We are using the feline immunodeficiency virus (FIV)-infected cat to model HIV mother-to-child-transmission (MTCT). Vertical transmission of either virus may result not only in infected offspring, but also failed pregnancy.In HIV infections, maternal hematological and virological parameters predict MTCT.We hypothesized that such parameters would likewise be predictors of FIV vertical transfer. We inoculated ten cats with FIV-B-2542; 10 cats were uninoculated. Cats were allowed to breed naturally. Fetuses were delivered at approximately week 3 (early) gestation by cesarean section. Fetal and placental tissues were collected.Blood samples were collected from the day of inoculation through delivery. We quantified CD4:CD8 T cell ratios, proviral load, and plasma viremia, and monitored seroreactivity to FIV proteins in longitudinal sera from both groups of cats. We documented clinical and reproductive outcome. The infected group produced reduced litter size and more failed pregnancies; CD4:CD8 ratios were depressed by 3.5 months p.i.Proviral DNA was detected in 14 of 14 (100%) placentas tested and 12 of 14 (86%) fetuses. However, the parameters assessed were not predictive of reproductive outcome and suggested a role for placental immunopathology in compromised pregnancy.Regulatory T cells (Treg) are anti-inflammatory and essential in maintaining pregnancy.Th17 cells are pro-inflammatory and associated with pregnancy failure. The activation of these cell populations is regulated by the cytokines TGF-? and IL-6. We hypothesized that placental immunology may result from altered dynamics of these cell populations.Using immunofluorescence confocal microscopy to measure Treg and Th17 markers FoxP3 and ROR ? , respectively, we quantified these cells in placental specimens from FIV-infected and control cats at early and late (week 8) gestation.Significantly higher levels of ROR ? were measured in FIV-infected placentas at early pregnancy; these cells co-localized at the maternaletal interface. We quantified the expression of Treg immunomodulators by quantitative PCR, noting higher expression of TGF-? in infected queens.A positive correlation of ROR ? with IL-6 occurred in control placentas, as predicted, but not in infected placentas.Collectively, the data suggest that an inflammatory placental microenvironment at early pregnancy in infected queens may result, in part, from dysregulation of the Treg/Th17 balance.

Surgical technique was taught to third-year medical students using the placenta as tissue for practice and to illustrate the proper use of surgical tools, using principles that promote accuracy, security and efficiency of manual control.

