A baixa ingestão de colostro compromete a sobrevivência e o desempenho dos leitões. O objetivo do estudo foi avaliar a produção de colostro de matrizes submetidas à indução ao parto aos 114 dias de idade gestacional e sua influência na taxa de sobrevivência na maternidade e desempenho dos leitões. A produção de colostro foi estimada em 96 matrizes de ordem de parto 3 a 7, divididas em dois grupos: Controle (n=48) composto por fêmeas com parto espontâneo e Induzido (n=48) composto por fêmeas induzidas ao parto aos 114 dias de gestação com análogo da PGF2α. Todos os leitões foram pesados ao nascer e 24 horas depois para estimar a produção de colostro através do ganho de peso individual. Amostras de sangue foram coletadas dos leitões no primeiro dia após o nascimento. Foram coletadas amostras de colostro e sangue de cada fêmea no momento do parto e 24 horas depois. Para avaliar o desempenho dos leitões durante a lactação foram acompanhadas 28 leitegadas de cada grupo contendo 12 leitões cada durante 20 dias após o nascimento. Durante o período lactacional todas as leitegadas foram pesadas nos dia 07, 14 e 20 após o nascimento. A duração da gestação do grupo Controle foi em torno de 12 horas mais longa do que no grupo Induzido (P=0,06). A indução ao parto não afetou (P>0,10) a duração do parto, o número total de leitões nascidos, o número de leitões nascidos vivos, o percentual de natimortos, o peso médio dos leitões, o peso médio da leitegada ao nascimento e a produção de colostro em relação ao grupo controle. Não houve diferença entre os grupos (P>0,05) no percentual de fêmeas com intervenção obstétrica. As concentrações de IgG no soro das fêmeas, dos leitões e do colostro foram similares (P>0,10) entre os dois grupos. Para acompanhar o desempenho dos leitões durante a lactação foram uniformizadas leitegadas com peso inicial e consumo de colostro semelhante (P>0,10) em fêmeas adotivas com semelhante produção de colostro (P>0,10). Não foram observadas diferenças (P>0,10) no peso médio dos leitões e das leitegadas nos dias 07, 14 e 20, bem como não houve diferença (P>0,10) na sobrevivência dos animais no mesmo período. A indução ao parto aos 114 dias de gestação não prejudica a produção de colostro e não altera a quantidade de imunoglobulinas G tanto no colostro quanto no soro dos leitões. O desempenho na fase de lactação de leitegadas de fêmeas induzidas ao parto foi semelhante ao de leitegadas de fêmeas com parto espontâneo. / Low colostrum intake influences piglet survival and performance. The aim of this study was to evaluate colostrum production by sows submitted to farrowing induction and its influence on pre-weaning survival rate and piglet performance. A total of 96 sows of parities three to seven were assigned into two groups: Control (n = 48) composed of sows with spontaneous labor and Induced (n = 48) composed of sows induced on day 114 with PGF2α analogue. Colostrum production was estimated by piglet individual weight gain in the first day of life, which was measured by weighing piglets at birth and 24 hours later. Blood samples were collected from piglets on the first day after birth. Colostrum and blood samples were collected from each sow at farrowing and 24 hours later. To evaluate piglet performance during lactation 28 litters from each group containing 12 piglets each were followed for 20 days after birth. During lactation period, all piglets were weighed on days 07, 14 and 20 after birth. Gestation length of control group was about 12 hours longer than induced group (P = 0.06). Farrowing duration, total born, born alive, stillborn rate, average piglet weight, average litter weight at birth and colostrum production were not significantly affected (P > 0.10) by induction of farrowing compared to control group. There was no difference (P > 0.05) in the percentage of sows with obstetric intervention between groups. IgG concentration in sow and piglet serum and colostrum were similar (P > 0.10) between groups. To monitor piglet performance during lactation litters with similar initial weight and colostrum intake (P > 0.10) were cross-fostered in sows with similar colostrum production (P > 0.10). No differences (P > 0.10) were observed in the average piglet and litter weight on days 07, 14 and 20, and on survival rate in the same period. Induction of labor at 114 days of gestation doesn’t affect colostrum production nor influence colostrum and piglet serum immunoglobulin G level. Litter performance during lactation was similar in sows with induced parturition and spontaneous labor.

