<p>  </p> <p>Little is known about pullet stress and welfare and even less is known about pullets in cage-free housing. The studies included in this dissertation evaluated bird responses to stocking density, feeder space, and ACTH stressors in pullets. The project and procedures were reviewed and approved by the Purdue University Animal Care and Use Committee (Protocol#:2001002002 and # 1908001938).  </p> <p>Two studies of external stressors and one study of an internal stressor were applied to Lohmann LB-Lite (Brown) and Lohmann LSL-Lite (White) pullets. For both studies of an external stressor, bird-based welfare parameters included evaluation of bird appearance through feather coverage, foot condition, and keel condition; blood parameters, i.e., cholesterol, corticosterone, cortisol, NDV/IBV IgG/IgY titer, and differential WBC (%) and heterophil to lymphocyte ratio; measurements of the spleen, bursa, thymus, and liver; and production values measured through body weight, body weight uniformity, shank length, feed consumption, and feed conversion ratio (FCR). Birds were followed from 2-16 weeks of age (WOA) for the external stressors and were housed on the floor.</p> <p>The first external stressor was applied through a 2 x 2 factorial randomized complete block design (RCBD) using Brown and White strains and two stocking densities- high (HSD) and low (LSD). Stocking densities were adjusted over time, with the ending density of 670 cm2/bird for HSD and 1352 cm2/bird for LSD at 16 WOA. Density alone affected relative bursal weight (%), with LSD generating heavier bursas. LSD also generated lower, better FCR for both white and brown strains though this difference was not statistically significant. Within a density, opposing strain effects were seen for body weight uniformity and feather coverage. White was improved at HSD while Brown was improved for LSD. </p> <p>The second external stressor study was a 2 x 2 x 3 factorial RCBD with Brown and White, 2 feeder space allocations (3.5 or 7 cm/bird), and 3 stocking densities. The ending stocking densities were 154.45, 518.76, and 1,247.38 cm2 for the high (HSD), medium (MSD), and low (LSD) stocking density treatments, respectively, at 16 WOA. In this study, less space per bird was associated with poorer feather condition but improved FCR. Reduced feeder space was associated with elevated or worse FCR.</p> <p>In a study of internal stressors, ten each of Lohmann LSL-Lite hens received either a saline-mannitol or synthetic human ACTH injection at 6, 17, and 26 weeks of age. Blood was collected prior to injection and 1- and 2-hours post-injection. H:L ratio, WBC differential counts, corticosterone, and cortisol were measured in blood. Albumen samples were collected to measure corticosterone and cortisol at 26 WOA only but were unaffected by treatment. Serum corticosterone and H:L ratio were also unaffected by treatment. Serum cortisol was increased 1 hour after ACTH injection at 17 and 26 WOA and percent heterophils and lymphocytes were affected by ACTH at 26 WOA indicating potential utility as a measure of stress for adult birds only. </p> <p>The results of these studies show that pullets are resilient and resistant to several stressors. Both acute and chronic as well as internal and external stressors were tested in these studies with limited effects on the birds’ physiology, immunity, condition, or production. Feather coverage and bursal size were improved at lower stocking densities and FCR was improved at greater feeder space but the implications of these differences are unclear. Further research is needed to identify why there are reduced stress responses in pullets; it is possible that domestication or preserved evolutionary pathways explain some of the differences between pullet and adult stress responses. And while the majority of parameters were unaffected by the stressors used, the results of this study presented baseline values for Lohmann LB-Lite and Lohmann LSL-Lite pullets that will help guide future research. Additionally, the results of these studies left perhaps more questions than answers but illustrate perfectly that more research will be critical to an understanding of pullet stress and welfare.</p>

