Pendant la grossesse, les hormones stéroïdes jouent un rôle indispensable dans la régulation des principales manifestations physiologiques telles que la reconnaissance maternelle de la gestation, la réceptivité de l'endomètre, le début du développement embryonnaire ainsi que le maintien de la gestation. Cependant, on sait très peu sur la production de ces hormones et les principaux facteurs des voies intracellulaires impliqués dans le processus de stéroïdogenèse dans le placenta bovin pendant les stades initiaux et plus avancés de la gestation. Par ailleurs, certaines anomalies du placenta chez les bovins suite à une mauvaise production de stéroïdes n'ont pas encore été démontrées. Les objectifs de cette thèse étaient donc de : 1) déterminer la présence et la localisation des principales protéines stéroïdiennes dans le placenta de bovins provenant de gestations de 50 à 120 jours, 2) comparer l'expression placentaire d'une série de gènes et de protéines stéroïdiennes entre une gestation impliquant un transfert de noyaux de cellules somatiques (SCNT) et une gestation non-clonale; 3) étudier l'impact des hormones trophiques et des seconds messagers sur la stéroïdogenèse dans le placenta bovin à 140 +10 jours de gestation. L’utilisation de techniques d’immunohistochimie, d’immunobuvardage et de PCR quantitatif nous a permis d’évaluer la présence d'un large éventail de gènes stéroïdiens (STAR, CYP11A1, HSD3B1, CYP17A1 et SCARB1) qui participent au transport du cholestérol et dans la production de différents types de stéroïdes. Dans cette thèse, nous avons démontré la capacité du placenta bovin d’initier la stéroïdogenèse au début de la gestation et nous avons également déterminé les principales cellules impliquées dans ce processus. Nous avons constaté que les tissus maternels expriment les principaux marqueurs de stéroïdogenèse suggérant une plus grande capacité stéroïdogénique que les tissus fœtaux. En outre, un modèle d'expression des protéines complémentaires stéroïdogéniques entre la caroncule et le cotylédon a été observé, indiquant que la stéroïdogenèse placentaire exige une communication cellule à cellule entre les cellules de la mère et du fœtus. Après avoir démontré les principales cellules impliquées dans la synthèse des hormones stéroïdiennes dans le placenta bovin en début de gestation, nous avons ensuite étudié les modifications possibles de la stéroïdogenèse dans les tissus SCNT cotylédonaires à 40 jours de gestation. Nous avons identifié d'importantes modifications dans l'expression des gènes STAR, CYP11A1, HSD3B1, CYP17A1, et SULT1E1. Conséquemment, nous postulons que l'expression réduite des gènes stéroïdiens peut provoquer une insuffisance de la biosynthèse des hormones stéroïdiennes, ce qui pourrait contribuer à un développement anormal du placenta et du fœtus dans les gestations SCNT à court ou long terme. Finalement, nous avons développé un modèle efficace de culture d’explants de placentome qui nous a permis d'explorer les mécanismes sous-jacents spécifiques à la stéroïdogenèse placentaire. Nous avons exploré l'effet stimulant des hormones trophiques et différents messagers secondaires sur l'expression de différentes protéines stéroïdogéniques ainsi que le taux de progestérone (P4) dans les explants de placentome. En utilisant les techniques de RIA et de PCR quantitatif, nous avons constaté que même si les analogues de l'hormone lutéinisante (hCG) ont un effet stimulant sur plusieurs gènes stéroïdiens, le calcium ionophore est le principal modulateur dans la synthèse de la P4. Ces résultats suggèrent que dans le placenta bovin, la synthèse de la P4 est modulée principalement par l'afflux de calcium intracellulaire, et apparemment les nucléotides cycliques ne semblent pas contrôler ce processus. En conclusion, cette étude contribue de manière significative à une meilleure compréhension des mécanismes d'entraînement de la synthèse des stéroïdes placentaires au début de la gestation et permet aussi d’apporter de nouveaux éclairages sur l'importance des stéroïdes placentaires dans la régulation du développement du placenta et du fœtus. / During pregnancy, steroid hormones have essential roles in regulating key physiological events such as maternal recognition, endometrial receptivity, early embryonic development, and maintenance of pregnancy. However, very little is known about the production of these hormones nor about the principal factors and intracellular pathways implicated in the steroidogenic process in bovine placenta, during early and advanced pregnancy. In addition, placental abnormalities in cattle following an improper steroid production in bovine placenta have not been yet demonstrated. The aims of this thesis were to: 1) determine the occurrence and localization of the principal steroidogenic proteins in bovine placenta from day 50 to day 120 of pregnancy; 2) compare the placental expression of a series of steroidogenic genes and proteins between somatic cell nuclear transfer (SCNT) pregnancies and non-SCNT gestations; 3) investigate the impact of trophic hormone, and second messengers on steroidogenesis in bovine placenta at 140 +10 days of gestation. Using immunohistochemistry, western blot and qPCR techniques, we evaluated the presence of a wide range of steroidogenic genes (STAR, CYP11A1, HSD3B1, CYP17A1 and SCARB1), that participate in the cholesterol transport and in the production of different types of steroids. In this thesis, we demonstrated the capability of the early bovine placenta to initiate steroidogenesis, and we also determined the principal cells implicated in this process. We found that maternal tissue expresses the principal steroidogenic markers suggesting it has a greater steroidogenic capacity compared to fetal tissue. Moreover, a complementary pattern of steroidogenic protein expression between the caruncle and the cotyledon were found, indicating that placental steroidogenesis requires cell to cell communication between the maternal and fetal cells. Having shown the principal cells involved in the synthesis of steroid hormones in bovine placenta during early pregnancies, we then studied possible alterations in steroidogenesis in cotyledonary tissue in SCNT at 40 days of pregnancy. We identified significant alterations in the expression of STAR, CYP11A1, HSD3B1, CYP17A1 and SULT1E1 transcripts. Therefore, we postulate that reduced expression of steroidogenic genes may cause an insufficient local biosynthesis of steroid hormones, which might contribute to the abnormal placental and fetal development in SCNT gestations at short or long term. Finally, we developed an efficient placentome explants culture model that allowed us to explore the specific mechanisms underlying placental steroidogenesis. We explored the stimulatory effect of trophic hormones and different second messengers on the expression of various steroidogenic proteins and the progesterone levels in placentome explants. By RIA and qPCR techniques, we found that although LH-like hormones (hCG), had a stimulatory effect on multiple steroidogenic genes, the calcium ionophore was the principal modulator in the synthesis of progesterone. These results suggest that in bovine placenta, the synthesis of progesterone is modulated principally by intracellular calcium influx, and cyclic nucleotides do not seem to be controlling this process. In conclusion, these studies significantly contribute to a better understanding of the driving mechanisms of placental steroid synthesis in early gestations and also provide new insights into the importance of placental steroids in the regulation of placental and fetal development.