A baixa ingestão de colostro compromete a sobrevivência e o desempenho dos leitões. O objetivo do estudo foi avaliar a produção de colostro de matrizes submetidas à indução ao parto aos 114 dias de idade gestacional e sua influência na taxa de sobrevivência na maternidade e desempenho dos leitões. A produção de colostro foi estimada em 96 matrizes de ordem de parto 3 a 7, divididas em dois grupos: Controle (n=48) composto por fêmeas com parto espontâneo e Induzido (n=48) composto por fêmeas induzidas ao parto aos 114 dias de gestação com análogo da PGF2α. Todos os leitões foram pesados ao nascer e 24 horas depois para estimar a produção de colostro através do ganho de peso individual. Amostras de sangue foram coletadas dos leitões no primeiro dia após o nascimento. Foram coletadas amostras de colostro e sangue de cada fêmea no momento do parto e 24 horas depois. Para avaliar o desempenho dos leitões durante a lactação foram acompanhadas 28 leitegadas de cada grupo contendo 12 leitões cada durante 20 dias após o nascimento. Durante o período lactacional todas as leitegadas foram pesadas nos dia 07, 14 e 20 após o nascimento. A duração da gestação do grupo Controle foi em torno de 12 horas mais longa do que no grupo Induzido (P=0,06). A indução ao parto não afetou (P>0,10) a duração do parto, o número total de leitões nascidos, o número de leitões nascidos vivos, o percentual de natimortos, o peso médio dos leitões, o peso médio da leitegada ao nascimento e a produção de colostro em relação ao grupo controle. Não houve diferença entre os grupos (P>0,05) no percentual de fêmeas com intervenção obstétrica. As concentrações de IgG no soro das fêmeas, dos leitões e do colostro foram similares (P>0,10) entre os dois grupos. Para acompanhar o desempenho dos leitões durante a lactação foram uniformizadas leitegadas com peso inicial e consumo de colostro semelhante (P>0,10) em fêmeas adotivas com semelhante produção de colostro (P>0,10). Não foram observadas diferenças (P>0,10) no peso médio dos leitões e das leitegadas nos dias 07, 14 e 20, bem como não houve diferença (P>0,10) na sobrevivência dos animais no mesmo período. A indução ao parto aos 114 dias de gestação não prejudica a produção de colostro e não altera a quantidade de imunoglobulinas G tanto no colostro quanto no soro dos leitões. O desempenho na fase de lactação de leitegadas de fêmeas induzidas ao parto foi semelhante ao de leitegadas de fêmeas com parto espontâneo. / Low colostrum intake influences piglet survival and performance. The aim of this study was to evaluate colostrum production by sows submitted to farrowing induction and its influence on pre-weaning survival rate and piglet performance. A total of 96 sows of parities three to seven were assigned into two groups: Control (n = 48) composed of sows with spontaneous labor and Induced (n = 48) composed of sows induced on day 114 with PGF2α analogue. Colostrum production was estimated by piglet individual weight gain in the first day of life, which was measured by weighing piglets at birth and 24 hours later. Blood samples were collected from piglets on the first day after birth. Colostrum and blood samples were collected from each sow at farrowing and 24 hours later. To evaluate piglet performance during lactation 28 litters from each group containing 12 piglets each were followed for 20 days after birth. During lactation period, all piglets were weighed on days 07, 14 and 20 after birth. Gestation length of control group was about 12 hours longer than induced group (P = 0.06). Farrowing duration, total born, born alive, stillborn rate, average piglet weight, average litter weight at birth and colostrum production were not significantly affected (P > 0.10) by induction of farrowing compared to control group. There was no difference (P > 0.05) in the percentage of sows with obstetric intervention between groups. IgG concentration in sow and piglet serum and colostrum were similar (P > 0.10) between groups. To monitor piglet performance during lactation litters with similar initial weight and colostrum intake (P > 0.10) were cross-fostered in sows with similar colostrum production (P > 0.10). No differences (P > 0.10) were observed in the average piglet and litter weight on days 07, 14 and 20, and on survival rate in the same period. Induction of labor at 114 days of gestation doesn’t affect colostrum production nor influence colostrum and piglet serum immunoglobulin G level. Litter performance during lactation was similar in sows with induced parturition and spontaneous labor.

