianal truncated (rFgTPx_nt) sequence of FgTPx was subcloned into prokaryotic plasmid pET28a(+) with a non-fusion expression technique, respectively. The recombinant proteins of rFgTPx and rFgTPx_nt were purified by His-bind affinity column (Ni-NTA). rFgTPx and rFgTPx_nt were used in indirect ELISA to test the antibody response of the serum samples. Sera of 27 fascioliasis gigantica patients, 15 patients with schistosomaisis japonica, 15 clonorchiasis sinensis patients, and 32 healthy donors were tested by using the recombinant protein based ELISA. The TPx recombinant proteins were obtained through expression, purification and renaturation, the relative molecular mass of rFgTPx and rFgTPx_nt were Mr 30,000 and Mr 26,000, respectively. The total diagnostic coincidence rate, sensitivity and specificity of rFgTPx_nt-based ELISA was 87.6% (78/89), 66.7% (18/27), and 96.8% (60/62), respectively. The cross reaction with Schistosoma japonicum and Clonorchis sinensis was 0 and 1/15 for rFgTPx_nt, respectively. Before and after treatment, A450 value of the serum samples from fascioliasis patients was 0.233 ± 0.088 and 0.129 ± 0.072, respectively (t = 4.27, P < 0.01). The gene encoding TPx is expressed in the prokaryotic expression system. The recombinant protein shows proper sensitivity and high specificity for the serodiagnosis of Fasciola gigantica infection.

The Adenylate Cyclase Toxins of Bacillus anthracis and Bordetella pertussis Promote Th2 Cell Development by Shaping T Cell Antigen Receptor Signaling

PubMed Central

Rossi Paccani, Silvia; Benagiano, Marisa; Capitani, Nagaja; Zornetta, Irene; Ladant, Daniel; Montecucco, Cesare; D'Elios, Mario M.; Baldari, Cosima T.

2009-01-01

The adjuvanticity of bacterial adenylate cyclase toxins has been ascribed to their capacity, largely mediated by cAMP, to modulate APC activation, resulting in the expression of Th2–driving cytokines. On the other hand, cAMP has been demonstrated to induce a Th2 bias when present during T cell priming, suggesting that bacterial cAMP elevating toxins may directly affect the Th1/Th2 balance. Here we have investigated the effects on human CD4+ T cell differentiation of two adenylate cyclase toxins, Bacillus anthracis edema toxin (ET) and Bordetella pertussis CyaA, which differ in structure, mode of cell entry, and subcellular localization. We show that low concentrations of ET and CyaA, but not of their genetically detoxified adenylate cyclase defective counterparts, potently promote Th2 cell differentiation by inducing expression of the master Th2 transcription factors, c-maf and GATA-3. We also present evidence that the Th2–polarizing concentrations of ET and CyaA selectively inhibit TCR–dependent activation of Akt1, which is required for Th1 cell differentiation, while enhancing the activation of two TCR–signaling mediators, Vav1 and p38, implicated in Th2 cell differentiation. This is at variance from the immunosuppressive toxin concentrations, which interfere with the earliest step in TCR signaling, activation of the tyrosine kinase Lck, resulting in impaired CD3ζ phosphorylation and inhibition of TCR coupling to ZAP-70 and Erk activation. These results demonstrate that, notwithstanding their differences in their intracellular localization, which result in focalized cAMP production, both toxins directly affect the Th1/Th2 balance by interfering with the same steps in TCR signaling, and suggest that their adjuvanticity is likely to result from their combined effects on APC and CD4+ T cells. Furthermore, our results strongly support the key role of cAMP in the adjuvanticity of these toxins. PMID:19266022

Intranuclear interactomic inhibition of NF-κB suppresses LPS-induced severe sepsis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Park, Sung-Dong; Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749; Cheon, So Yeong

Suppression of nuclear factor-κB (NF-κB) activation, which is best known as a major regulator of innate and adaptive immune responses, is a potent strategy for the treatment of endotoxic sepsis. To inhibit NF-κB functions, we designed the intra-nuclear transducible form of transcription modulation domain (TMD) of RelA (p65), called nt-p65-TMD, which can be delivered effectively into the nucleus without influencing the cell viability, and work as interactomic inhibitors via disruption of the endogenous p65-mediated transcription complex. nt-p65-TMD effectively inhibited the secretion of pro-inflammatory cytokines, including TNF-α, IL-1β, or IL-6 from BV2 microglia cells stimulated by lipopolysaccharide (LPS). nt-p65-TMD did notmore » inhibit tyrosine phosphorylation of signaling mediators such as ZAP-70, p38, JNK, or ERK involved in T cell activation, but was capable of suppressing the transcriptional activity of NF-κB without the functional effect on that of NFAT upon T-cell receptor (TCR) stimulation. The transduced nt-p65-TMD in T cell did not affect the expression of CD69, however significantly inhibited the secretion of T cell-specific cytokines such as IL-2, IFN-γ, IL-4, IL-17A, or IL-10. Systemic administration of nt-p65-TMD showed a significant therapeutic effect on LPS-induced sepsis model by inhibiting pro-inflammatory cytokines secretion. Therefore, nt-p65-TMD can be a novel therapeutics for the treatment of various inflammatory diseases, including sepsis, where a transcription factor has a key role in pathogenesis, and further allows us to discover new functions of p65 under normal physiological condition without genetic alteration. - Highlights: • The nt-p65-TMD is intra-nuclear interactomic inhibitor of endogenous p65. • The nt-p65-TMD effectively inhibited the secretion of pro-inflammatory cytokines. • The excellent therapeutic potential of nt-p65-TMD was confirmed in sepsis model.« less

Aperture Photometry Tool

NASA Astrophysics Data System (ADS)

Laher, Russ R.; Gorjian, Varoujan; Rebull, Luisa M.; Masci, Frank J.; Fowler, John W.; Helou, George; Kulkarni, Shrinivas R.; Law, Nicholas M.

2012-07-01

Aperture Photometry Tool (APT) is software for astronomers and students interested in manually exploring the photometric qualities of astronomical images. It is a graphical user interface (GUI) designed to allow the image data associated with aperture photometry calculations for point and extended sources to be visualized and, therefore, more effectively analyzed. The finely tuned layout of the GUI, along with judicious use of color-coding and alerting, is intended to give maximal user utility and convenience. Simply mouse-clicking on a source in the displayed image will instantly draw a circular or elliptical aperture and sky annulus around the source and will compute the source intensity and its uncertainty, along with several commonly used measures of the local sky background and its variability. The results are displayed and can be optionally saved to an aperture-photometry-table file and plotted on graphs in various ways using functions available in the software. APT is geared toward processing sources in a small number of images and is not suitable for bulk processing a large number of images, unlike other aperture photometry packages (e.g., SExtractor). However, APT does have a convenient source-list tool that enables calculations for a large number of detections in a given image. The source-list tool can be run either in automatic mode to generate an aperture photometry table quickly or in manual mode to permit inspection and adjustment of the calculation for each individual detection. APT displays a variety of useful graphs with just the push of a button, including image histogram, x and y aperture slices, source scatter plot, sky scatter plot, sky histogram, radial profile, curve of growth, and aperture-photometry-table scatter plots and histograms. APT has many functions for customizing the calculations, including outlier rejection, pixel “picking” and “zapping,” and a selection of source and sky models. The radial-profile-interpolation source model, which is accessed via the radial-profile-plot panel, allows recovery of source intensity from pixels with missing data and can be especially beneficial in crowded fields.

Regulation of two loblolly pine (Pinus taeda L.) isocitrate lyase genes in megagametophytes of mature and stratified seeds and during postgerminative growth.

PubMed

Mullen, R T; Gifford, D J

1997-03-01

Two full-length cDNAs encoding the glyoxysomal enzyme isocitrate lyase (ICL) were isolated from a lambda ZAP cDNA library prepared from megagametophyte mRNAs extracted from seeds imbibed at 30 degrees C for 8 days. The cDNAs, designated Ptbs ICL 8 and Ptbs ICL 12, have open reading frames of 1740 and 1719 bp, with deduced amino acid sequences of 580 and 573 residues, respectively. The predicted amino acid sequences of Ptbs ICL 8 and Ptbs ICL 12 exhibit a 79% identity with each other, and have a greater than 75% identity with ICLs from various angiosperm species. The C-termini of Ptbs ICL 8 and Ptbs ICL 12 terminate with the tripeptide Ser-Arg-Met and Ala-Arg-Met, respectively, both being conserved variants of the type 1 peroxisomal targeting signal. RNA blot and slot analysis revealed that Ptbs ICL 8 and Ptbs ICL 12 mRNAs were present at low levels in the megagametophyte of the mature and stratified seeds, and that the level of both transcripts increased markedly upon seed germination. Protein blot analysis indicated that the steady-state level of ICL was low in the mature and stratified seed, then increased rapidly upon seed germination, peaking at around 8-10 days after imbibition (DAI). Changes in the level of ICL activity in cell-free extracts was similar to the steady-state protein content with the exception that ICL activity was not detected in megagametophyte extracts of mature or stratified seeds. From 10-12 DAI when the megagametophyte tissue senesced, ICL activity decreased rapidly to near undetectable levels. In contrast, steady-state levels of ICL protein and mRNA remained relatively constant during megagametophyte senescence. In vivo synthesis of ICL protein was measured to shed light on these differences. ICL immunoselected from [(35)S]-methionine labelled proteins indicated that ICL was synthesized at very low levels during megagametophyte senescence. Together, the results show that loblolly pine ICL gene expression is complex. While temporal regulation appears to be primarily transcriptional, it also involves a number of post-transcriptional processes including at least one translational and/or post-translational mechanism.

Identification and Characterization of an Immunodominant 28-Kilodalton Coxiella burnetii Outer Membrane Protein Specific to Isolates Associated with Acute Disease

PubMed Central

Zhang, Guoquan; To, Ho; Russell, Kasi E.; Hendrix, Laura R.; Yamaguchi, Tsuyoshi; Fukushi, Hideto; Hirai, Katsuya; Samuel, James E.

2005-01-01

Coxiella burnetii causes acute Q fever in humans and occasional chronic infections that typically manifest as endocarditis or hepatitis. Isolates associated with acute disease were found to be distinct from a group of chronic disease isolates by a variety of biochemical parameters and in a guinea pig fever model of acute disease, suggesting a difference in virulence potential. We compared antigenic polypeptides among C. burnetii isolates and found an immunodominant 28-kDa protein in acute group isolates but not in chronic group isolates (T. Ho, A. Hotta, G. Q. Zhang, S. V. Nguyen, M. Ogawa, T. Yamaguchi, H. Fukushi, and K. Hirai, Microbiol. Immunol. 42:81-85, 1998). In order to clone the adaA gene, the N-terminal amino acid sequence of adaA was determined and a 59-bp fragment was amplified from Nine Mile phase I DNA by PCR. The putative gene fragment was used to screen a lambda ZAP II genomic DNA library, and an open reading frame expressing a 28-kDa immunoreactive protein was identified. Sequence analysis predicted a gene encoding an ∼28-kDa mature protein with a typical signal sequence. The adaA (acute disease antigen A) gene was detected in acute group C. burnetii isolates but not identified in chronic group isolates by PCR and Southern blotting. A typical signal peptide was predicted in adaA, and specific antibody to adaA reacted with the purified membrane fraction of acute group isolates by Western blotting, suggesting that adaA is exposed on the outer surface of C. burnetii. adaA was overexpressed in pET23a as a fusion protein in Escherichia coli to develop anti-recombinant adaA (anti-radaA) specific antibody, which recognized a ∼28-kDa band in acute group isolates but not in chronic group isolates. In addition, immunoblotting indicates that radaA reacted with sera derived from animals infected with acute group isolates but did not react with sera from animals infected with chronic group isolates. These results support the idea that an adaA gene-targeted PCR assay and an radaA antigen-based serodiagnostic test may be useful for differential diagnosis of acute and chronic Q fever. PMID:15731054

Methodology or method? A critical review of qualitative case study reports.

PubMed

Hyett, Nerida; Kenny, Amanda; Dickson-Swift, Virginia

2014-01-01

Despite on-going debate about credibility, and reported limitations in comparison to other approaches, case study is an increasingly popular approach among qualitative researchers. We critically analysed the methodological descriptions of published case studies. Three high-impact qualitative methods journals were searched to locate case studies published in the past 5 years; 34 were selected for analysis. Articles were categorized as health and health services (n=12), social sciences and anthropology (n=7), or methods (n=15) case studies. The articles were reviewed using an adapted version of established criteria to determine whether adequate methodological justification was present, and if study aims, methods, and reported findings were consistent with a qualitative case study approach. Findings were grouped into five themes outlining key methodological issues: case study methodology or method, case of something particular and case selection, contextually bound case study, researcher and case interactions and triangulation, and study design inconsistent with methodology reported. Improved reporting of case studies by qualitative researchers will advance the methodology for the benefit of researchers and practitioners.

Integrating Human Factors into Crew Exploration Vehicle (CEV) Design

NASA Technical Reports Server (NTRS)

Whitmore, Mihriban; Holden, Kritina; Baggerman, Susan; Campbell, Paul

2007-01-01

The purpose of this design process is to apply Human Engineering (HE) requirements and guidelines to hardware/software and to provide HE design, analysis and evaluation of crew interfaces. The topics include: 1) Background/Purpose; 2) HE Activities; 3) CASE STUDY: Net Habitable Volume (NHV) Study; 4) CASE STUDY: Human Modeling Approach; 5) CASE STUDY: Human Modeling Results; 6) CASE STUDY: Human Modeling Conclusions; 7) CASE STUDY: Human-in-the-Loop Evaluation Approach; 8) CASE STUDY: Unsuited Evaluation Results; 9) CASE STUDY: Suited Evaluation Results; 10) CASE STUDY: Human-in-the-Loop Evaluation Conclusions; 11) Near-Term Plan; and 12) In Conclusion

Enhancing the quality of case studies in health services research.

PubMed Central

Yin, R K

1999-01-01

OBJECTIVE: To provide guidance on improving the quality of case studies in health services research. DATA SOURCES: Secondary data, drawing from previous case study research. RESEARCH DESIGN: Guidance is provided to two audiences: potential case study investigators (eight items) and reviewers of case study proposals (four additional items). PRINCIPAL FINDINGS: The guidance demonstrates that many operational steps can be undertaken to improve the quality of case studies. These steps have been a hallmark of high-quality case studies in related fields but have not necessarily been practiced in health services research. CONCLUSIONS: Given higher-quality case studies, the case study method can become a valuable tool for health services research. Images Figure 3 PMID:10591280

Developing the DESCARTE Model: The Design of Case Study Research in Health Care.

PubMed

Carolan, Clare M; Forbat, Liz; Smith, Annetta

2016-04-01

Case study is a long-established research tradition which predates the recent surge in mixed-methods research. Although a myriad of nuanced definitions of case study exist, seminal case study authors agree that the use of multiple data sources typify this research approach. The expansive case study literature demonstrates a lack of clarity and guidance in designing and reporting this approach to research. Informed by two reviews of the current health care literature, we posit that methodological description in case studies principally focuses on description of case study typology, which impedes the construction of methodologically clear and rigorous case studies. We draw from the case study and mixed-methods literature to develop the DESCARTE model as an innovative approach to the design, conduct, and reporting of case studies in health care. We examine how case study fits within the overall enterprise of qualitatively driven mixed-methods research, and the potential strengths of the model are considered. © The Author(s) 2015.

Case Study: Testing with Case Studies

ERIC Educational Resources Information Center

Herreid, Clyde Freeman

2015-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. This month's issue discusses using case studies to test for knowledge or lessons learned.

Methodology or method? A critical review of qualitative case study reports

PubMed Central

Hyett, Nerida; Kenny, Amanda; Dickson-Swift, Virginia

2014-01-01

Despite on-going debate about credibility, and reported limitations in comparison to other approaches, case study is an increasingly popular approach among qualitative researchers. We critically analysed the methodological descriptions of published case studies. Three high-impact qualitative methods journals were searched to locate case studies published in the past 5 years; 34 were selected for analysis. Articles were categorized as health and health services (n=12), social sciences and anthropology (n=7), or methods (n=15) case studies. The articles were reviewed using an adapted version of established criteria to determine whether adequate methodological justification was present, and if study aims, methods, and reported findings were consistent with a qualitative case study approach. Findings were grouped into five themes outlining key methodological issues: case study methodology or method, case of something particular and case selection, contextually bound case study, researcher and case interactions and triangulation, and study design inconsistent with methodology reported. Improved reporting of case studies by qualitative researchers will advance the methodology for the benefit of researchers and practitioners. PMID:24809980

Case Study: Writing a Journal Case Study

ERIC Educational Resources Information Center

Prud'homme-Genereux, Annie

2016-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. This month's issue describes incorporating a journal article into the classroom by first converting it into a case study.

Fostering Entrepreneurship through Business Incubation: The Role and Prospects of Postsecondary Vocational-Technical Education. Report 2: Case Studies.

ERIC Educational Resources Information Center

Hernandez-Gantes, Victor M.; And Others

This report presents nine case studies featuring efforts to foster community development through business incubation and entrepreneurial programs at two-year colleges. A background section discusses case description, and organization and format of case studies. The second section describes the case study design. Each case study is introduced with…

The Misconception of Case-Control Studies in the Plastic Surgery Literature: A Literature Audit.

PubMed

Hatchell, Alexandra C; Farrokhyar, Forough; Choi, Matthew

2017-06-01

Case-control study designs are commonly used. However, many published case-control studies are not true case-controls and are in fact mislabeled. The purpose of this study was to identify all case-control studies published in the top three plastic surgery journals over the past 10 years, assess which were truly case-control studies, clarify the actual design of the articles, and address common misconceptions. MEDLINE, Embase, and Web of Science databases were searched for case-control studies in the three highest-impact factor plastic surgery journals (2005 to 2015). Two independent reviewers screened the resulting titles, abstracts, and methods, if applicable, to identify articles labeled as case-control studies. These articles were appraised and classified as true case-control studies or non-case-control studies. The authors found 28 articles labeled as case-control studies. However, only six of these articles (21 percent) were truly case-control designs. Of the 22 incorrectly labeled studies, one (5 percent) was a randomized controlled trial, three (14 percent) were nonrandomized trials, two (9 percent) were prospective comparative cohort designs, 14 (64 percent) were retrospective comparative cohort designs, and two (9 percent) were cross-sectional designs. The mislabeling was worse in recent years, despite increases in evidence-based medicine awareness. The majority of published case-control studies are not in fact case-control studies. This misunderstanding is worsening with time. Most of these studies are actually comparative cohort designs. However, some studies are truly clinical trials and thus a higher level of evidence than originally proposed.

Case Study: Puttin' on the Ritz: How to Put Science into Cases

ERIC Educational Resources Information Center

Herreid, Clyde Freeman

2017-01-01

There are multiple ways to put science into a case. This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. This month's issue discusses different ways of presenting science in case studies.

Coffee, decaffeinated coffee, tea and cancer of the colon and rectum: a review of epidemiological studies, 1990-2003.

PubMed

Tavani, Alessandra; La Vecchia, Carlo

2004-10-01

The literature from 1990 to 2003 on the relation between coffee, decaffeinated coffee, tea and colorectal cancer risk has been reviewed. For the relation with coffee, three cohort (517 total cases) and nine case-control studies (7555 cases) analysed colon cancer; three cohort (307 cases) and four case-control studies (2704 cases) rectal cancer; six case-control studies (854 cases) colorectal cancer. For colon cancer most case-control studies found risk estimates below unity; the results are less clear for cohort studies. No relation emerged for rectal cancer. A meta-analysis, including five cohort and twelve case-control studies, reported a pooled relative risk of 0.76 (significant). Any methodological artefact is unlikely to account for the consistent inverse association in different countries and settings. Plausible biological explanations include coffee-related reductions of cholesterol, bile acids and neutral sterol secretion in the colon; antimutagenic properties of selected coffee components; increased colonic motility. Decaffeinated coffee was not related to either colon or rectal cancer in three case-control studies. No overall association between tea and either colon or rectal cancer risk emerged in seven cohort (1756 total cases of colon, 759 of rectal and 60 of colorectal cancer) and 12 case-control studies (8058 cases of colon, 4865 of rectal, 604 of colorectal cancer).

Al-Qaeda in Iraq (AQI): An Al-Qaeda Affiliate Case Study

DTIC Science & Technology

2017-10-01

a comparative methodology that included eight case studies on groups affiliated or associated with Al-Qaeda. These case studies were then used as a... methodology that included eight case studies on groups affiliated or associated with Al-Qaeda. These case studies were then used as a dataset for cross...Case Study Zack Gold With contributions from Pamela G. Faber October 2017 This work was performed under Federal Government

Discovering the Future of the Case Study Method in Evaluation Research.

ERIC Educational Resources Information Center

Yin, Robert K.

1994-01-01

It is assumed that evaluators of the future will still be interested in case study methodology. Scenarios that ignore a case study method, that look back to a distinctive case study method, and that see the case study method as an integrating force in the qualitative-quantitative debate are explored. (SLD)

Qualitative Case Study Research as Empirical Inquiry

ERIC Educational Resources Information Center

Ellinger, Andrea D.; McWhorter, Rochell

2016-01-01

This article introduces the concept of qualitative case study research as empirical inquiry. It defines and distinguishes what a case study is, the purposes, intentions, and types of case studies. It then describes how to determine if a qualitative case study is the preferred approach for conducting research. It overviews the essential steps in…

Assembling a Case Study Tool Kit: 10 Tools for Teaching with Cases

ERIC Educational Resources Information Center

Prud'homme-Généreux, Annie

2017-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. The author shares the strategies and tools that teachers can use to manage a case study classroom effectively.

The case-control design in veterinary sciences: A survey.