Les problèmes de santé liés à l’alimentation hyperlipidique chez l’humain sont en constante progression. Or, une perturbation de l’environnement fœtal induit chez la descendance une susceptibilité plus grande à développer des maladies à l’âge adulte (DOHad : Developmental Origins of Health and Disease). L’objectif de ce travail de Thèse est d’évaluer, chez le lapin, les conséquences d’une alimentation hypercholestérolémique et hyperlipidique sur le développement embryonnaire, fœtal, et la survenue de troubles métaboliques à long terme.Nous avons nourri des lapines ad libitum avec un régime hypercholéstérolémique (0,2%) et hyperlipidique (8%) (HH) ou un régime témoin (C) à partir de l'âge de 10 (expérience 1) ou de 18 semaines (age de la mise à la reproduction, expérience 2). A la naissance, les portées ont été équilibrées et des croisements effectués pour différencier l'effet de l'alimentation de la mère pendant la gestation et pendant la lactation. Ainsi des lapereaux nés de mères HH ont été allaités par des mères C (groupe HH-C) ou HH (groupe HH-HH) et des lapereaux nés de mère C ont été allaités par des mères C (groupe C-C) ou HH (groupe C-HH). Au cours de l’expérience 1, un retard de croissance intra utérin (RCIU) significatif a été mis en évidence dès 9 jours de gestation par échographie dans le groupe HH (P<0,05). A la naissance, les laperaux étaient significativement plus légers (P<0,05). En raison d’un rattrapage pondéral rapide, il n’existait plus de différence significative au sevrage. Tous les lapins ont alors reçu un aliment témoin distribué ad libitum. A J176, il n’y avait pas de différence de poids entre les groupes HH-HH et HH-C, mais les animaux de ces deux groupes étaient significativement plus lourds que les groupes C-C and C-HH (P<0.05). De plus, la tension artérielle était plus élevée dans le groupe HH-HH par rapport à tous les autres groupes (P<0.05). Au cours de l’expérience 2, de tels effets physiologiques n’ont pas été observés. Les effets physiologiques n'ayant été observés que lorsque le régime avait été commencé avant la gestation, nous avons émis l'hypothèse que l'environnement maternel précoce avait été modifié, entrainant une perturbation du développement embryonnaire à l'origine des conséquences à long terme. l’expression des gènes au moment de la mise en route du génome embryonnaire a été étudié à l’aide d’une puce dédiée. L’analyse transcriptomique a permis de suggérer que certains transcrits étaient présents en quantités différentes. Nous avons montré par qRT-PCR que le régime HH induit une augmentation transitoire de la quantité de transcrit de l’adipophiline (présente à J2 mais pas à J5,5). L’analyse immunohistochimique montre une quantité plus importante de gouttelettes lipidiques localisées près du noyau dans les embryons issus de mères nourries par le régime HH à J5,5 comparé aux témoins. Ces résultats illustrent l’importance de la nutrition avant et pendant la gestation pour la determination de la croissance in utero et postnatale, ainsi que pour le développement de maladies métaboliques à long terme. La nutrition maternelle avant la gestation peut engendrer des modifications d’expression de gènes au moment de la transmission materno embryonnaire. / The prevalence of human health problems associated with high-fat diets continues to rise, as does the number of such problems known to be associated with this diet. Disruption of the fetal environment induces in progeny a greater susceptibility to developing diseases in adulthood (DOHad: Developmental Origins of Health and Disease). The objective of the work for this thesis was to assess in rabbits the consequences of a high-cholesterol and high-fat diet on embryonic and fetal development and on the onset of metabolic disorders in the long term.We fed rabbits ad libitum with a high-cholesterol (0.2%) and high-fat (8%) (HH) diet or a control (C) diet, starting at the age of 10 (experiment 1) or 18 weeks (age at which reproduction began, experiment 2).The litters were balanced at birth, and crossings were performed to differentiate the effect of the mother's food during gestation and during lactation. Accordingly, rabbits born to HH mothers were nursed by C (HH-C group) or HH (HH-HH) mothers and those born to C mothers were nursed either by C (C-C) or HH (C-HH) mothers. During experiment 1, ultrasound clearly showed significant intrauterine growth restriction (IUGR) beginning at 9 days of gestation in the HH group (P<0.05). At birth, these rabbits weighed significantly less than their C counterparts (P<0.05). Because of their rapid weight catch-up, the significant difference had disappeared at weaning. All the rabbits thereafter received control food distributed ad libitum. At D176, there was no difference in weight between the HH-HH and HH-C groups but the animals in both these groups were significantly heavier than those in the C-C and C-HH groups (P<0.05). Moreover, blood pressure was higher in the HH-HH group than in any of the other groups (P<0.05). These physiological effects were not observed during experiment 2. Because the physiological effects were observed only when the diet began before gestation, we hypothesized that the early maternal environment been modified, a change that resulted in disruption of embryo development with long-term consequences. We then used a specially designed chip to study gene expression at the maternal to embryonic transition. Transcriptomic analysis suggested that some transcripts were present in different quantities. We showed with qRT-PCR that the HH diet induced a transient augmentation in the quantity of adipophilin transcripts (present at D2 but not at D5.5). The immunohistochemical analysis on D5.5 showed a higher quantity of lipid droplets localized near the nucleus of embryos from mothers fed with the HH diet than in embryos of control mothers. These results illustrate the importance of nutrition before and during pregnancy in the determination of in utero and postnatal growth as well as in the development of metabolic diseases over the long term. Maternal nutrition before conception can engender modifications in gene expression at the moment of the maternal to embryonic transition.