No / Human saphenous vein endothelial cells (HSVECs) derived from primary cultures of adult human veins constitute an excellent in vitro model for studying human endothelial metabolism. In this study we report the14C-labelled prostanoid profile of HSVECs under resting and stimulated conditions and the effect of the n-3 polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid on them. Results indicate that HSVECs while under resting conditions produce mainly prostaglandin F2 ¿(PGF2 ¿). After stimulation with calcium ionophore A23187, the cells were found to synthesise PGI2, PGE2and PGF2¿as major products and thromboxane B2and PGD2as minor products. Production of14C-labelled hydroxyeicosatetraenoic acids was not detected. Eicosapentaenoic acid was found to inhibit basal and stimulated prostanoid production whereas docosahexaenoic acid inhibited basal but strongly increased stimulated prostanoid production. These results may offer the basis for further studies aiming to investigate targets for pharmacological intervention in inflammatory conditions.

<p>In the present thesis dietary markers for intake of milk fat, associations between intake of milk fat and risk factors for coronary heart disease (CHD), and the effects of supplementation with conjugated linoleic acid (CLA) to healthy humans are investigated.</p><p>The dietary fat quality is one of the main lifestyle factors affecting risk for CHD. When studying the associations between diet and health it is important to have accurate dietary information. Objective dietary markers increase the possibilities to interpret dietary surveys.</p><p>In a study of 62 men we demonstrated that the milk fatty acid pentadecanoic acid (15:0) measured in serum lipids can be used as marker for intake of fat from milk products. In the same study we observed inverse correlations between intake of milk fat and certain risk factors for CHD, especially anthropometric variables.</p><p>To further investigate these findings we supplemented humans with CLA, naturally present in milk. CLA has in animals and <i>in vitro</i> been ascribed positive effects on adiposity and glucose and lipid metabolism. When supplementing humans with CLA we observed a slight decrease in body fat, but no effects on other anthropometric variables or serum lipids. However, markers of lipid peroxidation and inflammation increased. From a second supplementation study we concluded that CLA <i>trans </i>10, <i>cis </i>12 induced lipid peroxidation more than did a mixture of isomers.</p><p>We conclude that the inverse associations between milk fat intake and CHD risk factors, and the effects of CLA, are interesting and need further investigation. However, according to current knowledge, the general population is still advised to have a limited intake of total and saturated fat and to instead choose unsaturated fats. In addition, there is to date no medical reasons for humans to take CLA as supplements.</p>

In the present thesis dietary markers for intake of milk fat, associations between intake of milk fat and risk factors for coronary heart disease (CHD), and the effects of supplementation with conjugated linoleic acid (CLA) to healthy humans are investigated. The dietary fat quality is one of the main lifestyle factors affecting risk for CHD. When studying the associations between diet and health it is important to have accurate dietary information. Objective dietary markers increase the possibilities to interpret dietary surveys. In a study of 62 men we demonstrated that the milk fatty acid pentadecanoic acid (15:0) measured in serum lipids can be used as marker for intake of fat from milk products. In the same study we observed inverse correlations between intake of milk fat and certain risk factors for CHD, especially anthropometric variables. To further investigate these findings we supplemented humans with CLA, naturally present in milk. CLA has in animals and in vitro been ascribed positive effects on adiposity and glucose and lipid metabolism. When supplementing humans with CLA we observed a slight decrease in body fat, but no effects on other anthropometric variables or serum lipids. However, markers of lipid peroxidation and inflammation increased. From a second supplementation study we concluded that CLA trans 10, cis 12 induced lipid peroxidation more than did a mixture of isomers. We conclude that the inverse associations between milk fat intake and CHD risk factors, and the effects of CLA, are interesting and need further investigation. However, according to current knowledge, the general population is still advised to have a limited intake of total and saturated fat and to instead choose unsaturated fats. In addition, there is to date no medical reasons for humans to take CLA as supplements.