PubMed

Cullen, Jonah N; Sargeant, Jan M; Makielski, Kelly M; O'Connor, Annette M

2016-11-01

The case-control study design is deceptively simple. However, many design considerations influence the estimated effect measure. An investigation of case-control studies in the human health literature suggested that some of these considerations are not described in reports of case-control studies. Our hypothesis was that the majority of veterinary studies labeled as case-controls would be incident density designs, and many would not interpret the effect measure obtained from those studies as the rate ratio rather than the odds ratio. Reference databases were searched for author-designated case-control studies. A survey of 100 randomly selected studies was conducted to examine the different design options described and estimated effect measures. Of the 100 author-identified case-control studies, 83 assessed an exposure-outcome association and, of those, only 54 (65.1%) sampled the study population based on an outcome and would thus be considered case-control designs. Twelve studies were incidence density designs but none used this terminology. Of the studies that reported an odds ratio as the effect measure, none reported on additional considerations that would have enabled a more interpretable result. This survey indicated many case-control-labeled studies were not case-control designs and among case-control studies, key design aspects were not often described. The absence of information about study design elements and underlying assumptions in case-control studies limits the ability to establish the effect measured by the study and the evidentiary value of the study might be underestimated. Copyright © 2016 Elsevier B.V. All rights reserved.

Beyond Clinical Case Studies in Psychoanalysis: A Review of Psychoanalytic Empirical Single Case Studies Published in ISI-Ranked Journals.

PubMed

Meganck, Reitske; Inslegers, Ruth; Krivzov, Juri; Notaerts, Liza

2017-01-01

Single case studies are at the origin of both theory development and research in the field of psychoanalysis and psychotherapy. While clinical case studies are the hallmark of psychoanalytic theory and practice, their scientific value has been strongly criticized. To address problems with the subjective bias of retrospective therapist reports and uncontrollability of clinical case studies, systematic approaches to investigate psychotherapy process and outcome at the level of the single case have been developed. Such empirical case studies are also able to bridge the famous gap between academic research and clinical practice as they provide clinically relevant insights into how psychotherapy works. This study presents a review of psychoanalytic empirical case studies published in ISI-ranked journals and maps the characteristics of the study, therapist, patient en therapies that are investigated. Empirical case studies increased in quantity and quality (amount of information and systematization) over time. While future studies could pay more attention to providing contextual information on therapist characteristics and informed consent considerations, the available literature provides a basis to conduct meta-studies of single cases and as such contribute to knowledge aggregation.

Beyond Clinical Case Studies in Psychoanalysis: A Review of Psychoanalytic Empirical Single Case Studies Published in ISI-Ranked Journals

PubMed Central

Meganck, Reitske; Inslegers, Ruth; Krivzov, Juri; Notaerts, Liza

2017-01-01

Single case studies are at the origin of both theory development and research in the field of psychoanalysis and psychotherapy. While clinical case studies are the hallmark of psychoanalytic theory and practice, their scientific value has been strongly criticized. To address problems with the subjective bias of retrospective therapist reports and uncontrollability of clinical case studies, systematic approaches to investigate psychotherapy process and outcome at the level of the single case have been developed. Such empirical case studies are also able to bridge the famous gap between academic research and clinical practice as they provide clinically relevant insights into how psychotherapy works. This study presents a review of psychoanalytic empirical case studies published in ISI-ranked journals and maps the characteristics of the study, therapist, patient en therapies that are investigated. Empirical case studies increased in quantity and quality (amount of information and systematization) over time. While future studies could pay more attention to providing contextual information on therapist characteristics and informed consent considerations, the available literature provides a basis to conduct meta-studies of single cases and as such contribute to knowledge aggregation. PMID:29046660

Does Case-Study Methodology Lack Rigour? The Need for Quality Criteria for Sound Case-Study Research, as Illustrated by a Recent Case in Secondary and Higher Education

ERIC Educational Resources Information Center

Kyburz-Graber, Regula

2004-01-01

There is a tendency to use case-study research methodology for research issues aiming at simply describing a complex situation, and to draw conclusions with insufficient rigour. Sound case-study research, however, follows discriminate rules which can be described in all the dimensions of a full case-study research process. This paper examines…

An upper limit on Early Mars atmospheric pressure from small ancient craters

NASA Astrophysics Data System (ADS)

Kite, E. S.; Williams, J.; Lucas, A.; Aharonson, O.

2012-12-01

Planetary atmospheres brake, ablate, and disrupt small asteroids and comets, filtering out small hypervelocity surface impacts and causing fireballs, airblasts, meteors, and meteorites. Hypervelocity craters <1 km diameter on Earth are typically caused by irons (because stones are more likely to break up), and the smallest hypervelocity craters near sea-level on Earth are ~20 m in diameter. 'Zap pits' as small as 30 microns are known from the airless moon, but the other airy worlds show the effects of progressively thicker atmospheres:- the modern Mars atmosphere is marginally capable of removing >90% of the kinetic energy of >240 kg iron impactors; Titan's paucity of small craters is consistent with a model predicting atmospheric filtering of craters smaller than 6-8km; and on Venus, craters below ~20 km diameter are substantially depleted. Changes in atmospheric CO2 concentration are believed to be the single most important control on Mars climate evolution and habitability. Existing data requires an early epoch of massive atmospheric loss to space; suggests that the present-day rate of escape to space is small; and offers only limited evidence for carbonate formation. Existing evidence has not led to convergence of atmosphere-evolution models, which must balance poorly understood fluxes from volcanic degassing, surface weathering, and escape to space. More direct measurements are required in order to determine the history of CO2 concentrations. Wind erosion and tectonics exposes ancient surfaces on Mars, and the size-frequency distribution of impacts on these surfaces has been previously suggested as a proxy time series of Mars atmospheric thickness. We will present a new upper limit on Early Mars atmospheric pressure using the size-frequency distribution of 20-100m diameter ancient craters in Aeolis Dorsa, validated using HiRISE DTMs, in combination with Monte Carlo simulations of the effect of paleo-atmospheres of varying thickness on the crater flux. These craters are interbedded with river deposits, and so the atmospheric state they record corresponds to an era when Mars was substantially wetter than the present, probably >3.7 Ga. An important caveat is that our technique cannot exclude atmospheric collapse-reinflation cycles on timescales much shorter than the sedimentary basin-filling time, so it sets an upper limit on the density of a thick stable paleoatmosphere. We will discuss our results in relation to previous estimates of ancient atmospheric pressure, and place new constraints on models of Early Mars climate.

Autonomous Selection of a Rover Laser Target on Mars

NASA Image and Video Library

2016-07-21

NASA's Curiosity Mars rover autonomously selects some of the targets for the laser and telescopic camera of the rover's Chemistry and Camera (ChemCam) instrument. For example, on-board software analyzed the image on the left, chose the target highlighted with the yellow dot, and pointed ChemCam to acquire laser analysis and the image on the right. Most ChemCam targets are still selected by scientists discussing rocks or soil seen in images the rover has sent to Earth, but the autonomous targeting provides an added capability. It can offer a head start on acquiring composition information at a location just reached by a drive. The software for target selection and instrument pointing is called AEGIS, for Autonomous Exploration for Gathering Increased Science. The image on the left was taken by the left eye of Curiosity's stereo Navigation Camera (Navcam) a few minutes after the rover completed a drive of about 43 feet (13 meters) on July 14, 2016, during the 1,400th Martian day, or sol, of the rover's work on Mars. Using AEGIS for target selection and pointing based on the Navcam imagery, Curiosity's ChemCam zapped a grid of nine points on a rock chosen for meeting criteria set by the science team. In this run, parameters were set to find bright-toned outcrop rock rather than darker rocks, which in this area tend to be loose on the surface. Within less than 30 minutes after the Navcam image was taken, ChemCam had used its laser on all nine points and had taken before-and-after images of the target area with its remote micro-imager (RMI) camera. The image at right combines those two RMI exposures. The nine laser targets are marked in red at the center. On the Navcam image at left, the yellow dot identifies the selected target area, which is about 2.2 inches (5.6 centimeters) in diameter. An unannotated version of this Sol 1400 Navcam image is available. ChemCam records spectra of glowing plasma generated when the laser hits a target point. These spectra provide information about the chemical elements present in the target. The light-toned patch of bedrock identified by AEGIS on Sol 1400 appears, geochemically, to belong to the "Stimson" sandstone unit of lower Mount Sharp. In mid-2016, Curiosity typically uses AEGIS for selecting a ChemCam target more than once per week. http://photojournal.jpl.nasa.gov/catalog/PIA20762

[Application of nested case-control study on safe evaluation of post-marketing traditional Chinese medicine injection].

PubMed

Xiao, Ying; Zhao, Yubin; Xie, Yanming

2011-10-01

The nested case-control study design (or the case-control in a cohort study) is described here as a new study design used in safe evaluation of post-marketing traditional Chinese medicine injection. In the nested case-control study, cases of a disease that occur in a defined cohort are identified and, for each, a specified number of matched controls is selected from among those in the cohort who have not developed the disease by the time of disease occurrence in the case. For many research questions, the nested case-control design potentially offers impressive reductions in costs and efforts of data collection and analysis compared with the full cohort approach, with relatively minor loss in statistical efficiency. The nested case-control design is particularly advantageous for studies in safe evaluation of post-marketing traditional Chinese medicine injection. Some examples of the application of nested case-control study were given.

Qualitative case study methodology in nursing research: an integrative review.

PubMed

Anthony, Susan; Jack, Susan

2009-06-01

This paper is a report of an integrative review conducted to critically analyse the contemporary use of qualitative case study methodology in nursing research. Increasing complexity in health care and increasing use of case study in nursing research support the need for current examination of this methodology. In 2007, a search for case study research (published 2005-2007) indexed in the CINAHL, MEDLINE, EMBASE, PsychINFO, Sociological Abstracts and SCOPUS databases was conducted. A sample of 42 case study research papers met the inclusion criteria. Whittemore and Knafl's integrative review method guided the analysis. Confusion exists about the name, nature and use of case study. This methodology, including terminology and concepts, is often invisible in qualitative study titles and abstracts. Case study is an exclusive methodology and an adjunct to exploring particular aspects of phenomena under investigation in larger or mixed-methods studies. A high quality of case study exists in nursing research. Judicious selection and diligent application of literature review methods promote the development of nursing science. Case study is becoming entrenched in the nursing research lexicon as a well-accepted methodology for studying phenomena in health and social care, and its growing use warrants continued appraisal to promote nursing knowledge development. Attention to all case study elements, process and publication is important in promoting authenticity, methodological quality and visibility.

Online Learning and Teaching with Technology: Case Studies, Experience and Practice. Case Studies of Teaching in Higher Education.

ERIC Educational Resources Information Center

Murphy, David, Ed.; Walker, Rob, Ed.; Webb, Graham, Ed.

This book contains case studies that look at using technology in a wide range of situations, from fully online courses to more traditional face-to-face settings. The case studies deal with issues related to student interaction, teaching and assessment, planning and development, and policy. The following case studies are included: (1) "Flame…

Case Study: Mini-Case Studies: Small Infusions of Active Learning for Large-Lecture Courses

ERIC Educational Resources Information Center

Carloye, Lisa

2017-01-01

In this article, the author introduces the usage of case studies to be an excellent method for engaging students through stories. The author notes she developed a series of mini-case studies that can be implemented, with a little advance preparation, within a 10- to 15-minute window during lecture. What makes them "mini" case studies?…

Lively Discussions vs. Discussions of Substance: Making the Case for Guided Case Study.

ERIC Educational Resources Information Center

Woloshin/Lerman, Phyllis

1996-01-01

Discusses the history and purpose of the case study method used to stimulate in-class discussion. Advocates the use of the guided case study method to ensure that substantive discussions take place and provides nine steps for using the method. Includes a sample case study worksheet. (11 citations) (BCY)

Case Study: Guidelines for Producing Videos to Accompany Flipped Cases

ERIC Educational Resources Information Center

Prud'homme-Généreux, Annie; Schiller, Nancy A.; Wild, John H.; Herreid, Clyde Freeman

2017-01-01

Three years ago, the "National Center for Case Study Teaching in Science" (NCCSTS) was inspired to merge the case study and flipped classroom approaches. The resulting project aimed to create the materials required to teach a flipped course in introductory biology by assigning videos as homework and case studies in the classroom. Three…

Implementation of case studies in undergraduate didactic nursing courses: a qualitative study.

PubMed

Dutra, Danette K

2013-07-04

The implementation of unfolding scenario-based case studies in the didactic classroom is associated with learner-centered education. The utilization of learner-centered pedagogies, such as case studies, removes the focus from the instructor and instead places it on the student. Learner-centered pedagogies are believed to improve students' levels of cognition. The purpose of this study was to examine how nurse educators are implementing the pedagogies of case studies in their undergraduate didactic courses. The goal was to examine, document, report, and, ultimately, implement the strategies. Purposeful sampling was utilized in this qualitative, multisite-designed study. For each of the four participants, three separate site visits were completed. Observations and post-observational interviews took place at each site visit. Transcribed data from interviews, observations, and course documents were imported into the computer program Nvivo8. Repetitive comparative analysis was utilized to complete the data coding process. The guiding research question of this study sought to investigate the implementation strategies of case studies in didactic nursing courses. The implementation of case studies by the participants reflected two primary patterns: Formal Implementation (FI) and Informal Implementation (II) of case studies. The FI of case studies was further divided into two subcategories: Formal Implementation of case studies used Inside the Classroom setting (FIIC) and Formal Implementation of cases studies used Outside of the Classroom (FIOC). Results of this investigation have led to an increased understanding of implementation strategies of unfolding scenario-based case studies in undergraduate nursing didactic courses. Data collected were rich in the description of specific methodologies for utilization of case studies and may serve as a resource for faculty in development of creative strategies to enhance the didactic classroom experience.

Implementation of case studies in undergraduate didactic nursing courses: a qualitative study

PubMed Central

2013-01-01

Background The implementation of unfolding scenario-based case studies in the didactic classroom is associated with learner-centered education. The utilization of learner-centered pedagogies, such as case studies, removes the focus from the instructor and instead places it on the student. Learner-centered pedagogies are believed to improve students’ levels of cognition. The purpose of this study was to examine how nurse educators are implementing the pedagogies of case studies in their undergraduate didactic courses. The goal was to examine, document, report, and, ultimately, implement the strategies. Methods Purposeful sampling was utilized in this qualitative, multisite-designed study. For each of the four participants, three separate site visits were completed. Observations and post-observational interviews took place at each site visit. Transcribed data from interviews, observations, and course documents were imported into the computer program Nvivo8. Repetitive comparative analysis was utilized to complete the data coding process. Results The guiding research question of this study sought to investigate the implementation strategies of case studies in didactic nursing courses. The implementation of case studies by the participants reflected two primary patterns: Formal Implementation (FI) and Informal Implementation (II) of case studies. The FI of case studies was further divided into two subcategories: Formal Implementation of case studies used Inside the Classroom setting (FIIC) and Formal Implementation of cases studies used Outside of the Classroom (FIOC). Conclusion Results of this investigation have led to an increased understanding of implementation strategies of unfolding scenario-based case studies in undergraduate nursing didactic courses. Data collected were rich in the description of specific methodologies for utilization of case studies and may serve as a resource for faculty in development of creative strategies to enhance the didactic classroom experience. PMID:23826925

The Use of Case Studies in Teaching Undergraduate Neuroscience

PubMed Central

Meil, William M.

2007-01-01

Case studies have been the cornerstone of many discoveries in neurology and continue to be an indispensable source of knowledge. Attaching a name, face, and story to the study of neurological disorders makes them more “real” and memorable. This article describes the value of the case study methodology and its advantages as a pedagogical approach. It also illustrates how the seminal case of H.M. can be used to highlight the advantages and disadvantages of the case study methodology. Three exercises are described for incorporating case studies into neuroscience courses. The first exercise requires students to conduct a literature review regarding their assigned case and then design an experiment to address a lingering question regarding that neurological disorder. Survey results of 90 students provide quantitative and qualitative support for this approach. The vast majority of students indicated this exercise was a valuable learning experience; sparked interest in the topic and in biopsychology; increased their knowledge and stimulated critical thinking. The second exercise discusses how students might conduct their own case studies. The third exercise emphasizes the use of case studies as a platform to examine competing hypotheses regarding neurological conditions and their treatment. A table listing case studies appropriate for undergraduate neuroscience courses is included. Cases are categorized by the type of neurological disorder and notes regarding the nature of and content of each case are provided. PMID:23493154

Abu Sayyaf Group (ASG): An Al-Qaeda Associate Case Study

DTIC Science & Technology

2017-10-01

completed in August 2017. In order to conduct this assessment, CNA used a comparative methodology that included eight case studies on groups affiliated...assessment, CNA used a comparative methodology that included eight case studies on groups affiliated or associated with Al-Qaeda. These case studies ...Case Study P. Kathleen Hammerberg and Pamela G. Faber With contributions from Alexander Powell October 2017 This work was performed

OD in Schools: The State of the Art. Vol. IV: Case Studies. Final Report.

ERIC Educational Resources Information Center

Fullan, Michael; And Others

This volume, the fourth of a five-volume series, contains three onsite case studies of organization development (OD) selected from a sample of 76 school districts. The purpose of the case studies was to analyze in detail different types of OD programs and their use. Each case study represents a different OD focus: case study A involves a survey…

World Perspective Case Descriptions on Educational Programs for Adults: Cameroon.

ERIC Educational Resources Information Center

Nyemba, J. A.

This document contains two case studies of adult education programs in Cameroon. Both case studies begin with a "face sheet" on which is recorded basic information about the program and the case study itself. One case study is intended to evaluate the impact of the agricultural university center in the Dschang area from 1977-1987 and to…

Turning to Case Studies as a Mechanism for Learning in Action Learning

ERIC Educational Resources Information Center

O'Leary, Denise; Coughlan, Paul; Rigg, Clare; Coghlan, David

2017-01-01

Case studies are a useful means of capturing and sharing experiential knowledge by allowing researchers to explore the social, organisational and political contexts of a specific case. Although accounts of action learning are often reported using a case study approach, it is not common to see individual case studies being used as a learning…

Making a Case for a Blended Approach: The Need for The Design-Based Case Study

ERIC Educational Resources Information Center

Deaton, Cynthia C. M.; Malloy, Jacquelynn A.

2017-01-01

Design-based case studies address research questions that involve instructional innovations within a bounded system. This blend of case study and design-based research provides a systematic approach to examining instructional innovations that are bounded by perspective, context, and time. Design-based case studies provide a framework for engaging…

The Effect of Using Case Studies in Business Statistics

ERIC Educational Resources Information Center

Pariseau, Susan E.; Kezim, Boualem

2007-01-01

The authors evaluated the effect on learning of using case studies in business statistics courses. The authors divided students into 3 groups: a control group, a group that completed 1 case study, and a group that completed 3 case studies. Results evidenced that, on average, students whom the authors required to complete a case analysis received…

CASE STUDY CRITIQUE; UPPER CLINCH CASE STUDY

EPA Science Inventory

Case study critique: Upper Clinch case study (from Research on Methods for Integrating Ecological Economics and Ecological Risk Assessment: A Trade-off Weighted Index Approach to Integrating Economics and Ecological Risk Assessment). This critique answers the questions: 1) does ...

Pathology informatics fellowship retreats: The use of interactive scenarios and case studies as pathology informatics teaching tools.

PubMed

Lee, Roy E; McClintock, David S; Balis, Ulysses J; Baron, Jason M; Becich, Michael J; Beckwith, Bruce A; Brodsky, Victor B; Carter, Alexis B; Dighe, Anand S; Haghighi, Mehrvash; Hipp, Jason D; Henricks, Walter H; Kim, Jiyeon Y; Klepseis, Veronica E; Kuo, Frank C; Lane, William J; Levy, Bruce P; Onozato, Maristela L; Park, Seung L; Sinard, John H; Tuthill, Mark J; Gilbertson, John R

2012-01-01

Last year, our pathology informatics fellowship added informatics-based interactive case studies to its existing educational platform of operational and research rotations, clinical conferences, a common core curriculum with an accompanying didactic course, and national meetings. The structure of the informatics case studies was based on the traditional business school case study format. Three different formats were used, varying in length from short, 15-minute scenarios to more formal multiple hour-long case studies. Case studies were presented over the course of three retreats (Fall 2011, Winter 2012, and Spring 2012) and involved both local and visiting faculty and fellows. Both faculty and fellows found the case studies and the retreats educational, valuable, and enjoyable. From this positive feedback, we plan to incorporate the retreats in future academic years as an educational component of our fellowship program. Interactive case studies appear to be valuable in teaching several aspects of pathology informatics that are difficult to teach in more traditional venues (rotations and didactic class sessions). Case studies have become an important component of our fellowship's educational platform.

[Case-non case studies: Principles, methods, bias and interpretation].

PubMed

Faillie, Jean-Luc

2017-10-31

Case-non case studies belongs to the methods assessing drug safety by analyzing the disproportionality of notifications of adverse drug reactions in pharmacovigilance databases. Used for the first time in the 1980s, the last few decades have seen a significant increase in the use of this design. The principle of the case-non case study is to compare drug exposure in cases of a studied adverse reaction with that of cases of other reported adverse reactions and called "non cases". Results are presented in the form of a reporting odds ratio (ROR), the interpretation of which makes it possible to identify drug safety signals. This article describes the principle of the case-non case study, the method of calculating the ROR and its confidence interval, the different modalities of analysis and how to interpret its results with regard to the advantages and limitations of this design. Copyright © 2017 Société française de pharmacologie et de thérapeutique. Published by Elsevier Masson SAS. All rights reserved.

Environmental Technology Verification (ETV) Program Case Studies: Demonstrating Program Outcomes, Volume III

EPA Science Inventory

This booklet, ETV Program Case Studies: Demonstrating Program Outcomes, Volume III contains two case studies, addressing verified environmental technologies for decentalized wastewater treatment and converting animal waste to energy. Each case study contains a brief description ...

Joint Forces Command - Operation United Assistance Case Study: Lessons and Best Practices

DTIC Science & Technology

2016-07-01

additional and prioritized computers and access in the operations center for these mission requirements are essential. 127 JFC-OUA CASE STUDY Issue...this publication is welcomed and highly encouraged. Joint Forces Command – Operation United Assistance Case Study JFC-OUA CASE STUDY iii Foreword...Based on information drawn from various sources including after action reports, lessons learned, case studies , umbrella-week visits, and key-leader

Case Study Teaching

ERIC Educational Resources Information Center

Herreid, Clyde Freeman

2011-01-01

This chapter describes the history of case study teaching, types of cases, and experimental data supporting their effectiveness. It also describes a model for comparing the efficacy of the various case study methods. (Contains 1 figure.)

