Factors Affecting Hampton Roads, Virginia, Elected Official Emergency Management Recovery Policy Decisions

For many regions and local governments, budgetary restraints limit funds appropriated for emergency management activities to inadequate levels, and little guidance exists related to decision factors used by elected officials in identifying budget and ordinance priorities. Using Kwon, Choi, and Bae\u27s conceptualization of punctuated equilibrium theory, the purpose of this case study was to examine how decision factors influenced Hampton Roads, Virginia, elected official disaster recovery policy between 2003 and 2012. Data were collected through review of 1,310 city documents and 10 semistructured interviews with elected officials. Data were inductively coded and analyzed using a thematic analysis procedure. Data analysis resulted in the identification of 3 decision factor themes that guided post disaster recovery in Hampton Roads: (a) establishing a sense of normalcy in terms of budget appropriations and ordinances for security, safety and quality of services short-term recovery policy, (b) budgetary resiliency to encourage the restoration of infrastructure related to long-term recovery policy, and (c) the development of self-sufficient processes that lead to an anticipatory mindset with issuance of mitigation ordinances and capital improvement appropriations policy. The findings confirmed punctuated equilibrium theory, as man-made disasters triggered short-term recovery policy decisions. Results of the study may affect positive social change by providing local elected representatives with a \u27tool kit\u27 of decision factors to consistently address post disaster recovery policy for public safety, security, and stability via the governance mechanisms of strategic planning, appropriation decisions, and assessment

Stress hormone-induced immunomodulation and interplay between immune cells and bacteria in response to stress hormones in domestic pigs