Pendant la grossesse, les hormones stéroïdes jouent un rôle indispensable dans la régulation des principales manifestations physiologiques telles que la reconnaissance maternelle de la gestation, la réceptivité de l'endomètre, le début du développement embryonnaire ainsi que le maintien de la gestation. Cependant, on sait très peu sur la production de ces hormones et les principaux facteurs des voies intracellulaires impliqués dans le processus de stéroïdogenèse dans le placenta bovin pendant les stades initiaux et plus avancés de la gestation. Par ailleurs, certaines anomalies du placenta chez les bovins suite à une mauvaise production de stéroïdes n'ont pas encore été démontrées. Les objectifs de cette thèse étaient donc de : 1) déterminer la présence et la localisation des principales protéines stéroïdiennes dans le placenta de bovins provenant de gestations de 50 à 120 jours, 2) comparer l'expression placentaire d'une série de gènes et de protéines stéroïdiennes entre une gestation impliquant un transfert de noyaux de cellules somatiques (SCNT) et une gestation non-clonale; 3) étudier l'impact des hormones trophiques et des seconds messagers sur la stéroïdogenèse dans le placenta bovin à 140 +10 jours de gestation. L’utilisation de techniques d’immunohistochimie, d’immunobuvardage et de PCR quantitatif nous a permis d’évaluer la présence d'un large éventail de gènes stéroïdiens (STAR, CYP11A1, HSD3B1, CYP17A1 et SCARB1) qui participent au transport du cholestérol et dans la production de différents types de stéroïdes. Dans cette thèse, nous avons démontré la capacité du placenta bovin d’initier la stéroïdogenèse au début de la gestation et nous avons également déterminé les principales cellules impliquées dans ce processus. Nous avons constaté que les tissus maternels expriment les principaux marqueurs de stéroïdogenèse suggérant une plus grande capacité stéroïdogénique que les tissus fœtaux. En outre, un modèle d'expression des protéines complémentaires stéroïdogéniques entre la caroncule et le cotylédon a été observé, indiquant que la stéroïdogenèse placentaire exige une communication cellule à cellule entre les cellules de la mère et du fœtus. Après avoir démontré les principales cellules impliquées dans la synthèse des hormones stéroïdiennes dans le placenta bovin en début de gestation, nous avons ensuite étudié les modifications possibles de la stéroïdogenèse dans les tissus SCNT cotylédonaires à 40 jours de gestation. Nous avons identifié d'importantes modifications dans l'expression des gènes STAR, CYP11A1, HSD3B1, CYP17A1, et SULT1E1. Conséquemment, nous postulons que l'expression réduite des gènes stéroïdiens peut provoquer une insuffisance de la biosynthèse des hormones stéroïdiennes, ce qui pourrait contribuer à un développement anormal du placenta et du fœtus dans les gestations SCNT à court ou long terme. Finalement, nous avons développé un modèle efficace de culture d’explants de placentome qui nous a permis d'explorer les mécanismes sous-jacents spécifiques à la stéroïdogenèse placentaire. Nous avons exploré l'effet stimulant des hormones trophiques et différents messagers secondaires sur l'expression de différentes protéines stéroïdogéniques ainsi que le taux de progestérone (P4) dans les explants de placentome. En utilisant les techniques de RIA et de PCR quantitatif, nous avons constaté que même si les analogues de l'hormone lutéinisante (hCG) ont un effet stimulant sur plusieurs gènes stéroïdiens, le calcium ionophore est le principal modulateur dans la synthèse de la P4. Ces résultats suggèrent que dans le placenta bovin, la synthèse de la P4 est modulée principalement par l'afflux de calcium intracellulaire, et apparemment les nucléotides cycliques ne semblent pas contrôler ce processus. En conclusion, cette étude contribue de manière significative à une meilleure compréhension des mécanismes d'entraînement de la synthèse des stéroïdes placentaires au début de la gestation et permet aussi d’apporter de nouveaux éclairages sur l'importance des stéroïdes placentaires dans la régulation du développement du placenta et du fœtus. / During pregnancy, steroid hormones have essential roles in regulating key physiological events such as maternal recognition, endometrial receptivity, early embryonic development, and maintenance of pregnancy. However, very little is known about the production of these hormones nor about the principal factors and intracellular pathways implicated in the steroidogenic process in bovine placenta, during early and advanced pregnancy. In addition, placental abnormalities in cattle following an improper steroid production in bovine placenta have not been yet demonstrated. The aims of this thesis were to: 1) determine the occurrence and localization of the principal steroidogenic proteins in bovine placenta from day 50 to day 120 of pregnancy; 2) compare the placental expression of a series of steroidogenic genes and proteins between somatic cell nuclear transfer (SCNT) pregnancies and non-SCNT gestations; 3) investigate the impact of trophic hormone, and second messengers on steroidogenesis in bovine placenta at 140 +10 days of gestation. Using immunohistochemistry, western blot and qPCR techniques, we evaluated the presence of a wide range of steroidogenic genes (STAR, CYP11A1, HSD3B1, CYP17A1 and SCARB1), that participate in the cholesterol transport and in the production of different types of steroids. In this thesis, we demonstrated the capability of the early bovine placenta to initiate steroidogenesis, and we also determined the principal cells implicated in this process. We found that maternal tissue expresses the principal steroidogenic markers suggesting it has a greater steroidogenic capacity compared to fetal tissue. Moreover, a complementary pattern of steroidogenic protein expression between the caruncle and the cotyledon were found, indicating that placental steroidogenesis requires cell to cell communication between the maternal and fetal cells. Having shown the principal cells involved in the synthesis of steroid hormones in bovine placenta during early pregnancies, we then studied possible alterations in steroidogenesis in cotyledonary tissue in SCNT at 40 days of pregnancy. We identified significant alterations in the expression of STAR, CYP11A1, HSD3B1, CYP17A1 and SULT1E1 transcripts. Therefore, we postulate that reduced expression of steroidogenic genes may cause an insufficient local biosynthesis of steroid hormones, which might contribute to the abnormal placental and fetal development in SCNT gestations at short or long term. Finally, we developed an efficient placentome explants culture model that allowed us to explore the specific mechanisms underlying placental steroidogenesis. We explored the stimulatory effect of trophic hormones and different second messengers on the expression of various steroidogenic proteins and the progesterone levels in placentome explants. By RIA and qPCR techniques, we found that although LH-like hormones (hCG), had a stimulatory effect on multiple steroidogenic genes, the calcium ionophore was the principal modulator in the synthesis of progesterone. These results suggest that in bovine placenta, the synthesis of progesterone is modulated principally by intracellular calcium influx, and cyclic nucleotides do not seem to be controlling this process. In conclusion, these studies significantly contribute to a better understanding of the driving mechanisms of placental steroid synthesis in early gestations and also provide new insights into the importance of placental steroids in the regulation of placental and fetal development.

