Dissertations / Theses on the topic 'Stressful conditions'

Lipid droplets (LDs) are phylogenetically conserved and ubiquitous organelles with many cellular functions. In the last two decades, our understanding of LD biology and of their roles in physiological processes has increased dramatically. In addition, increasing evidence suggests that LDs are highly involved in inflammatory processes, and in metabolic disorders such as type 2 diabetes mellitus (T2DM). Despite such advancement, many aspects of LD biology and of their roles in health and disease remain unknown.The core of LDs is highly enriched with neutral lipids and these can be mobilized to provide metabolic energy. The phospholipid monolayer surrounding the LD core is associated with a wide variety of proteins, including structural and signaling proteins, as well as metabolic enzymes. While LDs may be induced by physiological stimuli such as dietary fatty acids, they can also be formed under stressful conditions, in the absence of such fatty acids. However, exactly how cellular stress leads to LD accumulation remains unclear. Our main objective is to understand the regulation of LD formation under stressful conditions, specifically oxidative stress, inflammation, and metabolic stress. We first investigated LDs in cells exposed to environmental stressors, namely cytotoxic metallic nanoparticles and reactive oxygen species. LD formation and expression of perilipin-2, a key structural LD protein, were highly increased in rodent cells exposed to these stress agents. Interestingly, supplementation with antioxidant N-acetyl cysteine or pharmacological inhibition of p38 mitogen activated protein kinase (MAPK) reduced stress-induced LD accumulation, suggesting that oxidative stress and p38 MAPK activation play a role in the induction of LD formation. Inflammatory leukocytes and macrophages contain a large number of LDs. While this phenomenon has been widely investigated in peripheral immune cells, its explanation remains elusive in immune cells of the central nervous system. We therefore investigated LD dynamics and regulation in microglia, the resident immune cells in the brain. We found that stimulation of microglia with toll-like receptor 4 (TLR4) agonist, lipopolysaccharides (LPS), increased LD formation and perilipin-2 expression in an Akt and p38 MAPK-dependent manner. Interestingly, LPS-induced LDs extensively colocalized with cytosolic phospholipase A2-α (cPLA2-α), a key enzyme involved in the synthesis of eicosanoids, which are inflammatory lipid mediators. Collectively, these findings imply that LD formation may contribute to increased eicosanoid synthesis in activated microglia and could be microglial biomarkers of inflammation in the central nervous system. To gain a better insight into the role of LDs in human pathology, we sought to examine alterations in LD metabolism in pancreatic tissue obtained from T2DM and obese individuals. Immunohistochemical studies revealed increased islet and extra-islet perilipin-2 expression in tissues from lean or obese T2DM donors, but not in non-T2DM obese donors, suggesting that the diabetic status, but not the obesity status, is a requirement for increasing perilipin-2 expression and LD formation. Gene expression analysis by RT-qPCR confirmed the increase in perilipin-2 expression and revealed significant alterations in several genes related to islet function, metabolism and antioxidant defense. These alterations seem to be consistently associated with obesity and T2DM and imply an adaptive and compensatory response to insulin resistance and metabolic stress. In summary, our studies show that LDs are an integral part of the adaptive cellular response to oxidative, inflammatory and metabolic stress. Perhaps, the most important challenge in LD research in the upcoming decade will be to determine how the subcellular lipid and protein composition of this organelle affects its function in different cells.