The two main endocrine systems involved in the regulation of stress reactions are the HPA axis, leading to the synthesis of glucocorticoids like cortisol or corticosterone, and the SAM axis, whose activation is associated with the release of the catecholamines adrenaline and noradrenaline. These stress hormones modulate the function of the immune system. Although pigs in modern husbandry systems face many stressors, the consequences of elevated plasma stress hormone levels on porcine immune cell numbers and functionality are insufficiently resolved. While some research on glucocorticoid effects has been conducted, data on many parameters are missing and catecholamines have not been studied systematically in the pig, yet. It is known that stress can negatively affect pigs resistance to infections like salmonellosis, but the underlying mechanisms are still subject to intense research efforts, with new perspectives arising since the discovery of interkingdom-signalling and microbial catecholamine perception. The aim of this thesis was to determine the effects of cortisol, adrenaline and noradrenaline on porcine immune cell functionality and the blood numbers of different leukocyte subsets. Furthermore, the interplay of immune cells and Salmonella Typhimurium under the influence of catecholamines was investigated. Adult male castrated pigs were surgically equipped with indwelling catheters to enable stress-free blood collection and intravenous application of hormones. In an initial experiment, the effects of in vitro stress hormone treatment on lymphocyte proliferation and the production of the proinflammatory cytokine TNFa were described. Cortisol reduced both proliferation and number of TNFa producers. Both catecholamines caused an increased lymphocyte proliferation at low concentrations whereas noradrenaline drastically decreased proliferation at high concentrations. While noradrenaline had no impact on TNFa producers, they were reduced in gd T cells and monocytes upon adrenaline addition. Overall, the effects were comparable to humans in terms of direction and dose but there were some disparities regarding adrenaline. In the second part of the project, the impact of in vivo stress hormone administration on immune cell numbers and functionality was examined by infusion for 48h. Cortisol and noradrenaline led to a decreased lymphocyte proliferation but to a variable extent and all three hormones promoted phagocytic function of innate immune cells. Cortisol caused a marked increase of neutrophil numbers while almost all other cell types declined strongly. For most cell types, noradrenaline exerted similar effects but solely after 2h whereas cortisol-induced alterations lasted the whole treatment period. Adrenaline effects were mostly reduced to CD8- T cells, which were reduced at first but increased after 24h. A sharp peak in NK cell numbers after 2h adrenaline infusion is particularly noteworthy and resembles findings from rodent and human studies. Overall, both hormone groups led to a shift from adaptive to innate immunity, underpinning the picture of a promotion of fast and unspecific defence systems to respond to threats in stressful situations. In a third study, S. Typhimurium was grown in the presence of catecholamines to determine the effects of supernatants from these cultures on porcine immune cell function. Both lymphocyte proliferation and TNFα production were hampered substantially, as opposed to the findings on catecholamine effects in the first experiment. It was demonstrated that these effects were not caused by catecholamines or their oxidation products and the formation of a so-far unknown immunosuppressive substance by catecholamine-primed bacteria was assumed. The results contribute to a better understanding of the increased susceptibility to infection in stressed animals and reveal a new dimension of cross-species communication. Finally, the results of the present thesis were discussed regarding their comparability to studies in humans and rodents and previous stress experiments in pigs. Furthermore, the effects of acute and chronic stress as well as different coping styles that are characterised by a SAM or HPA predominance on animal welfare and pig health were discussed, based on the endocrine mechanisms investigated in the present thesis. Possible implications of enhanced glucocorticoid and catecholamine levels for practical pig husbandry were given. Lastly, suggestions for future research to further elucidate the impact of stress hormones on the porcine immune system and the interplay with pathogenic bacteria were made.Die beiden wichtigsten endokrinen Systeme, die an der Steuerung von Stressreaktionen beteiligt sind, sind die HPA-Achse, die zur Synthese von Glukokortikoiden wie Cortisol oder Corticosteron führt, und die SAM-Achse, deren Aktivierung mit der Freisetzung der Katecholamine Adrenalin und Noradrenalin verbunden ist. Diese Stresshormone modulieren die Funktion des Immunsystems. Obwohl Schweine in modernen Haltungssystemen vielen potenziellen Stressoren ausgesetzt sind, sind die Folgen erhöhter Stresshormonspiegel auf die Anzahl und Funktionalität der Immunzellen des Schweins nicht ausreichend geklärt. Zwar wurden einige Untersuchungen zu den Effekten von Glukokortikoiden durchgeführt, jedoch fehlen noch Daten zu vielen Parametern, und Katecholamineffekte wurden bislang nicht systematisch untersucht. Es ist bekannt, dass Stress die Empfänglichkeit von Schweinen gegen Infektionen wie die Salmonellose negativ beeinflussen kann, aber die zugrundeliegenden Mechanismen sind noch Gegenstand intensiver Forschungsbemühungen. Dabei haben sich seit der Entdeckung des Interkingdom-Signalling und der Wahrnehmung von Katecholaminen durch Mikroorganismen neue Perspektiven ergeben. Ziel dieser Arbeit war es, die Effekte von Cortisol, Adrenalin und Noradrenalin auf die Funktionalität von Schweineimmunzellen und die Zellzahlen verschiedener Leukozyten-Subpopulationen im Blut zu bestimmen. Darüber hinaus wurde das Zusammenspiel von Immunzellen und Salmonella Typhimurium unter dem Einfluss von Katecholaminen untersucht. Dafür wurden adulte Kastraten chirurgisch mit Venenverweilkathetern ausgestattet, um eine stressfreie Blutentnahme sowie intravenöse Hormonapplikation zu ermöglichen. In einem ersten Experiment wurden die Auswirkungen einer in vitro-Zugabe von Stresshormonen auf Lymphozytenproliferation und Produktion des proinflammatorischen Zytokins TNFa beschrieben. Cortisol führte zu einer Reduktion sowohl der Proliferation als auch der Anzahl von TNFa-Produzenten. Beide Katecholamine bewirkten eine erhöhte Lymphozytenproliferation bei niedrigen Konzentrationen, wohingegen Noradrenalin die Proliferation bei hohen Konzentrationen drastisch verringerte. Während Noradrenalin keinen Einfluss auf TNFa-produzierende Zellen hatte, waren sie nach Zugabe von Adrenalin unter den gd-T-Zellen und Monozyten reduziert. Insgesamt waren die Hormoneffekte in Richtung und Dosis mit den beim Menschen beschriebenen vergleichbar mit einigen Unterschieden bei Adrenalin. Im zweiten Teil des Projekts wurden die Auswirkungen einer in vivo-Gabe von Stresshormonen auf die Anzahl und Funktionalität von Immunzellen mittels 48-stündiger Infusion untersucht. Cortisol und Noradrenalin führten zu einer verminderten Lymphozytenproliferation und alle drei Hormone förderten die Phagozytosefunktion angeborener Immunzellen. Cortisol verursachte einen Anstieg der Neutrophilenzahl, wohingegen fast alle anderen Zelltypen zurückgingen. Bei den meisten Zelltypen übte Noradrenalin ähnliche Effekte aus, jedoch nur nach 2 Stunden, wohingegen die Cortisol-induzierten Veränderungen die gesamte Behandlungsdauer anhielten. Die Adrenalin-Effekte waren größtenteils auf CD8-negative T-Zellen begrenzt, deren Anzahl zunächst reduziert, aber nach 24 Stunden erhöht war. Ein starker Anstieg der NK-Zellzahl nach 2-stündiger Adrenalin-Infusion ist besonders erwähnenswert und spiegelt Ergebnisse aus Nager- und Humanstudien wider. Insgesamt betrachtet führten beide Hormongruppen zu einer Verschiebung von adaptiver zu angeborener Immunität. In einer dritten Studie wurden S. Typhimurium-Kulturen unter Zugabe von Katecholaminen angelegt, um die Wirkung von Überständen aus diesen Kulturen auf die Funktion von Schweineimmunzellen zu bestimmen. Sowohl Lymphozytenproliferation als auch TNFa-Produktion waren deutlich verringert. Es konnte gezeigt werden, dass diese Effekte nicht durch Katecholamine oder deren Oxidationsprodukte verursacht wurden, sodass die Bildung einer bislang unbekannten immunsuppressiven Substanz durch Katecholamin-behandelte Bakterien angenommen wird. Die Ergebnisse tragen zu einem besseren Verständnis der erhöhten Infektionsanfälligkeit gestresster Tiere bei und zeigen eine neue Dimension der artübergreifenden Kommunikation auf. Schließlich wurden die Ergebnisse der vorliegenden Arbeit hinsichtlich ihrer Vergleichbarkeit mit Studien an Menschen und Nagern sowie früheren Stressexperimenten an Schweinen diskutiert. Darüber hinaus wurden die Auswirkungen von akutem und chronischem Stress sowie unterschiedlicher Coping-Strategien, die sich durch eine SAM- oder HPA-Dominanz auszeichnen, auf das Tierwohl und die Schweinegesundheit diskutiert. Es wurden mögliche Auswirkungen von erhöhten Glukokortikoid- und Katecholaminwerten auf die praktische Schweinehaltung aufgezeigt. Schließlich wurden Vorschläge für zukünftige Forschung gemacht, um den Einfluss von Stresshormonen auf das Immunsystem von Schweinen und die Wechselwirkungen mit pathogenen Bakterien weiter aufzuklären

