The role of proestrus on fertility and postovulatory uterine function in the cow

Modern protocols to synchronize ovulations for timed artificial insemination and timed embryo transfer that include manipulations in the proestrus period (i.e., between luteolysis and estrus) affect fertility in cattle. Specifically, stimulating pre-ovulatory follicle growth and exposure to estrogens after CL regression increase the proportion of cows pregnant and decrease late embryo mortality. Such effects may be due to both preovulatory actions of estrogens and post-ovulatory actions of progesterone, as concentrations of the later hormone may be changed in response to manipulations conducted during proestrus. In the first portion of this paper we describe strategies used recently to manipulate the proestrus period in protocols for synchronization of ovulation, and to present evidence of their effects on fertility. Manipulations of timing and prominence of sex steroids during the proestrus and early diestrus that affect fertility may act on targets such as the endometrium. This tissue expresses receptors for both estrogens and progesterone and these hormones change endometrial function to support conceptus growth and pregnancy maintenance. However, specific cellular and molecular mechanisms through which fertility is affected via manipulations of the proestrus are poorly understood. In the second portion of this paper we describe a well-defined animal model to study changes in endometrial function induced by manipulations conducted during the proestrus. Such manipulations induced endometrial changes on sex steroid receptors expression, cell proliferation, oxidative metabolism and eicosanoid synthesis in the uterus, but not on glucose transport to uterine lumen. In summary, evidence is accumulating to support a positive role of increasing duration and estrogen availability during the proestrus on fertility to synchronization protocols. Such positive effects may be through changes in endometrial function to stimulate conceptus growth and survival.

Developmental aspects of early pregnancy in mares

In mares, the embryo migrates to the uterus between days 5 and 6 postovulation beginning its mobility through all uterine segments, which is essential for the maternal recognition of pregnancy. During the mobility phase, the embryonic vesicle shows a linear growth rate until its fixation between days 15 and 17, when the orientation phenomenon occurs. From fixation to day 28 of pregnancy, the embryonic growth is less evident (plateau) by cross-section ultrasound examination. After this period the linear growth rate is reestablished until day 46. This plateau is attributed to the increased uterine tone that compresses the vesicle and to volume expansion, making it difficult to detect the conceptus growth only by the cross-section diameter. Around day 20, the embryo proper is visualized as an echogenic spot in the ventral aspect of the vesicle. Additionally, development of allantoic sac, embryonic heartbeat, yolk sac regression and posterior umbilical cord formation also can be visualized from days 20 to 40. An intimate interaction between uterus and conceptus is essential for the normal pregnancy development. Color-and spectral-Doppler ultrasonography can be useful for the evaluation of this interface. A gradual increase on uterine vascularity during the early pregnancy and transient changes in endometrial vascularity accompanying the vesicle location during the mobility phase have been described. Around day 38 of gestation, the formation of the endometrial cups begins and, consequently, the synthesis of the equine chorionic gonadotropin (eCG) induces the formation and development of supplementary corpora lutea, which are important to secrete progesterone and to maintain pregnancy until around day 120.

The intricate role of mast cell proteases and the annexin A1-FPR1 system in abdominal wall endometriosis

Endometriosis is a continuous and progressive disease with a poorly understood aetiology, pathophysiology and natural history. This study evaluated the histological differences between eutopic and ectopic endometria (abdominal wall endometriosis) and the expression of mast cell proteases (tryptase and chymase), annexin A1 (ANXA1) and formyl peptide receptor 1 (FPR1). Ectopic endometrium from 18 women with abdominal wall endometriosis and eutopic endometrium from 10 women without endometriosis were obtained. The endometrial samples were analysed by histopathology, immunohistochemistry and ultrastructural immunogold labeling to determine mast cell heterogeneity (tryptase and chymase positive cells) and the expression levels of ANXA1 and FPR1. Histopathological analysis of the endometriotic lesions showed a glandular pattern of mixed differentiation and an undifferentiated morphology with a significant influx of inflammatory cells and a change in mast cell heterogeneity, as evidenced by a significant increase in the number of chymase-positive cells and endogenous chymase expression. The undifferentiated glandular pattern of endometriotic lesions was positively associated with a marked increase and co-localization of ANXA1 and FPR1 in the epithelial cells. In conclusion, the co-upregulated expression of mast cell chymase and ANXA1–FPR1 system in ectopic endometrium suggests their involvement in the development of endometriotic lesions.