Case Studies for School Administrators: Managing Change in Education.

ERIC Educational Resources Information Center

Benham, Maenette K. P. Ah Nee

This book examines case-based learning in educational leadership courses, discusses case-based learning as an educational tool, exemplifies methods of writing a case study, and contains 14 case-studies by teachers and administrators. "Stakeholders in a House of Cards," by Audrey Burgher, discusses integrating technology with innovation.…

How To Set Up Your Own Small Business. Service Company Case Study. Manufacturing Firm Case Study. Retail Store Case Study.

ERIC Educational Resources Information Center

Fallek, Max

This collection of case studies is intended for use in a course in setting up a small business. The first, a case study of the process of setting up a service company, covers analyzing the pros and cons of starting one's own business, assessing the competition and local market, and selecting a site for and financing the business. The principal…

[The case-case-time-control study design].

PubMed

Wang, Jing; Zhuo, Lin; Zhan, Siyan

2014-12-01

Although the 'self-matched case-only studies' (such as the case-cross-over or self-controlled case-series method) can control the time-invariant confounders (measured or unmeasured) through design of the study, however, they can not control those confounders that vary with time. A bidirectional case-crossover design can be used to adjust the exposure-time trends. In the areas of pharmaco-epidemiology, illness often influence the future use of medications, making a bidirectional study design problematic. Suissa's case-time-control design combines the case-crossover and the case-control design which could adjust for exposure-trend bias, but the control group may reintroduce selection bias, if the matching does not go well. We propose a "case-case-time-control" design which is an extension of the case-time-control design. However, rather than using a sample of external controls, we choose those future cases as controls for current cases to counter the bias that arising from temporal trends caused by exposure to the target of interest. In the end of this article we will discuss the strength and limitations of this design based on an applied example.

Real-Life Case Studies for Teachers.

ERIC Educational Resources Information Center

Hayes, William

Case studies described in this book reflect conditions present in today's public schools. Situations described in these case studies are intended to introduce education students to the variety of problems existing in today's schools. The 38 case studies highlight: student cheating; teacher's observation by administrator; inclusion; contract…

The Case Study as Research Heuristic: Lessons from the R&D Value Mapping Project.

ERIC Educational Resources Information Center

Bozeman, Barry; Klein, Hans K.

1999-01-01

Examines the role of prototype case studies as the foundation for later evaluation through two studies from the "R&D Value Mapping Project," a study that will involve more than 30 cases. Explores the usefulness of case studies in defining and assessing subsequent research efforts. (SLD)

A Careful Look at Modern Case Selection Methods

ERIC Educational Resources Information Center

Herron, Michael C.; Quinn, Kevin M.

2016-01-01

Case studies appear prominently in political science, sociology, and other social science fields. A scholar employing a case study research design in an effort to estimate causal effects must confront the question, how should cases be selected for analysis? This question is important because the results derived from a case study research program…

Case-control studies in neurosurgery.

PubMed

Nesvick, Cody L; Thompson, Clinton J; Boop, Frederick A; Klimo, Paul

2014-08-01

Observational studies, such as cohort and case-control studies, are valuable instruments in evidence-based medicine. Case-control studies, in particular, are becoming increasingly popular in the neurosurgical literature due to their low cost and relative ease of execution; however, no one has yet systematically assessed these types of studies for quality in methodology and reporting. The authors performed a literature search using PubMed/MEDLINE to identify all studies that explicitly identified themselves as "case-control" and were published in the JNS Publishing Group journals (Journal of Neurosurgery, Journal of Neurosurgery: Pediatrics, Journal of Neurosurgery: Spine, and Neurosurgical Focus) or Neurosurgery. Each paper was evaluated for 22 descriptive variables and then categorized as having either met or missed the basic definition of a case-control study. All studies that evaluated risk factors for a well-defined outcome were considered true case-control studies. The authors sought to identify key features or phrases that were or were not predictive of a true case-control study. Those papers that satisfied the definition were further evaluated using the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist. The search detected 67 papers that met the inclusion criteria, of which 32 (48%) represented true case-control studies. The frequency of true case-control studies has not changed with time. Use of odds ratios (ORs) and logistic regression (LR) analysis were strong positive predictors of true case-control studies (for odds ratios, OR 15.33 and 95% CI 4.52-51.97; for logistic regression analysis, OR 8.77 and 95% CI 2.69-28.56). Conversely, negative predictors included focus on a procedure/intervention (OR 0.35, 95% CI 0.13-0.998) and use of the word "outcome" in the Results section (OR 0.23, 95% CI 0.082-0.65). After exclusion of nested case-control studies, the negative correlation between focus on a procedure/intervention and true case-control studies was strengthened (OR 0.053, 95% CI 0.0064-0.44). There was a trend toward a negative association between the use of survival analysis or Kaplan-Meier curves and true case-control studies (OR 0.13, 95% CI 0.015-1.12). True case-control studies were no more likely than their counterparts to use a potential study design "expert" (OR 1.50, 95% CI 0.57-3.95). The overall average STROBE score was 72% (range 50-86%). Examples of reporting deficiencies were reporting of bias (28%), missing data (55%), and funding (44%). The results of this analysis show that the majority of studies in the neurosurgical literature that identify themselves as "case-control" studies are, in fact, labeled incorrectly. Positive and negative predictors were identified. The authors provide several recommendations that may reverse the incorrect and inappropriate use of the term "case-control" and improve the quality of design and reporting of true case-control studies in neurosurgery.

Methodology Series Module 2: Case-control Studies.

PubMed

Setia, Maninder Singh

2016-01-01

Case-Control study design is a type of observational study. In this design, participants are selected for the study based on their outcome status. Thus, some participants have the outcome of interest (referred to as cases), whereas others do not have the outcome of interest (referred to as controls). The investigator then assesses the exposure in both these groups. The investigator should define the cases as specifically as possible. Sometimes, definition of a disease may be based on multiple criteria; thus, all these points should be explicitly stated in case definition. An important aspect of selecting a control is that they should be from the same 'study base' as that of the cases. We can select controls from a variety of groups. Some of them are: General population; relatives or friends; and hospital patients. Matching is often used in case-control control studies to ensure that the cases and controls are similar in certain characteristics, and it is a useful technique to increase the efficiency of the study. Case-Control studies can usually be conducted relatively faster and are inexpensive - particularly when compared with cohort studies (prospective). It is useful to study rare outcomes and outcomes with long latent periods. This design is not very useful to study rare exposures. Furthermore, they may also be prone to certain biases - selection bias and recall bias.

Arsenic Removal - Adsorptive Media and Coagulation/Filtration Case Studies - Slides

EPA Science Inventory

This presentation provides information on the results of three case studies from USEPA arsenic demonstration program. The first case study presented is on the Rimrock, AZ project that used an adsorptive media technology (E33 media) to remove arsenic. The second case study is o...

Arsenic Removal: Adsorptive Media and Coagulation/Filtration Case Studies

EPA Science Inventory

This presentation provides information on the results of three case studies from USEPA arsenic demonstration program. The first case study presented is on the Rimrock, AZ project that used an adsorptive media technology (E33 media) to remove arsenic. The second case study is on...

Putting Words in Their Mouth: Writing Dialogue for Case Studies

ERIC Educational Resources Information Center

Herreid, Clyde Freeman

2018-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. This issue discusses dialogue writing guidelines most relevant to case writing.

Comparison of Estimates between Cohort and Case-Control Studies in Meta-Analyses of Therapeutic Interventions: A Meta-Epidemiological Study.

PubMed

Lanza, Amy; Ravaud, Philippe; Riveros, Carolina; Dechartres, Agnes

2016-01-01

Observational studies are increasingly being used for assessing therapeutic interventions. Case-control studies are generally considered to have greater risk of bias than cohort studies, but we lack evidence of differences in effect estimates between the 2 study types. We aimed to compare estimates between cohort and case-control studies in meta-analyses of observational studies of therapeutic interventions by using a meta-epidemiological study. We used a random sample of meta-analyses of therapeutic interventions published in 2013 that included both cohort and case-control studies assessing a binary outcome. For each meta-analysis, the ratio of estimates (RE) was calculated by comparing the estimate in case-control studies to that in cohort studies. Then, we used random-effects meta-analysis to estimate a combined RE across meta-analyses. An RE < 1 indicated that case-control studies yielded larger estimates than cohort studies. The final analysis included 23 meta-analyses: 138 cohort and 133 case-control studies. Treatment effect estimates did not significantly differ between case-control and cohort studies (combined RE 0.97 [95% CI 0.86-1.09]). Heterogeneity was low, with between-meta-analysis variance τ2 = 0.0049. Estimates did not differ between case-control and prospective or retrospective cohort studies (RE = 1.05 [95% CI 0.96-1.15] and RE = 0.99 [95% CI, 0.83-1.19], respectively). Sensitivity analysis of studies reporting adjusted estimates also revealed no significant difference (RE = 1.03 [95% CI 0.91-1.16]). Heterogeneity was also low for these analyses. We found no significant difference in treatment effect estimates between case-control and cohort studies assessing therapeutic interventions.

Global dengue death before and after the new World Health Organization 2009 case classification: A systematic review and meta-regression analysis.

PubMed

Low, Gary Kim-Kuan; Ogston, Simon A; Yong, Mun-Hin; Gan, Seng-Chiew; Chee, Hui-Yee

2018-06-01

Since the introduction of 2009 WHO dengue case classification, no literature was found regarding its effect on dengue death. This study was to evaluate the effect of 2009 WHO dengue case classification towards dengue case fatality rate. Various databases were used to search relevant articles since 1995. Studies included were cohort and cross-sectional studies, all patients with dengue infection and must report the number of death or case fatality rate. The Joanna Briggs Institute appraisal checklist was used to evaluate the risk of bias of the full-texts. The studies were grouped according to the classification adopted: WHO 1997 and WHO 2009. Meta-regression was employed using a logistic transformation (log-odds) of the case fatality rate. The result of the meta-regression was the adjusted case fatality rate and odds ratio on the explanatory variables. A total of 77 studies were included in the meta-regression analysis. The case fatality rate for all studies combined was 1.14% with 95% confidence interval (CI) of 0.82-1.58%. The combined (unadjusted) case fatality rate for 69 studies which adopted WHO 1997 dengue case classification was 1.09% with 95% CI of 0.77-1.55%; and for eight studies with WHO 2009 was 1.62% with 95% CI of 0.64-4.02%. The unadjusted and adjusted odds ratio of case fatality using WHO 2009 dengue case classification was 1.49 (95% CI: 0.52, 4.24) and 0.83 (95% CI: 0.26, 2.63) respectively, compared to WHO 1997 dengue case classification. There was an apparent increase in trend of case fatality rate from the year 1992-2016. Neither was statistically significant. The WHO 2009 dengue case classification might have no effect towards the case fatality rate although the adjusted results indicated a lower case fatality rate. Future studies are required for an update in the meta-regression analysis to confirm the findings. Copyright © 2018 Elsevier B.V. All rights reserved.

Business and Consumer Education Case Studies.

ERIC Educational Resources Information Center

Delta Pi Epsilon, Minneapolis, Minn. Phi Chapter.

This publication contains 58 case studies for classroom use in teaching various business and consumer education subjects at the high school level. A supplement to a previous Phi Chapter publication, "Office Education Case Studies" (1973), the case studies are intended to create class discussions and help students acquire the ability to analyze…

75 FR 11183 - Proposed Data Collections Submitted for Public Comment and Recommendations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-03-10

... technology. Written comments should be received within 60 days of this notice. Proposed Project Case Studies... case studies of six CPPW-funded states and 15 CPPW-funded communities. The case study sites will be... region, and targeted population. Case study information will be collected by conducting personal...

75 FR 51806 - Climate Change Vulnerability Assessment: Four Case Studies of Water Utility Practices

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-23

... case studies describing the approaches currently being taken by four water utilities to assess their... series of case studies describing the approaches currently being taken by four water utilities in the... Vulnerability Assessment: Four Case Studies of Water Utility Practices AGENCY: Environmental Protection Agency...

Natural Learning Case Study Archives

ERIC Educational Resources Information Center

Lawler, Robert W.

2015-01-01

Natural Learning Case Study Archives (NLCSA) is a research facility for those interested in using case study analysis to deepen their understanding of common sense knowledge and natural learning (how the mind interacts with everyday experiences to develop common sense knowledge). The database comprises three case study corpora based on experiences…

Paradise nearly Gained. Volume 2: Case Studies of Impact and Diversity for Frontline Management Initiative Practice.

ERIC Educational Resources Information Center

Barratt-Pugh, Llandis; Soutar, Geoffrey N.

This document presents the case studies from a multi-phase study of the impact of Australia's Frontline Management Initiative (FMI), which provides a framework for competency-based development of frontline managers in Australian enterprises. Nineteen organizational case studies and one individual case study of the FMI's impacts are included. The…

Developing and pretesting case studies in dental and dental hygiene education: using the diffusion of innovations model.

PubMed

Cragun, Deborah L; DeBate, Rita DiGioacchino; Severson, Herbert H; Shaw, Tracy; Christiansen, Steve; Koerber, Anne; Tomar, Scott L; Brown, Kelli McCormack; Tedesco, Lisa A; Hendricson, William D

2012-05-01

Case-based learning offers exposure to clinical situations that health professions students may not encounter in their training. The purposes of this study were to apply the Diffusion of Innovations conceptual framework to 1) identify characteristics of case studies that would increase their adoption among dental and dental hygiene faculty members and 2) develop and pretest interactive web-based case studies on sensitive oral-systemic health issues. The formative study spanned two phases using mixed methods (Phase 1: eight focus groups and four interviews; Phase 2: ten interviews and satisfaction surveys). Triangulation of quantitative and qualitative data revealed the following positive attributes of the developed case studies: relative advantage of active learning and modeling; compatibility with a variety of courses; observability of case-related knowledge and skills; independent learning; and modifiability for use with other oral-systemic health issues. These positive attributes are expected to increase the likelihood that dental and dental hygiene faculty members will adopt the developed case study once it is available for use. The themes identified in this study could be applied to the development of future case studies and may provide broader insight that might prove useful for exploring differences in case study use across dental and dental hygiene curricula.

Case Study: What Makes a Good Case, Revisited: The Survey Monkey Tells All

ERIC Educational Resources Information Center

Herried, Clyde Freeman; Prud'homme-Genereux, Annie; Schiller, Nancy A.; Herreid, Ky F.; Wright, Carolyn

2016-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. In this month's issue the authors provide a more definitive answer to the "What Makes a Good Case?" question based on a just-completed Survey Monkey survey given to NCCSTS teachers.

Theoretical pluralism in psychoanalytic case studies.

PubMed

Willemsen, Jochem; Cornelis, Shana; Geerardyn, Filip M; Desmet, Mattias; Meganck, Reitske; Inslegers, Ruth; Cauwe, Joachim M B D

2015-01-01

The aim of this study is to provide an overview of the scientific activity of different psychoanalytic schools of thought in terms of the content and production of case studies published on ISI Web of Knowledge. Between March 2013 and November 2013, we contacted all case study authors included in the online archive of psychoanalytic and psychodynamic case studies (www.singlecasearchive.com) to inquire about their psychoanalytic orientation during their work with the patient. The response rate for this study was 45%. It appears that the two oldest psychoanalytic schools, Object-relations psychoanalysis and Ego psychology or "Classical psychoanalysis" dominate the literature of published case studies. However, most authors stated that they feel attached to two or more psychoanalytic schools of thought. This confirms that the theoretical pluralism in psychoanalysis stretches to the field of single case studies. The single case studies of each psychoanalytic school are described separately in terms of methodology, patient, therapist, or treatment features. We conclude that published case studies features are fairly similar across different psychoanalytic schools. The results of this study are not representative of all psychoanalytic schools, as some do not publish their work in ISI ranked journals.

Theoretical pluralism in psychoanalytic case studies

PubMed Central

Willemsen, Jochem; Cornelis, Shana; Geerardyn, Filip M.; Desmet, Mattias; Meganck, Reitske; Inslegers, Ruth; Cauwe, Joachim M. B. D.

2015-01-01

The aim of this study is to provide an overview of the scientific activity of different psychoanalytic schools of thought in terms of the content and production of case studies published on ISI Web of Knowledge. Between March 2013 and November 2013, we contacted all case study authors included in the online archive of psychoanalytic and psychodynamic case studies (www.singlecasearchive.com) to inquire about their psychoanalytic orientation during their work with the patient. The response rate for this study was 45%. It appears that the two oldest psychoanalytic schools, Object-relations psychoanalysis and Ego psychology or “Classical psychoanalysis” dominate the literature of published case studies. However, most authors stated that they feel attached to two or more psychoanalytic schools of thought. This confirms that the theoretical pluralism in psychoanalysis stretches to the field of single case studies. The single case studies of each psychoanalytic school are described separately in terms of methodology, patient, therapist, or treatment features. We conclude that published case studies features are fairly similar across different psychoanalytic schools. The results of this study are not representative of all psychoanalytic schools, as some do not publish their work in ISI ranked journals. PMID:26483725

[A clinical study of PC-904 in pediatrics (author's transl)].

PubMed

Hori, M; Kono, S; Hashimoto, F

1978-07-01

A clinical study of PC-904 was performed in children. The results were as follows; 1) Serum levels of PC-904 after single administration of 20 mg/kg were studied in patients by one-shot intravenous injection or drip infusion for 1 hour or 2 hours. The results obtained in this fundamental study led to the suggestion that the proper dosage in bacterial infections would be 50 approximately 100 mg/kg intravenously in 3 approximately 4 divided doses daily. 2) A clinical study of PC-904 was performed in 16 patients including acute pharyngitis and bronchitis (3 cases), acute bronchopneumonia (3 cases), pertussis pneumonia (1 case), lymphadenitis purulenta (2 cases), acute bronchopneumonia (3 cases), pertussis pneumonia (1 case), lymphadenitis purulenta (2 cases), abscess with chronic granuloma (2 cases) and urinary tract infection (5 cases). About 50 approximately 100 mg/kg of PC-904 were administered in 3 approximately 4 divided doses daily by one-shot intravenous injection. The clinical effects of 2 cases out of 16 were impossible to be evaluated, and thses cases were excluded from the clinical evaluation. Ten cases out of 14 showed excellent or good responses (efficacy rate: 71.4%). 3) The bacteriological examinations were performed and the causative or isolated organisms were determined in 11 cases. The responses were "disappeared" in 6 cases, "decreased" in 2 cases and "not disappeared" in 3 cases. 4) No side effects were observed. Abnormalities of laboratory findings were not also noticed.

Pathology analysis for mesothelioma study in the United Kingdom: Current practice and historical development.

PubMed

Case, B W

2016-01-01

Following up on the largest case-control study of malignant mesothelioma yet performed, investigators at the London School of Hygiene and Tropical Medicine assessed 1732 male and 670 female cases as of May 2013. Epidemiological findings of a subset of these were published previously, excluding patients who died or who refused to be interviewed. Pathology reports were collected for subjects, including those both eligible and ineligible for epidemiology study based on vital status. The current investigation examined 860 cases having pathology reports available. Sixty-one cases were diagnosed using cytology only, often with equivocal diagnoses, while 799 reported at least a biopsy of the tumor. Of these, 748 had pathology sufficiently detailed for evaluation. These reports were examined for basis of diagnosis, differences between study cases and ineligible cases, pathology characteristics, and immunohistochemical and other tests used. The most prominent subtype was epithelioid (64% of study cases but only 49% of ineligible cases). Biphasic subtype was present in 10% of study cases and 16% of those ineligible. Sarcomatoid subtype was present in 7% of study cases and 19% of ineligible cases, most of whom died. Twelve percent of study cases displayed no specified subtype, versus 7% of ineligible cases. Of recorded immunohistochemical stains specific for mesothelial cell origin, calretinin (95%) and CK 5/6 or CK5 alone (84%) were by far the most common. Calretinin and CK 5/6 or CK 5 alone were also most sensitive and positive in 92% of cases presenting with surgical pathology report. Ninety percent of cases had at least one immunohistochemical marker for possible lung carcinoma applied, with BER-Ep4 and TTF-1 the most frequent at 68% and CEA at 58%. TTF-1 and CEA were positive in 1% or less of cases. Patterns of use and positive and negative results for each of these as well as other immunohistochemical stains are presented and discussed, along with a brief historical description of their development and use. Possible effects of the pathologic analysis on the results of previously published and future epidemiological studies are discussed.

CRIS Case Study Materials in Ethical Decision Making.

ERIC Educational Resources Information Center

Blanning, James R.

Designed for secondary-level classroom discussion, these materials contain nine, short case studies of ethical dilemmas. The cast studies focus mainly on incidents and issues relevant to high school students. Discussion questions for each case study require students to examine the case, discuss the issue, and make an ethical decision about how…

Case Study: The Chemistry of Cocaine

ERIC Educational Resources Information Center

Dewprashad, Brahmadeo

2011-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. This month's case study focuses on the chemistry of cocaine to teach a number of core concepts in organic chemistry. It also requires that students read and analyze an original research paper on…

Interpretive Research Aiming at Theory Building: Adopting and Adapting the Case Study Design

ERIC Educational Resources Information Center

Diaz Andrade, Antonio

2009-01-01

Although the advantages of case study design are widely recognised, its original positivist underlying assumptions may mislead interpretive researchers aiming at theory building. The paper discusses the limitations of the case study design for theory building and explains how grounded theory systemic process adds to the case study design. The…

Using Case Studies in the Introductory Public Relations Course.

ERIC Educational Resources Information Center

Adams, William C.

The case study method has received increased attention at both the graduate and undergraduate levels in a number of public relations programs. Unlike the Harvard managerial-oriented case studies, the approach useful in large, introductory public relations courses stems from a simplified team approach to classroom projects, case studies in the…

Peace Corps Water/Sanitation Case Studies and Analyses. Appropriate Technologies for Development. Case Study CS-4.

