No description available.

This study was conducted to test the hypothesis that dietary supplementation with L-arginine during early pregnancy will ameliorate embryonic loss in pigs. Gilts were bred at the second estrus, and housed individually in pens and fed twice daily 1 kg of a corn- and soybean meal-based diet supplemented with 0.0%, 0.4%, or 0.8% L-arginine (w/w) between d 0 and 25 of gestation (Experiment 1) or between d 14 and 25 of gestation (Experiments 2 and 3). At d 25 (Experiment 1 and 2) or d 60 (Experiment 3) of gestation, gilts were hysterectomized to obtain uteri and conceptuses. Total RNA and protein were extracted from the frozen tissues. Quantitative RT-PCR, western blotting, and microarray analyses were performed to determine the changes of gene expression at mRNA and protein levels. Dietary supplementation with 0.8% L-arginine between d 0 and 25 of gestation decreased uterine weight, total number of fetuses, number of corpora lutea (CL), total fetal weight, total volume of allantoic and amniotic fluids, concentrations of progesterone in maternal plasma and allantoic fluid, compared to the control group. However, dietary supplementation with 0.4% or 0.8% L-arginine between d 14 and 25 of gestation increased total volume of amniotic fluid, total amounts of arginine in allantoic and amniotic fluids, total amounts of fructose and most amino acids in amniotic fluid, placental growth, and the number of viable fetuses per litter by 2. Dietary supplementation with 0.4% or 0.8% L-arginine between d 14 and 25 of gestation increased the total number of fetuses and number of live fetuses, rate of embryonic survival, and volumes of allantoic and amniotic fluids in gilts with 15 to 18 CL on d 60 of gestation compared with the control group. The abundance of placental protein and expression of mRNA related to the genes for arginine transport and metabolism, including cationic amino acid transporter 1, endothelial nitric oxide synthase (NOS3), phosphorylated-NOS3, ornithine decarboxylase, and guanosine triphosphate cyclohydrolase-I was increased by dietary supplementation with 0.8% L-arginine between d 0 and 25 of gestation. The abundance of total and phosphorylated mechanistic target of rapamycin was also enhanced by dietary 0.8% L-arginine supplementation between d 0 and 25 of gestation. Microarray analysis revealed that supplementation with 0.8% arginine between d 14 and 25 of gestation affected placental expression of 575 genes. Findings from the current study not only advance basic knowledge of mammalian reproductive biology, but also have important implications for developing practical means to enhance fertility in female pigs.

El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / Purpose We sought to evaluate the extent to which childhood physical and/or sexual abuse history is associated with post-traumatic stress disorder (PTSD) during early pregnancy and to explore the extent to which the childhood abuse-PTSD association is mediated through, or modified by, adult experiences of intimate partner violence (IPV). Methods In-person interviews collected information regarding history of childhood abuse and IPV from 2,928 women aged 18-49 years old prior to 16 weeks of gestation. PTSD was assessed using the PTSD Checklist-Civilian Version. Multivariate logistic regressions were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). Results Compared to women with no childhood abuse, the odds of PTSD were increased 4.31-fold for those who reported physical abuse only (95% CI, 2.18–8.49), 5.33-fold for sexual abuse only (95% CI, 2.38–11.98), and 8.03-fold for those who reported physical and sexual abuse (95% CI, 4.10–15.74). Mediation analysis showed 13% of the childhood abuse-PTSD association was mediated by IPV. Furthermore, high odds of PTSD were noted among women with histories of childhood abuse and IPV compared with women who were not exposed to either (OR = 20.20; 95% CI, 8.18–49.85). Conclusions Childhood abuse is associated with increased odds of PTSD during early pregnancy. The odds of PTSD were particularly elevated among women with a history of childhood abuse and IPV. Efforts should be made to prevent childhood abuse and mitigate its effects on women's mental health. / Revisión por pares