In vitro produced (IVP) bovine embryos are useful for investigating the mechanisms affecting early embryonic failure. The work in this thesis explored how interleukin-6 (IL6), an embryokine that increases inner cell mass (ICM) influences post-transfer embryonic survival and development of the embryo-proper and fetus. Four replicates of slaughterhouse-derived cumulus oocyte complexes underwent in vitro maturation and fertilization. On day 5 post-fertilization, embryos were treated with either 1% Bovine Serum Albumin (BSA) (CONT) or 100ng/mL recombinant bovine IL6 with 1% BSA (TRT). On day 7.5 post-fertilization, individual blastocysts were loaded into transfer straws. Beef and dairy cow recipients were synchronized with the day of in vitro fertilization using a 7-d CO-Synch protocol. A subset of cows from each group underwent fixed-time artificial insemination (AI) (n=37). The remaining cows underwent embryo transfer (ET) in the uterine horn ipsilateral to a corpus luteum (CL) (IL6 n=35; CONT n=51). Embryo and fetal measurements were performed via transrectal ultrasonography weekly from days 28-56 post-insemination, respectively. Overall pregnancy rates were 40.0% IL6; 19.6% CONT; and 32.4% AI. Crown-rump lengths (CRL) were reduced (P<0.05) in CONT pregnancies when compared with IL6 and AI at days 28, 35, 42, and 56. A tendency (P=0.057) for larger abdominal diameters was detected between IL6 and CONT groups. Also, IL6 had larger crown-nose lengths than CONT (P<0.05) and tended to be larger than AI (P=0.07). In summary, IL6 treatment produced pregnancies resembling AI-generated pregnancies more so than conventionally cultured embryos, supporting the hypothesis that IL6 improves developmental competency of IVP embryos. / Master of Science / The incidences of pregnancy loss in both beef and dairy cattle industries are profound and are upwards of 60 percent. Financial stability for both of these industries revolves closely around the ability of cattle to give birth to a live calf annually. While artificial insemination (AI) has been heavily adopted and utilized widely in the dairy industry, the use of in vitro produced (IVP) bovine embryos has shown promise in lessening some of the stresses placed on impregnating cattle. The IVP of bovine embryos serves as a strong model to understand how pregnancy losses occur. Briefly, IVP involves the collection of eggs from donor animals, and subsequent fertilization to mimic what occurs within the animal naturally. A disadvantage of in vitro produced embryos is their reduced likelihood to establish pregnancy after transfer into recipient animals. Interleukin-6 (IL6) was recently identified as a pro-developmental factor that may improve the quality and post-transfer competency of in vitro produced embryos. The objective of this work was to determine if IL6 supplementation during in vitro culture improves post-transfer fetal development. Oocytes (i.e. eggs) were retrieved from slaughterhouse-derived ovaries and subjected to in vitro maturation and fertilization. On day 5 post-fertilization, embryos were treated with either 0 (CONT; 1% BSA) or 100ng/mL recombinant bovine IL6. On day 7.5 post-fertilization, individual embryos (blastocyst stage) were loaded into transfer straws. Estrous synchronized beef (n = ) and dairy (n = ) cow recipients were allocated into treatment groups in the following manner. A subset of cows from each group underwent fixed-time AI (n=37). Remaining cows underwent embryo transfer (ET) in the uterine horn ipsilateral to a corpus luteum; 51 of these cows received a CONT embryo and the remaining 35 cows received an IL6 embryo. Thus, there were three treatment groups: AI, CONT, and IL6. Embryo and fetal measurements were performed via transrectal ultrasonography weekly from day 28 to 56, these included crown-rump length, crown-nose length, abdominal diameter, and amniotic vesicle. Pregnancies that remained throughout the entirety of the experiment 40.0% for IL6 (14/35); 19.6% for CONT (10/51); and 32.4% for AI (12/37). In summary, IL6 treatment of embryos produced pregnancies with characteristics more similar to the current industry standard of AI, rather than conventionally cultured embryos (CONT), supporting the hypothesis that IL6 supplementation to bovine embryos on day 5 post-fertilization improves developmental competency of in vitro produced embryos.

Objectifs : Vérifier s'il existe une corrélation entre le volume placentaire, évalué par échographie tridimensionnelle au 1er trimestre de la grossesse et le poids de naissance. Méthodologie : Une étude de cohorte prospective de 543 femmes enceintes recrutées entre 11 et 13 semaines de grossesse. Nous avons évalué l’association entre le volume placentaire mesuré par échographie tridimensionnelle (rapportée en multiple de la médiane pour l’âge gestationnel) et le poids du nouveau-né à la naissance (rapporté en percentile). Nous avons utilisé des analyses non paramétriques (test de corrélation de Spearman, Mann-Whitney, et Kruskal-Wallis) avec une valeur de p significative si inférieure à 0.05. Résultats : Parmi 554 femmes recrutées et admissibles, nous avons observé une corrélation significative entre le volume placentaire au 1er trimestre et le poids de naissance (Coefficient corrélation(CC): 0,15; p&lt; 0.0001). Le volume placentaire n’était pas associé au risque de prééclampsie. Conclusion : Il existe une corrélation significative entre le volume placentaire évalué au 1er trimestre de la grossesse et le poids de naissance, mais aucune corrélation ne fut observée avec le développement d’une prééclampsie au cours de la grossesse. / Objectives: To assess the correlation between placental volume, measured by three dimensional ultrasound in the first trimester of pregnancy and birth weight. Methodology: A prospective cohort recruiting 543 women between 11 and 13 weeks gestation was conducted. We evaluated the correlation between placental volume measured by three- dimensional ultrasound (reported as multiple of the median for gestational age), and birth weight (reported in percentile). Data were analyzed using non-parametric tests (Spearman, Mann-Whitney, Kruskal- Wallis), with a significant p value under 0,05. Results: A total of 554 women were recruited. The placental volume showed significant correlation with birth weight (Correlation coefficient (CC); 0,15; p&lt; 0.0001). The placental volume was not associated with preeclampsia. Conclusion: There is a significant correlation between placental volume and birth weight. In this study, no correlation was found between placental volume and preeclampsia.