Trypanosoma cruzi est un parasite protozoaire à multiplication intracellulaire, agent de la maladie de Chagas, infectant 16 à 18 millions de personnes en Amérique latine. Il peut être transmis de la mère infectée au fœtus dans 2 à 10 % des cas, mais ses autres effets sur la gestation ont été peu étudiés. Par ailleurs, les cytokines ont des effets sur la gestation. Certaines d’entre elles, comme l’interleukine-1, l’IL-4, l’IL-5, l’IL-10, le GM-CSF et le TGF-b2, sont bénéfiques pour la gestation, tandis que d’autres, comme l’IL-2, l’IL-12, l’IFN-g et le TNF-a ont des effets nocifs sur celle-ci. L’impact de l’infection à T. cruzi, stimulant la production de TNF-a et d’IFN-g, sur l'implantation et la croissance fœtale n’a pas été étudié. Le but de notre travail était d’étudier les effets de l’infection aiguë à T. cruzi sur la capacité de reproduction de la souris. Nous avons ainsi évalué les effets de cette infection sur la fertilité, le développement et la viabilité des fœtus de souris et le rôle de l’IFN-g et du TNF produits au cours de l’infection sur le développement de la gestation. Nous avons montré que l’infection aiguë à T. cruzi : i) diminue la capacité de reproduction de la souris ; ii) provoque une mortalité fœtale massive précoce (résorptions), tardive et néonatale associée à un retard de croissance intra-utérin, et ce, iii) en dehors de toute transmission congénitale du parasite. Par ailleurs nos travaux montrent que la mortalité fœtale/néonatale est associée à une invasion parasitaire massive du placenta qui présente d’importantes lésions à type d’infiltrats inflammatoires, de nécrose ischémique, de dépôts de fibrine et de thromboses vasculaires. Nous avons noté qu’il existe une relation inverse entre la charge parasitaire des unités utéro-placentaires et la viabilité du conceptus, suggérant que ces lésions placentaires contribuent à la mortalité fœtale en limitant les échanges materno-fœtaux. Enfin, nous avons également étudié le rôle de cytokines abortogènes comme le TNF et l’IFN-g, produites abondamment pendant l’infection aiguë de la souris par T. cruzi. Les taux sanguins maternels d’IFN-g étaient augmentés au 9ième mais pas aux 17ième et 19ième jours de gestation, alors que les taux de TNF sanguin et la production placentaire de cette cytokine augmentaient aux 17ième et 19ième jours de gestation. Afin d’évaluer le rôle de ces deux cytokines dans la mortalité fœtale, des souris ont été traitées par la pentoxifylline, pour inhiber la transcription du gène de TNF-a et diminuer la production d’IFN-g. Ces souris montraient une réduction de la mortalité fœtale à mi-gestation, associée à une diminution de la production du TNF placentaire, sans modifications des taux systémiques et sans effets sur l’IFN-g, suggérant la contribution du TNF dans la mortalité fœtale associée à l’infection aiguë par T. cruzi. En conclusion, notre travail montre que l’infection aiguë à Trypanosoma cruzi exerce un effet particulièrement néfaste sur la capacité de reproduction et le développement de la gestation chez la souris et que les lésions placentaires liées à l’infection et la production de TNF par le placenta infecté contribuent à cet effet.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Orientador: José Carlos Peraçoli / Resumo: Introdução: Nos últimos 20 anos, o conceito de near miss (risco de morte iminente) é abordado na saúde materna como adjuvante dos inquéritos confidenciais de morte materna. No sistema de saúde, o Hospital Terciário/Quaternário é centro de referência para pacientes em situações graves que as colocam em risco de morte. A Maternidade do HC-FMB-Unesp é um dos centros terciário/quaternário de referência para patologias obstétricas do DRS-VI do Estado de São Paulo. A análise da frequência de situações de risco que chegam à maternidadegestantes portadoras de hipertensão arterial, bem com a assistência que receberam até serem referenciadas e ao chegarem à maternidade, poderá identificar deficiências e propor o seu aprimoramento, reduzindo-se assim o risco do binômio mãe-feto. Objetivo: Determinar a frequência de casos de morbidade materna grave não near missque evoluíram para situação de near miss, associados à hipertensão arterial e propor protocolo clínico de assistência pré-natal, bem como aprimorar o protocolo de assistência praticada no centro terciário com finalidade de reduzir o risco do binômio mãe-feto. Sujeitos e métodos: Foram identificados todos os casos de gestantes ou puérperas com diagnóstico de morbidade materna grave e de near miss,relacionados à hipertensão arterial, que receberam assistência obstétrica durante os anos de 2015 e 2016.De forma descritiva os resultados foram apresentados em porcentagem sob a forma de tabelas. O projeto foi aprovado pelo Comitê de Étic... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Between 10% and 15% of maternal deaths are related to hypertensive gestational diseases, and most of these deaths are avoided by effective and timely care. Over the past 20 years, the concept of near miss has been addressed in maternal health as an adjunct to confidential maternal death surveys. Objective: To determine the frequency of evolution of cases of severe maternal morbidity not near miss (SMM not MNM) for maternal near miss (MNM) associated with hypertension in the reference center of the Polo Cuesta and Jurumirim Vale subareas of the Regional Health Division - VI (DRS-VI) of the State of São Paulo. Methods: A cross-sectional retrospective study was carried out between 2015 and 2016 at a tertiary/quaternary reference center for the Polo Cuesta and Jurumirim Vale sub-areas of DRS-VI in the state of São Paulo. A total of 167 pregnant women or postpartum women with a diagnosis of hypertension with signs of severity were included. In addition to the frequency of near miss, demographic, clinical and origin data were obtained according to the subarea of the regional health study population. The project was approved by the Research Ethics Committee of the Faculty of Medicine of Botucatu - Unesp (Opinion No. 2,309,947). The categorical variables were analyzed by the statistical program SPSS. Results: The demographic characteristics of the population studied were predominantly between 20-35 years old (68%), Caucasian (75.4%), stable union (71.9%), and no paid oc... (Complete abstract click electronic access below) / Mestre

Com o intuito de estabelecerem-se os valores de referência do lipidograma, por meio da determinação de colesterol, triglicérides, ácidos graxos livres não esterificados (NEFA), &beta;-Hidroxibutirato (&beta;-HBO) e glicose de bovinos, da raça Holandesa, criados no Estado de São Paulo, bem como avaliar a influência dos fatores etários, sexuais e da gestação nos referidos parâmetros bioquímicos, foram colhidas e examinadas amostras de soro e plasma sangüíneo de 413 bovinos considerados clinicamente sadios. Para o estabelecimento dos valores de referência e avaliação da influência dos fatores etários foram colhidas 273 amostras de soro e plasma distribuídas em 7 subgrupos experimentais: G1 - 20 bezerras com até 3 meses; G2 - 20 bezerras entre 3 e 6 meses; G3 - 20 bezerras entre 6 e 12 meses; G4 - 40 novilhas entre 12 e 24 meses; G5 - 73 vacas entre 24 e 48 meses; G6 - 55 vacas entre 48 e 72 meses; G7 - 45 vacas com mais de 72 meses de idade. Para avaliar a influência dos fatores sexuais foram colhidas amostras de soro e plasma de 30 machos adultos e 30 fêmeas adultas. Para avaliar a influência da gestação e do puerpério foram colhidas 75 amostras de soro e plasma sangüíneo divididos em 5 subgrupos experimentais: G1 - 16 fêmeas não prenhes; G2 - 18 fêmeas na fase inicial da gestação, com até 3 meses de gestação; G3 -15 fêmeas prenhes, entre 3 e 6 meses de gestação; G4 - 15 fêmeas na fase final da gestação, entre 6 e 9 meses de gestação; G5 - 16 fêmeas recém-paridas, com até 30 dias de puerpério. Em decorrência à amamentação, observou-se que nos primeiros três meses de vida, o lipidograma dos bezerros apresentaram particularidades em alguns dos seus constituintes, pois os valores de colesterol e de glicose foram maiores, enquanto os de &beta;-HBO foram menores do que os observados nos bezerros desmamados com idade variando entre 3 e 6 meses. Após a desmama, o lipidograma continuou a sofrer influência dos fatores etários, pois com o início da puberdade, após os 12 meses de idade, os teores séricos de colesterol aumentaram gradativamente atingindo o valor máximo entre 24 e 48 meses de idade, sendo que a partir desta faixa etária o colesterol deixou de sofrer influência dos fatores etários. Com relação à influência dos fatores etários observou-se, ainda, que os teores séricos de triglicérides encontrados em animais com idade maior do que 48 meses foram menores do que nos animais mais jovens e que os teores plasmáticos de glicose diminuíram gradualmente com o evoluir da idade. O lipidograma de bovinos sadios da raça Holandesa sofreu influência dos fatores sexuais pois os teores séricos de colesterol e &beta;-HBO para machos adultos foram menores do que os teores encontrados para fêmeas adultas enquanto os teores plasmáticos de glicose obtidos para machos adultos foram maiores que os teores encontrados para fêmeas adultas. O lipidograma de fêmeas bovinas sadias da raça Holandesa sofreu influência da gestação pois os teores séricos de NEFA foram maiores no terço final de gestação enquanto os teores séricos de &beta; - HBO foram maiores durante o terço inicial de gestação. Durante o puerpério observou-se a significativa influência desse fator nos constituintes do lipidograma, pois nos primeiros 30 dias após a parição verificou-se que os teores séricos de NEFA e &beta; - HBO foram maiores enquanto que os teores de triglicérides foram menores nos animais paridos em relação àqueles que estavam em gestação. Diante dos resultados obtidos na presente pesquisa e considerando-se os seus fatores de variabilidade, recomenda-se, baseado nos intervalos de confiança, a adoção dos seguintes valores de referência para o lipidograma de bovinos da raça Holandesa, criados no Estado de São Paulo: teores séricos de colesterol - entre 86,5 e 120,8 mg/dL para bezerros lactentes com até 3 meses de idade; entre 46,3 e 79,7 mg/dL para bezerros desmamados com 3 a 12 meses de idade; entre 86,4 e 105,0 mg/dL para novilhas com 12 a 24 meses de idade; entre 116,0 e 147,9 mg/dL para vacas adultas com mais de 24 meses de idade; teores séricos de triglicérides - entre 16,3 e 34,8 mg/dL para animais com até 48 meses de idade; entre 14,9 e 24,0 mg/dL para animais com mais de 48 meses de idade; teores séricos de NEFA - entre 91,3 e 294,0 &mu;M/L independente da faixa etária; teores séricos de &beta;-HBO - entre 1,1 e 2,1 mg/dL para bezerros com até 3 meses de idade; entre 3,37 e 6,2 mg/dL para animais com mais de 3 meses de idade; teores plasmáticos de glicose - entre 75,1 e 88,3 mg/dL para bezerros lactentes com até 3 meses de idade; entre 64,0 