There is increasing recognition of the relationship between depression and obesity in the pediatric population and recently, there has been a focus on inflammation as a potential link. Both conditions are considered to be pro-inflammatory states, and certain inflammatory markers are linked to depression in obese adults and vice versa. Leptin has also been implicated in depression as a potential mediator between inflammation and depression. Brain derived neurotrophic factor (BDNF), which is associated with depression and obesity, is influenced by inflammation and leptin in animal models as well. Few studies have examined the interactions between depression, adiposity, and biological markers in obese youth and therefore, our objective was to explore the determinants of depression in obese youth in a clinical setting. We studied 244 youth aged 8-17 years (125 girls, 119 boys) at the time of entry to a weight management program, as part of a prospective, longitudinal study. The CES-DC depression-screening tool was used to assess depressive symptoms, and a participant was classified as having high depressive symptoms if the CES-DC score ≥15 or taking antidepressants. Questionnaires assessed socio-demographic factors and puberty while adiposity was measured using dual-energy X-ray absorptiometry (DXA). Inflammatory markers (IL-6, TNFα, CRP, IL-10), leptin, and BDNF were quantified by immunoassays. Of the 244 participants, 8 were on antidepressants and 88 (36.4%) met the criteria for high depressive symptoms. We confirmed previous findings that household income and body fat were important determinants of depressive symptoms. However for the first time, it was identified that leptin levels predicted CES-DC score independent of body fat. Neither inflammatory markers nor BDNF were significantly related to depression scores. Our findings suggest that leptin may mediate the relationship of adiposity and depression but it is uncertain if this is related to direct action or to the phenomenon of leptin resistance. / Thesis / Master of Science (MSc) / Obesity has a significant impact on depression in children and adolescents. Inflammation – the body’s response to injury – is measured through markers in the blood and leptin – the marker of body fat – have shown to be related to depression. Research indicates that depression influences these factors to act on obesity. However, research on the interactions of biological and socio-demographic factors with depression in youth with obesity is lacking. Therefore, our objective was to explore the impact of these factors on depression in obese youth entering into a weight management program. Using a depression-screening tool, we studied 244 youth under 18 years and confirmed that household income and body fat were important factors of depression. However for the first time, we found leptin influenced depression regardless of the amount of fat present suggesting that depression acts on obesity through leptin but it is uncertain how this occurs and further research is warranted.

No description available.

No description available.

<p>Salivary cortisol and electrocardiogram data was collected at baseline and after a stress-anticipation task in extremely low birth weight (ELBW; < 1000 grams) survivors and normal birth weight (NBW) controls, in order to examine the moderating influence of emotion regulation on the relationship between being born at ELBW and internalizing problems in adulthood. The stress manipulation was an adapted Trier Social Stress Task. The participants were told they would have three minutes to create a speech on one of three predetermined topics (i.e., gun control, same sex marriage, or abortion). After three minutes passed, the particpants were told that there would be no speech. All participants showed a decrease in salivary cortisol levels throughout the day, and an increase in heart rate during the stress anticipation task. When a median split was used to create high and low stress reactive cortisol and heart rate groups, an interaction was found between birth status (ELBW vs. NBW) and group (High vs. Low Stress Reactive Cortisol) on self-reported internalizing problems (anxiety, depression, withdrawal). Those born at ELBW who had high stress reactive cortisol self-reported significantly higher levels of internalizing problems compared to ELBWs with low stress reactive cortisol. Those born at NBW did not differ on self-reported internalizing problems based on their stress reactive cortisol levels. When the moderating effect was probed with a linear regression analysis, the ELBW group was driving the relation between stress reactive cortisol levels and internalizing problems. Taken together, the results suggest that emotion regulation, as indexed by the neuroendocrine system, is moderating the relation between being born at ELBW and internalizing problems in adulthood. This is indicative of a differential susceptibility of risk and resilency in ELBW survivors depending on their ability to regulate their emotions, specifically during periods of stress.</p> / Master of Science (MSc)