ERIC Educational Resources Information Center

Talbert, Diana E., Comp.

This document provides an overview of Peace Corps water and sanitation activities, five case studies (Thailand, Yemen, Paraguay, Sierra Leone, and Togo), programming guidelines, and training information. Each case study includes: (1) background information on the country's geography, population, and economics; (2) information on the country's…

Role of matching in case-control studies of antimicrobial resistance.

PubMed

Cerceo, Elizabeth; Lautenbach, Ebbing; Linkin, Darren R; Bilker, Warren B; Lee, Ingi

2009-05-01

Of 57 case-control studies of antimicrobial resistance, matching was used in 23 (40%). Matched variables differed substantially across studies. Of these 23 matched case-control studies, 12 (52%) justified the use of matching, and 9 (39%) noted the strengths or limitations of this approach. Analysis that accounted for matching was performed in only 52% of the case-control studies.

Evaluating Creative Thinking of Rn-Bsn Students in the Course of Clinical Case Study and Practicum

ERIC Educational Resources Information Center

Ku, Ya-Lie

2015-01-01

This case study evaluated creative thinking of RN-BSN students in the course of clinical case study and practicum. Study design used quantitative and qualitative evaluations of creative thinking of RN-BSN students by triangulation method in the course of clinical case study and practicum. Sixty RN-BSN students self-perceived the changing levels of…

Schematic representation of case study research designs.

PubMed

Rosenberg, John P; Yates, Patsy M

2007-11-01

The paper is a report of a study to demonstrate how the use of schematics can provide procedural clarity and promote rigour in the conduct of case study research. Case study research is a methodologically flexible approach to research design that focuses on a particular case - whether an individual, a collective or a phenomenon of interest. It is known as the 'study of the particular' for its thorough investigation of particular, real-life situations and is gaining increased attention in nursing and social research. However, the methodological flexibility it offers can leave the novice researcher uncertain of suitable procedural steps required to ensure methodological rigour. This article provides a real example of a case study research design that utilizes schematic representation drawn from a doctoral study of the integration of health promotion principles and practices into a palliative care organization. The issues discussed are: (1) the definition and application of case study research design; (2) the application of schematics in research; (3) the procedural steps and their contribution to the maintenance of rigour; and (4) the benefits and risks of schematics in case study research. The inclusion of visual representations of design with accompanying explanatory text is recommended in reporting case study research methods.

Case Reports, Case Series - From Clinical Practice to Evidence-Based Medicine in Graduate Medical Education.

PubMed

Sayre, Jerry W; Toklu, Hale Z; Ye, Fan; Mazza, Joseph; Yale, Steven

2017-08-07

Case reports and case series or case study research are descriptive studies that are prepared for illustrating novel, unusual, or atypical features identified in patients in medical practice, and they potentially generate new research questions. They are empirical inquiries or investigations of a patient or a group of patients in a natural, real-world clinical setting. Case study research is a method that focuses on the contextual analysis of a number of events or conditions and their relationships. There is disagreement among physicians on the value of case studies in the medical literature, particularly for educators focused on teaching evidence-based medicine (EBM) for student learners in graduate medical education. Despite their limitations, case study research is a beneficial tool and learning experience in graduate medical education and among novice researchers. The preparation and presentation of case studies can help students and graduate medical education programs evaluate and apply the six American College of Graduate Medical Education (ACGME) competencies in the areas of medical knowledge, patient care, practice-based learning, professionalism, systems-based practice, and communication. A goal in graduate medical education should be to assist residents to expand their critical thinking, problem-solving, and decision-making skills. These attributes are required in the teaching and practice of EBM. In this aspect, case studies provide a platform for developing clinical skills and problem-based learning methods. Hence, graduate medical education programs should encourage, assist, and support residents in the publication of clinical case studies; and clinical teachers should encourage graduate students to publish case reports during their graduate medical education.

Methodology Series Module 2: Case-control Studies

PubMed Central

Setia, Maninder Singh

2016-01-01

Case-Control study design is a type of observational study. In this design, participants are selected for the study based on their outcome status. Thus, some participants have the outcome of interest (referred to as cases), whereas others do not have the outcome of interest (referred to as controls). The investigator then assesses the exposure in both these groups. The investigator should define the cases as specifically as possible. Sometimes, definition of a disease may be based on multiple criteria; thus, all these points should be explicitly stated in case definition. An important aspect of selecting a control is that they should be from the same ‘study base’ as that of the cases. We can select controls from a variety of groups. Some of them are: General population; relatives or friends; and hospital patients. Matching is often used in case-control control studies to ensure that the cases and controls are similar in certain characteristics, and it is a useful technique to increase the efficiency of the study. Case-Control studies can usually be conducted relatively faster and are inexpensive – particularly when compared with cohort studies (prospective). It is useful to study rare outcomes and outcomes with long latent periods. This design is not very useful to study rare exposures. Furthermore, they may also be prone to certain biases – selection bias and recall bias. PMID:27057012

Case-Control Studies of Sporadic Enteric Infections: A Review and Discussion of Studies Conducted Internationally from 1990 to 2009

PubMed Central

Fullerton, Kathleen E.; Scallan, Elaine; Kirk, Martyn D.; Mahon, Barbara E.; Angulo, Frederick J.; de Valk, Henriette; van Pelt, Wilfrid; Gauci, Charmaine; Hauri, Anja M.; Majowicz, Shannon; O’Brien, Sarah J.

2015-01-01

Epidemiologists have used case-control studies to investigate enteric disease outbreaks for many decades. Increasingly, case-control studies are also used to investigate risk factors for sporadic (not outbreak-associated) disease. While the same basic approach is used, there are important differences between outbreak and sporadic disease settings that need to be considered in the design and implementation of the case-control study for sporadic disease. Through the International Collaboration on Enteric Disease “Burden of Illness” Studies (the International Collaboration), we reviewed 79 case-control studies of sporadic enteric infections caused by nine pathogens that were conducted in 22 countries and published from 1990 through to 2009. We highlight important methodological and study design issues (including case definition, control selection, and exposure assessment) and discuss how approaches to the study of sporadic enteric disease have changed over the last 20 years (e.g., making use of more sensitive case definitions, databases of controls, and computer-assisted interviewing). As our understanding of sporadic enteric infections grows, methods and topics for case-control studies are expected to continue to evolve; for example, advances in understanding of the role of immunity can be used to improve control selection, the apparent protective effects of certain foods can be further explored, and case-control studies can be used to provide population-based measures of the burden of disease. PMID:22443481

Case studies in pathophysiology: The development and evaluation of an interactive online learning environment to develop higher order thinking and argumentation

NASA Astrophysics Data System (ADS)

Titterington, Lynda C.

2007-12-01

This study presents a framework for examining the effects of higher order thinking on the achievement of allied health students enrolled in a pathophysiology course. A series of clinical case studies was developed and published in an enriched online environment that guided students through the process of developing a solution and supporting it through data analysis and interpretation. The series of case study modules scaffolded argumentation through question prompts. The modules began with a simple, direct problem and they became progressively more complex throughout the quarter. A control group was assigned a pencil-and-paper case study based upon recall. The case studies were scored for content accuracy and evidence of higher order thinking skills. Higher order thinking was measured using a rubric based upon the Toulmin argumentation pattern. The results indicated implementing a case study of either online or traditional format was associated with significant gains in achievement. The Web-enhanced case studies were associated with modest gains in knowledge acquisition. The argumentation scores across the series followed two trends: directed case studies were associated with higher levels of argumentation than ill-structured case studies, and there appeared to be an inverse relationship between the students' argumentation and content scores. The protocols developed for this study can serve as a template for a larger, extended investigation into student learning in the online environment.

[Clinical decision making: Fostering critical thinking in the nursing diagnostic process through case studies].

PubMed

Müller-Staub, Maria; Stuker-Studer, Ursula

2006-10-01

Case studies, based on actual patients' situations, provide a method of clinical decision making to foster critical thinking in nurses. This paper describes the method and process of group case studies applied in continuous education settings. This method bases on Balints' case supervision and was further developed and combined with the nursing diagnostic process. A case study contains different phases: Pre-phase, selection phase, case delineation and case work. The case provider narratively tells the situation of a patient. This allows the group to analyze and cluster signs and symptoms, to state nursing diagnoses and to derive nursing interventions. Results of the case study are validated by applying the theoretical background and critical appraisal of the case provider. Learning effects of the case studies were evaluated by means of qualitative questionnaires and analyzed according to Mayring. Findings revealed the following categories: a) Patients' problems are perceived in a patient centred way, accurate nursing diagnoses are stated and effective nursing interventions implemented. b) Professional nursing tasks are more purposefully perceived and named more precise. c) Professional nursing relationship, communication and respectful behaviour with patients were perceived in differentiated ways. The theoretical framework is described in the paper "Clinical decision making and critical thinking in the nursing diagnostic process". (Müller-Staub, 2006).

A Review of Case-Based Learning Practices in an Online MBA Program: A Program-Level Case Study

ERIC Educational Resources Information Center

Lee, Seung-hee; Lee, Jieun; Liu, Xiaojing; Bonk, Curt J.; Magjuka, Richard J.

2009-01-01

This study examines how a case-based learning approach was used and facilitated in online business education. Perceptions of students and instructors regarding the practices of case-based learning in online environments are explored in terms of instructional design, facilitation, and technology support. This study finds case-based learning to be a…

In Case You Are Interested: Results of a Survey of Case Study Teachers

ERIC Educational Resources Information Center

Herreid, Clyde Freeman; Schiller, Nancy A.; Herreid, Ky F.; Wright, Carolyn

2011-01-01

Case study teaching had a long tradition in law and business before it made the jump to medical school education in the form of Problem-Based Learning (PBL) in the 1970s. Today, both the University of Delaware's Clearinghouse and the University of Buffalo's National Center for Case Study Teaching in Science (NCCSTS) have hundreds of cases and…

Teaching Business French through Case Studies: Presentation of a Marketing Case.

ERIC Educational Resources Information Center

Federico, Salvatore; Moore, Catherine

The use of case studies as a means for teaching business French is discussed. The approach is advocated because of the realism of case studies, which are based on actual occurrences. Characteristics of a good case are noted: it tells a story, focuses on interest-arousing issues, is set in the past 10 years, permits empathy with the main…

Cases for the Net Generation: An Empirical Examination of Students' Attitude toward Multimedia Case Studies

ERIC Educational Resources Information Center

Sheppard, Michael; Vibert, Conor

2016-01-01

Case studies have been an important tool in business, legal, and medical education for generations of students. Traditional text-based cases tend to be self-contained and structured in such a way as to teach a particular concept. The multimedia cases introduced in this study feature unscripted web-hosted video interviews with business owners and…

Swiss Armed Forces Organizational Level Leader Development: A Qualitative Case Study

DTIC Science & Technology

2017-06-09

chapter, divided in five distinct parts, describes the chosen research methodology , explain why the qualitative case study is appropriate to conduct...research study uses a qualitative methodology by performing a qualitative case study on the organizational level leader’s development process within...develop an in-depth understsanding of the phenomen.”82 Summary This research study uses a qualitative methodology by performing a case study on the

Exchangeability in the case-crossover design

PubMed Central

Mittleman, Murray A; Mostofsky, Elizabeth

2014-01-01

In cohort and case-control studies, confounding that arises as a result of differences in the distribution of determinants of the outcome between exposure groups leading to non-exchangeability are addressed by restriction, matching or with statistical models. In case-only studies, this issue is addressed by comparing each individual with his/herself. Although case-only designs use self-matching and only include individuals who develop the outcome of interest, issues of non-exchangeability are identical to those that arise in traditional case-control and cohort studies. In this review, we describe one type of case-only design, the case-crossover design, and discuss how the concept of exchangeability can be used to understand issues of confounding, carryover effects, period effects and selection bias in case-crossover studies. PMID:24756878

Transportation planning effectiveness: twenty case studies

DOT National Transportation Integrated Search

1977-12-31

This document consists of 20 site-specific case studies of urban area experience with the Technical Study Grant Program of the Urban Mass Transportation Administration (UMTA). The objective of the case studies is to determine how the Technical Study ...

An alternative approach for socio-hydrology: case study research

NASA Astrophysics Data System (ADS)

Mostert, Erik

2018-01-01

Currently the most popular approach in socio hydrology is to develop coupled human-water models. This article proposes an alternative approach, qualitative case study research, involving a systematic review of (1) the human activities affecting the hydrology in the case, (2) the main human actors, and (3) the main factors influencing the actors and their activities. Moreover, this article presents a case study of the Dommel Basin in Belgium and the Netherlands, and compares this with a coupled model of the Kissimmee Basin in Florida. In both basins a pendulum swing from water resources development and control to protection and restoration can be observed. The Dommel case study moreover points to the importance of institutional and financial arrangements, community values, and broader social, economic, and technical developments. These factors are missing from the Kissimmee model. Generally, case studies can result in a more complete understanding of individual cases than coupled models, and if the cases are selected carefully and compared with previous studies, it is possible to generalize on the basis of them. Case studies also offer more levers for management and facilitate interdisciplinary cooperation. Coupled models, on the other hand, can be used to generate possible explanations of past developments and quantitative scenarios for future developments. The article concludes that, given the limited attention they currently get and their potential benefits, case studies deserve more attention in socio-hydrology.

Human exploration mission studies

NASA Technical Reports Server (NTRS)

Cataldo, Robert L.

1989-01-01

The Office of Exploration has established a process whereby all NASA field centers and other NASA Headquarters offices participate in the formulation and analysis of a wide range of mission strategies. These strategies were manifested into specific scenarios or candidate case studies. The case studies provided a systematic approach into analyzing each mission element. First, each case study must address several major themes and rationale including: national pride and international prestige, advancement of scientific knowledge, a catalyst for technology, economic benefits, space enterprise, international cooperation, and education and excellence. Second, the set of candidate case studies are formulated to encompass the technology requirement limits in the life sciences, launch capabilities, space transfer, automation, and robotics in space operations, power, and propulsion. The first set of reference case studies identify three major strategies: human expeditions, science outposts, and evolutionary expansion. During the past year, four case studies were examined to explore these strategies. The expeditionary missions include the Human Expedition to Phobos and Human Expedition to Mars case studies. The Lunar Observatory and Lunar Outpost to Early Mars Evolution case studies examined the later two strategies. This set of case studies established the framework to perform detailed mission analysis and system engineering to define a host of concepts and requirements for various space systems and advanced technologies. The details of each mission are described and, specifically, the results affecting the advanced technologies required to accomplish each mission scenario are presented.

How informative are case studies of spider bites in the medical literature?

PubMed

Stuber, Marielle; Nentwig, Wolfgang

2016-05-01

We analyzed the reliability and information content of 134 medical case studies on spider bites, published in 91 journal articles. Overall, we found that only 22% of these studies fulfilled the criteria for a verified spider bite. This means that the majority of such case studies cannot be attributed to a given spider species and usually not even to a spider. Their scientific value is negligible, moreover, such publications are even dangerous because they suggest incorrect conclusions. Secondly, we found that such case studies usually do not follow an obvious structure and many details on the development of symptoms, therapy and healing process are widely lacking. So even for verified spider bites, the comparability of case studies is limited. We discuss the obvious failure of a reviewing process for case studies and give recommendations how to increase the currently low information content of medical case studies on spider bites. Copyright © 2016 Elsevier Ltd. All rights reserved.

Case Study Observational Research: A Framework for Conducting Case Study Research Where Observation Data Are the Focus.

PubMed

Morgan, Sonya J; Pullon, Susan R H; Macdonald, Lindsay M; McKinlay, Eileen M; Gray, Ben V

2017-06-01

Case study research is a comprehensive method that incorporates multiple sources of data to provide detailed accounts of complex research phenomena in real-life contexts. However, current models of case study research do not particularly distinguish the unique contribution observation data can make. Observation methods have the potential to reach beyond other methods that rely largely or solely on self-report. This article describes the distinctive characteristics of case study observational research, a modified form of Yin's 2014 model of case study research the authors used in a study exploring interprofessional collaboration in primary care. In this approach, observation data are positioned as the central component of the research design. Case study observational research offers a promising approach for researchers in a wide range of health care settings seeking more complete understandings of complex topics, where contextual influences are of primary concern. Future research is needed to refine and evaluate the approach.

Statin-Associated Polymyalgia Rheumatica. An Analysis Using WHO Global Individual Case Safety Database: A Case/Non-Case Approach

PubMed Central

de Jong, Hilda J. I.; Saldi, Siti R. F.; Klungel, Olaf H.; Vandebriel, Rob J.; Souverein, Patrick C.; Meyboom, Ronald H. B.; Passier, J. L. M. (Anneke); van Loveren, Henk; Tervaert, Jan Willem Cohen

2012-01-01

Objective To assess whether there is an association between statin use and the occurrence of polymyalgia rheumatic (PMR) in the spontaneous reporting database of the World Health Organisation (WHO). Methods We conducted a case/non-case study based on individual case safety reports (ICSR) in the WHO global ICSR database (VigiBase). Case reports containing the adverse event term polymyalgia rheumatica (WHOART or MedDRA Preferred Term) were defined as cases. Non-cases were all case reports containing other adverse event terms. Each case was matched to five non-cases by age, gender, and time of reporting. Case reports regarding a statin as suspected or concomitant drug were identified using the Anatomical Therapeutic Chemical (ATC) classification. Multivariate logistic regression was used to calculate reporting odds ratios (RORs) with 95% confidence intervals (CI). Results We identified 327 reports of PMR as cases and 1635 reports of other ADRs as non-cases. Among cases, statins were more frequently reported as suspected agent (29.4%) compared to non-cases (2.9%). After adjustment for several covariates, statins were significantly associated with reports of PMR (ROR 14.21; 95% CI 9.89–20.85). Conclusion The results of this study lends support to previous anecdotal case reports in the literature suggesting that the use of a statin may be associated with the occurrence of PMR. Further studies are needed to study the strength of the association in more detail and to elucidate the underlying mechanism. PMID:22844450

An Arizona Border Wall Case Study

DTIC Science & Technology

2017-12-01

WALL CASE STUDY by Justin Alexander Bristow December 2017 Thesis Advisor: Erik Dahl Second Reader: Jorge Gonzalez THIS PAGE...4. TITLE AND SUBTITLE AN ARIZONA BORDER WALL CASE STUDY 5. FUNDING NUMBERS 6. AUTHOR(S) Justin Alexander Bristow 7. PERFORMING ORGANIZATION...PAGE INTENTIONALLY LEFT BLANK iii Approved for public release. Distribution is unlimited. AN ARIZONA BORDER WALL CASE STUDY Justin Alexander

22 CFR 96.47 - Preparation of home studies in incoming cases.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Preparation of home studies in incoming cases... Cases) § 96.47 Preparation of home studies in incoming cases. (a) The agency or person ensures that a home study on the prospective adoptive parent(s) (which for purposes of this section includes the...

22 CFR 96.47 - Preparation of home studies in incoming cases.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 22 Foreign Relations 1 2012-04-01 2012-04-01 false Preparation of home studies in incoming cases... Cases) § 96.47 Preparation of home studies in incoming cases. (a) The agency or person ensures that a home study on the prospective adoptive parent(s) (which for purposes of this section includes the...

22 CFR 96.47 - Preparation of home studies in incoming cases.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 22 Foreign Relations 1 2011-04-01 2011-04-01 false Preparation of home studies in incoming cases... Cases) § 96.47 Preparation of home studies in incoming cases. (a) The agency or person ensures that a home study on the prospective adoptive parent(s) (which for purposes of this section includes the...

22 CFR 96.47 - Preparation of home studies in incoming cases.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 22 Foreign Relations 1 2013-04-01 2013-04-01 false Preparation of home studies in incoming cases... Cases) § 96.47 Preparation of home studies in incoming cases. (a) The agency or person ensures that a home study on the prospective adoptive parent(s) (which for purposes of this section includes the...

22 CFR 96.47 - Preparation of home studies in incoming cases.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 22 Foreign Relations 1 2014-04-01 2014-04-01 false Preparation of home studies in incoming cases... Cases) § 96.47 Preparation of home studies in incoming cases. (a) The agency or person ensures that a home study on the prospective adoptive parent(s) (which for purposes of this section includes the...

Security Quality Requirements Engineering (SQUARE): Case Study Phase III

DTIC Science & Technology

2006-05-01

Security Quality Requirements Engineering (SQUARE): Case Study Phase III Lydia Chung Frank Hung Eric Hough Don Ojoko-Adams Advisor...Engineering (SQUARE): Case Study Phase III CMU/SEI-2006-SR-003 Lydia Chung Frank Hung Eric Hough Don Ojoko-Adams Advisor Nancy R. Mead...1 1.1 The SQUARE Process ............................................................................... 1 1.2 Case Study Clients

Executive Headteachers: What's in a Name? Case Study Compendium

ERIC Educational Resources Information Center

Wespieser, Karen, Ed.

2016-01-01

This Case Study Compendium provides an overview of the 12 cases that were investigated as part of the study "Executive Headteachers: What's in a Name?'" (Lord et al., 2016). The case study overviews are based on in-depth analysis and research as described in the full report (ibid) and the Technical Appendix (Harland and Bernardinelli,…

Guidelines for Using Case Studies in the Teaching-Learning Process

ERIC Educational Resources Information Center

McFarlane, Donovan A.

2015-01-01

In this paper, the author expresses concerns regarding the extensive use of case studies in the college classroom and advises college and university faculties to be more careful in their selection and use of case studies as an effective­-based method of teaching. The author communicates the important role that case studies play in teaching and…

Bewitching Ideas Influence Learning: An Evaluation of an Interdisciplinary Teaching Experience

ERIC Educational Resources Information Center

Nava-Whitehead, Susan M.; Augusto, Kerri W.; Gow, Joan-Beth

2011-01-01

This column provides original articles on innovations in case study teaching, assessment of the method, as well as case studies with teaching notes. In this month's issue the authors describe an interdisciplinary approach to case study teaching that addresses the demand to balance the goals of process and content. The case study, Salem's Secrets…

78 FR 39769 - National Register of Historic Places; Notification of Pending Nominations and Related Actions

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-02

... Chautauqua Blvd., Los Angeles, 13000513 Community Clubhouse, 1200 N. Vista St., West Hollywood, 13000510... Study House No. 23A, (Case Study House Program MPS) 2342 Rue de Anne, La Jolla, 13000520 Case Study House No. 23C, (Case Study House Program MPS) 2339 Rue de Anne, La Jolla, 13000521 Ventura County Case...