Background: : Few investigators have evaluated the association between early pregnancy social support and depression; however, increased social support may improve mental health during pregnancy. Our objective is to examine whether in early pregnancy there is an association between social support and maternal depression among women in Peru. Methods: : 2,062 pregnant women participated in structured interviews. Early pregnancy social support was measured using the Social Support Questionnaire (SSQ-6). We evaluated the number of individuals that participants could turn to in different situations (Social Support Number Score; SSQN) and their satisfaction with support received (Social Support Satisfaction Score; SSQS). Median SSQN and SSQS characterized participants according to high and low levels of support. SSQN family vs. non-family support were also evaluated separately. Antepartum depression was assessed using the Patient Health Questionnaire-9. Multivariable logistic regression procedures were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). Results: : 39.6% of women reported high SSQN and 45.5% reported high SSQS. Approximately 25% had antepartum depression. Women with high SSQN had 22% lower odds of antepartum depression (OR = 0.78; 95%CI: 0.63–0.97). Similarly, women with high SSQS scores had 45% lower odds of antepartum depression (OR = 0.55; 95%CI: 0.45–0.68). Women with high SSQN non-family scores had 30% lower odds antepartum depression compared to those with low SSQN non-family scores (OR = 0.70; 95%CI: 0.57–0.86). The association between SSQN family scores and antepartum depression did not reach statistical significance. Conclusion: : Increased social support may improve maternal mental health during pregnancy and this association should be assessed in longitudinal studies. / This research was supported by awards from the National Institutes of Health ( NIH ), National Institute of Minority Health and Health Disparities ( T37-MD-001449 ) and Eunice Kennedy Shriver National Institute of Child Health and Human Development ( R01-HD-059835 ). The NIH had no further role in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication. / Revisión por pares

Maternal contributions from the uterine endometrial luminal (LE) and glandular (GE) epithelia are unequivocally required to support ruminant conceptus growth and development, elongation and implantation. Therefore, studies were conducted to examine expression of endometrial genes hypothesized to regulate conceptus development. The first study investigated two genes specifically expressed in the LE and superficial GE of the ovine uterus. Insulin-like growth factor binding protein (IGFBP1) and (IGFBP3) expression was coordinate with ovine conceptus elongation. Treatment with P4 induced and IFNT stimulated IGFBP1, but not IGFBP3; however, IFNT only moderately stimulated IGFBP1, indicating that another conceptus-derived factor stimulates endometrial IGFBP1 expression. IGFBP1 did not affect proliferation of ovine trophectoderm (oTr) cells in vitro, but stimulated their migration and attachment. Results indicated that IGFBP1, but not IGFBP3 is a marker of conceptus elongation in ruminants and stimulates cell migration and attachment. The second study evaluated the effects of pregnancy, P4 and IFNT on expression of hydroxysteroid (11-beta) dehydrogenases (HSD11B1 and HSD11B2), nuclear receptor subfamily 3, group C, member 1 (NR3C1), and prostaglandin-endoperoxide synthase 2 (PTGS2) in the ovine uterus. Expression of HSD11B1 mRNA and PTGS2 protein in endometrial LE and sGE were coordinate with conceptus elongation, while HSD11B2 mRNA was expressed primarily in the conceptus. Further, P4 induced, but IFNT only moderately stimulated HSD11B1. Thus, HSD11B1 expression may be regulated by prostaglandins (PGs) during early pregnancy. The presence of NR3C1 in the ovine uterus implicates cortisol, the main product of HSD11B1, in peri-implantation period events that include elongation of the ovine conceptus. The third study determined in vivo effects of PGs on ovine conceptus elongation and endometrial gene expression. Compared to control ewes, intrauterine infusions of a PTGS2 inhibitor, meloxicam, retarded elongation and decreased expression of elongation-related genes including IGFBP1, IGFBP3, HSD11B1, galectin 15 (LGALS15), solute carrier family 2, member 1 (SLC2A1), gastrin-releasing peptide (GRP), cystatin C (CST3), radical S-adenosyl methionine domain containing 2 (RSAD2), and ISG15 ubiquitin-like modifer (ISG15). Collectively, these studies assessed the effects of pregnancy, P4, IFNT, and PGs on endometrial genes implicated in conceptus growth. These results indicate that IGFBP1 is a marker of conceptus elongation in ruminants and provide novel roles for both cortisol and PGs in endometrial gene expression and conceptus elongation.