[pt] As emissões de combustíveis fósseis derivados do petróleo têm um impacto significativo no meio ambiente, nas mudanças climáticas, na poluição do ar e outros. Por isso, existe um grande interesse por fontes alternativas de energia, principalmente aquelas que podem reduzir as emissões de poluentes atmosféricos. Este trabalho teve por objetivo avaliar os perfis das concentrações de poluentes regulamentados (material particulado e CO2) e não regulamentados (benzeno, tolueno, etilbenzeno, xilenos, HPA, hidrocarbonetos alifáticos, aldeídos e metais) originados da queima de diferentes combustíveis em um motor estacionário do ciclo diesel e verificar os possíveis efeitos toxicológicos utilizando células epiteliais brônquica. Os combustíveis utilizados foram: mistura binária diesel com 5 por cento de biodiesel (B5), biodiesel de soja (B100), biodiesel de soja aditivado (B100 adt) e etanol aditivado. Para obter uma maior confiabilidade dos resultados realizou-se a verificação do método para a determinação de monoaromáticos e aldeídos por meio de cromatografia gasosa com detector de ionização de chama (CG-DIC) e cromatografia líquida de alta eficiência (CLAE), respectivamente. O MP foi determinado por gravimetria, Os HPA associados ao MP foram determinados por cromatografia gasosa acoplada a espectrometria de massa (CG-EM), hidrocarbonetos alifáticos (HA) por CG-DIC e metais foram analisados por espectrometria de massa com plasma indutivamente acoplado (ICP-MS). Os perfis das concentrações foram avaliados considerando: o tempo de queima de combustível (15, 30 e 60 min) e, o tempo de armazenamento (7, 14 e 21 dias) a 40 graus Celsius, para avaliar os processos oxidativos que podem ocorrer durante o armazenamento, especialmente no caso do biodiesel. A operação do motor quando ele ainda está frio, por exemplo, em 15 minutos, apresentou maior emissão de MP, benzeno e etilbenzeno para o biodiesel de soja (B100) em comparação com a mistura de diesel com 5 por cento de biodiesel (B5). O estudo dos perfis das concentrações dos compostos mono aromáticos, hidrocarbonetos alifáticos e hidrocarbonetos policíclicos aromáticos (HPA) mostraram resultados semelhantes aos descritos na literatura, em que o combustível B5 emiti mais poluentes do que o biodiesel. No entanto, comparando alguns poluentes individualmente, as emissões de benzeno e etilbenzeno foram maiores para B100 e B100 adt. O B5 apresentou o maior perfil de concentração para a soma HPA na fase gasosa do que na forma particulada em períodos de funcionamento do motor mais longos. O tempo de armazenamento reduziu as emissões de MP em 40 por cento para B100, 20 por cento para B100 adt e 3 por cento para B5. Em relação à concentração de compostos aromáticos, a redução foi de aproximadamente 60 por cento desses poluentes para B100 e B100 adt. A menor emissão de poluentes foi durante a queima do etanol aditivado. / [en] The emissions of fossil fuels derived from oil have a significant impact on the environment, climate change, air pollution, and others. Therefore, there is great interest in alternative energy sources, especially those that can reduce emissions of air pollutants. The aim of this study was the evaluation of concentration profiles of criteria pollutants, particulate matter (PM) and CO2 and unregulated pollutants (i.e., benzene, toluene, ethylbenzene, xylenes, polycyclic aromatic hydrocarbons (PAH), aliphatic hydrocarbons, aldehyde and metals) originated from burning of different fuels: The evaluated fuels were the binary mixture of fossil diesel with 5 percent biodiesel (B5), biodiesel soybean (B100), additivated biodiesel soybean (B100 adt) and additivated ethanol using a stationary diesel engine cycle, operating at the speed of 1800 rpm and 0 percent load. For a larger reliability of results, the method validation for determination the monoaromatic and aldehydes by gas chromatography flame ionization detection (GC-FID) and high-performance liquid chromatography (HPLC), respectively. PM mass was determinate by gravimetry, PAH associated to the PM were determined by gas chromatography coupled to mass spectrometry (GC-MS), aliphatic hydrocarbons were determined by GC-FID, and metals were determined by inductively coupled plasma mass spectrometry (ICP-MS). The concentration profiles were evaluated considering: 1) burning time (15, 30, and 60 minutes) and; 2) storage time (7, 14, and 21 days) at 40 Celsius Degrees to evaluate the oxidative processes that occur during storage, especially for biodiesel. When it is still cold (i.e., 15 minutes), the operation of the engine showed negative effect on particulate matter (PM), benzene, and ethylbenzene emissions for B100 compared to B5. In this study, the concentration profiles of the monoaromatic, aliphatic hydrocarbons, and PAH showed similar results to those reported in the literature, where B5 fuel emits more pollutants than pure biodiesel. However, comparing some pollutants individually, the benzene and ethylbenzene emissions were higher for B100 and B100 adt. For long engine operation periods, B5 showed the highest concentration profile for the PAH sum in the gaseous in comparison to PAH sum in the particulate phase. The storage time reduced the PM emissions in 40 percent (B100), 20 percent (B100 adt), and 3 percent (B5). Regarding the concentration of aromatics, the reduction was circa 60 percent for B100 and B100 adt. The lowest emission of pollutants was observed for additived ethanol burning.

