Estudo de metodologias para as criopreservação de sêmen de jumento(Equus Asinus) por meio de testes laboratoriais e fertilidade

Pós-graduação em Medicina Veterinária - FMVZ

Analysis of systemic inflammatory response in the carcinogenic process of uterine cervical neoplasia

The inflammatory response is an active process in cervical cancer and may act in the progression and/or regression of the lesion. At the site of inflammation, macrophages and neutrophils are present as well as cytokines such as TNF-alpha and IFN-gamma. This study aims to evaluate the inflammatory response levels in women with cervical intraepithelial lesions (CIN) and with squamous cell carcinoma (SCC) of the cervix. Serum samples obtained from women without evidence of disease (n = 30), with CIN (n = 30) and with SCC of the cervix (n = 30) were analyzed for the activities of N-acetylglucosaminidase (NAG) and myeloperoxidase (MPO) by enzymatic assay and the serum levels of TNF-alpha and IFN-gamma by ELISA assay. The activities of NAG and MPO and the level of TNF-alpha were higher in women with CIN compared to the women with SCC. The levels of IFN-gamma were lower in the group of women with CIN compared to the group with SCC. There was not a significant association between the degree of the CIN and the staging of the SCC of the cervix and the degree of inflammation as assessed by the levels of inflammatory markers. The inflammatory response was inversely correlated with the progression of the carcinogenic process. In the three groups, the control group, women with CIN and women with invasive SCC, there was no association between the degree of preinvasive lesions and staging of the SCC of the cervix. (C) 2011 Elsevier Masson SAS. All rights reserved.

Use of corticosteroid therapy on the modulation of uterine inflammatory response in mares after artificial insemination with frozen semen

In order to modulate uterine inflammatory response and evaluate the effect of corticosteroid therapy on fertility, 90 cycles of 45 mares were used for artificial insemination with frozen semen, using three different protocols: G1 - inseminated with frozen semen (800 x 10(6) viable spermatozoa pre-freezing) + 20 mL of seminal plasma; G2 - inseminated with frozen semen (800 x 10(6) viable spermatozoa pre-freezing) + corticosteroid therapy; G3 - inseminated with frozen semen (800 x 10(6) viable spermatozoa pre-freezing) + 20 mL of seminal plasma + corticosteroid therapy. Corticosteroid therapy consisted on one administration of prednisolone acetate (0.1 mg/Kg - Predef (R)) when mares presented 35mm follicles and uterine edema, concomitantly with the unique dose of hCG (human chorionic gonadotropin), then repeated each 12 hours until ovulation. on first fertility trial, with normal mares, there was no difference between control and treated groups (p>0.05), using seminal plasma associated with corticosteroid therapy (40 vs. 38%, respectively) or corticosteroid therapy alone (40 vs. 45% respectively). The second fertility trial, performed with mares with previous history of post-insemination endometritis, demonstrated a significant increase of pregnancy rate when mares were submitted to corticosteroid therapy (0.0 vs. 64.5%, respectively; p<0.05). Corticosteroid therapy was shown to be safe, with no physical or reproductive alterations on treated mares, demonstrating to be an adequate option to those animals with history of post-breeding or post-insemination endometritis. Further clinical research is necessary to confirm these results and contribute to the establishment of preventive therapy for cases of post-insemination endometritis.

Uterine and luteal hemodynamic evaluation of the non pregnant mare

Background: Studies with Doppler ultrassonography started at the end of the 90s for the determination of physiological and pathological alterations in the reproductive tract of the mare. Uterine alterations caused by inflammation, response from seminal plasma infusion, hormonal variations during estrous and diestrus, pregnancy and action of various vasoactive factors influence on the vascular perfusion detected by Doppler ultrasound. The development of efficient methods for uterine quality evaluation is of big importance for field equine reproduction veterinarians, once uterine environment is responsible for pregnancy maintenance. Review: Nowadays, the most used methods of uterine evaluation are the mode B ultrassonography, cytology, culture and biopsy. Hemodynamic evaluation of the uterus can be done by spectral data collected from large vessels, as A. uterine and its ramifications, or from subjective or objective evaluations from endometrium, miometrium and mesometrium attachment, which provide data referent to local and specific alterations of the evaluated area. Alterations in uterine vascular perfusion has been detected during estrous cycle, during pregnancy and in cases of infusion of inflammatory substances. These alterations happen because of vasoactive substances that act in the uterus during these events, however, most of these vasoactive substances are probably not even known. Also, important hemodynamic alterations in old mares, as an increase in vascular resistance, have been described. This increase might result from fibrosis of the uterus and in women it is considered to be a cause of infertility. In mares, periglandular fibrosis of the endometrium is considered to be the major diagnosable cause of embryonic and fetal loss in older mares. For the CL, ovarian artery of the mare supplies the ovary as well as the oviduct and therefore can be used for evaluation of these areas. The CL evaluation can also be done by the percentage of luteum area with colored signals as an indicator of the extent of blood flow. The percentage of the CL area with colored signals is determined subjectively by images observations in real time and/or by a freezing Power Doppler cross-section image with the maximum number of color pixels taped and the total number of color pixels is assessed by a computer analysis system. Therefore, a high correlation between plasmatic progesterone and CL vascularization also allows the CL evalution by this technique. In a first report, CL circulation reached its maximum on D5, the progesterone concentration in peripheral blood increased until D7 and in a posterior report, maximum perfusion was achieved two days after the maximum progesterone concentration (D8). Blood flow reduced between D10-D14 some days before the plasma progesterone decrease and, during the luteolytic period (D15-D17), the decline in CL blood-flow area was greater than blood flow decrease. Conclusion: Doppler ultrassonography add knowledge about uterine viability and CL functionality can be easily used by veterinarians in the field. It is a noninvasive method that provides real time results. However, because of the short time studies in this area have been done, many other answers still need to be found until normal and pathological patterns will be established.