Les gouttelettes lipidiques (GL) sont des organites phylogénétiquement conservées et impliquées dans plusieurs fonctions cellulaires. Durant les deux dernières décennies, notre compréhension des rôles biologiques et physiologiques des GL a augmenté de manière draconienne. Plusieurs observations suggèrent fortement que les GL jouent un rôle important dans l'inflammation, ainsi que dans les désordres métaboliques tels que le diabète de type 2 (DT2). Malgré cette avancée, plusieurs aspects de la biologie des GL et de leurs rôles dans des maladies demeurent méconnus.Le centre des GL est riche en lipides neutres qui peuvent se mobiliser et servir comme source d'énergie. La couche phospholipidique entourant le centre de la GL est associée à plusieurs protéines et enzymes métaboliques. Bien que les GL puissent être induites par des acides gras, elles peuvent aussi l'être dans des conditions de stress. Par contre, les mécanismes de l'accumulation de GL par des conditions de stress ne sont pas encore bien compris. Notre objectif principal est de comprendre la régulation de la formation de GL par le stress oxydatif, l'inflammation et le stress métabolique. Premièrement, nous avons investigué les GL dans des cellules exposées à des stresseurs tels que des nanocrystaux métalliques et des dérivés réactifs d'oxygène. La formation de GL et l'expression de perilipin-2, qui est une protéine structurelle des GL, ont tous deux augmenté dans les cellules stressées. De plus, une supplémentation en antioxydant (n-acétylcystéine) ou un traitement avec un inhibiteur de p38 MAPK a réduit l'accumulation de GL causée par le stress. Ces observations suggèrent que le stress oxydatif et p38 MAPK jouent un rôle dans l'accumulation de GL dans des cellules stressées. Il est bien connu que les leucocytes et macrophages qui sont engagés dans l'inflammation contiennent une grande quantité de GL. Même si ce phénomène a bien été exploré dans les cellules immunitaires périphériques, il reste inexploré dans le système nerveux central (SNC). Ce faisant, nous avons investigué la dynamique et la régulation des GL dans les microglies, les cellules résidentes immunitaires dans le cerveau. Nous avons trouvé que dans les microglies stimulées avec les lipopolysaccharides (LPS), les GL et l'expression de perilipin-2 ont augmenté d'une manière dépendante de l'activation de l'Akt et p38 MAPK. Dans ces cellules activées, la phospholipase cytosolique A2-α (PLC A2-α), une enzyme fonctionnant dans la synthèse d'éicosanoides, des médiateurs lipidiques inflammatoires, colocalisait avec les GL. Ensemble, ces résultats indiquent que la formation de GL pourrait contribuer à la synthèse d'éicosanoides dans les microglies activées et servir de biomarqueurs d'inflammation dans le SNC.Pour mieux comprendre le rôle des GL dans la pathologie humaine, nous les avons examinées dans des tissues pancréatiques provenant de patients obèses ou diabétiques T2. Nos études immunohistochimiques ont révélé une augmentation de perilipin-2 dans les îlots de Langerhans chez les patients diabétiques obèses ou maigres, mais pas dans ceux de patients non-diabétiques. Ceci suggère que le DT2, mais non l'obésité, est requis pour une augmentation de perilipin-2 dans le pancréas. L'analyse d'expression de gènes par RT-PCR a confirmé l'augmentation de perilipin-2 observé antérieurement dans les îlots et a également révélé des altérations dans des gènes reliés aux fonctions des îlots, au métabolisme, et aux défenses anti-oxydantes. Ces changements, qui sont souvent associés à l'obésité et au DT2, constituent un mécanisme d'adaptation à la résistance à l'insuline et au stress métabolique.Pour résumer, nos études démontrent que l'accumulation de GL fait partie intégrante de l'adaptation des cellules au stress. Durant la prochaine décennie, le plus grand obstacle dans la recherche sur les GL sera de déterminer comment la composition lipidique ou protéinique de ces organites affecte leurs fonctions biologiques.