L'objectif de cette étude cas-témoin est d'examiner la relation entre les maladies parodontales et la prématurité à moins de 34 semaines, et de définir leur association à l'inflammation foetale ou placentaire. Dans cette étude, 50 femmes ayant accouché prématurément à la suite d'un travail spontané ou une rupture prématurée des membranes, ont été comparées à 100 femmes ayant accouché à terme. L'examen parodontal post-partum, la mesure d'IL-6 dans le sang du cordon ombilical et l'examen histopathologique des membranes foetales ont été effectués. Les résultats de cette étude n'ont pas révélé d'association entre les naissances prématurées, accompagnée ou non de chorioamnionite, et la présence de maladie parodontale. Nous avons par contre observé que la prématurité était plus souvent associée à une éducation moins avancée (p=0,046) et à la présence de chorioamnionite pathologique (p[plus petit]0.01). En conclusion, la prématurité est fortement associée à la présence de chorioamnionite, mais pas à la présence de maladies parodontales.

CRIPTO 1 (CR1) é membro da família de fatores de crescimento epidermal-CRIPTO-1/FRL-1/Cryptic com funções específicas na embriogênese e tumores. Este estudo verificou a presença de CR1 em placentas saudáveis e com distúrbios de invasividade. Nossos resultados mostraram a expressão de CR3 e CR1 em placentas de primeiro e terceiro trimestre de gestação, respectivamente. Placentas acretas e coriocarcinomas também foram imunorreativos para CR1, principalmente em células citotrofoblásticas invasivas e malignas, respectivamente. A expressão de CR1 em células citotrofoblásticas extravilosas de primeiro trimestre prevaleceu em células em diferenciação e em células invasivas, sugerindo sua participação nesses processos. CR1 recombinante aumentou significativamente a invasão e migração em células HTR8/SV-neo, o que foi abolido na presença de RNA de interferência. Esses achados corroboram a ação dessa proteína sobre as células trofoblásticas. / CRYPTO 1 (CR1) is member of the family of epidermal growth factor-CRYPTO-1 / FRL-1 / Cryptic with specific roles in embryogenesis and tumors. This study investigated the presence of CR1 in healthy placentas and in placentas with invasive disorders. Our results showed the expression of CR3 and CR1 in the first and third trimester placentas, respectively. Placental choriocarcinoma and accreta placentas were also immunoreactive for CR1, especially in malignant and invasive cytotrophoblast cells, respectively. CR1 expression in first trimester extravillous cytotrophoblast cells prevailed in differentiating and invasive cells, suggesting a role in these processes. Recombinant CR1 significantly increased invasion and migration HTR8 / SV-Neo cells, which was abolished in the presence of interference RNA. These findings corroborate the action of this protein on the trophoblast cells.

Le retard de croissance intra-utérin (RCIU) se définit comme étant un échec dans la croissance du fœtus à atteindre son potentiel de croissance génétiquement déterminé. Il touche entre 7 à 10 % des grossesses. Alors que le dépistage commence seulement à partir de la vingtième semaine de grossesse, cette complication pourrait être détectable dès le premier trimestre. Un dépistage précoce des femmes à risque élevé permettrait de débuter un traitement préventif. L’échographie 3D Doppler de puissance a démontré des résultats encourageants pour le dépistage précoce de la pré-éclampsie. La biopsie virtuelle du placenta et ses indices de vascularisation pourraient possiblement permettre la détection précoce de cas de RCIU dès le premier trimestre. Les objectifs de l’étude étaient d’évaluer la valeur de la biopsie virtuelle, du volume placentaire et des indices de vascularisation placentaires au 1er trimestre dans la prédiction du RCIU stratifié selon la sévérité. Nous avons observé que le volume placentaire est un bon prédicteur du RCIU. Cependant, la biopsie virtuelle du placenta et les indices de vascularisation placentaire ne prédisent pas bien le RCIU. En conclusion, la biopsie placentaire et sa vascularisation déterminées par échographie 3D Doppler de puissance ne sont pas des modalités utiles en clinique pour le dépistage précoce du RCIU au premier trimestre de la grossesse. / Fetal growth restriction (FGR) is a pathological process defined as the failure of the fetus to reach its biologically determined growth potential. It affects 7 to 10% pregnancies. Screening for FGR begins only at the 20th weeks of pregnancy but this complication could be detectable during the first-trimester of gestation. Early detection of high-risk pregnancies could allow to start a preventive treatment as soon as possible. Threedimensional Power Doppler ultrasound has shown some encouraging results for early identification of women at high risk of preeclampsia. Placental volume and ultrasonographic sphere biopsy (USSB) with their vascularization indices could possibly detect FGR fetuses during the first-trimester. Our objective was to estimate the performance of the whole placental volume and USSB vascularization in the prediction of severe and non-severe FGR fetuses in the first-trimester. Our results showed that placental volume was a good predictor of severe and non-severe FGR. However, USSB displayed a poor predictive ability. Therefore, first-trimester USSB and its vascularization indices are not good modalities for clinical practice for the early screening of FGR pregnancies.