Integrated Approaches to Testing and Assessment Under ...

EPA Pesticide Factsheets

Presentation on IATA case study for phenols which is joint project with Health Canada and NCCT, presented at the OECD IATA Case Study Meeting No. 2 in Paris, France. Presentation on IATA case study for phenols which is joint project with Health Canada and NCCT, presented at the OECD IATA Case Study Meeting No. 2 in Paris, France.

Effectiveness of Three Case Studies and Associated Teamwork in Stimulating Freshman Interest in an Introduction to Engineering Course

ERIC Educational Resources Information Center

McIntyre, Joseph S.

2011-01-01

The effectiveness of three case studies and associated teamwork to stimulate interest of college freshman in engineering was investigated by observing students. Case studies were assigned as laboratory team exercises in an introduction to engineering course at Auburn University. Student interest in the case studies was evaluated qualitatively…

A Case Study of Project ATHENA: Tactical Level Technological Innovation Aboard the USS Benfold

DTIC Science & Technology

2014-12-01

case studies , grounded theory research , phenomenological research and narrative research . 13 3. Qualitative Methods : Selection The researcher’s...12 B. THE CASE STUDY METHOD 1. Introduction The case study is a type of qualitative research that enables the researcher to chronicle and...objectives and the research environment ultimately determine the

76 FR 71341 - BASINS and WEPP Climate Assessment Tools: Case Study Guide to Potential Applications

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-17

... report presents a series of short case studies designed to illustrate the capabilities of these tools for... change impacts on water. This report presents a series of short case studies using the BASINS and WEPP climate assessment tools. The case studies are designed to illustrate the capabilities of these tools for...

Al-Qaeda-Syria (AQS): An Al-Qaeda Affiliate Case Study

DTIC Science & Technology

2017-10-01

comparative methodology that included eight case studies on groups affiliated or associated with Al-Qaeda. These case studies were then used as a...Case Study Zack Gold With contributions from Pamela G. Faber October 2017 This work was performed under Federal Government...for Stability and Development Center for Strategic Studies REPORT DOCUMENTATION PAGE Form Approved OMB No. 0704-0188 Public reporting burden

Silicosis in Sandblasters: A Case Study Adapted for Use in U.S. High Schools. NIOSH Case Study in Occupational Epidemiology.

ERIC Educational Resources Information Center

Malit, Bonita D.

This document presents a case study of silicosis in sandblasters that has been adapted for instructional use in U.S. high schools. The primary objective of the case study is to teach students about epidemiology by studying an occupational hazard, disease associated with the hazard, and methods for preventing the disease. The introduction offers…

Risk Management and Performance in the Balkans Support Contract

DTIC Science & Technology

2005-01-01

Defense contracts-United States-Case studies . 2. United States. Army- Procurement-Case studies . 3. Bosnia and Hercegovina-History, Military-20ts...24 3.1. Assessment of GAO Cost Study .................................. 72 3.2. Comparison of Potential Sources of Nonperformance...accepting inappropriate risks, what can it do about it? Case studies of CSS contracts can provide some answers. In this report, we present a case study of

Putting the Learning in Case Learning? The Effects of Case-Based Approaches on Student Knowledge, Attitudes, and Engagement

ERIC Educational Resources Information Center

Krain, Matthew

2016-01-01

This study revisits case learning's effects on student engagement and assesses student learning as a result of the use of case studies and problem-based learning. The author replicates a previous study that used indirect assessment techniques to get at case learning's impact, and then extends the analysis using a pre- and post-test experimental…

Case Study: A Picture Worth a Thousand Words? Making a Case for Video Case Studies

ERIC Educational Resources Information Center

Pai, Aditi

2014-01-01

A picture, they say, is worth a thousand words. If a mere picture is worth a thousand words, how much more are "moving pictures" or videos worth? The author poses this not merely as a rhetorical question, but because she wishes to make a case for using videos in the traditional case study method. She recommends four main approaches of…

Participative Case Studies: Integrating Case Writing and a Traditional Case Study Approach in a Marketing Context

ERIC Educational Resources Information Center

Forman, Howard

2006-01-01

Case-based pedagogy is a valuable tool for applying business concepts and theories to organizational contexts. Traditional case-based pedagogy offers such learning opportunities. What this pedagogy lacks, however, is an element of real-time experiential learning opportunities. This research focuses on the advantages of incorporating a case-writing…

In Case You Teach English: Case Studies in the English Classroom.

ERIC Educational Resources Information Center

Johannessen, Larry R.

Case studies can be a powerful strategy to use for helping students learn critical thinking processes that are key to interpreting and responding to literature and writing. Some of the major benefits of applying case methods are: cases provide an environment for active learning; they encourage the creation of a community of learners; cases help…

Using Case Studies to Enrich Field Experience.

ERIC Educational Resources Information Center

Florio-Ruane, Susan; Clark, Christopher M.

1990-01-01

This paper discusses the use of field experience in teacher education and how it can be augmented by phenomenological case studies. It summarizes a particular case study involving three teacher education classes, noting that reflective analysis of cases can prepare students to observe in the field. (SM)

Pathological characteristics and clinical specifications in gastroenteropancreatic neuroendocrine tumors: a study of 68 cases.

PubMed

Stoica-Mustafa, Elena; Pechianu, C; Iorgescu, Andreea; Hortopan, Monica; Dima, Simona Olimpia; Tomulescu, V; Dumitraşcu, T; Ungureanu, C; Andronesi, D; Popescu, I; Herlea, V

2012-01-01

Gastroenteropancreatic neuroendocrine tumors (GEP-NETs) represent a group of tumors, having their origin in cells of diffuse endocrine system, with particular clinical course, diagnosis and treatment. In our study, were included 68 patients with neuroendocrine digestive tumors admitted, diagnosed and treated in Fundeni Clinical Institute, Bucharest, in the last ten years--2000-2010 (retrospective study). Thirty-three (49%) patients were males, 35 (51%) females, and the main age was 58.9 years. In 62 (90.3%) cases was possible to find the primary tumor. The examined tumors had different localizations: pancreas--32 (47.04%) cases (head--17 (24.99%) cases, and body and tail--15 (22.05%) cases), stomach--7 (10.29%) cases, small intestine--7 (10.29%) cases, 6 (8.82%) cases--unknown primary site (diagnosis was established on metastases), right colon--6 (8.82%) cases, liver--6 (8.82%) cases, rectum--2 (2.94%) cases, and retroperitoneum--2 (2.94%) cases. Microscopic examination revealed 59 (86.8%) malignant tumors and 9 (13.2%) benign tumors. Using WHO 2000 Classification, 28 cases of malignant tumors were well-differentiated neuroendocrine carcinomas, and 31 cases were poor differentiated neuroendocrine carcinomas. From malignant cases, 25 (42.3%) have distant metastases and 15 (25.9%) lymph node metastases. Cases of gastroenteropancreatic neuroendocrine tumors included in our study had clinical and histopathological features in correspondence with data from literature--slight predominance in women, predominance in 5th and 6th decades of life, the most frequent localizations were at pancreatic level--both head and body and tail, but the rarest were in colon and retroperitoneum. Most of the cases studied, were malignant tumors, from these more than a half were poor differentiated, and a quarter of them having lymph node or distant metastases.

What Case Managers Should Know About Their Roles and Functions: A National Study From the Commission for Case Manager Certification: Part 1.

PubMed

Tahan, Hussein M; Watson, Annette C; Sminkey, Patrice V

2015-01-01

The purpose of this national role and function study was to identify the essential activities and necessary knowledge areas for effective case manager practice from the perspective of those currently functioning in various care settings and across diverse professional disciplines. The national study covered all case management practices and/or work settings across the full continuum of health care. This cross-sectional descriptive study used the practice analysis method and online survey research design. It employed a purposive sample of case managers, in which 52,370 individuals received an invitation to participate. Data collection completed over a 4-week period, resulting in 7,668 useable survey responses (nearly a 15% response rate). The study identified the common activities and knowledge areas necessary for competent and effective performance of case managers, as is highlighted in this article, which is the first of a 2-part series on the role and function study. The results informed the needed update of the test specifications for the Certified Case Manager (CCM) certification examination, as will be delineated in Part 2 of the article series, to ensure that it continues to be substantiated in current practice. Of special note are the emergence of specific activity and knowledge domains in the area of case management ethical, legal, and practice standards, and an increase in the number of employers requiring certified case managers to fill vacant positions and compensating them financially for such qualifications. This study helps keep the CCM credentialing examination evidence-based and maintain its validity for evaluating competency of case managers. Specifically, the study identified essential activities and knowledge domains that define competent case management practice. Findings can be used for developing programs and curricula for the training and development of case managers. The study instrument also can be used for further research of case management practice.

The South Carolina National Guard Secure Area Duty Officer Program: A Reserve Component Active Shooter Contingency Case Study

DTIC Science & Technology

2017-12-01

SADOP) was authorized as SCNG policy on October 3, 2015. This research constitutes a case study of the SCNG SADOP and catalogs the program from...concealed firearms carry law. C. METHODOLOGY This thesis is a single case study of SADOP, which is an exceptional case and the only one of its kind...CAROLINA NATIONAL GUARD SECURE AREA DUTY OFFICER PROGRAM: A RESERVE COMPONENT ACTIVE SHOOTER CONTINGENCY CASE STUDY by Barry N. Ramey December

Reliability of medical records in diagnosing inflammatory breast cancer in Egypt.

PubMed

Le, Lynne; Schairer, Catherine; Hablas, Ahmed; Meza, Jane; Watanabe-Galloway, Shinobu; Ramadan, Mohamed; Merajver, Sofia D; Seifeldin, Ibrahim A; Soliman, Amr S

2017-03-16

Inflammatory breast cancer (IBC) is a rare, aggressive breast cancer diagnosed clinically by the presence of diffuse erythema, peau d'orange, and edema that arise quickly in the affected breast. This study evaluated the validity of medical records in Gharbiah, Egypt in identifying clinical signs/symptoms of IBC. For 34 IBC cases enrolled in a case-control study at the Gharbiah Cancer Society and Tanta Cancer Center, Egypt (2009-2010), we compared signs/symptoms of IBC noted in medical records to those recorded on a standardized form at the time of IBC diagnosis by clinicians participating in the case-control study. We calculated the sensitivity and specificity of medical records as compared to the case-control study for recording these signs/symptoms. We also performed McNemar's tests. In the case-control study, 32 (94.1%) IBC cases presented with peau d'orange, 30 (88.2%) with erythema, and 31 (91.2%) with edema. The sensitivities of the medical records as compared to the case-control study were 0.8, 0.5, and 0.2 for peau d'orange, erythema, and edema, respectively. Corresponding specificities were 1.0, 0.5, and 1.0. p values for McNemar's test were <0.05 for all signs. Medical records had data on the extent and duration of signs for at most 27% of cases for which this information was recorded in the case-control study. Twenty-three of the 34 cases (67.6%) had confirmed diagnosis of IBC in their medical records. Medical records lacked information on signs/symptoms of IBC, especially erythema and edema, when compared to the case-control study. Deficient medical records could have implications for diagnosis and treatment of IBC and proper documentation of cases in cancer registries.

Psychological Intervention: Case Studies in School Psychological Services, Volume 3, 1979.

ERIC Educational Resources Information Center

Iowa State Dept. of Public Instruction, Des Moines. Div. of Pupil Personnel Services.

The book presents 27 case studies illustrating psychological interventions with behavior problem school children. Studies ususally introduce the target population, describe the method of psychological evaluation, report the results of treatment, and discuss the case's implications. Among cases reported are investigations of stimulant medication on…

Problems of Practice: Canadian Cases in Leadership and Policy

ERIC Educational Resources Information Center

Pollock, Katina; Ryan, James

2013-01-01

One way to support aspiring and current administrators is through the use of case study teaching (Barnes, Christensen, & Hansen, 1994). Using case studies as a teaching and learning tool helps bring real-life situations into principal preparation and support programs. Case study approaches provide students with time to interrogate contemporary…

Report on Federal Productivity. Volume 2, Productivity Case Studies.

ERIC Educational Resources Information Center

Joint Financial Management Improvement Program, Washington, DC.

Volume 2 contains 15 productivity case studies which illustrate and expand on the causal factors mentioned in volume 1. The cases illustrate many different approaches to productivity measurement improvement. The case studies are: Development of an Output-Productivity Measure for the Air Force Medical Service; Measuring Effectiveness and Efficiency…

Using full-cohort data in nested case-control and case-cohort studies by multiple imputation.

PubMed

Keogh, Ruth H; White, Ian R

2013-10-15

In many large prospective cohorts, expensive exposure measurements cannot be obtained for all individuals. Exposure-disease association studies are therefore often based on nested case-control or case-cohort studies in which complete information is obtained only for sampled individuals. However, in the full cohort, there may be a large amount of information on cheaply available covariates and possibly a surrogate of the main exposure(s), which typically goes unused. We view the nested case-control or case-cohort study plus the remainder of the cohort as a full-cohort study with missing data. Hence, we propose using multiple imputation (MI) to utilise information in the full cohort when data from the sub-studies are analysed. We use the fully observed data to fit the imputation models. We consider using approximate imputation models and also using rejection sampling to draw imputed values from the true distribution of the missing values given the observed data. Simulation studies show that using MI to utilise full-cohort information in the analysis of nested case-control and case-cohort studies can result in important gains in efficiency, particularly when a surrogate of the main exposure is available in the full cohort. In simulations, this method outperforms counter-matching in nested case-control studies and a weighted analysis for case-cohort studies, both of which use some full-cohort information. Approximate imputation models perform well except when there are interactions or non-linear terms in the outcome model, where imputation using rejection sampling works well. Copyright © 2013 John Wiley & Sons, Ltd.

Is routine postoperative gastrografin study needed after laparoscopic sleeve gastrectomy? Experience of 712 cases.

PubMed

Wahby, M; Salama, A F; Elezaby, A F; Belgrami, F; Abd Ellatif, M E; El-Kaffas, H F; Al-Katary, M

2013-11-01

The current standard of care is to perform a postoperative gastrografin study following laparoscopic sleeve gastrectomy (LSG) to detect leakage or obstruction. This study evaluated the usefulness of this routine procedure. A retrospective chart review was performed in December 2012. All patients had routine intraoperative methylene blue testing to check for possible leakage from the staple line, and any leaking points were oversewn. We also performed postoperative contrast study (gastrografin) routinely in the first 24-48 h for all patients. From June 2007 to December 2012, 712 cases underwent LSG during the study period. Patients included in this study were 556 women (78.1%) and 156 men (21.9%). The mean age was 35 years. The mean BMI was 48 kg/m2. The operative time was 107 ± 29 min, and there were no conversions to open surgery. Intraoperative methylene blue test detected leakage in 28 cases (3.93%). Postoperative contrast study (gastrografin) was negative for leakage in all cases. Computed tomography (CT) scan with oral contrast study detected leakage in 1.4% (ten cases); none of these cases were detected by regular contrast study. Our study showed that intraoperative methylene blue test for leakage is a very sensitive and effective method for detecting leakage during sleeve gastrectomy and should be done routinely in all cases. Routine postoperative contrast study is not needed to detect leakage unless clinically indicated in selected cases, and in such cases contrast-enhanced CT scans are the modality of choice.

Impact of an Engineering Case Study in a High School Pre-Engineering Course

ERIC Educational Resources Information Center

Rutz, Eugene; Shafer, Michelle

2011-01-01

Students at an all-girls high school who were enrolled in an introduction to engineering course were presented an engineering case study to determine if the case study affected their attitudes toward engineering and their abilities to solve engineering problems. A case study on power plants was implemented during a unit on electrical engineering.…

The Effectiveness of a Case Study-Based First-Year Biology Class at a Black Women's College

ERIC Educational Resources Information Center

Pai, Aditi; Benning, Tracy; Woods, Natasha; McGinnis, Gene; Chu, Joanne; Netherton, Josh; Bauerle, Cynthia

2010-01-01

The authors used a case study-based approach in the introductory biology course at Spelman College. The course taught to entering freshmen was divided into three modules--ecology, evolution, and biodiversity, each designed around a case study. They noted that (1) case study teaching was dramatically more effective than the traditional lecture…

Flawed Execution: A Case Study on Operational Contract Support

DTIC Science & Technology

2016-06-01

NAVAL POSTGRADUATE SCHOOL MONTEREY, CALIFORNIA JOINT APPLIED PROJECT FLAWED EXECUTION: A CASE STUDY ON OPERATIONAL CONTRACT SUPPORT June 2016...applied project 4. TITLE AND SUBTITLE FLAWED EXECUTION: A CASE STUDY ON OPERATIONAL CONTRACT SUPPORT 5. FUNDING NUMBERS 6. AUTHOR(S) Scott F...unlimited FLAWED EXECUTION: A CASE STUDY ON OPERATIONAL CONTRACT SUPPORT Scott F. Taggart, Captain, United States Marine Corps Jacob Ledford

Finding The Limit: The Strategic Potential Of The Network Based Actor

DTIC Science & Technology

2016-12-01

6  E.  CASE STUDY SELECTION AND PROCESS .......................................6  1.  Selection of Baseline Case Studies: Pierre-Charles...Pathé and the Trust ..................................................................................7  2.  Selection of Test Case Studies...academic debate, which argues that non-state, network-based actors can have a strategic planning capacity. E. CASE STUDY SELECTION AND PROCESS This

Hindsight Is 20/20: A Case Study of Vision and Reading Issues Sheds Light for Teacher Education Programs

ERIC Educational Resources Information Center

Chandler, Kristie B.; Box, Jean A.

2012-01-01

This paper presents a case study designed to educate students in pre-service teacher education programs about the importance of a comprehensive eye exam. The case study chronicles a family's multi-year search for solutions to their child's reading difficulties. The research supporting the case study explores the connection between vision…

Case Studies in the Organizational Communication Course: Applying Textbook Concepts to Real Life Organizations.

ERIC Educational Resources Information Center

Byers, Peggy Yuhas

This paper urges the use of case study discussions in the organizational communication class as an effective instructional technique. The paper presents a variety of formats for bringing case studies to life for students in the organizational communication course. It first discusses sources for case studies such as the World Wide Web, local and…

WeSaySo Case Study: Designing and Implementing a Case Study for Use in an Instructional Design Class.

ERIC Educational Resources Information Center

Graves, Rick; Barnett, Mardee; Gamble, Yolanda; Kolak, Mike

A case study was used in an instructional design class to facilitate the transfer of conceptual knowledge to concrete concerns and to aid instructional technology graduate students' understanding of the steps involved in designing, analyzing, and implementing an effective needs analysis. The case study involved real events at fictitious company…

The Fish Kill Mystery: Using Case Studies in the Middle School Classroom

ERIC Educational Resources Information Center

Heid, Christy; Biglan, Barbara; Ritson, Margaret

2008-01-01

Case studies are an excellent method for engaging middle school students in the current work of scientists. Students learn to think like scientists as they decide how to investigate the dilemma presented in the case study. This article describes one such case study, the Fish Kill Mystery, which takes place at a popular vacation spot--the beaches…

Benefits from Funding the MSD Engineering List: A Fiscal Year 1999 Case Study

DTIC Science & Technology

2004-03-01

BENEFITS FROM FUNDING THE MSD ENGINEERING LIST: A FISCAL YEAR 1999 CASE STUDY THESIS...States Government. AFIT/GLM/ENS/04-03 BENEFITS FROM FUNDING THE MSD ENGINEERING LIST: A FISCAL YEAR 1999 CASE STUDY...ENS/04-03 BENEFITS FROM FUNDING THE MSD ENGINEERING LIST: A FISCAL YEAR 1999 CASE STUDY David L. Gehrich, BS Captain, USAF

Recall bias in melanoma risk factors and measurement error effects: a nested case-control study within the Norwegian Women and Cancer Study.

PubMed

Parr, Christine L; Hjartåker, Anette; Laake, Petter; Lund, Eiliv; Veierød, Marit B

2009-02-01

Case-control studies of melanoma have the potential for recall bias after much public information about the relation with ultraviolet radiation. Recall bias has been investigated in few studies and only for some risk factors. A nested case-control study of recall bias was conducted in 2004 within the Norwegian Women and Cancer Study: 208 melanoma cases and 2,080 matched controls were invited. Data were analyzed for 162 cases (response, 78%) and 1,242 controls (response, 77%). Questionnaire responses to several host factors and ultraviolet exposures collected at enrollment in 1991-1997 and in 2004 were compared stratified on case-control status. Shifts in responses were observed among both cases and controls, but a shift in cases was observed only for skin color after chronic sun exposure, and a larger shift in cases was observed for nevi. Weighted kappa was lower for cases than for controls for most age intervals of sunburn, sunbathing vacations, and solarium use. Differences in odds ratio estimates of melanoma based on prospective and retrospective measurements indicate measurement error that is difficult to characterize. The authors conclude that indications of recall bias were found in this sample of Norwegian women, but that the results were inconsistent for the different exposures.

Rethinking Case Study Methodology in Poststructural Research.

PubMed

Mohammed, Shan; Peter, Elizabeth; Gastaldo, Denise; Howell, Doris

2015-03-01

Little consideration has been given to how case study might be used in poststructural research to explore power relations that constitute a phenomenon. Many case study scholars, most notably Robert Yin, adopt a postpositivist perspective that assumes the "truth" can be accessed through applying prescriptive and rigid research techniques. Using a discussion of Michel Foucault's key theoretical ideas and the insights gained through a Foucauldian case study of people with advanced cancer who continue to receive curative treatment, the authors argue for the expansion of case study in poststructural inquiry. They propose that the use of poststructuralist case study is valuable because of the flexibility and comprehensiveness of the methodology, which allows for the exploration of a deeper understanding of the broader discourses that shape a phenomenon, as well as how power/knowledge relations shape the behaviours and perceptions of people. They also introduce the reflexive implications of poststructural case study research. Copyright© by Ingram School of Nursing, McGill University.