e 76,1 mg/dL para animais jovens entre 3 e 24 meses de idade; entre: 60,6 e 67,2 mg/dL para vacas adultas com mais de 24 meses. Para machos adultos, com mais de 24 meses de idade, recomenda-se a adoção de valores de referência diferenciados para os seguintes constituintes do lipidograma: teores séricos de colesterol entre 73,9 e 90,2 mg/dL; teores séricos de &beta;-HBO entre 3,1 e 3,6 mg/dL e teores plasmáticos de glicose entre 71,9 e 76,5 mg/dL. Os valores de referência apresentados anteriormente não devem ser utilizados para vacas que estejam no último mês de gestação e nos primeiros 45 dias de lactação, recomenda-se para este período a adoção de valores de referência específicos para animais na fase de periparto. / In order to establish the reference values for the lipid profile of Holstein cattle, bred in the state of São Paulo, we collected and analyzed samples of blood serum and plasma of 413 clinically healthy animals. We determined the concentrations of cholesterol, triglycerides, nonesterified fatty acids (NEFA), &beta;-hydroxibutyrate (&beta;-HBO) and glucose and analyzed the influence of age, gender and gestation on those biochemical parameters. The influence of age was studied through 273 samples of serum and plasma, divided in 7 experimental sub-groups: G1 - 20 heifer calves up to 3 months old; G2 - 20 heifer calves between 3 and 6 months; G3 - 20 heifer calves between 6 and 12 months; G4 - 40 heifers between 12 and 24 months; G5 - 73 cows between 24 and 48 months; G6 - 55 cows between 48 and 72 months; G7 - 45 cows with more than 72 months of age. The influence of gender was studied through serum and plasma of blood samples collected from 30 adult males and 30 adult females. The influence of gestation and puerperium was studied through serum and plasma from 75 animals divided into 5 experimental subgroups: G1 - 16 non pregnant females; G2 - 18 females in the initial phase of gestation, up to 3 months; G3 -15 pregnant females, with gestation between 3 and 6 months; G4 - 15 females in the last stage of gestation, between 6 and 9 months; G5 - 16 females soon after delivery, up to 30 days in puerperium. Due to lactation, we observed that in the first 3 months of life, the calves&#39; lipid profile has particularities in some of its constituents, such as higher levels of cholesterol and glucose and lower levels of &beta;-HBO, if compared to those observed in weaned calves between 3 and 6 months of age. After weaning, the lipid profile continued to be influenced by age; in the early stages of puberty, after 12 months of age, the serum cholesterol concentration increased gradually reaching its peak between 24 and 48 months - after this stage, the serum cholesterol concentration was no longer influenced by the animals&#39; age. Also concerning the age, we observed that the serum triglyceride concentration in animals older than 48 months was lower than that of younger animals and the glucose concentration in the plasma diminished gradually with age. The lipid profile of healthy Holstein cattle was influenced by the gender of the animals. The serum concentrations of cholesterol and &beta;-HBO for adult males were lower than those for adult females. The plasma glucose in adult males was higher than that in adult females. The lipid profile of healthy Holstein cows was influenced by gestation. The serum concentration of NEFA was higher in the last trimester of gestation while the serum concentration of &beta;-HBO was higher in the first trimester of gestation. The puerperium had a significant influence on the lipid profile. In the first 30 days after delivery, the serum concentrations of NEFA and &beta;-HBO were higher, while the triglycerides were lower when compared to those of cows during gestation. According to the results of this research and considering its variability factors, we suggest, based on the confidence interval, the use of the following reference values for the lipid profile of Holstein cattle bred in the state of São Paulo: serum cholesterol concentration - between 86.5 and 120.8 mg/dL for lactating calves up to 3 months old; between 46.3 and 79.7 mg/dL for weaned calves between 3 and 12 months; between 86.4 and 105.0 mg/dL for heifers between 12 and 24 months; between 116.0 and 147.9 mg/dL for adult cows older than 24 months; serum triglycerides concentration - between 16.3 and 34.8 mg/dL for animals up to 48 months; between 14.9 and 24.0 mg/dL for animals older than 48 months; serum NEFA concentration - between 91.3 and 294.0 &mu;M/L regardless of the age; serum &beta;-HBO concentration - between 1.1 and 2.1 mg/dL for calves up to 3 months old; between 3.37 and 6.2 mg/dL for animals older than 3 months; plasma glucose concentration - between 75.1 and 88.3 mg/dL for lactating calves up to 3 months old; between 64.0 and 76.1 mg/dL for animals between 3 and 24 months; between 60.6 and 67.2 mg/dL for adult cows older than 24 months. For adult males, older than 24 months, we suggest different reference values for the following constituents of the lipid profile: Serum cholesterol concentration between 73.9 and 90.2 mg/dL; serum &beta;-HBO concentration between 3.1 and 3.6 mg/dL and plasma glucose concentration between 71.9 and 76.5 mg/dL. The reference values that were previously presented should not be used for cows in the last month of gestation and in the first 45 days of lactation. In this case, the reference values should be specific for animals around parturition.