O decorim e o biglicam são membros da família dos pequenos proteoglicanos ricos em leucina e possuem importantes funções no controle da proliferação, migração e invasão do citotrofoblasto extraviloso (TEV). O objetivo deste trabalho foi de caracterizar a expressão diferencial e a imunolocalização de decorim e biglicam em placentas humanas normais a termo (PNT), na placenta acreta (PA), na mola invasora (MI) e no coriocarcinoma (CO). Na PNT, as células deciduais apresentaram positividade para o decorim, enquanto o TEV foi negativo. O decorim foi fracamente expresso na matriz endometrial, mas negativo no fibrinoide do tipo matriz, enquanto foi positivo para biglicam. Na PA e na MI, o TEV mostrou positividade para decorim e biglicam. No CO, somente o citotrofoblasto foi positivo para ambos proteoglicanos. Portanto, o decorim e o biglicam são expressos diferencialmente em placentas normais e patológicas, sugerindo que os padrões de expressão desses proteoglicanos nas patologias estudadas indicam um papel na modulação da migração e da invasão do trofoblasto. / Decorin and biglycan are family members of the small leucine-rich proteoglycans family, and they have many functions as controlling proliferation, migration and invasion of extravillous trophoblast cells (EVT). The aim of this study was to characterize decorin and biglycan differential expression and immunolocalization in human normal term placenta (NTP), in placenta accreta (PA), in invasive mole (IM), and in choriocarcinoma (CH) samples. In PNT, deciduas cells were positive to decorin whereas EVT was negative. Decorin was faintly stained at endometrial matrix, but negative at matrix-type fibrinoid, although it was positive for biglycan. In PA and IM, the EVT was positive for decorin and biglycan. In CH, only cytotrophoblast cells were positive for both proteoglycans. Therefore, decorin and biglycan are differentially expressed in normal placenta and in placenta pathologies, suggesting that the expression patterns of the proteoglycans in studied pathologies indicate a role in modulating trophoblast migration and invasion.

O decorim e o biglicam são membros da família dos pequenos proteoglicanos ricos em leucina e possuem importantes funções no controle da proliferação, migração e invasão do citotrofoblasto extraviloso (TEV). O objetivo deste trabalho foi de caracterizar a expressão diferencial e a imunolocalização de decorim e biglicam em placentas humanas normais a termo (PNT), na placenta acreta (PA), na mola invasora (MI) e no coriocarcinoma (CO). Na PNT, as células deciduais apresentaram positividade para o decorim, enquanto o TEV foi negativo. O decorim foi fracamente expresso na matriz endometrial, mas negativo no fibrinoide do tipo matriz, enquanto foi positivo para biglicam. Na PA e na MI, o TEV mostrou positividade para decorim e biglicam. No CO, somente o citotrofoblasto foi positivo para ambos proteoglicanos. Portanto, o decorim e o biglicam são expressos diferencialmente em placentas normais e patológicas, sugerindo que os padrões de expressão desses proteoglicanos nas patologias estudadas indicam um papel na modulação da migração e da invasão do trofoblasto. / Decorin and biglycan are family members of the small leucine-rich proteoglycans family, and they have many functions as controlling proliferation, migration and invasion of extravillous trophoblast cells (EVT). The aim of this study was to characterize decorin and biglycan differential expression and immunolocalization in human normal term placenta (NTP), in placenta accreta (PA), in invasive mole (IM), and in choriocarcinoma (CH) samples. In PNT, deciduas cells were positive to decorin whereas EVT was negative. Decorin was faintly stained at endometrial matrix, but negative at matrix-type fibrinoid, although it was positive for biglycan. In PA and IM, the EVT was positive for decorin and biglycan. In CH, only cytotrophoblast cells were positive for both proteoglycans. Therefore, decorin and biglycan are differentially expressed in normal placenta and in placenta pathologies, suggesting that the expression patterns of the proteoglycans in studied pathologies indicate a role in modulating trophoblast migration and invasion.

Call number: LD2668 .T4 1964 H84 / Master of Science