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The speed and efficacy with which front-line warfighters in stressful conditions can submit resource requests, such as a casualty evacuation, could mean the difference between life and death. Traditional methods to call for resources require training, are error-prone and can be sluggish. The Handheld Emergency Logistics Program (HELP) was developed by the authors of this thesis to assist both trained and untrained persons in requesting resources from supporting agencies. HELP was developed to prove the concept that off-the-shelf mobile technology can significantly improve the speed and efficacy of resource requests. This thesis aims to allow HELP to exploit built-in sensors in modern commercial off-the-shelf handheld smart devices and their computation and communication capability to reduce the chance of error, reduce the need to pull information from memory, reduce manual data entry, and provide multiple redundant modalities for performing the same action. Our findings indicate that with the assistance of HELP, users submitting resource requests committed half as many errors and completed the request in half the amount of time as compared to a control group using traditional methods. We recommend that the concept of using smart devices to call for resources be further developed into a program of record.

Background: The common concern surrounding the poor level of displaced life is that it might cause harmful psychological conditions. Another concern of displacement is the impact of conflict and the ability to survive in adverse situations. The reasons for conducting this thesis were developed through the literature review, and noticing a lack of studies that assess psychological issues such as well-being, resilience, prejudice and forgiveness among Syrian refugees and Iraqi displaced persons comprehensively. Research aims: In particular, the studies address three issues that are fundamental to understanding how the thesis is structured. The first issue is essential in examining how war has affected individuals’ well-being in short and long term of life engagement of the refugees. The second issue investigates the impact of the displacement situation on prejudices and the possibility of forgiveness after conflict and violence between groups of people. The final issue involves identifying the factors that might help individuals to survive and face any potentially harmful situation. Methods: The overall research sample includes 1,256 individuals of both genders. For collecting the data, self-report questionnaires and objective measures were used. Results: The results show that the research samples indicated poor levels of quality of life, well-being, forgiveness and resilience. Importantly, these findings reveal the significant role played by positive relation domains of psychological well-being in improving resilience and forgiveness. The research has also has shown that the psychological health domain of quality of life recorded the highest significant associations with resilience and psychological well-being. To Sum up, this thesis illustrates the importance of positive social relations, lower prejudice and greater resilience in predicting mental health states.

Thesis (M.S.)--West Virginia University, 2004.
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The intestine and associated bacterial microbiota have a central role the physiology and homoeostasis of the host. The understanding of how farming conditions affect the intestine and associated microbiota of fish is the high importance to counteract the potential threats to health and welfare. Thus, this thesis aims to understand the role of stressful husbandry conditions on the intestine and associated microbiota of rainbow trout and Atlantic salmon. Within this context, the role of Pediococcus acidilactici as health promoter was also investigated Chapter 3 investigated the replacement of fishmeal by different plant protein ingredients in rainbow trout. The results of this chapter revealed that the effect of P. acidilactici on the microbiota of distal intestine in rainbow trout was dependent on the ingredients of the diet. The results also showed that the FM substitution induced major changes in the intestinal microbiota. Moreover, the modulation induced by plant-based diets on the microbiota varied according to the ingredients used. Chapter 4 studied the effect of dietary oxytetracycline in the distal intestinal microbiota of rainbow trout and the role of P. acidilactici to ameliorate the impact of antibiotic therapy. Experimental groups fed the diets with oxytetracycline had substantial changes in the distal intestinal microbiota including a decrease in the bacterial diversity. P. acidilactici did not ameliorate the effect of antibiotic therapy in the intestinal microbiota. Chapter 5 used Atlantic salmon during smoltification to study the changes in the microbiota of distal intestine and the role of P. acidilactici to promote intestinal health. The results showed that bacterial communities in the mucosa differed from the digesta. Seawater transfer and P. acidilactici had significant changes in the intestinal microbiota of both mucosa and digesta. However, the modulatory effect of both factors evaluated was larger in the mucosa-associated microbiota than in the digesta-associated microbiota. Furthermore, P. acidilactici induced a significant increase in antiviral-related genes. Chapter 6 investigated the replacement of fish oil by rapeseed oil alone or combined with P. acidilactici on the intestinal health and microbiota of two intestinal regions in Atlantic salmon. Replacement of fish oil by rapeseed oil alone or in combination with P. acidilactici supplementation did not induce major changes in the intestinal health and microbiota. The bacterial communities found were significantly different between the pyloric caeca and mid-intestine. In conclusion, this thesis contributes to new knowledge regarding the effect of dietary supplementation of P. acidilactici and the impact of different potential challenging factors in the health and intestinal microbiota of farmed salmonid species.