Corpus luteum is a transient endocrine structure formed from the ruptured ovarian follicle. Its main function is to secrete P4, a pro-gestational hormone, essential for establishment and maintenance of pregnancy in mammals. The modulators of CL structure and function are classified as trophic and lytic factors. The luteotrophic factors include pituitary hormones, growth factors, intra luteal factors and cytokines, while luteolytic factors include PGF2α and oxytocin. The interplay between luteotrophic and luteolytic factors regulates luteal steroidogenesis. The precise timing of expression of various enzymes/proteins required for synthesis and metabolism of P4 constitutes an important process in the overall regulation of CL function. The three hormones LH/CG, E2 and PRL are regarded as luteotrophic factors crucial for control of CL function in mammals. Depending on species, either individually or all three hormones in the form of luteotrophic complex have been shown to participate in the regulation of CL function. In addition to the well-established endocrine role of E2, its secretion is the hallmark of the ovulating follicle, has an important role in the intraovarian growth, differentiation and survival of cells. Chapter I provides a comprehensive review of literature on CL structure and function with emphasis on factors that influence its growth, development, function and demise in bovines and rodents. In Chapter II, studies have been carried out to examine 20α-HSD expression and its activity in the CL of buffalo cow. During induced and spontaneous luteolysis, rapid fall in circulating P4 is one of the early signs of initiation of luteolytic process in several species. In rodents, it is well recognized that during luteolysis, P4 is catabolized into inactive metabolite, 20α-OHP by the reaction of 20α-HSD enzyme during luteolysis. Experiments were carried out to determine 20α-HSD expression and activity throughout the luteal phase and during induced luteolysis in the buffalo cow. Circulating P4 concentration declined rapidly in response to PGF2α treatment, but HPLC analysis of serum samples did not reveal changes in circulating 20α-OHP levels in buffalo cows. In contrast, pseudo pregnant rats receiving PGF2α treatment showed higher 20α-OHP levels at 24 h post treatment. qPCR expression of 20α-HSD in CL during different stages of luteal phase and PGF2α-treated buffalo cows was carried out and higher expression of 20α-HSD was observed at 3 and 18 h post treatment, but its activity was not altered post PGF2α treatment at other time points examined. The expression of the transcription factor Nurr77 which is involved in increased expression of 20α-HSD increased several fold 3 h post PGF2α treatment similar to the observation in PGF2α-treated pseudo pregnant rats. The results suggested that the synthesis rather than catabolism of P4 appears to be primarily affected by PGF2α treatment in buffalo cows in contrast to increased metabolism of P4 as seen in rodents. In bovines, to date no luteotropic actions for E2 has been demonstrated and whether E2 has direct effect on CL function has also not been reported. Expression of CYP19A1 gene that encodes aromatase enzyme although gets down regulated post ovulation but its expression recovers in the CL and also E2 biosynthesis has been reported in the bovine CL. Recently it was observed that CYP19A1 expression was consistently down regulated following administration of luteolytic dose of PGF2α. Experiments were conducted to examine the expression of ERα and ERβ in the CL throughout the buffalo estrous cycle as well as examined the luteal E2 levels post PGF2α treatment. The results indicated that ER expression was detectable during different stages of CL and that circulating and luteal E2 levels declined post PGF2α treatment. It was hypothesized that decrease in luteal E2 levels leads to down regulation of ER signaling and changes in expression of E2 responsive genes in the CL. To test the hypothesis, 89 genes which were regarded as E2 responsive genes were selected and the previously published global gene expression data of the buffalo CL was mined for E2 responsive genes. It was observed that 57 of 89 genes regarded as E2 responsive genes were found to be differentially expressed. Since non pregnant buffalo CL is not regarded as major site of E2 production, to validate the