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23569号 / 医博第4783号 / 新制||医||1054(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 川口 義弥, 教授 篠原 隆司, 教授 近藤 玄 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Thesis (MMed)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: Background - Preeclampsia is a multi-system disease characterized by hypertension and proteinuria in pregnant women at greater than 20 weeks of gestational age. It remains one of the leading causes of maternal and foetal mortality and morbidity. While the cause of preeclampsia is essentially unknown, the important theories strongly implicate disturbed placental function in early pregnancy. Additionally, some researchers have investigated the possibility of a seasonal relationship with the incidence of preeclampsia. Differences in incidences of preeclampsia, examined exclusively on the basis of delivery timing, have also been noted to have seasonal variation, but results have been inconsistent. Objective - Our primary objective was to investigate the seasonal variation in preeclampsia in relation to the timing of conception and the timing of presentation with clinical disease over the period of one year. Methods - We performed a retrospective descriptive study of all women with preeclampsia who delivered at Tygerberg Hospital in 2010. Preeclampsia was diagnosed as hypertension associated with proteinuria after the 20th week of gestation. Names of patients were identified from labour ward records and data was collected and recorded on a data-sheet. Data were primarily analysed in relation to the season of delivery and also the season of the last menstruation. Summer was diagnosed as lasting from summer solstice to autumn equinox, autumn as lasting from autumn equinox until winter solstice, winter as lasting from winter solstice until spring equinox and spring as lasting from spring equinox until summer solstice. The data was analysed using the SPSS software (Statistical Package for Social Science). Discrete data was compared by calculating relative risks with 95% confidence limits, as well as the chi2 test. Fisher‘s exact test was used to compare ratios where the expected value in any cell of a two-by-two table is less than five. The means of normally distributed continuous data was compared by analysis of variance, while the medians of continuous data which are not distributed normally, where calculated using the non-parametric Mann Whitney u test. A p-value of < 0.05 was considered to be statistically significant, where applicable. Results - The peak incidence of preeclampsia was during winter with 32.2% of all cases occurring during this season. This was significantly higher than during the summer when only 169 (17.17%) cases of preeclampsia were delivered. When we analysed the data looking at the timing of menstruation (and therefore conception), we found that 292 (29.7%) patients that developed preeclampsia had their last menstrual period in the spring, with November the month of peak incidence. The lowest incidence was found in winter, with only 218 (22.2%) patients. Conclusion - We have confirmed a previous finding of a seasonal variation in the occurence of preeclampsia in Tygerberg Hospital. We have also confirmed that this seasonal variation is not only influenced by the timing of delivery, but also by the timing of conception. / AFRIKAANSE OPSOMMING: Agtergrond - Preeklampsie is ‘n multi-sisteem siekte wat gekenmerk word deur hipertensie en proteinurie. Dit word slegs gedurende swangerskap aangetref - gewoonlik na 20 weke. Dit is steeds een van die voorste oorsake van moederlike en fetale morbiditeit en mortaliteit. Terwyl die oorsaak van preeklampsie steeds onbekend is, dui die belangrikste teorië op versteurde plasentale ontwikkeling en funksionering vroeg in swangerskap. Sekere navorsers het ook die moontlikheid van ‘n seisonale patroon in die voorkoms van preeklampsie ondersoek. ‘n Seisonale patroon is wel identifiseer, maar dit is slegs gebasseer op die datum van verlossing en die resultate tussen studies wissel. Doel - Ons primêre doel was om die seisonale patroon in die insidensie van preeklampsie te ondersoek oor ‘n tydperk van een jaar en dan die datum van bevrugting te vergelyk met die datum van diagnose en verlossing. Metodiek - Ons het ‘n retrospektiewe beskrywende studie gedoen oor al die pasiënte met preeklampsie wat tydens 2010 by Tygerberg Hospitaal verlos is. Preeklampsie is gediagnoseer as hipertensie met geassosieerde proteinurie met aankoms na 20 weke. Die name van die pasiënte is verkry uit die kraamsaal-registers en data is versamel en op ‘n datastel aangebring. Data is primer geanaliseer in terme van die seisoen van verlossing en die seisoen waartydens die laaste maandstonde plaasgevind het. Die seisoene is as volg geklassifiseer : somer vanaf die summer solstice to autumn equinox, autumn as lasting from autumn equinox until winter solstice, winter as lasting from winter solstice until spring equinox and spring as lasting from spring equinox until summer solstice. Die data is geanaliseer met die SPSS sagteware (Statistical Package for Social