Tuberculosis (TB) disease, caused by the pathogen Mycobacterium tuberculosis (Mtb), remains a major global health problem claiming 1.5-2 million lives annually. One of the major factors contributing towards Mtb's success as a pathogen is its unique cell wall and its ability to counteract various arms of the host's immune response. Understanding these survival mechanisms will help us develop new therapeutic interventions that can enhance the capacity of the immune system to kill the pathogen. A recent genome scale study profiled a list of candidate genes that are predicted to be essential for Mtb survival of host mediated responses. One candidate was ftsEX, a protein complex comprised of an ATP binding domain, FtsE, and a transmembrane domain, FtsX. FtsEX functions through interaction with a periplasmic hydrolase, RipC. FtsEX homologs in other bacteria have been linked to a key role in regulation of PG hydrolysis during elongation and division. Using M. smegmatis as a model, we hypothesised that FtsEX and RipC are required in the regulation of PG hydrolysis during normal cell wall elongation and division under stressful conditions in vitro. Antibiotic sensitivity was confirmed using Alamar blue MIC determination assays, which showed that ftsEX and ripC had increased sensitivity to chloramphenicol and not to rifampicin, isoniazid and ethambutol. Our growth curve analysis showed that ftsEX and ripC are not essential for survival in normal growth conditions. However, ftsEX and ripC are conditionally essential for M. smegmatis in low salt media. Growth defects in this condition were characterized by short and bulgy cells, as well as elongated filamentous cells with visible chaining. Major morphological changes were seen under nitrosative stress. A higher proportion of cells struggled to divide normally and formed chains. Lateral branching was also observed in ΔftsE, ΔftsX and ΔftsEX but not in ΔripC. The protein complex was also required for survival in media containing rifampicin. Treatment with the drug exacerbated growth defects of all the mutants, which were much shorter than WT cells, indicating impairment in the elongation process. Collectively, mutants are much shorter in length with an exception of a few extremely lengthy cells, suggesting that ftsEX and ripC are required for both normal cell elongation and division and ultimately for survival in stressful conditions.

The purification and biochemical characterization of the substance P (SP) hydrolyzing enzyme, substance P endopeptidase (SPE), have been carried out; with subsequent orientation in neurobiological fundamental processes involved in opioid dependence, withdrawal, and heat-stress.

SPE was purified from rat spinal cord, human spinal cord and cerebrospinal fluid (CSF), rat ventral tegemental area (VTA), and rat hippocampus. The enzyme activity was found to release the biologically active fragments SP(1-7) and SP(1-8) as major products. The purified enzymes were characterized with regard to their biochemical and kinetic properties. The typical SPE is neither inhibited by phosphoramidon nor captopril nor phenylmethanesulfonylflourid (PMSF). In comparison to other known proteases SPE differed in characteristics regarding substrate specificity, inhibition-profile, cleavage pattern, and other kinetic parameters. The technically very delicate approach of micro purification of SPE from the rat ventral tegemental area (VTA) (this is a very small tissue), turned out to be possible with the ÄKTA™-purifier system. Studies revealed a crucial role of SPE in a series of clinically important neuropathological conditions, such as opioid tolerance, and withdrawal (SPE, increased); and heat-stress (SPE, increased). These findings emerged from assessment of enzyme activity in hypothalamus, nucleus accumbens (NAc) periaqueductal gray (PAG), pituitary, striatum, substantia nigra (SN), VTA, spinal cord. Viewing the role of SPE in morphine tolerance, it was possible to note regional differences with a decrease in PAG, and striatum, whereas an increase was seen in SN, and VTA. After heat-stress treatment, SPE was raised in several regions (cerebral cortex, hippocampus, diencephalon, cerebellum, spinal cord), and the most precise observation of this was located to the hippocampus structure.