Durante a gestação, a interação entre os organismos materno e embrionário é mediada por diversas moléculas reguladoras produzidas no ambiente uterino que são fundamentais para o estabelecimento de um ambiente tolerante ao feto. A expressão de uma grande variedade de mediadores inflamatórios também participa deste processo influenciando o ambiente uterino e a própria placenta. As citocina, potentes mediadores celulares, regulam as respostas imunológicas, integrando a rede sinalizadora que controlam a gestação. Desbalanço nos perfis de citocinas, sistêmica ou localmente no ambiente placentário tem sido descrito em muitos distúrbios gestacionais que podem culminar com perdas fetais. Evidências recentes mostraram que a expressão de uma nova citocina, a Interleucina 25, está alterada em pacientes que apresentam abortos recorrentes, sugerindo um papel imuno-regulador da IL-25 nas funções reprodutivas. Neste contexto, torna-se bastante interessante avaliar a presença da IL-25 na interface materno-fetal durante toda a gestação. Neste estudo objetivamos caracterizar a expressão de IL-25 e seu receptor IL-17BR em células placentárias e no sangue materno ao longo da gestação em camundongos. A análise da expressão gênica de IL-25 e seu receptor IL-17BR por qRT-PCR, foi realizada em sítios de implantação e em porções fetais e materna de placentas nos dias 10,5, 13,5, 16,5 e 19,5 de gestação (dg). Células mononucleares do sangue (CMNs) e tecidos placentários dissecados foram utilizados para ensaios de Citometria de fluxo nos mesmos dias de gestação. A imunolocalização de citocina e receptor foi avaliada em cortes histológicos nos diversos dias gestacionais estudados. Nossos dados demonstraram que a expressão gênica de IL-25 se intensifica após o período implantacional, aumentando os níveis de mRNA gradativamente a partir de 10,5dg até 16,5dg. Citocina e receptor são expressos de forma diferenciada pelos compartimentos placentários. A análise da expressão proteica por citometria de fluxo confirmam os dados de expressão gênica e as marcações observadas nas reações de imunohistoquímica. IL-25 e IL-17BR são expressos em toda a interface materno-fetal, sendo a citocina prevalente na placenta fetal, enquanto o receptor se expressa de forma intensa na porção materna. Entre as populações celulares que poderiam contribuir para os níveis de citocina na placenta fetal, observa-se a presença de células trofoblásticas, fenotípicamente caracterizadas, produtoras de IL-25 e células leucocitárias (CD45+) expressando o receptor IL-17BR na região da decídua materna. Entre o período gestacional estudado, a expressão de IL-25 e IL-17BR ocorreu principalmente nos dias 13,5 e 16,5 dg. O significado funcional destes achados, no entanto, merece investigações mais detalhadas, entretanto, devido ao caráter imunoregulatório atribuído a citocina IL-25, está parece estar associada ao controle das funções placentárias contribuindo para a formação de um ambiente imunológicamente tolerante ao desenvolvimento fetal. / During pregnancy, embryo and maternal organisms interaction is mediated by different regulatory molecules produced in the uterine environment that are essential to the establishment of a tolerant environment to fetus development. The expression of a variety of inflammatory mediators influence the uterine and placenta environment. Cytokine are potent cellular mediators, regulating the immune responses by signaling and controlling pregnancy. Cytokine imbalance systemic or locally in the placental environment has been described in many pregnancy disorders, leading to fetal loss. Recent evidence showed that the expression of a novel cytokine, Interleukin 25 is decreased in patients with recurrent abortions, suggesting for IL-25 a immunoregulatory role in reproductive function. In this context, it is interesting to evaluate the presence of IL-25 in the maternal-fetal interface throughout pregnancy. This study aimed to characterize the IL-25 expression and its receptor IL-17BR in placental cells and maternal blood during pregnancy in mice. The analysis of gene expression of IL-25 and its receptor IL-17BR by qRT-PCR was performed on implantation sites and fetal and maternal portions of placentas in the days 10.5, 13.5, 16.5 and 19, 5 of gestation (dg). Peripheral blood mononuclear cell (PBMCs) and placental dissected tissues were used for flow cytometry analyses on the same days of gestation. The immunolocalization for cytokine and receptor was evaluated in histological sections for different gestational days. These data shows that IL-25 gene expression is intensified after the implantation period, increasing mRNA levels gradually from 10,5dg to 16,5dg. Cytokine and receptor are expressed differently by placental compartments. Analysis of protein expression by flow cytometry confirm the gene expression data and reactivity observed in the immunohistochemistry reactions. IL-25 and IL-17BR are expressed throughout the maternal-fetal interface, and the cytokine prevalent in the fetal placenta, while the receptor is major expressed in the maternal portion. Among the cell populations could contribute to the cytokine levels in fetal placenta is observed the presence of trophoblast cells phenotypically characterized producing IL-25 and leukocytes (CD45 +) expressing the IL-17BR in the maternal decidua. Among the period studied, IL-25 and IL-17BR expression mainly occurred on days 13.5 and 16.5 dg. The functional significance of these findings needs further investigation, however, due to immunoregulatory character attributed to IL-25 cytokine, it is seems to be associated with the control of placental functions contributing to the formation of an immunologically tolerant environment for fetal development.

O presente estudo verificou a prevalência de anemia em gestantes atendidas em 13 Unidades Básicas de Saúde da Supervisão Técnica de Saúde do Butantã-SP. Trata-se de estudo transversal com dados secundários de prontuários de 772 gestantes, à primeira consulta do pré-natal, nos anos de 2006 (387) e de 2008 (385). Foram incluídas no estudo as gestantes cujos prontuários apresentavam os seguintes dados: idade, peso, trimestre gestacional, concentração de hemoglobina e RDW na primeira consulta de pré-natal e ausência de doenças crônicas. De acordo com o estado nutricional pré-gestacional, observaram-se maiores prevalências de eutrofia (61 %), seguidas por 21,5 % de sobrepeso e por 10,8 % de obesidade. A prevalência de anemia foi de 10,0 % em 2006 e de 8,8 % em 2008, sem diferença estatisticamente significativa entre os valores (p > 0,05). Como esperado, a prevalência aumentou com a evolução da gestação. Em 2006 as médias de Hb diminuíram de 12,6 (1,0) g/dL no I trimestre para 12,2 (1,1) g/dL no II trimestre e 11,5 (1,0) g/dL no III trimestre gestacional. Os valores foram similares no ano de 2008. A distribuição das médias de Hb e RDW nos dois anos não mostrou diferenças estatísticas significativas (p > 0,05). As gestantes que estavam no II trimestre apresentaram risco de anisocitose maior em 41%, quando comparadas às que fizeram sua primeira consulta no primeiro trimestre. Esse nível de prevalência de anemia é classificado como associado a um risco populacional leve, segundo a Organização Mundial de Saúde. / The present study verified the prevalence of anemia among pregnant women assisted in 13 Health Basic Units of the Health Technical Supervision in Butantã-SP. This is a cross-sectional study with secondary data from medical charts of 772 pregnant women, upon the first prenatal visit to the doctor, in 2006 (387 women) and 2008 (385 women). The study included pregnant women whose medical charts contained the following data: age, weight, gestational quarter, hemoglobin concentration and RDW upon the first prenatal visit and lack of chronic diseases. According to the gestational nutritional status, the largest prevalence of eutrophic women (61 %) was observed, followed by 21.5 % overweight and 10.8 % obese. The prevalence of anemia was 10.0 % in 2006 and 8.8 % in 2008, with no statistically significant difference between the values (p > 0.05). As expected, the prevalence increased with the evolution of gestation. In 2006, the mean Hb values decreased from 12.6 (1.0) g/dL in the first quarter to 12.2 (1.1) g/dL in the second quarter and 11.5 (1.0) g/dL in the third quarter of pregnancy. The values were similar in 2008. The mean Hb distribution and RDW in both years showed no statistically significant differences (p > 0.05). The pregnant women who were in the second quarter of pregnancy showed a 41 % larger risk of anisocytosis when compared to the ones who attended the first medical visit in the first quarter of pregnancy. This level of prevalence for anemia is classified as associated to a slight risk for the population, according to the World Health Organization.