Science). Diskrete data is vergelyk deur die relatiewe risiko’s te bereken met vertrouensintervalle van 95%, sowel as die chi2 toets. Fisher se eksakte toets is gebruik om ratios te vergelyk waar die verwagte waarde van enige sel in ‘n 2-by-2 tabel minder as 5 is. Die gemiddeldes van normaal-verspreide aaneenlopende data is vergelyk deur die analise van variance. Die mediane van aaneenlopende data wat nie normaal versprei was nie, is bereken met die non-parametriese Mann-Whitney-U-toets. ‘n P-warde van <0,05 is beskou as statisties betekenisvol, waar van toepassing. Resultate - Die piek –insidensie van preeklampsie was gedurende die wintermaande, met 32.2% van alle gevalle. Dit was betekenisvol hoër as die gedurende die somer, waar slegs 169 (17.17%) van gevalle verlos is. Toe ons die data analiseer na gelang van die datum van laaste menstruasie (en gevolglik bevrugting), het ons gevind dat 292 (29.7%) van die pasiënte wat preeklampsie ontwikkel het, het hul laaste maandstonde gedurende die lente ervaar. Die piek-insidensie was gedurende November. Daarteenoor is die laagste insidensie gevind in pasiënte met ‘n laaste menstruasie in die winter, met slegs 218 (22.2%) pasiënte. Gevolgtrekking - Ons het die vorige bevinding by Tygerberg Hospitaal van ‘n seisonale patroon in die ontwikkeling van preeklampsie bevestig. Ons het ook bevestig dat dit nie die datum van diagnose en verlossing is wat’n rol speel nie, maar wel die datum van laaste maandstonde en bevrugting.

Early pregnancy is characterized by complex interactions between blood vessels, leukocytes, and conceptus-derived trophoblasts within the gestational uterus. Uterine Natural Killer (uNK) cells become the most abundant leukocyte during decidualization and produce a wide array of angiogenic factors, yet little is known regarding their early pregnancy functions. To characterize the role(s) of uNK cells, whole mount in situ immunohistochemistry of live early implant sites was performed. A timecourse examination of murine early pregnancy (virgin, and gd4.5-9.5) implantation sites was performed. Comparison of Gd6.5, 8.5 and 9.5 implant sites from BALB/c+/+ controls (BALB/c) and BALB/c-Rag2-/-Il2rg-/- (alymphoid) identified anomalies that result from the absence of lymphocytes. In alymphoid decidua basalis, mesometrial angiogenesis was widespread but pruning of nascent vessels within alymphoid decidua basalis was deficient. As early gestation progressed, vessels of alymphoid decidua basalis showed no evidence for remodeling. Alymphoid implantation sites showed ~24h delay in uterine lumen closure and embryonic development. To determine if uNK cells would normalize the anomalies observed in alymphoid implantation sites, adoptive cell transfer of NK+ B- T- marrow to alymphoid mice was performed. All of the above anomalies were reversed by adoptive transfer of NK+B-T- marrow. My results suggest that uNK cells support vascular growth and development which ensures the decidua can support the growing conceptus early in pregnancy prior to formation and function of the placenta. Human decidual NK cells may fill similar roles and be important targets for strategies designed to correct intra-uterine growth restriction. / Thesis (Master, Anatomy & Cell Biology) -- Queen's University, 2013-08-02 08:42:06.487

RÉSUMÉ Les protéines associées à la gestation (PAGs) constituent une grande famille de protéinases aspartiques exprimées dans les cellules épithéliales de la couche superficielle du placenta des artiodactyles et plus précisément dans les granules des cellules binucléées. Des investigations par biologie moléculaire (clonage et séquençage) ont permis de montrer que les protéines de la gestation font partie de la grande famille des protéases aspartiques tout comme le pepsinogène, la pepsine, la chymosine, les cathepsines D et E, et la rénine. La détermination des concentrations en PAG par radioimmunoassay dans le sérum ou dans le plasma est actuellement employée comme méthode sérologique spécifique pour le diagnostic de gestation chez le bovin dès le 28e jour après la conception. Au-delà de ce délai, les dosages des PAGs peuvent également être utilisés pour assurer le suivi de la gestation notamment dans le cadre de létude de la mortalité embryonnaire précoce ou tardive et de la mortalité ftale. Au moment dentreprendre nos investigations expérimentales, une attention particulière a été portée à notre maîtrise des systèmes de dosages radioimmunologiques notamment dans les zones des faibles concentrations de PAG caractérisant le premier trimestre de la gestation chez les bovins. Cest ainsi que les deux premières études sur lesquelles repose ce travail de thèse ont visé à tester de nouveaux antisérums dirigés contre la PAG et à comparer leurs performances en