Leadership in stressful and dangerous situations is vitally important in terms of lives, property, and national strategic objectives. But our understanding of effective leadership in these and other contexts is limited. Part of the problem is that interactionist theoretical perspectives are not reflected in contemporary leadership thinking. In addition, the impact of individual differences on leadership is often misrepresented or hidden by linear correlations and regressions conducted on continuous scores. This study employed new, innovative, indirect conditional reasoning measures to assess the personalities of 627 leaders entering the militaryâ s most challenging and stressful combat leader development course (the US Army Ranger School). These innovative measures predicted compelling differences in leadership, attrition, and in the peer evaluations made during the training. Analyses conducted on the continuous personality scores demonstrate that these findings are misrepresented or hidden by linear correlations and regressions. As an alternative, I present a configural scoring scheme, couched in a poker analogy, to explain how these individual differences combine to predict the odds of success for each of the 18 personality types studied.

This thesis evaluates how regulation on capital requirements is associated with cost and profit efficiency of banks. It allows an international comparison on banking regulation, capturing changes on public policies towards banks over time further, it encompasses one of the major financial crises since 1929. The impact of regulatory capital requirements on banking efficiency is assessed using the stochastic frontier production function for panel data to estimate inefficiency effects. It is used a panel dataset of 865 observations from 156 publicly listed commercial banks operating in 30 countries and covering a nine year time-horizon (2004-2012). Besides the impact on capital regulation, we also controlled for the influence of other regulatory variables, macroeconomic conditions, market structure characteristics and the state of financial development. Our results suggest that capital requirements negatively affect inefficiency, meaning that, for our sample, an increase on regulatory equity ratios will lead to an efficiency improvement. Regarding other regulatory control variables, we would say that regulation should aim to increase market discipline, while restricting banks activities. The power of supervisory agencies seems to negatively impact banks’ efficiency, but in in stressful conditions, it may help banks being more cost efficient.

L’objectif principal de ce mémoire est de déterminer l’effet modérateur des évènements stressants subis lors de l’enfance sur la relation entre les conditions de l’organisation du travail et le niveau de détresse psychologique vécue par les policiers et les employés de bureau de la ville de Montréal (SPVM). Les objectifs secondaires sont de déterminer le rôle direct des conditions de l’organisation du travail et des évènements stressants subis lors de l’enfance sur le niveau de détresse psychologique de cette population. Des données secondaires découlant d’une enquête réalisée auprès de 410 employés du SPVM nous ont permis de réaliser cette étude et de faire les analyses statistiques nécessaires. Cette recherche nous a permis de déceler quelques facteurs pouvant influencer le niveau de détresse psychologique des employés du SPVM. Entre autres, les résultats nous indiquent un niveau de détresse psychologique assez faible pour notre échantillon. De plus, ceux-ci nous démontrent que parmi les conditions de l’organisation du travail, deux variables indépendantes sont associées à une variation du niveau de détresse psychologique, c’est-à-dire les demandes psychologiques et le soutien social des superviseurs. En effet, les demandes psychologiques font augmenter le niveau de détresse psychologique, tandis que le soutien des superviseurs le fait diminuer. De plus, nos résultats soutiennent que le fait d’être un homme, de vivre en couple, de ne pas subir de tensions avec son enfant et son conjoint, de pratiquer des activités physiques, de ne pas consommer de tabac et de posséder un centre de contrôle interne sont associés à un faible niveau de détresse psychologique. Parallèlement, cette étude nous a permis d’avoir une compréhension plus exhaustive du rôle spécifique des évènements de vie stressants vécus lors de l’enfance en ce qui concerne la détresse psychologique vécue en milieu de travail. Ainsi, nos analyses multivariées nous ont permis d’identifier le fait qu’il n’y avait pas présence d’un effet modérateur des évènements stressants subis lors de l’enfance sur la relation entre les conditions de l’organisation du travail et le niveau de détresse psychologique. Pour ce qui est de la relation des évènements stressants subis lors de l’enfance sur le niveau de détresse psychologique, aucun lien n’a été décelé lors des analyses.
The main objective of this master thesis was to verify the moderating effects of stressful life events from childhood on the relation between the work organization’s conditions and the level of psychological distress lived by workers from the Service de Police de la Ville de Montréal (SPVM). As a secondary objective, this thesis determined the direct effect of work organization conditions on the level of psychological distress. To complete this study, data from an investigation of 410 SPVM workers was used. This thesis demonstrated a low level of distress in this population. Also, the completed multilevel analysis confirmed that psychological demands and social support from the superiors were associated at the level of psychological distress. In fact, psychological demands contribute to increase it and the social support from the superior decrease the level of psychological distress. In addition, our results indicate that distress is to be less pronounced among men, couples, life habits (physical activities and no tobacco use),no tensions with children, no tensions with the spouse and the personality trait sense of control. Finally, this thesis hasn’t found any moderating effects of the children’s stressful life events on the relation between the work organization’s conditions and the level of psychological distress. Moreover, the multilevel analyses haven’t demonstrated the direct effect of children’s stressful life events psychological on the level of psychological distress.