A ractopamina (Rac) tem sido utilizada durante a gestação de fêmeas suínas gestantes, com objetivo de melhorar o aporte nutricional ao feto, sendo considerada como repartidor de nutrientes, promovendo a lipólise e a retenção de nitrogênio, e a síntese proteica. O aumento no fluxo de nutrientes pode apresentar impactos positivos na condição corporal da fêmea gestante e o desenvolvimento do feto. O presente trabalho avaliou os efeitos da inclusão de 10 e 20 mg/kg de ractopamina na ração de fêmeas suínas gestantes, entre os dias 25 a 50 da gestação, sobre o desempenho produtivo, reprodutivo, parâmetros fisiológicos, hemograma e bioquímica sanguínea das fêmeas e sobre a vitalidade, hemograma, bioquímica sanguínea, número de fibras musculares no músculo semitendinosus e estado metabólico dos leitões. Foram utilizadas 41 fêmeas de linhagem comercial híbrida, DB 90 (DB-Genética Suína, Patos de Minas, MG), divididas de acordo com a ordem de parto em 3 tratamentos: Controle: sem ractopamina; Rac10: 10 mg/kg de ractopamina; Rac20: 20 mg/kg de ractopamina. O tratamento foi administrado diariamente via top dressed durante os 25 a 50 dias de gestação. As avaliações e as coletas de amostras foram realizadas nos 25, 37, 50, 107 dias de gestação e nos 1, 7, 14 e 21 dias da lactação. Para a avaliação da vitalidade foi utilizado o teste de pontuação Apgar modificado por Motta-Rojas (2005). A suplementação de ractopamina durante a gestação não influenciou o desempenho reprodutivo das fêmeas, dos leitões até o desmame e no número de fibras musculares no músculo semitendinosus ao nascimento. Houve um aumento na frequência cardíaca e de leitões com vitalidade baixa nas fêmeas suplementadas com Rac. Houve alterações no hemograma, bioquímica sanguínea das fêmeas, mas não nos leitões. Os resultados obtidos a partir das análises do hemograma e bioquímica sanguínea indicam ação da ractopamina sobre o metabolismo lipídico e proteico das fêmeas sem reflexos importantes nos valores observados nos leitões. O presente trabalho concluiu que a suplementação de Rac durante os 25 a 50 dias da gestação não influencia negativamente no metabolismo das fêmeas e sua progênie até o desmame. Por outro lado, a suplementação de Rac aumenta a frequência cardíaca sem aumento de frequência respiratória e temperatura retal das fêmeas, no entanto, pode ser considerado ainda dentro dos parâmetros fisiológicos normais durante a gestação. O aumento no número de leitões de vitalidade baixa 10 pode estar relacionado a um aumento numérico do peso ao nascimento dos leitões. Contudo, a suplementação de Rac durante os 25 50 dias da gestação não influencia negativamente no desempenho produtivo das fêmeas e sua progênie até o desmame. / The ractopamine (Rac) has been used in pregnant sows, aiming to improve the nutritional contribution to the fetus, being considered as nutrient delivery, promoting lipolysis and retention of nitrogen and protein synthesis. Increased nutrient flow may have positive impacts on the body condition of sows and fetal development. The present study evaluated the effects of the inclusion of 10 and 20 mg / kg of ractopamine in the diet sows from 25 to 50 days of pregnancy. Assessing possible effects on productive, reproductive performance, physiological parameters, complete blood count and blood serum biochemistry of females and on vitality, complete blood count, blood serum biochemistry, number of muscle fibers in the semitendinosus muscle and metabolic status of their piglets. Forty one sows of hybrid commercial line, DB 90 (DB-Genetics Swine, Patos de Minas, Minas Gerais),were used and divided according to the order of parity in 3 treatments: Control: No ractopamine; Rac10: 10 mg / kg ractopamine; Rac20: 20 mg / kg ractopamine. The treatment was administered daily via top dressed during the 25 to 50 days of gestation. The evaluations and sample collections were performed at 25, 37, 50, 107 days of gestation and at 1, 7, 14 and 21 days of lactation. Assessment of vitality was used the apgar score test modified by Motta-Rojas (2005). The supplementation of ractopamine during gestation did not influence the female reproductive performance, piglets performance until weaning and muscle fibers in the semitendinosus muscle at birth. There was an increase in heart rate and low vitality in females supplemented with Rac. There were alterations in the complete blood count, blood serum biochemistry of the females, but not in the piglets. The results obtained from the hemogram and blood biochemistry analysis indicate the action of ractopamine on the lipid and protein metabolism of the females without significant reflexes in the piglets values. The present study concluded that the supplementation of Rac during the 25 to 50 days of gestation does not negatively influence the metabolism of the females and their progeny until weaning. On the other hand, supplementation of Rac increases the heart rate without increasing respiratory rate and rectal temperature of the females, however, this can still be considered within the normal physiological parameters during gestation. The increase in the numbers of low-vitality piglets may be related to a numerical increase in the birth weight of the piglets. However, Rac 12 supplementation during the 25-50 days of gestation does not negatively influence the productive performance of the females and their progeny until weaning.