milieu sérique ou plasmatique. Ensuite, cinq différents systèmes de dosages (utilisant différents antisérums) ont été comparés quant à leur sensibilité, spécificité, exactitude des diagnostics positifs et exactitude des diagnostics négatifs, sur des séries de plasmas provenant de femelles non gestantes et de femelles gestantes durant le premier trimestre de la gestation. Dans une troisième étude, les concentrations de PAG, LH et prolactine ont été interprétées en fonction dune classification basée sur le niveau de progestérone mesuré chez les femelles à jour 21. Dans la première étude, lobjectif a été de déterminer les paramètres de limite minimale de détection, reproductibilité, exactitude, spécificité et parallélisme des dilutions caractérisant les cinq systèmes radioimmunologiques différents : RIA-497, RIA-706, RIA-780, RIA-809 et RIA-Pool. Ensuite, leur capacité à distinguer les femelles non gestantes et gestantes prélevées au 30e jour suivant linsémination artificielle a été analysée dans le détail. Létude a été menée sur un effectif de 40 femelles de race Holstein-Frisonne dâge et de parité mélangés dont la gestation a été confirmée par exploration rectale réalisée entre le 2e et le 3e mois après linsémination. Un groupe contrôle composé de dix vaches maintenues en stabulation entravée confirmées non gestantes par examen échographique et exploration rectale a été utilisé simultanément. Les caractéristiques générales des différents systèmes étaient satisfaisantes. Leur spécificité était excellente à lexception du système RIA-809 ayant montré une très légère interférence de lacide syalique et de lhCG. Les concentrations mesurées dans le plasma des femelles non gestantes sont restées inférieures à 0,5 ng/mL excepté celles qui étaient mesurées dans les systèmes RIA-780 et RIA-809. Seuls les systèmes RIA-497, RIA-706 et RIA-Pool ont fourni des valeurs de sensibilité, spécificité, exactitude des diagnostics positifs, exactitude des diagnostics négatifs égales à 100%. Dans la deuxième étude, les cinq systèmes de dosage ont été utilisés pour déterminer les concentrations de PAG dans le plasma aux jours 30, 45, 60 et 80. Les corrélations ont été calculées entre les systèmes ainsi que les rapports de concentrations mesurées par chacun deux : les systèmes RIA-706, RIA-780, RIA-809 et RIA-Pool ont révélé des valeurs plus élevées que le RIA-497 à tous les stades étudiés. La troisième étude a visé à vérifier une hypothèse formulée à plusieurs reprises par différents auteurs : le niveau de concentration en progestérone atteint en début de gestation est-il susceptible dinteragir avec les concentrations de protéines trophoblastiques, de LH et de prolactine mesurées plus tard durant le premier trimestre de la gestation ? Dans cette étude, nous avons déterminé les concentrations de lensemble des paramètres sur les prélèvements correspondant aux jours 0, 21, 30, 45, 60 et 80 de 37 femelles gestantes. Les femelles ont été réparties en deux sous groupes : celles dont la progestérone au jour 21 était inférieure à la moyenne (Low-P4) et celles dont la même progestérone était supérieure à la moyenne au jour 21 (High-P4). Lorsque les deux sous groupes ont été suivis aux périodes ultérieures (jours 30, 45, 60 et 80), il est apparu que les concentrations de PAG ont eu tendance à être plus élevées dans tous les systèmes chez les vaches du groupe High-P4. Une différence significative est atteinte en utilisant le RIA-497 au jour 80. Les concentrations en LH et prolactine ont eu tendance à être plus faibles à toutes les périodes chez les vaches faisant partie du groupe High-P4. La différence a été significative pour la LH à jour 60 et pour la prolactine à jour 21. Dans leur ensemble, les résultats de ce travail montrent que le diagnostic de gestation peut être amélioré (sensibilité, spécificité, exactitude des diagnostics positifs, exactitude des diagnostics négatifs) en sélectionnant soigneusement les antisérums et en les mélangeant de façon appropriée. Ces systèmes sont utilisables pour suivre les concentrations de PAG durant le premier trimestre de la gestation, les résultats étant fortement corrélés tout en montrant des profils différents. Associées au dosage de la progestérone, de la LH et de la prolactine, les investigations sur les PAGs permettent de progresser dans létude des régulations qui relient lhypophyse, les gonades et le trophoblaste durant le premier trimestre de la gestation. SUMMARY Pregnancy Associated Glycoproteins (PAGs) constitute a large family of aspartic proteinases expressed in the outer epithelial cells layer of the placenta of eutherian species and more precisely in the granules of the binucleate cells. Investigations by molecular biology (cloning and sequencing) showed that the PAGs