Case Studies in Wilderness Medicine.

ERIC Educational Resources Information Center

Gray, Melissa; Tarter, Shana Lee

Five case studies explore issues in wilderness medicine, with emphasis on evacuation decision making. The cases describe medical problems encountered during wilderness trips involving college or high school students. In each case, the situation and facts of the case are outlined, including the patient's medical history and vital signs, and at…

A Study on the Failure of Conventional Deterrence: The Case Study of the Bombardment of Yeonpyeong in 2010

DTIC Science & Technology

2016-12-01

December 2016 3. REPORT TYPE AND DATES COVERED Master’s thesis 4. TITLE AND SUBTITLE A STUDY ON THE FAILURE OF CONVENTIONAL DETERRENCE: THE CASE ...Approved for public release. Distribution is unlimited A STUDY ON THE FAILURE OF CONVENTIONAL DETERRENCE: THE CASE STUDY OF THE BOMBARDMENT OF...necessary to analyze these diverse elements comprehensively. E. RESEARCH DESIGN This thesis focuses on a single case study about the bombardment of

Issues of Noncompliance. NCABR 2017 | Science Inventory ...

EPA Pesticide Factsheets

This presentation is a case study to be presented at an animal welfare conference. The purpose of the case study is to allow other animal welfare professionals a chance to work through and discuss an potential noncompliance situation. It is for practice, and to encourage people to learn from each other. Members of oversight organizations will be present to offer their perspectives on the case study as well. The purpose of this case study is to help educate other members of the animal welfare profession. This presentation is a case study to be presented at an animal welfare conference. The purpose of the case study is to allow other animal welfare professionals a chance to work through and discuss an potential noncompliance situation. It is for practice, and to encourage people to learn from each other.

Study designs may influence results: the problems with questionnaire-based case-control studies on the epidemiology of glioma.

PubMed

Johansen, Christoffer; Schüz, Joachim; Andreasen, Anne-Marie Serena; Dalton, Susanne Oksbjerg

2017-03-28

Glioma is a rare brain tumour with a very poor prognosis and the search for modifiable factors is intense. We reviewed the literature concerning risk factors for glioma obtained in case-control designed epidemiological studies in order to discuss the influence of this methodology on the observed results. When reviewing the association between three exposures, medical radiation, exogenous hormone use and allergy, we critically appraised the evidence from both case-control and cohort studies. For medical radiation and hormone replacement therapy (HRT), questionnaire-based case-control studies appeared to show an inverse association, whereas nested case-control and cohort studies showed no association. For allergies, the inverse association was observed irrespective of study design. We recommend that the questionnaire-based case-control design be placed lower in the hierarchy of studies for establishing cause-and-effect for diseases such as glioma. We suggest that a state-of-the-art case-control study should, as a minimum, be accompanied by extensive validation of the exposure assessment methods and the representativeness of the study sample with regard to the exposures of interest. Otherwise, such studies cannot be regarded as 'hypothesis testing' but only 'hypothesis generating'. We consider that this holds true for all questionnaire-based case-control studies on cancer and other chronic diseases, although perhaps not to the same extent for each exposure-outcome combination.

Comparing Management Approaches for Automatic Test Systems: A Strategic Missile Case Study

DTIC Science & Technology

2005-03-01

ground up, and is commonly conducted following five methods : ethnography , grounded theory , case study , phenomenological study , and biography...traditions frequently used (Creswell, 1998:5). The five traditions are biography, phenomenological study , grounded theory study , ethnography , and... Ethnography Biography Case Study Grounded Theory

Conceptualizing juvenile prostitution as child maltreatment: findings from the National Juvenile Prostitution Study.

PubMed

Mitchell, Kimberly J; Finkelhor, David; Wolak, Janis

2010-02-01

Two studies were conducted to identify the incidence (Study 1) and characteristics (Study 2) of juvenile prostitution cases known to law enforcement agencies in the United States. Study 1 revealed a national estimate of 1,450 arrests or detentions (95% confidence interval [CI]: 1,287-1,614) in cases involving juvenile prostitution during a 1-year period. In Study 2, exploratory data were collected from a subsample of 138 cases from police records in 2005. The cases are broadly categorized into three main types: (a) third-party exploiters, (b) solo prostitution, and (c) conventional child sexual abuse (CSA) with payment. Cases were classified into three initial categories based on police orientation toward the juvenile: (a) juveniles as victims (53%), (b) juveniles as delinquents (31%), and (c) juvenile as both victims and delinquents (16%). When examining the status of the juveniles by case type, the authors found that all the juveniles in CSA with payment cases were treated as victims, 66% in third-party exploiters cases, and 11% in solo cases. Findings indicate law enforcement responses to juvenile prostitution are influential in determining whether such youth are viewed as victims of commercial sexual exploitation or as delinquents.

Vitamin C Intake and Pancreatic Cancer Risk: A Meta-Analysis of Published Case-Control and Cohort Studies.

PubMed

Hua, Yong-Fei; Wang, Gao-Qing; Jiang, Wei; Huang, Jing; Chen, Guo-Chong; Lu, Cai-De

2016-01-01

Observational studies inconsistently reported the relationship between vitamin C intake and risk of pancreatic cancer. We conducted a meta-analysis of published case-control and cohort studies to quantify the association. Potentially eligible studies were found on PubMed and EMBASE databases through May 31, 2015. A random-effects model was assigned to compute summary point estimates with corresponding 95% confidence intervals (CIs). Subgroup and meta-regression analyses were also performed to explore sources of heterogeneity. Our final analyses included 20 observational studies comprising nearly 5 thousand cases of pancreatic cancer. When comparing the highest with the lowest categories of vitamin C intake, the summary odds ratio/relative risk for case-control studies (14 studies), cohort studies (6 studies) and all studies combined was 0.58 (95% CI: 0.52-0.66), 0.93 (95% CI: 0.78-1.11) and 0.66 (95% CI: 0.58-0.75), respectively. The difference in the findings between case-control and cohort studies was statistically significant (P < .001). Possible publication bias was shown in the meta-analysis of case-control studies. There is insufficient evidence to conclude any relationship between vitamin C intake and risk of pancreatic cancer. The strong inverse association observed in case-control studies may be affected by biases (eg, recall and selection biases) that particularly affect case-control studies and/or potential publication bias. Future prospective studies of vitamin C intake and pancreatic cancer are needed.

Using Case Studies in Calculus-based Physics

NASA Astrophysics Data System (ADS)

Katz, Debora M.

2006-12-01

Do your students believe that the physics only works in your classroom or laboratory? Or do they see that physics underlies their everyday experience? Case studies in physics help students connect physics principles to their everyday experience. For decades, case studies have been used to teach law, medicine and biology, but they are rarely used in physics. I am working on a calculus-based physics textbook for scientists and engineers. Case studies are woven into each chapter. Stop by and get a case study to test out in your classroom. I would love to get your feedback.

The lack of selection bias in a snowball sampled case-control study on drug abuse.

PubMed

Lopes, C S; Rodrigues, L C; Sichieri, R

1996-12-01

Friend controls in matched case-control studies can be a potential source of bias based on the assumption that friends are more likely to share exposure factors. This study evaluates the role of selection bias in a case-control study that used the snowball sampling method based on friendship for the selection of cases and controls. The cases selected fro the study were drug abusers located in the community. Exposure was defined by the presence of at least one psychiatric diagnosis. Psychiatric and drug abuse/dependence diagnoses were made according to the Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R) criteria. Cases and controls were matched on sex, age and friendship. The measurement of selection bias was made through the comparison of the proportion of exposed controls selected by exposed cases (p1) with the proportion of exposed controls selected by unexposed cases (p2). If p1 = p2 then, selection bias should not occur. The observed distribution of the 185 matched pairs having at least one psychiatric disorder showed a p1 value of 0.52 and a p2 value of 0.51, indicating no selection bias in this study. Our findings support the idea that the use of friend controls can produce a valid basis for a case-control study.

Optimal design of studies of influenza transmission in households. II: comparison between cohort and case-ascertained studies.

PubMed

Klick, B; Nishiura, H; Leung, G M; Cowling, B J

2014-04-01

Both case-ascertained household studies, in which households are recruited after an 'index case' is identified, and household cohort studies, where a household is enrolled before the start of the epidemic, may be used to test and estimate the protective effect of interventions used to prevent influenza transmission. A simulation approach parameterized with empirical data from household studies was used to evaluate and compare the statistical power of four study designs: a cohort study with routine virological testing of household contacts of infected index case, a cohort study where only household contacts with acute respiratory illness (ARI) are sampled for virological testing, a case-ascertained study with routine virological testing of household contacts, and a case-ascertained study where only household contacts with ARI are sampled for virological testing. We found that a case-ascertained study with ARI-triggered testing would be the most powerful design while a cohort design only testing household contacts with ARI was the least powerful. Sensitivity analysis demonstrated that these conclusions varied by model parameters including the serial interval and the risk of influenza virus infection from outside the household.

Evaluating psychiatric case-control studies using the STROBE (STrengthening the Reporting of OBservational Studies in Epidemiology) statement.

PubMed

Goi, Pedro Domingues; Goi, Julia Domingues; Cordini, Kariny Larissa; Ceresér, Keila Mendes; Rocha, Neusa Sica da

2014-01-01

Case-control studies are important in developing clinical and public health knowledge. The STROBE statement (STrengthening the Reporting of OBservational Studies in Epidemiology) was developed to establish a checklist of items that should be included in articles reporting observational studies. Our aim was to analyze whether the psychiatric case-control articles published in Brazilian journals with CAPES Qualis rating B1/B2 in 2009 conformed with the STROBE statement. Descriptive study on psychiatric papers published in Brazilian journals, within the Postgraduate Medical Program on Psychiatry, at Universidade Federal do Rio Grande do Sul. All psychiatric case-control studies from Brazilian Qualis B1/B2 journals of psychiatry, neurology and public health in 2009 were analyzed. The four most specific items of the STROBE statement were used to evaluate whether these studies fitted within the case-control parameters: 1) selection of cases and controls; 2) controlling for bias; 3) statistical analysis; and 4) presentation of results. Sixteen case-control studies were identified, of which eleven (68.75%) were in psychiatry-focused journals. From analysis using the STROBE statement, all of the articles conformed with item 1; two (12.5%) completely conformed with item 2; none completely conformed with item 3; and only three (18.8%) conformed with item 4. The case-control studies analyzed here did not completely conform with the four STROBE statement items for case-control design. In view of the inadequate methodology of the published studies, these findings justify focusing on research and methodology and expanding the investigations on adherence of studies to their designs.

Case Studies for Effective Business Instruction.

ERIC Educational Resources Information Center

McAlister-Kizzier, Donna

This book is designed as a resource for educators who teach business content in a variety of instructional settings. It contains case studies representing all functional areas of business, including corporate training, for grades 7 through graduate education. Chapter 1 provides an overview of the case study method. The history of the case method,…

Using Case-Based Pedagogy in the Philippines: A Narrative Inquiry

NASA Astrophysics Data System (ADS)

Arellano, Elvira L.; Barcenal, Tessie L.; Bilbao, Purita P.; Castellano, Merilin A.; Nichols, Sharon E.; Tippins, Deborah J.

2001-04-01

This study features use of case-based pedagogy as a context for exploring science teacher education reform. A central premise of the study is that science teacher education reform is a complex process characterised by the individual and collective narrative histories of teachers. The study was conducted at West Visayas State University in the Philippines as a collaborative research endeavour involving two US and four Filipino science teacher educators. The centrepiece of this study was a case experience prompted by reflection on prospective and practicing elementary teachers' written cases. Results of the study are presented here as a layered narrative. An initial layer explores how a case, written by a prospective teacher, was interwoven with her past and future life history. A second layer looks at sociocultural and professional tensions that mediated the participants' case experience. A third layer represents how the case experience provided a community that supported critical reflection among the research team members. Implications from the study emphasise that case-based pedagogy can serve as a basis for critical reflection to inform community-based approaches to science teacher education pedagogical and research reform.

A Case Study of the Degree of Collaboration Between Various Levels in the Reparable Chain in the United States Air Force

DTIC Science & Technology

2005-03-01

qualitative research methods , a case study approach was selected to conduct this research . “A case study can be defined as an empirical study ... qualitative in nature, and also described the qualitative research method chosen as a case study . From 49 there, data collection was focused upon... qualitative nature of the research , a qualitative design was used to conduct the

Response rates in case-control studies of cancer by era of fieldwork and by characteristics of study design.

PubMed

Xu, Mengting; Richardson, Lesley; Campbell, Sally; Pintos, Javier; Siemiatycki, Jack

2018-04-09

The purpose of this study was to describe time trends in response rates in case-control studies of cancer and identify study design factors that influence response rate. We reviewed 370 case-control studies of cancer published in 12 journals during indicator years in each of the last four decades. We estimated time trends of response rates and reasons for nonresponse in each of the following types of study subjects: cases, medical source controls, and population controls. We also estimated response rates according to characteristics of study context. Median response rates among cases and population controls were between 75% and 80% in the 1970s. Between 1971 and 2010, study response rates declined by 0.31% per year for cases and 0.78% for population controls. Only a minority of studies reported reasons for nonparticipation; subject refusal was the most common reported reason. Studies conducted in North America had lower median response rates than studies conducted in Europe. In-person and telephone interviews elicited higher response rates than mail questionnaires. Response rates from case-control studies of cancer have declined, and this could threaten the validity of results derived from these studies. Copyright © 2018 Elsevier Inc. All rights reserved.

Case Studies in the Field of Marketing Education: Learner Impact, Case Performance, and Cost Efficiency

ERIC Educational Resources Information Center

Spais, George S.

2005-01-01

The major objective of this study is to identify a methodology that will help educators in marketing to efficiently manage the design, impact, and cost of case studies. It is my intention is to examine the impact of case study characteristics in relation to the degree of learner involvement in the learning process. The author proposes that…

Case Studies for Management Development in Bangladesh. Fourth Book. Parepared for the World Bank/Bangladesh Management Education and Training Project.

ERIC Educational Resources Information Center

McLean, Gary N.

This document contains five case studies of management and finance situations in Bangladesh, which are intended to foster classroom discussion and thereby advance management development in that country. One to four discussion questions follow each case study. The titles and authors of the case studies on management are: "Azir Ahmed & Co.,…

Case Studies for Management Development in Bangladesh. Third Book. Prepared for the World Bank/Bangladesh Management Education and Training Project.

ERIC Educational Resources Information Center

McLean, Gary N.

This document contains 10 case studies of management situations in Bangladesh, which are intended to foster classroom discussion and thereby advance management development in that country. Two to four discussion questions follow each case study. Three case studies on the subject of pay and compensation were prepared by Abu Hossain Siddique; the…

Colombia - A Case Study in Smart Power

DTIC Science & Technology

2014-05-22

COLOMBIA–A CASE STUDY IN SMART POWER A Monograph by John P. Brady Foreign Service Officer United States Agency for...Colombia–A Case Study in Smart Power 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) John P. Brady...of Candidate: John P. Brady Monograph Title: Colombia–A Case Study in Smart Power Approved by: , Monograph Director Robert W

Incorporating ITS into transportation improvement planning : the Seattle Case Study using PRUEVIIN

DOT National Transportation Integrated Search

1998-01-01

This project explored methods to analyze ITS strategies within Major Investment Study (MIS) studies and to apply them in a case study. The case study developed methods to define alternatives, and to estimate impacts and costs at the level required fo...

Building Language Educators: The Implications of Case-Based Pedagogy Using Practicum-Based Student-Teacher-Generated Cases for Pre-Service Language Teacher Education

ERIC Educational Resources Information Center

Cournoyer, Amy Beth

2014-01-01

This case study investigated case-based pedagogy using student-teacher-generated cases as an instructional tool in the preparation of 12 pre-service ESL, Bilingual, and Modern Foreign Language teachers enrolled in a Student Teaching Seminar at a post-secondary institution. In the fall methods course, each participant generated a case study based…

Biomass production in agroforestry and forestry systems on salt-affected soils in South Asia: exploration of the GHG balance and economic performance of three case studies.

PubMed

Wicke, Birka; Smeets, Edward M W; Akanda, Razzaque; Stille, Leon; Singh, Ranjay K; Awan, Abdul Rasul; Mahmood, Khalid; Faaij, Andre P C

2013-09-30

This study explores the greenhouse gas balance and the economic performance (i.e. net present value (NPV) and production costs) of agroforestry and forestry systems on salt-affected soils (biosaline (agro)forestry) based on three case studies in South Asia. The economic impact of trading carbon credits generated by biosaline (agro)forestry is also assessed as a potential additional source of income. The greenhouse gas balance shows carbon sequestration over the plantation lifetime of 24 Mg CO2-eq. ha(-1) in a rice-Eucalyptus camaldulensis agroforestry system on moderately saline soils in coastal Bangladesh (case study 1), 6 Mg CO2-eq. ha(-1) in the rice-wheat- Eucalyptus tereticornis agroforestry system on sodic/saline-sodic soils in Haryana state, India (case study 2), and 96 Mg CO2-eq. ha(-1) in the compact tree (Acacia nilotica) plantation on saline-sodic soils in Punjab province of Pakistan. The NPV at a discount rate of 10% is 1.1 k€ ha(-1) for case study 1, 4.8 k€ ha(-1) for case study 2, and 2.8 k€ ha(-1) for case study 3. Carbon sequestration translates into economic values that increase the NPV by 1-12% in case study 1, 0.1-1% in case study 2, and 2-24% in case study 3 depending on the carbon credit price (1-15 € Mg(-1) CO2-eq.). The analysis of the three cases indicates that the economic performance strongly depends on the type and severity of salt-affectedness (which affect the type and setup of the agroforestry system, the tree species and the biomass yield), markets for wood products, possibility of trading carbon credits, and discount rate. Copyright © 2013 Elsevier Ltd. All rights reserved.

[Case and studies].

PubMed

Schubert, András

2015-11-15

Case studies and case reports form an important and ever growing part of scientific and scholarly literature. The paper deals with the share and citation rate of these publication types on different fields of research. In general, evidence seems to support the opinion that an excessive number of such publications may negatively influence the impact factor of the journal. In the literature of scientometrics, case studies (at least the presence of the term "case study" in the titles of the papers) have a moderate share, but their citation rate is practically equal to that of other publication types.

Combining epidemiology and biomechanics in sports injury prevention research: a new approach for selecting suitable controls.

PubMed

Finch, Caroline F; Ullah, Shahid; McIntosh, Andrew S

2011-01-01

Several important methodological issues need to be considered when designing sports injury case-control studies. Major design goals for case-control studies include the accounting for prior injury risk exposure, and optimal definitions of both cases and suitable controls are needed to ensure this. This article reviews methodological aspects of published sports injury case-control studies, particularly with regard to the selection of controls. It argues for a new approach towards selecting controls for case-control studies that draws on an interface between epidemiological and biomechanical concepts. A review was conducted to identify sport injury case-control studies published in the peer-review literature during 1985-2008. Overall, 32 articles were identified, of which the majority related to upper or lower extremity injuries. Matching considerations were used for control selection in 16 studies. Specific mention of application of biomechanical principles in the selection of appropriate controls was absent from all studies, including those purporting to evaluate the benefits of personal protective equipment to protect against impact injury. This is a problem because it could lead to biased conclusions, as cases and controls are not fully comparable in terms of similar biomechanical impact profiles relating to the injury incident, such as site of the impact on the body. The strength of the conclusions drawn from case-control studies, and the extent to which results can be generalized, is directly influenced by the definition and recruitment of cases and appropriate controls. Future studies should consider the interface between epidemiological and biomechanical concepts when choosing appropriate controls to ensure that proper adjustment of prior exposure to injury risk is made. To provide necessary guidance for the optimal selection of controls in case-control studies of interventions to prevent sports-related impact injury, this review outlines a new case-control selection strategy that reflects the importance of biomechanical considerations, which ensures that controls are selected based on the presence of the same global injury mechanism as the cases. To summarize, the general biomechanical principles that should apply to the selection of controls in future case-control studies are as follows: (i) each control must have been exposed to the same global injury mechanism as the case, (e.g. head impact, fall onto outstretched arm); and (ii) intrinsic (individual) factors (e.g. age, sex, skill level) that might modify the person's response to the relevant biomechanical loads are adjusted when either selecting the controls or are in the analysis phase. The same considerations for control selection apply to other study designs such as matched cohort studies or case-crossover studies.

Improving the Effectiveness of Professional Education: Learning Managerial Accounting via a Complex Case.

ERIC Educational Resources Information Center

Carter, Melissa; And Others

To give students more experience with real situations, many professional schools use case studies in their courses. Creating complex cases, case experiences that immerse students in complex problems, rather than mere case studies that require armchair analysis should help students gain better and more integrated knowledge. Designing, implementing,…

A systematic review of occupational safety and health business cases.

PubMed

Verbeek, Jos; Pulliainen, Marjo; Kankaanpää, Eila

2009-12-01

Business cases are commonly developed as means to rationalize investment. We systematically reviewed 26 reported cases on occupational safety and health (OSH) interventions to assess if health and productivity arguments make a good business case. To be included in the review, studies had to analyze the costs and benefits, including productivity, of an OSH intervention at the enterprise level. We searched Medline and Embase for studies and used Google search in addition. Two reviewers independently selected studies and extracted data. The intervention profitability was calculated in euros (euro in 2008) as the first year's benefits minus the total intervention costs per worker. The payback period was calculated as the intervention costs divided by the first year's benefits. We found three ex-ante and 23 ex-post cases. In 20 cases, the study design was a before-after comparison without a control group. Generally a 100% reduction of injuries or sickness absence was assumed. In two cases, productivity and quality increases were very large. The main benefit was avoided sick leave. Depreciation or discounting was applied only in a minority of cases. The intervention profitability was negative in seven studies, up to euro 500 per employee in 12 studies and more than euro 500 per employee in seven studies. The payback period was less than half a year for 19 studies. Only a few studies included sensitivity analyses. Few ex-ante business cases for management decisions on OSH are reported. Guidelines for reporting and evaluation are needed. Business cases need more sound assumptions on the effectiveness of interventions and should incorporate greater uncertainty into their design. Ex-post evaluation should be based preferably on study designs that control for trends at a time different from that of the intervention.