Kiri tundmatule, Leipzig

http://tartu.ester.ee/record=b1890562~S1*es

Kiri J. G. Walch `ile, Lipsiae

http://tartu.ester.ee/record=b1880403~S1*es

Targeting Pyk2 to β1-Integrin–Containing Focal Contacts Rescues Fibronectin-Stimulated Signaling and Haptotactic Motility Defects of Focal Adhesion Kinase–Null Cells

Focal adhesion kinase–null (FAK−/−) fibroblasts exhibit morphological and motility defects that are reversed by focal adhesion kinase (FAK) reexpression. The FAK-related kinase, proline-rich tyrosine kinase 2 (Pyk2), is expressed in FAK−/− cells, yet it exhibits a perinuclear distribution and does not functionally substitute for FAK. Chimeric Pyk2/FAK proteins were created and expressed in FAK−/− cells to determine the impact of Pyk2 localization to focal contacts. Whereas an FAK/Pyk2 COOH-terminal (CT) domain chimera was perinuclear distributed, stable expression of a Pyk2 chimera with the FAK-CT domain (Pyk2/FAK-CT) localized to focal contact sites and enhanced fibronectin (FN)-stimulated haptotactic cell migration equal to FAK-reconstituted cells. Disruption of paxillin binding to the FAK-CT domain (S-1034) inhibited Pyk2/FAK-CT localization to focal contacts and its capacity to promote cell motility. Paxillin binding to the FAK-CT was necessary but not sufficient to mediate the indirect association of FAK or Pyk2/FAK-CT with a β1-integrin–containing complex. Both FAK and Pyk2/FAK-CT but not Pyk2/FAK-CT S-1034 reconstituted FAK−/− cells, exhibit elevated FN-stimulated extracellular signal–regulated kinase 2 (ERK2) and c-Jun NH2-terminal kinase (JNK) kinase activation. FN-stimulated FAK or Pyk2/FAK-CT activation enhanced both the extent and duration of FN-stimulated ERK2 activity which was necessary for cell motility. Transient overexpression of the FAK-CT but not FAK-CT S-1034 domain inhibited both FN-stimulated ERK2 and JNK activation as well as FN-stimulated motility of Pyk2/FAK-CT reconstituted cells. These gain-of-function studies show that the NH2-terminal and kinase domains of Pyk2 can functionally substitute for FAK in promoting FN-stimulated signaling and motility events when localized to β-integrin–containing focal contact sites via interactions mediated by the FAK-CT domain