An Integrative Genomic and Transcriptomic Analysis Reveals Potential Targets Associated with Cell Proliferation in Uterine Leiomyomas

Background: Uterine Leiomyomas (ULs) are the most common benign tumours affecting women of reproductive age. ULs represent a major problem in public health, as they are the main indication for hysterectomy. Approximately 40-50% of ULs have non-random cytogenetic abnormalities, and half of ULs may have copy number alterations (CNAs). Gene expression microarrays studies have demonstrated that cell proliferation genes act in response to growth factors and steroids. However, only a few genes mapping to CNAs regions were found to be associated with ULs. Methodology: We applied an integrative analysis using genomic and transcriptomic data to identify the pathways and molecular markers associated with ULs. Fifty-one fresh frozen specimens were evaluated by array CGH (JISTIC) and gene expression microarrays (SAM). The CONEXIC algorithm was applied to integrate the data. Principal Findings: The integrated analysis identified the top 30 significant genes (P<0.01), which comprised genes associated with cancer, whereas the protein-protein interaction analysis indicated a strong association between FANCA and BRCA1. Functional in silico analysis revealed target molecules for drugs involved in cell proliferation, including FGFR1 and IGFBP5. Transcriptional and protein analyses showed that FGFR1 (P = 0.006 and P<0.01, respectively) and IGFBP5 (P = 0.0002 and P = 0.006, respectively) were up-regulated in the tumours when compared with the adjacent normal myometrium. Conclusions: The integrative genomic and transcriptomic approach indicated that FGFR1 and IGFBP5 amplification, as well as the consequent up-regulation of the protein products, plays an important role in the aetiology of ULs and thus provides data for potential drug therapies development to target genes associated with cellular proliferation in ULs. © 2013 Cirilo et al.

Dose-Dependent Effects and Reversibility of the Injuries Caused by Nandrolone Decanoate in Uterine Tissue and Fertility of Rats

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

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.

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)

Perinatal exposure to androgen excess and the effects on the rat uterine estradiol responsiveness

Androgen exposure during sexual development induces alterations in steroidal target tissues. The objective of this study was to evaluate the uterine responsiveness to estradiol after perinatal androgenization. Pregnant Wistar rats were exposed to corn oil or testosterone propionate at 0.05, 0.1, or 0.2 mg/kg from gestational day 12 until postnatal day 21. Female offspring was challenged with estradiol (E2 ) after weaning (0.4 mg/kg) and at adulthood (10 or 100 µg/day), when the pituitary response was also evaluated. At adulthood, control and 0.05 mg/kg groups presented a uterine weight increment when exposed to 100 µg/day of E2 , 0.1 mg/kg group only responded to 10 µg/day of E2 , and the 0.2 mg/kg group showed increased uterine weight at both doses. The pituitary weight was similarly increased after estradiol stimulation in all experimental groups. In conclusion, testosterone propionate exposure induced an abnormal stimulation of uterine tissue growth by estrogen stimulus without affecting pituitary response. More studies are needed to clarify whether these alterations are capable of impairing the reproductive capacity of the female tract. © 2015 Wiley Periodicals, Inc. Environ Toxicol, 2015.

Acute phase response following ovariohysterectomy in female dogs

The concentrations of acute phase proteins are correlated with the severity of the surgical trauma, being elevated when severe tissue injury is produced. The aim of this study was to evaluate the leukogram and the acute phase proteins concentrations in healthy female dogs submitted to minimally invasive ovariohysterectomy, with the use of nylon cable ties (G1), and conventional (laparotomy) ovariohysterectomy, with the use of nylon suture (G2), as methods to ligate the ovarian pedicles and uterine body, respectively. Blood samples from 30 adult healthy female dogs (15 for each group) were obtained before surgery, and at 24 and 48 h and 7 days after surgery. Serum protein fractions were determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis. Neutrophilia was observed 24 h after both surgical procedures but did not differ (P ≥ 0.05) between the groups. In G1, the estimated concentration of ceruloplasmin increased (P ≤ 0.05) 48 h after the surgical procedure. The estimated ceruloplasmin concentration was significantly higher in G1 (P ≤ 0.05) when compared to G2 48 h postoperatively, and the estimated haptoglobin concentration was also significantly greater in G1 (P ≤ 0.05) than in G2 from 24 h to 7 days after surgery. Through the ceruloplasmin and haptoglobin estimated concentrations, we conclude that the minimally invasive ovariohysterectomy caused a more intense inflammatory response, which was not reflected in the white blood cell count.