The Use of Interrupted Case Studies to Enhance Critical Thinking Skills in Biology

PubMed Central

White, Tracy K.; Whitaker, Paul; Gonya, Terri; Hein, Richard; Kroening, Dubear; Lee, Kevin; Lee, Laura; Lukowiak, Andrea; Hayes, Elizabeth

2009-01-01

There has been a dramatic increase in the availability of case studies for use in the biology classroom, and perceptions of the effectiveness of case-study-based learning are overwhelmingly positive. Here we report the results of a study in which we evaluated the ability of interrupted case studies to improve critical thinking in the context of experimental design and the conventions of data interpretation. Students were assessed using further case studies designed to evaluate their ability to recognize and articulate problematic approaches to these elements of experimentation. Our work reveals that case studies have broad utility in the classroom. In addition to demonstrating a small but statistically significant increase in the number of students capable of critically evaluating selected aspects of experimental design, we also observed increased student engagement and documented widespread misconceptions regarding the conventions of data acquisition and analysis. PMID:23653687

Better cancer biomarker discovery through better study design.

PubMed

Rundle, Andrew; Ahsan, Habibul; Vineis, Paolo

2012-12-01

High-throughput laboratory technologies coupled with sophisticated bioinformatics algorithms have tremendous potential for discovering novel biomarkers, or profiles of biomarkers, that could serve as predictors of disease risk, response to treatment or prognosis. We discuss methodological issues in wedding high-throughput approaches for biomarker discovery with the case-control study designs typically used in biomarker discovery studies, especially focusing on nested case-control designs. We review principles for nested case-control study design in relation to biomarker discovery studies and describe how the efficiency of biomarker discovery can be effected by study design choices. We develop a simulated prostate cancer cohort data set and a series of biomarker discovery case-control studies nested within the cohort to illustrate how study design choices can influence biomarker discovery process. Common elements of nested case-control design, incidence density sampling and matching of controls to cases are not typically factored correctly into biomarker discovery analyses, inducing bias in the discovery process. We illustrate how incidence density sampling and matching of controls to cases reduce the apparent specificity of truly valid biomarkers 'discovered' in a nested case-control study. We also propose and demonstrate a new case-control matching protocol, we call 'antimatching', that improves the efficiency of biomarker discovery studies. For a valid, but as yet undiscovered, biomarker(s) disjunctions between correctly designed epidemiologic studies and the practice of biomarker discovery reduce the likelihood that true biomarker(s) will be discovered and increases the false-positive discovery rate. © 2012 The Authors. European Journal of Clinical Investigation © 2012 Stichting European Society for Clinical Investigation Journal Foundation.

Case-control vaccine effectiveness studies: Preparation, design, and enrollment of cases and controls.

PubMed

Verani, Jennifer R; Baqui, Abdullah H; Broome, Claire V; Cherian, Thomas; Cohen, Cheryl; Farrar, Jennifer L; Feikin, Daniel R; Groome, Michelle J; Hajjeh, Rana A; Johnson, Hope L; Madhi, Shabir A; Mulholland, Kim; O'Brien, Katherine L; Parashar, Umesh D; Patel, Manish M; Rodrigues, Laura C; Santosham, Mathuram; Scott, J Anthony; Smith, Peter G; Sommerfelt, Halvor; Tate, Jacqueline E; Victor, J Chris; Whitney, Cynthia G; Zaidi, Anita K; Zell, Elizabeth R

2017-06-05

Case-control studies are commonly used to evaluate effectiveness of licensed vaccines after deployment in public health programs. Such studies can provide policy-relevant data on vaccine performance under 'real world' conditions, contributing to the evidence base to support and sustain introduction of new vaccines. However, case-control studies do not measure the impact of vaccine introduction on disease at a population level, and are subject to bias and confounding, which may lead to inaccurate results that can misinform policy decisions. In 2012, a group of experts met to review recent experience with case-control studies evaluating the effectiveness of several vaccines; here we summarize the recommendations of that group regarding best practices for planning, design and enrollment of cases and controls. Rigorous planning and preparation should focus on understanding the study context including healthcare-seeking and vaccination practices. Case-control vaccine effectiveness studies are best carried out soon after vaccine introduction because high coverage creates strong potential for confounding. Endpoints specific to the vaccine target are preferable to non-specific clinical syndromes since the proportion of non-specific outcomes preventable through vaccination may vary over time and place, leading to potentially confusing results. Controls should be representative of the source population from which cases arise, and are generally recruited from the community or health facilities where cases are enrolled. Matching of controls to cases for potential confounding factors is commonly used, although should be reserved for a limited number of key variables believed to be linked to both vaccination and disease. Case-control vaccine effectiveness studies can provide information useful to guide policy decisions and vaccine development, however rigorous preparation and design is essential. Published by Elsevier Ltd.

The Ethnic/Racial Variations of Intracerebral Hemorrhage (ERICH) Study Protocol

PubMed Central

Woo, Daniel; Rosand, Jonathan; Kidwell, Chelsea; McCauley, Jacob L.; Osborne, Jennifer; Brown, Mark W.; West, Sandra E.; Rademacher, Eric W.; Waddy, Salina; Roberts, Jamie N.; Koch, Sebastian; Gonzales, Nicole R.; Sung, Gene; Kittner, Steven J.; Birnbaum, Lee; Frankel, Michael; Daniel Testai, Fernando; Hall, Christiana E.; Elkind, Mitchell S. V.; Flaherty, Matthew; Coull, Bruce; Chong, Ji Y.; Warwick, Tanya; Malkoff, Marc; James, Michael L.; Ali, Latisha K.; Worrall, Bradford B.; Jones, Floyd; Watson, Tiffany; Leonard, Anne; Martinez, Rebecca; Sacco, Ralph I; Langefeld, Carl D.

2013-01-01

Background and Purpose Epidemiologic studies of intracerebral hemorrhage (ICH) have consistently demonstrated variation in incidence, location, age at presentation, and outcomes among non-Hispanic white, black, and Hispanic populations. We report here the design and methods for this large, prospective, multi-center case-control study of ICH. Methods The ERICH study is a multi-center, prospective case-control study of ICH. Cases are identified by hot-pursuit and enrolled using standard phenotype and risk factor information and include neuroimaging and blood sample collection. Controls are centrally identified by random digit dialing to match cases by age (+/−5 years), race, ethnicity, gender and metropolitan region. Results As of March 22, 2013, 1,655 cases of ICH had been recruited into the study which is 101.5% of the target for that date and 851 controls had been recruited which is 67.2% of the target for that date (1,267 controls) for a total of 2,506 subjects which is 86.5% of the target for that date (2,897 subjects). Of the 1,655 cases enrolled, 1,640 cases had the case interview entered into the database of which 628 (38%) were non-Hispanic black, 458 (28%) were non-Hispanic white and 554 (34%) were Hispanic. Of the 1,197 cases with imaging submitted, 876 (73.2%) had a 24 hour follow-up CT available In addition to CT imaging, 607 cases have had MRI evaluation. Conclusion The ERICH study is a large, case-control study of ICH with particular emphasis on recruitment of minority populations for the identification of genetic and epidemiologic risk factors for ICH and outcomes after ICH. PMID:24021679

Research participation experiences of informants of suicide and control cases: taken from a case-control psychological autopsy study of people who died by suicide.

PubMed

Wong, Paul W C; Chan, Wincy S C; Beh, Philip S L; Yau, Fiona W S; Yip, Paul S F; Hawton, Keith

2010-01-01

Ethical issues have been raised about using the psychological autopsy approach in the study of suicide. The impact on informants of control cases who participated in case-control psychological autopsy studies has not been investigated. (1) To investigate whether informants of suicide cases recruited by two approaches (coroners' court and public mortuaries) respond differently to the initial contact by the research team. (2) To explore the reactions, reasons for participation, and comments of both the informants of suicide and control cases to psychological autopsy interviews. (3) To investigate the impact of the interviews on informants of suicide cases about a month after the interviews. A self-report questionnaire was used for the informants of both suicide and control cases. Telephone follow-up interviews were conducted with the informants of suicide cases. The majority of the informants of suicide cases, regardless of the initial route of contact, as well as the control cases were positive about being approached to take part in the study. A minority of informants of suicide and control cases found the experience of talking about their family member to be more upsetting than expected. The telephone follow-up interviews showed that none of the informants of suicide cases reported being distressed by the psychological autopsy interviews. The acceptance rate for our original psychological autopsy study was modest. The findings of this study are useful for future participants and researchers in measuring the potential benefits and risks of participating in similar sensitive research. Psychological autopsy interviews may be utilized as an active engagement approach to reach out to the people bereaved by suicide, especially in places where the postvention work is underdeveloped.

Acute respiratory infection case definitions for young children: a systematic review of community-based epidemiologic studies in South Asia.

PubMed

Roth, Daniel E; Gaffey, Michelle F; Smith-Romero, Evelyn; Fitzpatrick, Tiffany; Morris, Shaun K

2015-12-01

To explore the variability in childhood acute respiratory infection case definitions for research in low-income settings where there is limited access to laboratory or radiologic investigations. We conducted a systematic review of community-based, longitudinal studies in South Asia published from January 1990 to August 2013, in which childhood acute respiratory infection outcomes were reported. Case definitions were classified by their label (e.g. pneumonia, acute lower respiratory infection) and clinical content 'signatures' (array of clinical features that would be always present, conditionally present or always absent among cases). Case definition heterogeneity was primarily assessed by the number of unique case definitions overall and by label. We also compared case definition-specific acute respiratory infection incidence rates for studies reporting incidence rates for multiple case definitions. In 56 eligible studies, we found 124 acute respiratory infection case definitions. Of 90 case definitions for which clinical content was explicitly defined, 66 (73%) were unique. There was a high degree of content heterogeneity among case definitions with the same label, and some content signatures were assigned multiple labels. Within studies for which incidence rates were reported for multiple case definitions, variation in content was always associated with a change in incidence rate, even when the content differed by a single clinical feature. There has been a wide variability in case definition label and content combinations to define acute upper and lower respiratory infections in children in community-based studies in South Asia over the past two decades. These inconsistencies have important implications for the synthesis and translation of knowledge regarding the prevention and treatment of childhood acute respiratory infection. © 2015 John Wiley & Sons Ltd.

Study design options in evaluating gene-environment interactions: practical considerations for a planned case-control study of pediatric leukemia.

PubMed

Goodman, Michael; Dana Flanders, W

2007-04-01

We compare methodological approaches for evaluating gene-environment interaction using a planned study of pediatric leukemia as a practical example. We considered three design options: a full case-control study (Option I), a case-only study (Option II), and a partial case-control study (Option III), in which information on controls is limited to environmental exposure only. For each design option we determined its ability to measure the main effects of environmental factor E and genetic factor G, and the interaction between E and G. Using the leukemia study example, we calculated sample sizes required to detect and odds ratio (OR) of 2.0 for E alone, an OR of 10 for G alone and an interaction G x E of 3. Option I allows measuring both main effects and interaction, but requires a total sample size of 1,500 cases and 1,500 controls. Option II allows measuring only interaction, but requires just 121 cases. Option III allows calculating the main effect of E, and interaction, but not the main effect of G, and requires a total of 156 cases and 133 controls. In this case, the partial case-control study (Option III) appears to be more efficient with respect to its ability to answer the research questions for the amount of resources required. The design options considered in this example are not limited to observational epidemiology and may be applicable in studies of pharmacogenomics, survivorship, and other areas of pediatric ALL research.

A Homogenization Approach for Design and Simulation of Blast Resistant Composites

NASA Astrophysics Data System (ADS)

Sheyka, Michael

Structural composites have been used in aerospace and structural engineering due to their high strength to weight ratio. Composite laminates have been successfully and extensively used in blast mitigation. This dissertation examines the use of the homogenization approach to design and simulate blast resistant composites. Three case studies are performed to examine the usefulness of different methods that may be used in designing and optimizing composite plates for blast resistance. The first case study utilizes a single degree of freedom system to simulate the blast and a reliability based approach. The first case study examines homogeneous plates and the optimal stacking sequence and plate thicknesses are determined. The second and third case studies use the homogenization method to calculate the properties of composite unit cell made of two different materials. The methods are integrated with dynamic simulation environments and advanced optimization algorithms. The second case study is 2-D and uses an implicit blast simulation, while the third case study is 3-D and simulates blast using the explicit blast method. Both case studies 2 and 3 rely on multi-objective genetic algorithms for the optimization process. Pareto optimal solutions are determined in case studies 2 and 3. Case study 3 is an integrative method for determining optimal stacking sequence, microstructure and plate thicknesses. The validity of the different methods such as homogenization, reliability, explicit blast modeling and multi-objective genetic algorithms are discussed. Possible extension of the methods to include strain rate effects and parallel computation is also examined.

The evaluation of complex interventions in palliative care: an exploration of the potential of case study research strategies.

PubMed

Walshe, Catherine

2011-12-01

Complex, incrementally changing, context dependent and variable palliative care services are difficult to evaluate. Case study research strategies may have potential to contribute to evaluating such complex interventions, and to develop this field of evaluation research. This paper explores definitions of case study (as a unit of study, a process, and a product) and examines the features of case study research strategies which are thought to confer benefits for the evaluation of complex interventions in palliative care settings. Ten features of case study that are thought to be beneficial in evaluating complex interventions in palliative care are discussed, drawing from exemplars of research in this field. Important features are related to a longitudinal approach, triangulation, purposive instance selection, comprehensive approach, multiple data sources, flexibility, concurrent data collection and analysis, search for proving-disproving evidence, pattern matching techniques and an engaging narrative. The limitations of case study approaches are discussed including the potential for subjectivity and their complex, time consuming and potentially expensive nature. Case study research strategies have great potential in evaluating complex interventions in palliative care settings. Three key features need to be exploited to develop this field: case selection, longitudinal designs, and the use of rival hypotheses. In particular, case study should be used in situations where there is interplay and interdependency between the intervention and its context, such that it is difficult to define or find relevant comparisons.

The contribution of case study design to supporting research on Clubhouse psychosocial rehabilitation.

PubMed

Raeburn, Toby; Schmied, Virginia; Hungerford, Catherine; Cleary, Michelle

2015-10-01

Psychosocial Clubhouses provide recovery-focused psychosocial rehabilitation to people with serious mental illness at over 300 sites in more than 30 countries worldwide. To deliver the services involved, Clubhouses employ a complex mix of theory, programs and relationships, with this complexity presenting a number of challenges to those undertaking Clubhouse research. This paper provides an overview of the usefulness of case study designs for Clubhouse researchers; and suggests ways in which the evaluation of Clubhouse models can be facilitated. The paper begins by providing a brief explanation of the Clubhouse model of psychosocial rehabilitation, and the need for ongoing evaluation of the services delivered. This explanation is followed by an introduction to case study design, with consideration given to the way in which case studies have been used in past Clubhouse research. It is posited that case study design provides a methodological framework that supports the analysis of either quantitative, qualitative or a mixture of both types of data to investigate complex phenomena in their everyday contexts, and thereby support the development of theory. As such, case study approaches to research are well suited to the Clubhouse environment. The paper concludes with recommendations for future Clubhouse researchers who choose to employ a case study design. While the quality of case study research that explores Clubhouses has been variable in the past, if applied in a diligent manner, case study design has a valuable contribution to make in future Clubhouse research.

Teaching Business Demography Using Case Studies.

PubMed

Swanson, David A; Morrison, Peter A

2010-02-01

Many faculty members consider using case studies but not all end up using them. We provide a brief review of what cases are intended to do and identify three ways in which they can be used. We then use an example to illustrate how we have used the case study method in teaching business demography. Among other benefits, we note that the case studies method not only encourages the acquisition of skills by students, but can be used to promote "deep structure learning," an approach naturally accommodates other features associated with the case studies method-the development of critical thinking skills, the use of real world problems, the emphasis of concepts over mechanics, writing and presentation skills, active cooperative learning and the "worthwhileness" of a course. As noted by others, we understand the limitations of the case study method. However, given its strengths, we believe it has a place in the instructional toolbox for courses in business demography. The fact that courses we teach is a testament to our perceived efficacy of this tool.

Rare variant association analysis in case-parents studies by allowing for missing parental genotypes.

PubMed

Li, Yumei; Xiang, Yang; Xu, Chao; Shen, Hui; Deng, Hongwen

2018-01-15

The development of next-generation sequencing technologies has facilitated the identification of rare variants. Family-based design is commonly used to effectively control for population admixture and substructure, which is more prominent for rare variants. Case-parents studies, as typical strategies in family-based design, are widely used in rare variant-disease association analysis. Current methods in case-parents studies are based on complete case-parents data; however, parental genotypes may be missing in case-parents trios, and removing these data may lead to a loss in statistical power. The present study focuses on testing for rare variant-disease association in case-parents study by allowing for missing parental genotypes. In this report, we extended the collapsing method for rare variant association analysis in case-parents studies to allow for missing parental genotypes, and investigated the performance of two methods by using the difference of genotypes between affected offspring and their corresponding "complements" in case-parent trios and TDT framework. Using simulations, we showed that, compared with the methods just only using complete case-parents data, the proposed strategy allowing for missing parental genotypes, or even adding unrelated affected individuals, can greatly improve the statistical power and meanwhile is not affected by population stratification. We conclude that adding case-parents data with missing parental genotypes to complete case-parents data set can greatly improve the power of our strategy for rare variant-disease association.

The Crucial Role of the Operational Artist: A Case Study of Operation Barbarossa

DTIC Science & Technology

2017-05-25

historical case study helps to develop a better understanding of operational art today. Future studies might focus on the limitations and opportunities...The Crucial Role of the Operational Artist: A Case Study of Operation Barbarossa A Monograph by LTC (GS) Hagen H. Ruppelt German Army...a Case Study of Operation Barbarossa 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) LTC (GS) Hagen H

A Case Study of ESD Implementation: Signs of Sustainable Leadership

ERIC Educational Resources Information Center

Iliško, Dzintra; Badyanova, Yelena

2014-01-01

This article presents a case study of two schools that were identified as a result of UNESCO associated schools survey as cases of sustainable leadership and governance. The aim of the study is to present the two cases that were crystalized in the survey carried out at end of the United Nations' "Decade of Education for Sustainable…

Two Case Studies of Beginning Teachers in State-Mandated Induction Programs: The Influence of Institutional Factors.

ERIC Educational Resources Information Center

Paulissen, Margaret O.; And Others

The Teacher Induction Study investigated 2 state-mandated beginning teacher programs and examined the translation of state policy by 4 school districts, 13 individual schools, and 32 classrooms. From the case histories of 16 teams, 2 case histories were selected for further study. One case illustrated how institutional factors influenced team…

Online Case-Based Discussions: Examining Coverage of the Afforded Problem Space

ERIC Educational Resources Information Center

Ertmer, Peggy A.; Koehler, Adrie A.

2014-01-01

Case studies hold great potential for engaging students in disciplinary content. However, little is known about the extent to which students actually cover the problem space afforded by a particular case study. In this research, we compared the problem space afforded by an instructional design case study with the actual content covered by 16…

World Perspective Case Descriptions on Educational Programs for Adults: Chile.

ERIC Educational Resources Information Center

Donoso, Patricio; Gajardo, Marcela

This document contains two case studies of adult education programs in Chile. Both case studies begin with a "face sheet" on which is recorded basic information about the program and the description. The first case study, prepared by Patricio Donoso, reports on Centro El Canelo de Nos, an inservice center for educators who work with…

Case Study: Let's Get Personal--Putting Personality into Your Cases

ERIC Educational Resources Information Center

Herreid, Clyde Freeman

2017-01-01

In this case study, the subject for discussion is the need to use teaching notes in order to add personality to case studies. Personality, it is said, is more important than policy, than demographics, than finances, or amorous peccadillos. Personality is essential. It is the interaction of people that capture our interest, yet, the question is…

78 FR 12764 - Draft Office of Health Assessment and Translation Approach for Systematic Review and Evidence...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-25

... Approach--February 2013 might be needed, OHAT plans to apply it to two case-study evaluations. One case... Availability: Draft OHAT Approach--February 2013 will be available by February 26, 2013, and case-study... framework, describe the contents in the case-study protocols, and respond to questions from the public on...

Problematising Ethnography and Case Study: Reflections on Using Ethnographic Techniques and Researcher Positioning

ERIC Educational Resources Information Center

Parker-Jenkins, Marie

2018-01-01

This paper was prompted by the question, what do we mean by conducting "ethnography"? Is it in fact "case study" drawing on ethnographic techniques? My contention is that in many cases, researchers are not actually conducting ethnography as understood within a traditional sense but rather are engaging in case study, drawing on…

A Case-Based Approach Increases Student Learning Outcomes and Comprehension of Cellular Respiration Concepts

ERIC Educational Resources Information Center

Rybarczyk, Brian J.; Baines, Antonio T.; McVey, Mitch; Thompson, Joseph T.; Wilkins, Heather

2007-01-01

This study investigated student learning outcomes using a case-based approach focused on cellular respiration. Students who used the case study, relative to students who did not use the case study, exhibited a significantly greater learning gain, and demonstrated use of higher-order thinking skills. Preliminary data indicate that after engaging…

The Analysis of a Murder, a Case Study

ERIC Educational Resources Information Center

Dinan, Frank J.; Szczepankiewicz, Steven H.; Carnaham, Melinda; Colvin, Michael T.