碩士
國立臺灣海洋大學
生物科技研究所
92
ABSTRACT A high expression of heat shock protein 70 (HSP70) was found to arise in zebrafish (Danio rerio) at 72 to 84 hr after fertilization [36, 37]. The spontaneously expressed HSP70 was later shown to comprise predominantly heat shock cognate 70 (HSC70), HSP70 synthesized under nonstressful condition, and very low level of stress-induced HSP 70 [53]. Owing to the constitutive expression of hsp70/hsc70 mRNA in developing zebrafish, the spontaneous production of HSC/HSP 70 was thought to be regulated at translational level [37]. A parallel up- and down-regulation of HSC/HSP 70 and their cochaperone DJ-2 in 72 to 108-hr-old zebrafish was also observed [53]. The objective of this research was to study the correlation between HSC/HSP 70 and DJ-2 gene expression in various tissues of developing zebrafish at both the mRNA and the protein level. Tissue-specific expression of heat shock factors 1 & 2 (HSF 1&2) that trigger the transcription of hsp/hsc 70 was also examined. Whole mount in situ hybridization using a riboprobe targeting hsp/hsc 70 mRNA detected a wide distribution of hsp70 mRNA in the brain and somite regions of 36-hr-old embryos. In contrast, the probe specifically hybridized with stress-induced hsp 70 mRNA gave hybridization signals only around the lens of zebrafish at different stages. Hence, hsc70 mRNA was produced in most zebrafish tissues. In 60 to 108-hr-old zebrafish late embryos and larvae, hsc70 mRNA was clearly detectable in their brains and pectoral fins, while a low level of this mRNA was found in somites. Tissue-specific HSC70 expression not closely associated with its corresponding mRNA was detected by whole mount immunocytochemistry The synthesis of HSC70 was found in the hindbrains, eye regions and the body surfaces of somites of 36-hr-old embryos In addition to its presence in the eyes and pectoral fins, HSC70 appeared as spots in some epithelial tissues of head regions in 60 to 108-hr-old zebrafish. The major fraction of dj-2 mRNA and DJ-2 molecules were shown to colocalize with hsc70 mRNA and HSC70 in developing zebrafish, suggesting the interaction between HSC70 and DJ-2 for assisting protein folding during zebrafish development. The appearance of hsf1 and hsf2 mRNA with hsc70 mRNA was seldom correlated in 36-hr-old zebrafish, yet colocalization of these transcripts was frequently observed in more mature zebrafish.