Repeat testing of low-level HIV-1 RNA: Assay performance and implementation in clinical trials

Objective: Assess the performance of HIV-1 RNA repeat testing of stored samples in cases of low-level viremia during clinical trials. Design: Prospective and retrospective analysis of randomized clinical trial samples and reference standards. Methods: To evaluate assay variability of the Cobas AmpliPrep/Cobas TaqMan HIV-1 Test, v2.0, three separate sources of samples were utilized: the World Health Organization (WHO) HIV reference standard (assayed using 50 independent measurements at six viral loads <200 copies/ml), retrospective analysis of four to six aliquots of plasma samples from four clinical trial participants, and prospective repeat testing of 120 samples from participants in randomized trials with low-level viremia. Results: The TaqMan assay on the WHO HIV-1 RNA standards at viral loads <200 copies/ml performed within the expected variability according to assay specifications. However, standards with low viral loads of 36 and 18 copies/ml reported values of ≥ 50 copies/ml in 66 and 18% of tests, respectively. In participants treated with antiretrovirals who had unexpected viremia of 50-200 copies/ml after achieving <50 copies/ml, retesting of multiple aliquots of stored plasma found <50 copies/ml in nearly all cases upon retesting (14/15; 93%). Repeat testing was prospectively implemented in four clinical trials for all samples with virologic rebound of 50-200 copies/ml (n = 120 samples from 92 participants) from which 42% (50/120) had a retest result of less than 50 copies/ml and 58% (70/120) retested ≥ 50 copies/ml. Conclusion: The TaqMan HIV-1 RNA assay shows variability around 50 copies/ml that affects clinical trial results and may impact clinical practice. In participants with a history of viral load suppression, unexpected low-level viremia may be because of assay variability rather than low drug adherence or true virologic failure. Retesting a stored aliquot of the same sample may differentiate between assay variability and virologic failure as the source of viremia. This retesting strategy could save time, money, and anxiety for patients and their providers, as well as decrease follow-up clinic visits without increasing the risk of virologic failure and resistance development

Integrin signaling through FAK in the regulation of mammary stem cells and breast cancer

Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase identified as a key mediator of intracellular signaling by integrins, a major family of cell surface receptors for extracellular matrix, in the regulation of different cellular functions in a variety of cells. Upon activation by integrins through disruption of an autoinhibitory mechanism, FAK undergoes autophosphorylation and forms a complex with Src and other cellular proteins to trigger downstream signaling through its kinase activity or scaffolding function. A number of integrins are identified as surface markers for mammary stem cells (MaSCs), and both integrins and FAK are found to play crucial roles in the maintenance of MaSCs in studies using mouse models, suggesting that integrin signaling through FAK may serve as a functional marker for MaSCs. Consistent with previous studies linking increased expression and activation of FAK to human breast cancer, these findings suggest a novel cellular mechanism of FAK promotion of mammary tumorigenesis by maintaining the pools of MaSCs as targets of oncogenic transformation. Furthermore, FAK inactivation in mouse models of breast cancer also reduced the pool of mammary cancer stem cells (MaCSCs), decreased their self-renewal in vitro , and compromised their tumorigenicity and maintenance in vivo , suggesting a potential role of integrin signaling through FAK in breast cancer growth and progression through its functions in MaCSCs. This review discusses these recent advances and future studies into the mechanism of integrin signaling through FAK in breast cancer through regulation of MaCSCs that may lead to development of novel therapies for this deadly disease. © 2010 IUBMB IUBMB Life, 62(4): 268–276, 2010Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/69164/1/303_ftp.pd