2007-01-01

The analysis of a Murder case study can be used in general chemistry or nonscience major chemistry courses to teach data handling and analysis in a non-laboratory context. This case study will help students to respond more enthusiastically and with more interest to a set of material placed before them related to a real case rather than…

Pershing in Mexico: a case study in limited contingency operations

DTIC Science & Technology

2016-05-26

Pershing in Mexico: A Case Study in Limited Contingency Operations A Monograph by MAJ Timothy J. Lawrence United States Army School...DATE (00-MM-YYYY) REPORT TYPE 12. 31-03-2016 SAMS Monograph 4. TITLE AND SUBTITLE Pershing in Mexico: A Case Study in Limited Contingency Operations...Monograph Approval Page Name of Candidate: MAJ Timothy J. Lawrence Monograph Title: Pershing in Mexico: A Case Study in Limited Contingency Operations

Reexamining Ground SOF Command and Control: Does One Size Fit All

DTIC Science & Technology

2016-12-01

Case studies covering varying methods of employment of special operations forces in limited contingencies will be utilized. The case studies focus on ...operating environment, much like an open organization. This research examines three case studies including operation JUST CAUSE in Panama, RESTORE HOPE in... case studies , the HQ elements formed around existing homogenous commands and not a Theater Special Operations Command (TSOC) staff with limited

[Clinical studies on flomoxef in neonates].

PubMed

Tabuki, K; Nishimura, T

1993-07-01

Clinical studies on flomoxef (FMOX) were performed in neonates and the results obtained are summarized as follows. Treatment with FMOX was made in 4 cases of neonatal bacterial infections; 2 cases of sepsis (suspected) and 1 case each of infection of umbilicus and staphylococcal scalded skin syndrome. Results obtained were excellent in 1 case, good in 3 cases. No significant side effects due to the drug were observed in any cases.

Comparison of Success of Clomiphene citrate and Letrozole in Ovulation Induction.

PubMed

Saha, J; Akhter, S; Prasad, I; Siddiq, S

2016-01-01

The study was carried out to evaluate which drug is better in ovulation induction between clomiphene citrate and letrozole. The study was carried out in the infertility unit of Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka and Centre for Assisted Reproduction (CARE) at Bangladesh Institute of Research and Rehabilitation in Diabetes Endocrine and Metabolic Disorders (BIRDEM), Dhaka from January 2007 to December 2007. One hundred and sixty five cases were taken for the study. It was a prospective interventional comparative study of clomiphene citrate and letrozole in infertile cases. The patients were divided into three groups. Group I--newly detected cases of sub fertility studied with clomiphene citrate. Group II--clomiphene citrate resistant cases studied with letrozole, Group III--newly detected cases of sub fertility studied with letrozole. The cases were followed up for outcome; (ovulation). The TVS was done on 12th or 13th day of menstruation and level of serum progesterone on 21st day of menstrual cycle to see the evidence of ovulation. Endometrial thickness was also measured. The data was collected on a predesigned questionnaire. The variables that influenced the study were-age, occupation, socioeconomic status, menstrual cycle, marital age, parity, history of MR, history of abortion, past medical and surgical history. In the current study it was observed that the signs of ovulation were significantly (p<0.05) higher in Group I treated with clomiphene citrate in comparison to Group II clomiphene citrate resistant cases treated with letrozole. The rate of ovulation was higher in Group I than that of Group III treated with letrozole, but the difference was not statistically significant (p>0.05). The signs of ovulation were present in 45(81.8%) cases in Group I, 33(60.0%) cases in Group II and 37(67.3%) cases in Group III. This findings of the study suggested that clomiphene citrate is higher successful than letrozole though not statistically significant. Letrozole can be preferred in clomiphene citrate resistant cases before starting other expensive therapies.

Reverse case study: to think like a nurse.

PubMed

Beyer, Deborah A

2011-01-01

Reverse case study is a collaborative, innovative, active learning strategy that nurse educators can use in the classroom. Groups of students develop a case study and a care plan from a list of medications and a short two- to three-sentence scenario. The students apply the nursing process to thoroughly develop a complete case study written as a concept map. The strategy builds on previous learned information and applies the information to new content, thus promoting critical thinking and problem solving. Reverse case study has been used in both associate and baccalaureate nursing degree theory courses to generate discussion and assist students in thinking like a nurse. 2011, SLACK Incorporated.

IVHS institutional issues and case studies : ADVANCE case study

DOT National Transportation Integrated Search

1994-04-01

This operational test case study is one of six performed in response to a Volpe National Transportation Systems Center technical task directive (TTD) to Science Applications International Corporation (SAIC) entitled, IVHS Institutional Issues and ...

Alternative Fuels Data Center: E85 (Flex Fuel)

Science.gov Websites

. Alternative Fueling Stations by Fuel Type More Ethanol Data | All Maps & Data Case Studies Municipality More Ethanol Case Studies | All Case Studies Publications Ethanol Strong; 2018 Ethanol Industry Outlook

Social media campaigns that make a difference: what can public health learn from the corporate sector and other social change marketers?

PubMed

Freeman, Becky; Potente, Sofia; Rock, Vanessa; McIver, Jacqueline

2015-03-30

A great deal of enthusiasm and interest exists in using social media for public health communications, but few research studies have examined its success in promoting and adopting protective health behaviours. To begin to understand how best to develop effective online social marketing campaigns, this paper provides a summary of success factors and key lessons learnt from selected social media campaign case studies. Case study review Methods: A selection of case studies was reviewed for lessons in campaign development, delivery and evaluation from both the corporate and public health sectors. Information about the objective of the campaign, the tactics used and the lessons learnt was extracted from each case study. Lessons learnt from across the case studies were then sorted according to themes. Lessons from the nine case studies selected were categorised into eight themes: planning, use of social media tools, community, content, personal benefits, promotion, costs and challenges. Outcome evaluation data were lacking in the case studies. Overall, the nine case studies show that social media hold promise in changing user behaviours and that social media are highly effective in recruiting participants and motivating them to take small, concrete actions. The case studies also demonstrate that there is room in social media for targeted, inexpensive, small-scale projects, as well as large, well-funded, mass-reach marketing blitzes. Social media campaign process and impact evaluation measures are readily available. Outcome evaluation models and measures are needed to better assess the effectiveness of social media campaigns in changing health behaviours.

Find a Physician from the Society for Vascular Medicine

MedlinePlus

... by SVM_tweets About SVM Event Calendar Practice Tools Case Study Education Journal Scientific Sessions Website FAQ Copyright © ... Choosing Wisely DVT Toolkit A-Fib Decision Making Tool Job Bank Case Study Current Case Case Archive Submission Guidelines Education ...

Prevalence and spectrum of residual symptoms in Lyme neuroborreliosis after pharmacological treatment: a systematic review.

PubMed

Dersch, R; Sommer, H; Rauer, S; Meerpohl, J J

2016-01-01

Controversy exists about residual symptoms after pharmacological treatment of Lyme neuroborreliosis. Reports of disabling long-term sequels lead to concerns in patients and health care providers. We systematically reviewed the available evidence from studies reporting treatment of Lyme neuroborreliosis to assess the prevalence and spectrum of residual symptoms after treatment. A literature search was performed in three databases and three clinical trial registers to find eligible studies reporting on residual symptoms in patients after pharmacological treatment of LNB. Diagnosis must have been performed according to consensus-derived case definitions. No restrictions regarding study design or language were set. Symptom prevalence was pooled using a random-effects model. Forty-four eligible clinical trials and studies were found: 8 RCTs, 17 cohort studies, 2 case-control studies, and 17 case series. The follow-up period in the eligible studies ranged from 7 days to 20 years. The weighted mean proportion of residual symptoms was 28 % (95 % CI 23-34 %, n = 34 studies) for the latest reported time point. Prevalence of residual symptoms was statistically significantly higher in studies using the "possible" case definition (p = 0.0048). Cranial neuropathy, pain, paresis, cognitive disturbances, headache, and fatigue were statistically significantly lower in studies using the "probable/definite" case definition. LNB patients may experience residual symptoms after treatment with a prevalence of approximately 28 %. The prevalence and spectrum of residual symptoms differ according to the applied case definition. Symptoms like fatigue are not reported in studies using the "probable/definite" case definition. As the "possible" case definition is more unspecific, patients with other conditions may be included. Reports of debilitating fatigue and cognitive impairment after LNB, a "post-Lyme syndrome", could therefore be an artifact of unspecific case definitions in single studies.

25-hydroxyvitamin D and breast cancer, colorectal cancer, and colorectal adenomas: case-control versus nested case-control studies.

PubMed

Grant, William B

2015-02-01

Existing literature includes concerns regarding reliability of case-control studies of breast cancer incidence with respect to 25-hydroxyvitamin D (25(OH)D) concentrations. For breast cancer, only case-control studies consistently find inverse correlations between 25(OH)D and breast cancer. However, for colorectal cancer, nested case-control studies find significant inverse correlations with respect to 25(OH)D concentrations at baseline for mean follow-up times of 7 years. This is a review of results currently existing in literature. I provide evidence that 25(OH)D concentration values are only useful for short follow-up times for breast cancer since it develops rapidly. To support the robust nature of breast cancer case-control studies, I show that results from 11 studies from seven countries align in a robust power-law fit to the odds ratio versus mean 25(OH)D concentrations. Case-control studies of breast cancer incidence rates provide reliable results. Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

Use of existing systematic reviews for evidence assessments in infectious disease prevention: a comparative case study.

PubMed

Harder, Thomas; Remschmidt, Cornelius; Haller, Sebastian; Eckmanns, Tim; Wichmann, Ole

2016-10-11

Given limited resources and time constraints, the use of existing systematic reviews (SR) for the development of evidence-based public health recommendations has become increasingly important. Recently, a five-step approach for identifying, analyzing, appraising and using existing SRs based on recent guidance by the US Agency for Healthcare Research and Quality (AHRQ) was proposed within the Project on a Framework for Rating Evidence in Public Health (PRECEPT). However, case studies are needed to test whether this approach is useful, what challenges arise and how problems can be solved. In two case studies, the five-step approach was applied to integrate existing SRs in the development of evidence-based public health recommendations. Case study A focused on the role of neonatal sepsis as a risk factor for adverse neurodevelopmental outcome. Case study B examined the efficacy, effectiveness and safety of influenza vaccination during pregnancy. For each step, we report the approach of the review team, discuss challenges and describe solutions. For case study A, one existing SR was identified, while in case study B four SRs were eligible for analysis. We found that comparison of inclusion criteria alone was sufficient to judge on relevance of SRs in case study A, but not B. Although methodological quality of all identified SRs was acceptable, risk of bias assessments of individual studies included in the SRs had to be repeated in both case studies. Particular challenges appeared in case study B where multiple SRs addressed the same research question. With the help of spreadsheets comparing the characteristics of the existing SR we decided to use the most comprehensive one for our evidence synthesis and supplemented the results with those from the other SRs. In both case studies using the complete SR was not possible. The five-step approach provided useful and structured guidance and should be routinely applied when using existing SRs as a basis for evidence-based recommendations in public health. In situations where more than one SR has to be considered, the development of spreadsheets comparing characteristics, inclusion criteria, risk of bias, included studies and outcomes seems useful.

Synthesis of Single-Case Experimental Data: A Comparison of Alternative Multilevel Approaches

ERIC Educational Resources Information Center

Ferron, John; Van den Noortgate, Wim; Beretvas, Tasha; Moeyaert, Mariola; Ugille, Maaike; Petit-Bois, Merlande; Baek, Eun Kyeng

2013-01-01

Single-case or single-subject experimental designs (SSED) are used to evaluate the effect of one or more treatments on a single case. Although SSED studies are growing in popularity, the results are in theory case-specific. One systematic and statistical approach for combining single-case data within and across studies is multilevel modeling. The…

Case Studies Behavior Modification.

ERIC Educational Resources Information Center

Wark, David M.

The case histories of five students enrolled in a university course in how to study are reported. The students ranged in age from 18 to 35, included two males and three females, and varied in school experience from no college in one case and some college in two cases to college degrees in two cases. Students were initially taught to chart their…

Creative Interactive Teaching: Case Method & Other Techniques. Selected Papers of the International Conference on Case Methods Research & Application (16th, Caceres, Spain, 1999).

ERIC Educational Resources Information Center

Klein, Hans E., Ed.

This book presents a selection of papers from the international, interdisciplinary conference of the World Association for Case Method Research & Application. Papers are categorized into seven areas: (1) "International Case Studies" (e.g., event-based entrepreneurship, case studies on consumer complaints, and strategic quality…

Liver histopathology of fatal phosphine poisoning.

PubMed

Saleki, Sepideh; Ardalan, Farid Azmoudeh; Javidan-Nejad, Abdullah

2007-03-02

Two commonly used pesticides in agriculture are phosphides of aluminium and zinc. Both of these metal phosphides act through elaboration of toxic phosphine gas. The poisoning in Iran is mostly oral and suicidal. Phosphine is rapidly absorbed throughout the gastrointestinal tract after ingestion and it is partly carried to the liver by the portal vein. In this study the liver histopathology of fatal poisoning is scrutinized. A descriptive, retrospective study was performed on 38 fatal phosphine poisonings. The slides of liver specimens of the cases were retrieved and studied separately by two pathologists. The poisoning was suicidal in 33 (86.5%) of cases. Portal inflammation was negligible in 37 cases and only in one of the cases, a moderate degree of chronic inflammation accompanied by granuloma formation was observed. Major histopathologic findings were as follows: mild sinusoidal congestion; 12 cases (31.6%), severe sinusoidal congestion; 25 cases (45.8%), central vein congestion; 23 cases (60.5%), centrilobular necrosis; 3 cases (7.9%), hepatocytes nuclear fragmentation; 6 cases (15.8%), sinusoidal clusters of polymorphonuclear leukocytes; 12 cases (31.6%), and mild macrovesicular steatosis; 5 cases (13.2%). Fine isomorphic cytoplasmic vacuoles were observed in 36 cases (94.7%). These vacuoles were distributed uniformly in all hepatic zones in the majority (75%) of cases. This study reveals that the main histopathologic findings of fatal phosphine poisoning in the liver are fine cytoplasmic vacuolization of hepatocytes and sinusoidal congestion.

Role of survivor bias in pancreatic cancer case-control studies.

PubMed

Hu, Zhen-Huan; Connett, John E; Yuan, Jian-Min; Anderson, Kristin E

2016-01-01

The purpose of this study was to evaluate the impact of survivor bias on pancreatic cancer case-control studies. The authors constructed five case-loss scenarios based on the Iowa Women's Health Study cohort to reflect how case recruitment in population-based studies varies by case survival time. Risk factors for disease incidence included smoking, body mass index (BMI), waist circumference, diabetes, and alcohol consumption. Odds ratios (ORs) were estimated by conditional logistic regression and quantitatively compared by the interactions between risk factors and 3-month survival time. Additionally, Kaplan-Meier estimates for overall survival were compared within the subset cohort of pancreatic cancer cases. BMI and waist circumference showed a significant inverse relationship with survival time. Decreasing trends in ORs for BMI and waist circumference were observed with increasing case survival time. The interaction between BMI and survival time based on a cutpoint of 3 months was significant (P < .01) as was the interaction between waist circumference and survival time (P < .01). The findings suggested that case losses could result in survivor bias causing underestimated odds ratios for both BMI and waist circumference, whereas other risk factors were not significantly affected by case losses. Copyright © 2016 Elsevier Inc. All rights reserved.

Effectiveness of integrating case studies in online and face-to-face instruction of pathophysiology: a comparative study.

PubMed

Saleh, Suha M; Asi, Yara M; Hamed, Kastro M

2013-06-01

Due to growing demand from students and facilitated by innovations in educational technology, institutions of higher learning are increasingly offering online courses. Subjects in the hard sciences, such as pathophysiology, have traditionally been taught in the face-to-face format, but growing demand for preclinical science courses has compelled educators to incorporate online components into their classes to promote comprehension. Learning tools such as case studies are being integrated into such courses to aid in student interaction, engagement, and critical thinking skills. Careful assessment of pedagogical techniques is essential; hence, this study aimed to evaluate and compare student perceptions of the use of case studies in face-to-face and fully online pathophysiology classes. A series of case studies was incorporated into the curriculum of a pathophysiology class for both class modes (online and face to face). At the end of the semester, students filled out a survey assessing the effectiveness of the case studies. Both groups offered positive responses about the incorporation of case studies in the curriculum of the pathophysiology class. This study supports the argument that with proper use of innovative teaching tools, such as case studies, online pathophysiology classes can foster a sense of community and interaction that is typically only seen with face-to-face classes, based on student responses. Students also indicated that regardless of class teaching modality, use of case studies facilitates student learning and comprehension as well as prepares them for their future careers in health fields.

Is case-chaos methodology an appropriate alternative to conventional case-control studies for investigating outbreaks?

PubMed

Edelstein, Michael; Wallensten, Anders; Kühlmann-Berenzon, Sharon

2014-08-15

Case-chaos methodology is a proposed alternative to case-control studies that simulates controls by randomly reshuffling the exposures of cases. We evaluated the method using data on outbreaks in Sweden. We identified 5 case-control studies from foodborne illness outbreaks that occurred between 2005 and 2012. Using case-chaos methodology, we calculated odds ratios 1,000 times for each exposure. We used the median as the point estimate and the 2.5th and 97.5th percentiles as the confidence interval. We compared case-chaos matched odds ratios with their respective case-control odds ratios in terms of statistical significance. Using Spearman's correlation, we estimated the correlation between matched odds ratios and the proportion of cases exposed to each exposure and quantified the relationship between the 2 using a normal linear mixed model. Each case-control study identified an outbreak vehicle (odds ratios = 4.9-45). Case-chaos methodology identified the outbreak vehicle 3 out of 5 times. It identified significant associations in 22 of 113 exposures that were not associated with outcome and 5 of 18 exposures that were significantly associated with outcome. Log matched odds ratios correlated with their respective proportion of cases exposed (Spearman ρ = 0.91) and increased significantly with the proportion of cases exposed (b = 0.054). Case-chaos methodology missed the outbreak source 2 of 5 times and identified spurious associations between a number of exposures and outcome. Measures of association correlated with the proportion of cases exposed. We recommended against using case-chaos analysis during outbreak investigations. © The Author 2014. Published by Oxford University Press on behalf of the Johns Hopkins Bloomberg School of Public Health. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Visualization case studies : a summary of three transportation applications of visualization

DOT National Transportation Integrated Search

2007-11-30

The three case studies presented in "Visualization Case Studies" are intended to be helpful to transportation agencies in identifying effective techniques for enhancing and streamlining the project development process, including public outreach activ...

Al Qaeda in the Indian Subcontinent (AQIS): An Al Qaeda Affiliate Case Study

DTIC Science & Technology

2017-10-01

conduct this assessment, CNA used a comparative methodology that included eight case studies on groups affiliated or associated with Al-Qaeda. These case...Case Study Pamela G. Faber and Alexander Powell October 2017 This work was performed under Federal Government Contract No. N00014-16-D-5003. Copyright...Director Center for Stability and Development Center for Strategic Studies REPORT DOCUMENTATION PAGE Form ApprovedOMB No. 0704-0188 Public reporting burden

The Modular need for the Division Signal Battalion

DTIC Science & Technology

2017-06-09

findings and analyzes them to expand on them. It is with these findings and subsequent analysis that the case studies shape the answer to the three...These case studies focus on the signal leadership development and how it occurred in the pre-modular force structure, during modularity, and the...the comparative case study research. The case studies focus on signal leader development in a pre-modular signal force, a modular signal force, and

Civil Wars Hijacked: A Case Study of the Lebanese Civil War

DTIC Science & Technology

2016-06-01

will focus on in-depth case studies of external state actors and their sponsored NSAs and outcomes. The state strategies of... case study because they did not employ NSA relationships on the scale of the three other state actors, and U.S. policy often complimented rather than...HIJACKED: A CASE STUDY OF THE LEBANESE CIVIL WAR by Kyle L. Greenheck June 2016 Thesis Advisor: Sean Everton Co-Advisor: Doowan Lee THIS PAGE

Military Hydrology. Report 12. Case Study Evaluation of Alternative Dam-Breach Flood Wave Methods. Volume 1. Main Report.

DTIC Science & Technology

1986-11-01

Report Organization. .................... 7 *PART 11: CASE STUDIES .......................... 9 Teton Dam Failure Flood. ...................... 9...channel, (3) Laurel Run Dam , and (4) Stillhouse Hollow Dam . The Laurel Run and Teton case studies involved field data sets from actual dam failures. The...hypothetical prismatic channel case study used the Teton reservoir and dam data but replaced the complex Teton Valley geometry with a prismatic channel

Evaluation of the United States Drug War Policy Abroad: A Case Study in Colombia

DTIC Science & Technology

2010-12-01

Case Study in Colombia By: Kevin T. Wright and Joseph S. Hamilton December 2010 Advisors: David R. Henderson, John Enns...States Drug War Policy Abroad: A Case Study in Colombia 6. AUTHOR(S) Kevin T. Wright, Joseph S. Hamilton 5. FUNDING NUMBERS 7. PERFORMING...public release; distribution is unlimited EVALUATION OF THE UNITED STATES DRUG WAR POLICY ABROAD: A CASE STUDY IN COLOMBIA Kevin T. Wright

"To Give an Outsider an Idea of What It Could Be Like": A Case Study of the Creative Representation of Hearing Voices

ERIC Educational Resources Information Center

Flavin, Michael; James, Bethany

2018-01-01

This paper reports on a case study which aims to recreate the hearing voices symptom in schizophrenia. The case study was submitted for a co-curricular module at King's College London by a first-year undergraduate Music student, Bethany James, and was created using the web application, Mahara. The core of the case study consists of a soundscape of…

CASE STUDY RESEARCH: THE VIEW FROM COMPLEXITY SCIENCE

PubMed Central

Anderson, Ruth; Crabtree, Benjamin F.; Steele, David J.; McDaniel, Reuben R.

2005-01-01

Many wonder why there has been so little change in care quality, despite substantial quality improvement efforts. Questioning why current approaches are not making true changes draws attention to the organization as a source of answers. We bring together the case study method and complexity science to suggest new ways to study health care organizations. The case study provides a method for studying systems. Complexity theory suggests that keys to understanding the system are contained in patterns of relationships and interactions among the system’s agents. We propose some of the “objects” of study that are implicated by complexity theory and discuss how studying these using case methods may provide useful maps of the system. We offer complexity theory, partnered with case study method, as a place to begin the daunting task of studying a system as an integrated whole. PMID:15802542