Evaluation of glucose metabolism in women with multiple ovarian follicles

Objective ：To investigate glucose metabolism in women with multiple ovarian follicles （MOF） and explore the relationship between glucose metabolism, insulin resistance and body weight. Methods：We evaluated 46 women with MFO and 30 normal women as controls. All the subjects were given 75g of glucose orally in order to perform the oral glucose tolerance test （OGTT） and insulin releasing test （IRT）, and they were also evaluated for insulin resistance using the insulin resistance index with homeostatic model assessment （HOMA）. Results：The occurrence of impaired glucose tolerance in women with MOF was 10.87%, which was significantly higher than that in the control group （3.33% ,P 〈 0.05）. The rate of insulin resistance was 30.43% in the study group as compared to 10.00% in the control group. The results showed that there was significant difference between the two groups（P 〈 0.05）. The levels of FSH,LH,PRL,E2,T and P between the two groups had no significant difference （P 〉 0.05）. BMI in women with impaired glucose tolerance was correlated positively to insulin resistance （r = 0.567, P 〈 0.05）. Conclusion：Abnormal glucose metabolism was observed in women with unitary multiple ovarian follicles, and this could be attributed to obesity and insulin resistance. Women with MOF and associated obesity should be subjected to OGTT so that their glucose levels can be monitored as a preventive measure.

Quercetin modulates ovarian autophagy-related molecules and stereological parameters in a rat model of PCOS

Objective:To examine the effect of quercetin on stereological parameters and autophagy-related genes in ovaries of polycystic ovary syndrome(PCOS)rats.Methods:Fifty female Sprague-Dawley rats were randomly divided into five groups:the control group,the ethanol group,the quercetin group(15 mg/kg/day),the PCOS group,as well as the PCOS+quercetin group.After the induction of PCOS,quercetin was administered orally for 30 days.Histological,stereological and real-time PCR analyses were carried out to evaluate the effect of quercetin on PCOS rats.Results:Stereological analysis revealed that quercetin significantly increased the number of ovarian follicles and the volume of corpus luteum and induced a significant decrease in atretic follicles in comparison to the PCOS group.In addition,quercetin markedly increased mTOR gene expression while decreasing Beclin-1 and LC3 gene expression.Conclusions:Quercetin strongly modulates the expression of ovarian autophagy-related genes and stereological parameters in PCOS rats.Therefore,it can be considered as an ameliorative component for ovarian follicular impairments.

Expression of VEGF165 and VEGF165b during ovarian follicular development

Objective:To investigate the role of vascular endothelial growth factor(VEGF)165a,VEGF165b,and VEGF receptor(VEGFR)in the development of bovine follicles.Methods:We cultured follicular cells that were collected from small,medium,and large sized bovine follicles with estrogen and measured the expression of VEGF,VEGFR2 and VEGF165b by Western blot analysis and immunofluorescence.Results:The expression of VEGF165 increased in all follicle sizes and the expression of VEGF165b was increased in the small and large follicles after culturing in an estrogen containing medium.The expression of VEGFR2 was increased in the medium and large follicles after culturing with estrogen for 96 h.VEGF165 was activated at 100 ng/mL estrogen in the large follicles for 96 h.In addition,VEGFR2 was upregulated in the medium and large follicles after treated with 100 ng/mL estrogen for 96 h.Conclusions:This evidence suggests that the expression of VEGF165 and VEGFR is associated with estrogen stimulation during the development of bovine follicles and in an autocrine or paracrine manner.This reveals an advantage during oocyte maturation in vitro.