authenticity of differentially E2 expressed genes post PGF2α, CL of another species, the macaque, which is known to secrete abundant E2 was included for the analysis. Incidentally, the global gene expression data for the PGF2 α treated macaques (in which CYP19A1 gene expression also gets down regulated) has previously been reported from the laboratory. Here again, it was observed that nearly 79 of 89 genes were identified to be differentially expressed. To further determine the consequences of decreased ER signaling, molecules associated with survival and apoptosis were examined. The results indicated decreased expression (both mRNA and protein levels) of Akt, Bax and Bcl-2 genes. The results suggested an important role for E2 on CL function in the buffalo cow. In Chapter III, several experiments were conducted in another model system, pregnant rat, in which aromatase expression and therefore E2 production is high in the CL. Experiments were conducted to examine the effects of E2 inhibition and E2 replacement on the expression of genes. For this purpose, pregnant rats were treated with a specific aromatase inhibitor on day 12-15 of pregnancy. Together with AI, exogenous E2 was administered to another group of pregnant rats. The CL collected from different groups of rats on day 16 of pregnancy was subjected to microarray analysis. The analysis post validation of microarray data has shown that clusters of genes could be segregated into various pathways involving luteal steroidogenesis, immune system, various growth factors and apoptotic processes, all directed towards the regulation of CL function. The involvement of E2 in luteal cell proliferation and lipid deposition well corroborated with protein levels for cyclin D1 and ki67 and the results of oil red O staining, respectively. There have been reports implicating PI3K/Akt signaling in cyclin D1 accumulation, but mechanism of action does not appear to involve transcriptional activation of cyclin D1. The results of the present study indicate a decrease in cyclin D1 protein levels due to inhibition of PI3K/Akt signaling by AI treatment which is prevented upon administration of E2 during AI treatment. The findings provide a comprehensive overview for the mechanisms associated with the cell survival, progression, etc. The bioinformatics approach provided complete landscape of functional changes affected by the upstream regulators of genes associated with survival and apoptosis. Also, the findings further strengthen the hypothesis of involvement of E2 in the regulation of CL function by way of activation of Akt, the primary mediator of PI3K signaling in the regulation of cellular component that affect cell survival. In the present study, IGFBP5 which was up regulated during luteal inhibition of E2 with AI treatment was selected for further studies. Although IGFBP5 is known to be associated with follicular atresia in the rat ovary, there is limited data for the involvement of IGFBP5 in either a growth stimulatory or inhibitory action on ovarian cells. Based on present findings, a causal link between reduced ERα transcriptional activities resulting in inhibition of Akt/PKB in the presence of IGFBP5 expression could be proposed. Further, the cellular hypertrophy mediated by E2 has been speculated due to increased proliferation of vascular endothelial cells, blood supply and thus nutrients. E2, together with PRL and placental lactogens, regulates steroidogenesis and cell hypertrophy in the rat CL of pregnancy. In CL, the prominent IGFBP5 mRNA expression in different types of luteal cells has not been reported. The mRNA expression for IGFBP5 across the two types of luteal cells showed higher expression in SLC. Hence, in the present study, it has been speculated that prevention of conversion of SLC to LLC due to lack of E2 biosynthesis in presence of AI might be acting as a source for the increased IGFBP5 levels during mid pregnancy in rat CL and brings about changes associated with lack of E2. Various receptor studies on rat CL have demonstrated the lack of progesterone receptor (PR) mRNA expression in the rat CL negating its involvement as an autocrine/paracrine regulator of CL function through an intracellular receptor, but the involvement of non-PR involvement in mediating such mechanism further strengthens the role of ERs. The luteotrophic complex formation in