Utiização de altrenogest injetável par o estabelecimento e manutençao da gestação em éguas receptoras de embriões acíclicas

Pós-graduação em Medicina Veterinária - FMVZ

Histopathologycal findings in the ovaries and uterus of albino female rats promoted by co-administration of synthetic steroids and nicotine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos da somatotropina recombinante bovina na taxa de concepção, composição celular do corpo lúteo e morfometria das glândulas endometriais em vacas de corte

Pós-graduação em Ciência Animal - FMVA

Caracterização das proteínas caveolinas -1 e -2 na placenta de conceptos bovinos clonados transgênicos

The transgenic application of green fluorescent protein (GFP) as fetal cell marker on cattle cloned placenta could provide an exclusive model for studying the morphologic and immunologic maternal-fetal interactions, providing information about its mapping, distinguishing the fetal from maternal cells. This model will have direct application, mainly because these animals present problems during its development. With this model's support, we intend to verify the substances transport between mother and fetus during endocytosis, through the immunolocalization of protein named caveolae. For these, we used 06 cloned bovine and 30 cattle samples of artificial insemination (AI) with 90 days of pregnancy, which had been their development interrupted by humanitarian slaughter of the recipient and recovery of the pregnant uterus. We collected the placentome and the chorion. A part of the samples was cut and fixed, by immersion, on a solution containing 4% of parafomaldehyde or 10% of formaldehyde on a sodium phosphate buffer (PBS), at 0,1 M pH 7.4, Zamboni solution (4% of paraformaldehyde, 15% of picric acid, on sodium phosphate buffer 0,1 M pH 7.4), metacarn (60% of metanol, 30% of chloroform, and 10% glacial acetic acid), for morphologic and immunohistochemistry verification for caveolinas proteins -1 and -2 (CAV -1 and CAV-2). The caveolins -1 were found in fetal and maternal villi, but its strongest staining was observed in the endometrial stroma. The caveolins -2 had positive staining in trophoblast and chorioallantoic membrane, and specifically in giant trophoblastic binucleated cell. Therefore the results were compared between cloned cattle and from AI or natural mating, for assisting on detection of the reason of many placental alterations, embryonic losses, spontaneous abortion, post-natal mortality and large offspring syndrome on laboratory-manipulated animals. The result suggests that the proteins caveolins -1 and -2 (CAV-1 and CAV-2) are part of the caveolae composition and important structures related to the molecule transfer to the fetus, nourish it through endocytosis and pinocytosis.

Real-time characterization of the uterine blood flow in mares before and after artificial insemination

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Investigação da evolução dos pólipos endometriais em câncer de endométrio

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histomorfometria do útero de gatas submetidas à ovariosalpingohisterectomia

Histomorphometric data were obtained from cats uterine horns: either nulliparous (n = 6), primiparous (n = 6), multiparous (n = 6) and treated with contraceptive (n = 6). The material was collected after surgery, fixed in paraformaldehyde and embedded in paraplast® resin to be sliced in a microtome. The obtained sections were stained with hematoxylin - eosin and measured under a light microscope: uterine wall total thickness (μm), endometrium total height, endometrial glands diameter and glandular epithelia height, total myometrium, internal and external myometrium and vascular layer thickness. It was concluded that: 1 - contraceptive use and number of pregnancies altered the uterine structure, 2 - one pregnancy does not appear to affect the uterine lining structures as occurs in multiparous cats, 3 – there was no variation on the evaluated structures between nulliparous and primiparous cats except for inner myometrium, 4 - the total myometrium hight and the endometrium hight showed similar variations except for the contraceptive treated cats group, 5 - the outer myometrium showed marked changes in the treated cats, 6 – the less marked morphological variations were for the endometrial glands diameter and glandular epithelium hight, 7 - the presence of dilated endometrial glands was found only in treated cats.