Thesis (MSc (Physiological Sciences))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Bi-directional communication exists between HPA-axis activation and interleukin-6 (IL-6). However, the relative contribution of centrally versus peripherally secreted IL- 6 remains unclear, especially under psychological stress conditions. We hypothesised that the HPA response to mild psychological stress is dependent on IL- 6, both centrally and peripherally. 120 male Wistar rats were divided into four groups, depending on whether they received an anti-IL-6 antibody (Ab) (2μg/ml/kg body weight) or a placebo (sterile saline) injection and whether or not they were subjected to 1 hour of restraint stress for 1, 2 or 3 days. Rats were euthanized 24 hours after stress exposure. Plasma corticosteroid (GC) levels remained significantly increased 24 hours after a single stress exposure (control placebo (CP) versus stress placebo (SP): p < 0.05). The undetectable plasma IL-6 levels evident across all groups may be explained by the short half-life of IL-6. Plasma IL-1β levels decreased when IL-6 was blocked in unstressed animals (CP versus CAb: p < 0.05), suggesting a role for IL-6 in the maintenance of IL-1β levels under tonic physiological conditions. At tissue level, pituitary gland mass increased significantly at time point 2, independently of stress when blocking IL-6 (CAb: p < 0.05). This suggests that when normal homeostasis is threatened, immediate adaption or at least compensation may occur. It was observed that GR, IL-1β, IL-1βR, IL-6, IL-6R and GABAARα1 showed no response to stress alone in the pituitary. It is therefore more likely that resistance to adaptation exists centrally. IL-1β and IL-1βR (p < 0.05) and GABAARα1 (p < 0.005) expression increased in the CAb group in the pituitary, again suggesting a role for IL-6 under control conditions. In terms of the adrenal, blocking IL-6 resulted in decreased glandular mass at time point 1, independent of stress (CAb and SAb: p < 0.005). The up-regulation in GR expression seen in CAb and SAb (p < 0.05) may be the effect of a compensatory mechanism to increase IL-6 dependent bioactivity of GCs. The fact that expression of IL-6, IL-6R, IL-1β and IL- 1βR consistently increased in the Ab groups, and mostly in the zona fasciculata and zona reticularis, suggests that lack of local direct negative cytokine feedback occurred in response to very low plasma IL-6 levels and that this contributes more than GCs in the down-regulation of inflammatory cytokine release. In conclusion, consistent effects of the Ab were apparent in the tissues investigated, even in control conditions, suggesting that IL-6 plays a role in the maintenance of basal homeostasis, including its regulation of the response to psychological stress. We found differential regulation in terms of cytokines and GCs when comparing peripheral versus central effects of stress and Ab, as well as the levels of cytokines in the blood compartment, compared to within tissues. / AFRIKAANSE OPSOMMING: Daar bestaan twee-rigting kommunikasie tussen HPA-as aktivering en interleukin-6 (IL-6), allhoewel die relatiewe bydrae van sentraal versus perifeer afgeskeide IL-6 nog onduidelik is, veral gedurende sielkundige strestoestande. Ons hipotese is dat die HPA reaksie tot sielkundige stres afhanklik van IL-6 is, beide sentraal en in die periferie. 120 manlike Wistar rotte is in vier groepe verdeel, afhangende van of hulle ‘n anti-IL- 6 teenliggaampie (Ab) (2μg/ml/kg liggaamsgewig) of ‘n plasebo (steriele soutoplossing) inspuiting gekry het, en of hulle onderworpe was aan 1 uur van vaskeer-stres vir 1, 2 of 3 dae. Rotte is 24 uur na blootstelling aan stres aan genadedood onderwerp. Bloed kortikosteroïed (GC) vlakke het beduidend toegeneem binne 24 uur na ‘n eenmalige stres blootstelling (kontrole plasebo (CP) versus stres plasebo (SP): p < 0.05). Die onmeetbaar lae vlakke van IL-6 regoor al die groepe, kan verduidelik word na aanleiding van die kort half-leeftyd van