are part of the great family of the aspartic proteases like pepsinogen, pepsin, chymosin, cathepsins D and E, and renin. The determination of the concentrations in PAG by radioimmunoassay in serum or plasma is currently used as specific serological method for the diagnosis of gestation at day 28 after conception. Beyond this time, PAGs assays can also be used to ensure the follow-up of gestation in particular during the study of early or late embryonic mortality and early foetal mortality. When we started our experimental investigations, a special attention was paid to our control of radioimmunoassay systems especially in the zones with low concentrations of PAG which characterize the first trimester of bovine gestation. Thus the first two studies on which this work was based, aimed testing new antisera raised against PAG, and comparing their performances in seric or plasmatic medium. Then, five various assays (using different antisera) were compared as for their sensitivity, specificity, exactitude of the positive diagnoses and exactitude of the negative diagnoses, on series of plasmas from nonpregnant and pregnant females during the first trimester of gestation. In a third study, the concentrations of PAG, LH and prolactin were interpreted according to a classification based on the progesterone level measured in females at day 21. In the first study, the objective was to determine the parameters of minimal limit of detection, reproducibility, exactitude, specificity and parallelism of dilutions characterizing the five different radioimmunoassay systems: RIA-497, RIA-706, RIA-780, RIA-809 and RIA-Pool. Then, their capacity to discriminate non-pregnant and pregnant females investigated at day 30 after artificial insemination was also analyzed in detail. The study was undertaken on 40 Holstein-Friesian females of different age and parity, and whose gestation was confirmed by rectal exploration carried out between the second and the third month after insemination. A control group of ten cows maintained in stabling, confirmed non-pregnant by ultrasonography and rectal exploration, was used simultaneously. The general characteristics of the various systems were satisfactory. Their specificity was excellent except for system RIA-809 which shown a small interference of syalic acid and hCG. The concentrations measured in the plasma of non-pregnant females remained lower than 0.5 ng/mL except those which were measured in systems RIA-780 and RIA-809. Only the systems RIA-497, RIA-706 and RIA-Pool provided values of sensitivity, specificity, exactitude of the positive diagnoses, exactitude of the negative diagnoses equal to 100%. In the second study, all five systems were used in order to determine the concentrations of PAG in plasma at days 30, 45, 60 and 80. The correlations were calculated between the systems as well as the ratios of concentrations measured by each one of them: systems RIA-706, RIA-780, RIA-809 and RIA-Pool revealed higher values than the RIA-497 at all the studied stages. The third study aimed to check a hypothesis formulated by various authors: is the concentration level of progesterone reached at beginning of gestation likely to interact with the concentrations of trophoblastic proteins, LH and prolactin measured later during the first trimester of gestation ? In this study, we determined concentrations of the whole parameters on samples corresponding to days 0, 21, 30, 45, 60 and 80 of 37 pregnant females. The females were divided into two groups: those whose progesterone at day 21 was lower than the average (Low-P4) and those whose progesterone was higher than the average at day 21 (High-P4). When both groups were investigated at later periods (days 30 and 80), it appeared that the concentrations of PAG tended to be higher in all systems in the cows of the High-P4 group. A significant difference was raised by using RIA-497 at day 80. The concentrations in LH and prolactin lead to be weaker during all the periods in the cows of the High-P4 group. The difference was significant for LH at day 60 and for prolactin at day 21. In conclusion, the results of this work show that the diagnosis of gestation can be improved (sensitivity, specificity, exactitude of the positive diagnoses, exactitude of the negative diagnoses) by selecting the antisera carefully and by mixing them in a suitable way. These systems are usable to follow the concentrations of PAG during the first trimester of gestation, the results are strongly correlated while showing different profiles. Combined to assays of progesterone, LH and prolactin, the investigations on PAGs make progress in the study of the regulations between pituitary gland, gonads and the trophoblast during the first trimester of gestation.