Análise hormonal, imunolocalização e quantificação dos receptores de andrógenos (AR) e estrógenos (ER-α E ER-β) em ovário e útero de ratas submetidas a diferentes doses de decanoato de nandrolona: avaliação nos períodos pós-tratamento e pós-recuperação

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Experimental model of intestinal endometriosis: pilot study

Aims: The objective of this study is to create an experimental model of intestinal endometriosis in pigs, which might allow better understanding of deep infiltrating endometriosis and development of new treatment techniques. As secondary objective, we intend to create endometrial implants accessible by transrectal ultrasonography (TRUS). Study Design: Surgical experimental study in swine. Place and Duration of Study: This study was performed at the Instituto de Ensino e Pesquisa do Hospital Sírio-Libanês, São Paulo, Brazil, between January 2012 and December 2012. Methodology: Two sexually mature female minipigBR pigs underwent two laparotomies (each animal). The first laparotomy was performed to implant two fragments of autologous endometrium in the rectal wall. The second one was performed thirty days later to visualize, measure and obtain tissue of the site of the implants for histopathology study. A TRUS study was performed prior to the second surgery. The Institution’s Animal Utilization Study Committee approved the study. Results: In the first laparotomy a 5-cm segment of right uterine horn was resected. The endometrium was separated from the myometrium through sub-endometrial saline injection. Two endometrial fragments (1.0 x 2.0 cm) were dissected and sutured in the intra peritoneal anterior rectal wall of the animals. Thirty days later, all implants were identified during preoperative TRUS. “En-bloc” resection of the intestinal segment with the implants was performed during the second surgery. The autologous implants of endometrium invaded the muscular layer in one of the two animals. Conclusion: We demonstrated that the creation of an animal model of deep infiltrating endometriosis with intestinal involvement is feasible through a simple surgical technique. We believe that this model can be applied in experimental and clinical studies but further studies are necessary to refine the technique.

Pólipos endometriais e seu risco de malignização: aspectos epidemiológicos, clínicos e imunoistoquímicos

Endometrial polyps are neoformations that result from focal hyperplasia of the endometrial basal layer associated with hormonal hyperstimulation. Their pathogenesis is still unclear, and there is no consensus on their natural history, actual relevance as pathologic entities, and relationship with endometrial neoplasia. Endometrial polyps are the most frequent indication of surgical hysteroscopy, but their optimal management remains controversial. Therefore, an overview of this condition may help choosing the most adequate treatment strategies.

Avaliação da cavidade uterina com histerossonografia, uma revisão

Sonohysterography was firstly described three decades ago. The saline solution infusion into the uterine cavity favors its use and provides excellent visualization of the anatomy and the inner cavity of the uterus better than the conventional transvaginal sonography To check the current role of sonohysterography in the uterine cavity assessment in women with abnormal uterine bleeding and asymptomatic, a literature review comparing sonohysterograph with conventional transvaginal sonography and/or ambulatory diagnostic hysteroscopy was carried out. To this end, relevant studies were researched in electronic databases Medline/PubMed, SciELO/LILACS. The sonohysterography is an ambulatory procedure, non-invasive, better cost-benefit, better sensitivity and specificity to identify uterine abnormalities, causing minimal discomfort and low complications rate. It was subject to revision which there is no more doubt about its accuracy. It can be concluded that the sonohysterography is a useful tool in the propedeutics to assess uterine cavity of symptomatic patients with abnormal uterine bleeding, infertility, recurrent miscarriages, and embryonic implantation failures in assisted reproduction treatment / in vitro fertilization and in any other intra and extra uterine cavity alteration. Hence, conventional transvaginal sonography is indicated as an initial method of assessment of the uterine cavity previously to ambulatory diagnostic hysteroscopy.

Detecção imunoistoquímica de receptores de estrógeno e progesterona no endométrio de vacas Nelore (Bos taurus indicus) durante o anestro pós-parto

Twenty-four postpartum anestrous Nelore purebred cows were used in the study. Anestrous was determined based on the reproductive history which was confirmed in each cow based on plasma progesterone concentration and by transrectal ultrasonography. Endometrial biopsies were collected. The animals were separated into two groups according to maximum follicular diameter- Group 1: follicles <6mm (n=12) and Group 2: follicles >-6mm follicles (n=12). The immunohistochemistry technique was employed to evaluate the presence of estrogen and progesterone receptors in the uterine glandular epithelium and stroma. High counts of positive nuclei and high intensity of immunostain for estrogen and progesterone receptors in the glandular epithelium and stroma were observed in the two groups. However, the immunostain intensity of progesterone receptors in the glandular epithelium was higher in Group 2 compared to Group 1. When glandular epithelium and stroma were compared within each group, the relative number of estrogen receptors in the Group 1 was higher in the glandular epithelium compared to stroma and the immunostain intensity for the progesterone receptor in Group 2 was higher in the glandular epithelium compared to stroma. The results suggest that the mechanisms that control the expression of endomerial receptors in the anestrus are similar to the ones observed during the estrus cycle.