pregnant rat principally by PRL and E2 has been discussed at length in Chapter III. PRL appears to maintain luteal ER content in the CL during rat pregnancy which further determines the luteotrophic and luteolytic actions of E2. Further, study on expression of E2 responsive genes would help in identifying E2 regulating molecules to get a clear picture on the role of E2 in understanding regulation of the CL function. The interaction of E2 with growth factor signaling including the IGF pathway has been well established in different species and this interaction is tightly linked to ERα expression, an observation interpreted as physiological coupling of growth factor and stress signaling pathways. Attempts were made towards understanding cross talk between the E2 signaling and the IGF1 signaling in few experiments carried out in Chapter IV. Based on the results, it can be proposed that a causal link exists between reduced ERα transcriptional activity and inhibition of Akt/PKB in the presence of IGFBP5. The present study has shown the activity of IGF on ERα activity mediated partly via PI3K/Akt pathway. Hence, the finding further speculates that inhibitory effect of IGFBP5 on E2 induced ERα function was due to sequestration of IGF1, possibly present in serum or produced within the cells. Another striking observation was the down regulation of glucocorticoid receptor (GR) gene, NR3C1, in the data of earlier studies [Priyanka, 2009, GEO accession number GSE8371 and Kunal, 2014, GEO accession number GSE27961] and the present study has been compared and discussed in this thesis. Glucocorticoids provide key signals for differentiation of fetal and placental tissues. Therefore, regulation of glucocorticoid access to the placenta and fetus is recognized as an important determinant of prognosis outcome and subsequent development of the postnatal phenotype. Differential regulation of these genes in CL post E2 deprivation and replacement further emphasize the regulation of CL via various biological, cellular and molecular functions. Interestingly, besides transcriptional regulation of IGF axis components, E2 activated ERα also rapidly influence the activity of IGF axis related to signaling proteins in a non-genomic manner, especially by the PI3K/Akt pathway. PI3K/Akt pathway analysis has been carried out in E2 inhibition and replacement experiments. To further confirm the observations of E2 and growth factor interaction, experiments have been set up with exogenous GH for increasing circulating levels of IGF in the system. The findings suggest that the non-genomic signaling pathway activated by the phosphorylation of ERα induced by E2 gets inhibited in the presence of AI result in increased expression of IGFBP5. The reduction in circulating IGF1 in pregnant rats may be associated with the effect on IGFBP, important for determining biological action of IGF1. The changes observed in the present study emphasize the exclusive effects of the IGFBP5 on the CL function brought about perturbations in luteal E2 content. The experiments described in the present thesis aim at understanding the mechanism responsible for decreased serum and luteal P4 post PGF2α treatment in buffalo cows, i.e. whether PGF2α acts on biosynthetic or catabolic process of P4. In the present study, experiments were designed to elucidate the role of E2 in regulation of CL function, since down regulation of CYP19A1 gene mRNA was one of the early events observed in buffalo cows post PGF2α treatment. This line of research work was extended to rodents, a species that secretes high levels of E2 during pregnancy. Genome wide transcriptional changes data revealed differential expression of several E2 responsive genes following E2 inhibition and replacement treatments. The results revealed importance of ER-mediated PI3K/Akt signaling essential for regulation of many transcriptional regulatory molecules in the CL and an interesting involvement of IGFBP5 as a link between E2 and IGF signaling. These findings further provide an insight into the role of IGFBP5 in E2-mediated actions in rat CL during pregnancy. In conclusion, the present findings suggest inhibitory effect of IGFBP5 on E2-induced ERα function and hence, its selection as a target molecule for regulation of CL function and for many beneficial processes involved in anti-carcinogenic properties can be thought of.