This research was conducted to test the hypothesis that select nutrients including glucose, leucine, arginine and glutamine stimulate conceptus development by activating mTOR (mammalian target of rapamycin; HGNC approved gene name: FRAP1, FK506 binding protein 12-rapamycin associated protein 1) signaling pathway. First, temporal changes in quantities of select nutrients (glucose, amino acids, glutathione, calcium, sodium and potassium) in uterine lumenal fluid from cyclic (Days 3 to 16) and pregnant (Days 10 to 16) ewes were determined. Total recoverable glucose, Arg, Gln, Leu, Asp, Glu, Asn, His, beta-Ala, Tyr, Trp, Met, Val, Phe, Ile, Lys, Cys, Pro, glutathione, calcium and sodium was greater in uterine fluid of pregnant compared to cyclic ewes between Days 10 and 16 after onset of estrus. Of note were remarkable increases in glucose, Arg, Leu and Gln in uterine flushings of pregnant ewes between Days 10 and 16 of pregnancy. Second, effects of the estrous cycle, pregnancy, progesterone (P4) and interferon tau (IFNT) on expression of both facilitative (SLC2A1, SLC2A3 and SLC2A4) and sodium-dependent (SLC5A1 and SLC5A11) glucose transporters, cationic amino acid transporters (SLC7A1, SLC7A2 and SLC7A3), neutral amino acid transporters (SLC1A4, SLC1A5, SLC3A1, SLC6A14, SLC6A19, SLC7A5, SLC7A6, SLC7A8, SLC38A3, SLC38A6 and SLC43A2) and acidic amino acid transporters (SLC1A1, SLC1A2 and SLC1A3) in ovine uterine endometria from Days 10 to 16 of the estrous cycle and Days 10 to 20 of pregnancy as well as in conceptuses from Days 13 to 18 of pregnancy were determined. Among these genes, SLC2A3 and SLC7A6 were detectable only in trophectoderm and endoderm of conceptuses. The abundance of mRNAs for SLC2A1, SLC2A4, SLC5A1, SLC5A11, SLC7A1, SLC7A2, SLC1A4, SLC1A5, SLC43A2 and SLC1A3 changed dynamically in ovine uterine endometria according to day of the estrous cycle and early pregnancy. Expression of mRNAs for SLC2A1, SLC5A11 and SLC7A1 in endometria was induced by P4 and further stimulated by IFNT with shortterm treatment (12 days), while expression of SLC7A1 and SLC1A5 in endometria required long-term treatment (20 days) with P4 and IFNT. Third, effects of the estrous cycle, pregnancy, P4 and IFNT on expression of nitric oxide synthase (NOS1, NOS2 and NOS3), GTP cyclohydrolase (GCH1), ornithine decarboxylase 1(ODC1), insulin-like growth factor II (IGF2), FRAP1 complexes (FRAP1, LST8, MAPKAP1, RAPTOR, RICTOR), regulators (TSC1, TSC2, RHEB) and an effector (EIF4EBP1) of FRAP1 signaling in ovine uterine endometria from Days 10 to 16 of the estrous cycle and Days 10 to 20 of pregnancy as well as in conceptuses from Days 13 to 18 of pregnancy were determined. All of these genes were expressed in ovine uterine endometrium and conceptuses. Among these genes, expression of NOS1, IGF2, RHEB and EIF4EBP1 changed dynamically due to day of the estrous cycle and early pregnancy. Progesterone stimulated NOS1 and GCH1 expression while IFNT inhibited NOS1 expression in uterine endometria, and P4 and IFNT stimulated expression of RHEB and EIF4EBP1 in uterine endometria. Collectively, these results indicate that: 1) the availability of select nutrients in the ovine uterine lumen increases to support the rapid growth and elongation of the conceptus during the peri-implantation stage of pregnancy; 2) P4 and/or IFNT stimulate(s) glucose and amino acid transporters to facilitate their transport from maternal tissues and/or blood into the uterine lumen during early pregnancy; 3) the FRAP1 cell signaling pathway mediates interactions between the maternal uterus and peri-implantation conceptus and both P4 and IFNT affect this pathway by regulating expression of RHEB and EIF4EBP1. Expression of NOS, ODC1 and IGF2 appear to be linked to FRAP1 signaling in both uteri and peri-implantation conceptuses.