IL-6. Bloed IL-1β vlakke het afgeneem in kontrole rotte wanneer IL-6 geblok is (CP versus CAb: p < 0.05). Dit kan beteken dat IL-6 noodsaaklik is vir die onderhoud van IL-1β vlakke gedurende basale toestande. Op weefselvlak het die hipofise massa toegeneem by tydpunt 2 toe IL-6 geblok is, onafhanklik van stres (CAb: p < 0.05). Dit dui aan dat wanneer normale homeostase bedreig word, daar onmiddelike aanpassing of kompensasie plaasvind. Dit is opvallend dat GR, IL-1β, IL-1βR, IL-6, IL-6R en GABAARα1 geen respons in terme van stres alleen in die hipofise getoon het nie. Na aanleiding daarvan is dit meer waarskynlik dat weerstand tot aanpassing sentraal bestaan. IL-1β and IL-1βR (p <0.05) en GABAARα1 (p < 0.005) uitdrukking in die hipofise het toegeneem in die CAb groep, wat weereens ‘n rol vir IL-6 onder kontrole toestande uitwys. In terme van die bynier, het die blok van IL-6 ‘n afname in massa veroorsaak by tydpunt 1, wat weer onafhanklik van stres was (CAb en SAb: p < 0.005). Die opregulering in die CAb en SAb groepe (p < 0.05), kan wees as gevolg van ‘n kompensasie meganisme om IL-6 afhanklike GC aktiwiteit te verhoog. Die feit dat die uitdrukking van IL-6, IL-6R, IL-1β and IL-1βR in die Ab groepe deurlopend verhoog was, en meeste in die zona fasciculata en zona reticularis, stel voor dat daar ‘n tekort aan plaaslike, direkte sitokien negatiewe terugvoering was, as gevolg van die merkwaardige lae bloed IL-6 vlakke en dat dit meer bydra as GCs in die afregulering van inflammatoriese sitokien vrystelling.

Thesis (MSc (Physiological Sciences))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: The hypothalamic-pituitary-adrenal-(HPA) axis has long been closely associated with psychological stress-induced activation of the adrenal cortex and subsequent glucocorticoid production. Another, less known peripheral limb of the psychological stress response, is the sympatho adrenal medullary pathway. We hypothesized that the sympatho-adrenal medullary system constitutes the primary response to acute psychological stress, with the HPA-axis functioning as a secondary response. We tested our hypothesis by manipulating a model of acute mild psychological stress (restraint) by blocking IL-6, a valuable constituent of the sympatho-adrenal medullary system. Serum corticosterone concentration increased in response to stress (7 ± 3 vs. 57 ± 4 ng/ml; P<0.0001), a response attenuated when IL-6 was blocked (17 ± 7 ng/ml). Stress increased pituitary mass only when IL-6 was blocked (38 ± 3 vs. 65 ± 6 mg; P <0.001). Stress increased left adrenal mass only in the presence of IL-6 (34 ± 1 vs. 73 ± 8 mg; P <0.00001). Stress did not influence the circulating levels of TNF-α, IL-1β or IL-6 significantly. IL-1β and TNF-α concentrations in the unstressed rats were lower when IL-6 was blocked. We then manipulated the stress model by administering S. frutescens extract to elucidate both the central and peripheral effects of acute S. frutescens administration on the psychological stress response. Restraint caused decreases in hippocampal GR levels when compared to respective controls. S. frutescens administration and exposure to restraint synergistically decreased hippocampal GABAAR levels. In addition, exposure to both stress and S. frutescens led to a noteworthy increase in pituitary mass (P = 0.078), as well as pituitary ACTH levels (P < 0.01). Similarly, differences in circulating ACTH levels showed an effect of stress on ACTH secretion only in the presence S. frutescens (P < 0.05). Adrenal mass was significantly increased in S. frutescenstreated animals that were also exposed to restraint (P < 0.05). Adrenal levels of ACTH showed a reciprocal trend to pituitary and circulating ACTH levels. No statistically significant differences were seen in adrenal IL-6 content. However, marked increases in IL-6 levels were seen at this level with administration of S. frutescens stress exposure and a cumulative increase seen with both S. frutescens-treatment and stress exposure. Hippocampal