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.

Comportamento da artéria oftálmica no peuerpério imediato de pacientes com pré-eclâmpsia grave

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

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)

Inducing ovulation early postpartum influences uterine health and fertility in dairy cows

The objective of the current study was to evaluate the effect of GnRH early postpartum on induction of ovulation, uterine health, and fertility in dairy cows. Holstein cows without a corpus luteum (CL) at 17 +/- 3 DIM were assigned randomly to receive i.m. GnRH (n = 245) at 17 +/- 3 and 20 +/- 3 DIM or remain as controls (n = 245). Ovaries were scanned by ultrasonography twice weekly totaling 4 examinations. Ovulation was characterized by the appearance of a CL >= 20 mm at any ultrasound or CL <20 mm in 2 consecutive examinations. Clinical and cytological endometritis were diagnosed at 35 DIM. Compared with control, GnRH increased ovulation up to 3.5 d after the last treatment (78.7 vs. 45.0%) and did not affect the prevalence of clinical endometritis (23.9 vs. 18.6%) or cytological endometritis (30.9 vs. 32.8%). Prevalence of clinical endometritis increased in cows that had calving problems (32.6 vs. 15.9%) and metritis (40.6 vs. 15.8%). Metritis increased prevalence of cytological endometritis (50.7 vs. 23.5%). Treatment with GnRH did not affect pregnancy per artificial insemination at 32 (37.6 vs. 38.6%) or 74 d after artificial insemination (35.0 vs. 31.5%), but reduced pregnancy loss (6.8 vs. 18.1%). No overall effect of GnRH treatment on hazard of pregnancy was observed; however, an interaction between GnRH treatment and ovulation showed that GnRH-treated cows that ovulated had increased hazard of pregnancy by 300 DIM compared with GnRH-treated and control cows that did not ovulate (hazard ratio = 2.0 and 1.3, respectively), but similar to control cows that ovulated (hazard ratio = 1.1). Gonadotropin-releasing hormone early postpartiim induced ovulation without affecting uterine health, but failed to improve pregnancy per artificial insemination or time to pregnancy, although it reduced pregnancy loss.

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.

Effect of age and endometrial degenerative changes on uterine blood flow during early gestation in mares

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

Uterine vascular perfusion and spectral-doppler measurements during early gestation in mares: new concepts of evaluation

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

HMGA2 overexpression is associated with differential expression of miRNAs in uterine leiomyomas

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

Ganho de peso de novilhas mestiças submetidas ou não à esterilização pela introdução intrauterina de esferas inoxidáveis e ao uso ou não de modificador orgânico

Avaliou-se o ganho de peso de novilhas mestiças, 1/4 Simental e 3/4 Nelore, empregando-se o método experimental de esterilização, que consiste na introdução intrauterina de esferas inoxidáveis. Foram utilizadas 100 novilhas nulíparas, destinadas ao abate, com idades entre 12 e 24 meses e com média de peso de 275kg. Todos os animais receberam o mesmo manejo alimentar, em sistema de pastejo em Brachiaria brizantha, com água e sal mineral ad libitum, e pesagens a intervalos de 28 dias, obedecido o jejum prévio de 16 horas. Os animais foram distribuídos aleatoriamente em quatro grupos (G) experimentais: G1 - composto por 25 novilhas testemunhas; G2 - por 25 novilhas esterilizadas e aplicação de um modificador orgânico; G3 - por 25 novilhas esterilizadas; e G4 - por 25 novilhas não esterilizadas e aplicação de um modificador orgânico. Foram introduzidas 12 esferas de aço inoxidável, previamente esterilizadas, no útero de cada novilha. Houve maior ganho de peso total e diário entre os animais do G2, 140,50kg e 0,578g/dia vs 108,58kg e 0,447g/dia (G1), 103,73kg e 0,427g/dia (G3), 102,68kg e 0,423g/dia (G4), respectivamente. Esta técnica pode ser recomendada aos criadores.

Morfologia do útero de cutias nulíparas e não nulíparas

Descreveu-se a morfologia do útero de cutias nulíparas e não nulíparas por meio de ovariossalpingoisterectomia. A análise macroscópica do útero foi realizada in loco e na peça retirada. Segmentos das porções proximal, média e distal do órgão foram colhidas, fixadas e preparadas histologicamente, sendo as amostras analisadas à microscopia de luz, e realizada a histomorfometria das camadas uterinas. Topograficamente, o útero localiza-se na região sublombar, caudalmente aos rins, em continuação aos ovários e às tubas uterinas, estendendo-se até à entrada da pelve, onde se posiciona dorsalmente à bexiga. Caracteriza-se como do tipo duplo, embora culmine em apenas um óstio cervical externo. Microscopicamente, a mucosa uterina é formada por variações do epitélio, de cilíndrico a pseudoestratificado, que se apoia em tecido conjuntivo frouxo, onde se observam glândulas endometriais revestidas por epitélio cilíndrico, além da presença de vasos sanguíneos. A camada muscular subdivide-se em interna ou submucosa, média ou vascular e externa ou subserosa. A camada serosa é composta por tecido conjuntivo e mesotélio. Na histomorfometria, verificou-se que a espessura uterina total e a espessura da camada mucosa, em média, foram maiores nas fêmeas não nulíparas.

Histologia e morfometria das glândulas das junções infundíbulo-magno e útero-vagina de codorna doméstica

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

Possible role of bovine trophoblast giant cells in transplacental transmission of Neospora caninum in cattle

Neospora caninum is an aplicomplexan parasite that has brought several concerns to cattle raisers worldwide due to its relationship to fetal loss. However, the mechanism of the parasite's transplacental infection and induced abortions are not completely understood. Bovine trophoblastic binucleated cells (BNC) play a major role in the maternal-fetal interactions, migrating during the entire pregnancy from chorionic connections to uterine epithelium. This study aimed to investigate the possible role of BNC as phagocytic cells and its participation in the bovine transplacental infection of N. caninum. BNC was isolated by discontinuous Percoll gradient, and characterized by Hoeschst 33342 nucleus-specific staining. Isolated BNC were cultured in DMEM supplemented with 10% bovine fetal serum, and infected with 10(4) tachyzoites of N. caninum NC-1 strain. Parasite invasion was visualized by indirect immunofluorescence and Giemsa technique. Multiplication of parasites took place in 2-3 day cycles. Healthy cows' placenta and normal and infected cultured BNC was immunostained with monoclonal antibodies against CD-163, MAC-387 and NOS, demonstrating their phagocyte capacity. Thus, BNC was characterized as cells with macrophagic activity, which may host N. caninum in vitro. Therefore, we may conclude that BNC could potentially participate in the transplacental infection of bovine neosporosis.