GABAAR, pituitary mass, pituitary ACTH and circulating ACTH levels showed a similar trend towards a synergistic effect of S. frutescens and restraint in activation of the psychological stress response, while adrenal ACTH levels showed an inverse trend. Hippocampal GR did not show any effect of stress or S. frutescens-treatment. The results from these two experiments indicate that the sympatho-adrenal medullary system constitutes the primary response to acute mild psychological stress and that the HPA-axis is only activated during an exacerbated stress response or when the sympatho-adrenal medullary contribution is inadequate. Furthermore, the acute administration of S. frutescens possibly led to a functional shift in GABAergic function, resulting in activation of the stress response. The anecdotal reports of a “docile” effect of S. frutescens most likely results from activation of the mesolimbic dopaminergic system by the hippocampus and amygdala. These results have dramatic consequence in GABA-based anxiety-treatments. / AFRIKAANSE OPSOMMING: Die hipotalamo-pituïtêre-adrenale (HPA)-as is lank bekend as ‘n primêre rolspeler in die respons op emosionele stres en daaropvolgende glukokortikoïed produksie. ‘n Ander, minder bekende arm van die sielkundige stres respons is die simpatiese bynier-medulla-sisteem. Ons hipotese was dat die laasgenoemde simpatiese bynier-medulla-sisteem die primêre respons tot sielkundige stres behartig terwyl die HPA-as ‘n sekondêre respons bied. Ons het ons hipotese getoets deur die manupilering van ‘n beproefde stres model waar ons IL-6, ‘n waardevolle rolspeler in die simpatiese bynier-medulla-sisteem, onderdruk het. In respons op stress, het serum kortikosteroon konsentrasies toegeneem slegs in die teenwoordigheid van IL-6 (7 ± 3 vs. 57 ± 4 ng/ml; P<0.0001), maar nie wanneer IL-6 onderdruk is nie (17 ± 7 ng/ml). Stres het ‘n verhoging in hipofise massa teweeggebring slegs tydens die onderdrukking van IL-6 (38 ± 3 vs. 65 ± 6 mg; P <0.001). Stres het ook linker-byniermassa verhoog slegs wanneer voldoende IL-6 beskikbaar was (34 ± 1 vs. 73 ± 8 mg; P <0.00001). Stres alleen het geen invloed gehad op serum IL-1β, IL-6 of TNF-α nie, maar die onderdrukking van IL-6 het wel ‘n inhiberende effek op basale IL-1β en TNF-α gehad. Daarna het ons weer eens die stresmodel manipuleer deur die rotte ‘n S. frutescens ekstrak te gee in ‘n poging om beide die sentrale en perifere effekte daarvan op die sielkundige stres respons te evalueer. Stres alleen het gelei tot ‘n afname in GR terwyl ‘n kombinasie van stres en S. frutescens administrasie tot ‘n afname in GABAARα1 in die hippokampus gelei het. Hierdie kombinasie het ook tot ‘n merkwaardige toename in hipofise massa (P = 0.078) sowel as ACTH-inhoud van die hipofise (P < 0.01) gelei. ‘n Soortgelyke patroon is waargeneem betreffende sirkulerende ACTH en byniermassa met P < 0.05 vir elk. Bynier ACTH inhoud, aan die ander kant, het ‘n omgekeerd eweredige verhouding met ACTH in die hipofise en in sirkulasie getoon. Bynier IL- 6 inhoud het geen statisties beduidende verskille getoon nie, maar ‘n merkwaardige verhoging is weereens gesien met ‘n kombinasie van stres en S. frutescens administrasie. Die soortgelyke tendens wat waargeneem word in GABAAR in die hippokampus, asook hipofise- en sirkulerende ACTH vlakke, en dui op ‘n samewerkende rol van stres en S. frutescens in die aktivering van die sielkundige stres respons. GR in die hippokampus toon geen veranderinge nie. Die resultate van die twee eksperimente dui op ‘n primêre rol van die simpatiese bynier-medulla-sisteem in die respons op ‘n akute stressor en dat die HPA-as net geaktiveer word tydens ‘n ooreiste stres reaksie of indien die simpatiese bynier-medulla-sisteem onderdruk word. Die waargenome “verdowings”-effek van S. frutescens word moontlik deur aktivering van die mesolimbiese dopamien pad deur die hippokampus en amigdala bewerkstellig. Die resultate mag ook lei tot die heroorweging van GABA-gebaseerde angs medikasies.

La lectine de Helix pomatia (HPA), extraite de la glande à albumine de l'escargot de Bourgogne et spécifique du résidu GalNAc, appartient à une nouvelle famille de lectine dite de type H. Elle est utilisée depuis plus de vingt ans comme marqueur d'adénocarcinomes (notamment du sein, du colon, du poumon) à fort pouvoir métastatique et donc faible pronostic vital. Son utilisation comme outil de routine en oncologie est, cependant, fortement limitée par son impossibilité à la produire sous forme recombinante. Afin de contourner ces difficultés, des protéines homologues ont été recherchées chez d'autres invertébrés. Deux lectines de type H ont été identifiées chez l'amibe Dictyostelium discoideum (discoidines) et une chez le corail Sinularia lochmodes (SLL-2). Les discoidines sont composées de deux domaines distincts, un domaine C-terminal, spécifique des résidus galactosylés et homologue à HPA et un domaine N-terminal, dit domaine discoidine, de fonction inconnue. Ces travaux de thèse portent, dans un premier temps, sur la poursuite de la caractérisation structurale de la discoidine 1 puis sur la production du domaine N-terminal de la discoidine 2 afin de confirmer la fonction lectine supposée. Dans un second temps, des expériences de microscopie confocale ont montrés que les discoidines ne possédaient pas la capacité d'HPA dans la discrimination des cellules métastatiques par rapport aux non métastatiques. La construction, par mutagenèse, d'une protéine chimérique entre la discoidine 2, très facilement produite dans E. coli, et HPA a alors été entreprise, le but étant de lui apporter la même spécificité qu'HPA. Enfin, la protéine SSL-2 a été clonée et de nombreux essais d'expression sous forme soluble et de purification ont été réalisés en vue de sa caractérisation biochimique et structurale pour sa possibilité d'utilisation comme marqueurs en histopathologie

Introduction: Prenatal maternal depression is associated with an increased risk of psychopathology in childhood. The understanding of the mechanisms underlying this association is limited. Further, despite high rates of prenatal depression in the developing world, no research investigating this issue exists from these settings. Objectives: The primary objectives of this thesis are to study the association between prenatal maternal depression and the following early offspring outcomes in a non-smoking, non-alcohol consuming prenatal sample from rural, South India: Foetal stress responsivity, measured through foetal heart rate (FHR). Infant stress responsivity, measured through infant cortisol response to immunisation. Infant temperament. Methods: 194 pregnant women from Solur, India were assessed for depression. The first 67 mothers with elevated symptoms of prenatal depression and the first 66 controls underwent FHR monitoring to study foetal stress responsivity. 58 mother-infant dyads returned at 1.5-3 months post birth. Infant salivary cortisol was measured before and after immunisation. Information on infant temperament and maternal postnatal depression (PND) was also collected. Results: Twenty nine mothers (14.9%) met a diagnosis of major depression during pregnancy while 67 (34.5%) had elevated symptoms of prenatal depression. Whilst there were no linear association between prenatal depression and foetal responsivity, a curvilinear (U shaped) association existed with the foetuses of mothers with very high and very low levels of prenatal depression having elevated stress responses compared to those with moderate levels of prenatal depression. Prenatal depression predicted infant cortisol responsivity independent of PND (B=13.08, p=0.02).The relationship between infant cortisol responsivity and prenatal depression was also U shaped. There was no association between prenatal depression and infant temperament. Conclusions: This is the first study from the developing world investigating the relationship between prenatal depression and offspring outcomes. It provides evidence suggestive of the programming influence of prenatal depression on the developing offspring.