Ovarian Cysts in Users of Two Kinds of Progestin-only Contraceptives Implants

Objective To evaluate the prevalence of functional ovarian cysts in users of two different types of contraceptive implants. Methods A total of 239 women were enrolled at 3 months of use of the etonogestrelreleasing implant （Implanon） and the levonorgestrel-releasing implant （Jadelle）. Bimanual pelvic examination and vaginal ultrasound were performed during routine 3, 6 and 12-month visits of asymptomatic women（control group）. Women with ovarian cysts （or enlarged ovarian follicles 〉25 mm） （cysts group） were assessed weekly until disappearence or reduction of the image （including estradiol （E2） and progesterone measurement and women with no ovarian enlargement underwent same evaluation for the same period of time. Results Ovarian cysts were detected in 5.1% and 13.0% of users of Implanon and Jadelle, respectively, at 3rd month. At the 6th month of use, prevalences were 7.1% and 7.8%, and at 12th month rates were 25.7% and 14.7% in the two groups, respectively. E2 levels were significantly higher in cysts group than in control group. The time until disappearance of the ovarian cyst was similar in Implanon and Jadelle group. There were more cases of menorrhagia in patients rveth ovarian cysts than in patients with no ovarian enlargement. Conclusions The finding of ovarian cysts or enlarged ovarian follicles during the first year of use of Implanon and Jadelle implants is common and transient and should not be interpreted as a pathologic ovarian cyst. No further medical interventions are necessary.

Extracellular matrix-templating fibrous hydrogels promote ovarian tissue remodeling and oocyte growth

Synthetic matrices which mimic the extracellular composition of native tissue create a comprehensive model for studying development and disease.Here,we have engineered a composite material which retains cell-secreted ECM for the culture of ovarian follicles by embedding electrospun dextran fibers functionalized with basement membrane binder(BMB)peptide in PEG hydrogels.In the presence of ECM-sequestering fibers,encapsulated immature primordial follicles and ovarian stromal cells aggregated into large organoid-like structures with dense deposition of laminin,perlecan,and collagen I,leading to steroidogenesis and significantly greater rates of oocyte survival and growth.We determined that cell aggregation restored key cell-cell interactions critical for oocyte survival,whereas oocyte growth was dependent on cell-matrix interactions achieved in the presence of BMB.Here we have shown that sequestration and retention of cell-secreted ECM along synthetic fibers mimics fibrous ECM structure and restores the cell-cell and cell-matrix interactions critical for engineering an artificial ovary.

Iodoacetic acid exposure alters the transcriptome in mouse ovarian antral follicles

Iodoacetic acid(IAA) is an unregulated water disinfection byproduct that is an ovarian toxicant. However, the mechanisms of action underlying IAA toxicity in ovarian follicles remain unclear. Thus, we determined whether IAA alters gene expression in ovarian follicles in mice. Adult female mice were dosed with water or IAA(10 or 500 mg/L) in the water for 35-40 days. Antral follicles were collected for RNA-sequencing analysis and sera were collected to measure estradiol. RNA-sequencing analysis identified 1063 differentially expressed genes(DEGs) in the 10 and 500 mg/L IAA groups(false discovery rate FDR < 0.1), respectively, compared to controls. Gene Ontology Enrichment analysis showed that DEGs were involved with RNA processing and regulation of angiogenesis(10 mg/L) and the cell cycle and cell division(500 mg/L). Pathway Enrichment analysis showed that DEGs were involved in the phosphatidylinositol 3-kinase and protein kinase B(PI3K-Akt), gonadotropin-releasing hormone(Gn RH), estrogen, and insulin signaling pathways(10 mg/L). Pathway Enrichment analysis showed that DEGs were involved in the oocyte meiosis, Gn RH, and oxytocin signaling pathways(500 mg/L). RNA-sequencing analysis identified 809 DEGs when comparing the 500 and 10 mg/L IAA groups(FDR < 0.1). DEGs were related to ribosome, translation, m RNA processing, oxidative phosphorylation, chromosome, cell cycle, cell division, protein folding, and the oxytocin signaling pathway. Moreover, IAA exposure significantly decreased estradiol levels(500 mg/L) compared to control. This study identified key candidate genes and pathways involved in IAA toxicity and can help to further understand the molecular mechanisms of IAA toxicity in ovarian follicles.

Apoptosis in Granulosa cells during follicular atresia:relationship with steroids and insulin-like growth factors

It is well known that during mammalian ovarian follicular development, the majority of follicles undergo atresia at various stages of their development. However, the mechanisms controlling this selection process remain unknown. In this study, we investigated apoptosis in granulosa cells during goat follicular atresia by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The changes in the levels of steroids, insulin-like growth factors (IGFs) and IGF receptors were studied by radioimmunoassay (RIA) and semi-quantitative reverse transcrip-tion-PCR. We found that the percentage of apoptotic granulosa cells in the atretic (A) follicles was significantly higher than that in the slightly atretic (SA) and healthy (H) follicles. The level of estradiol and the ratio of estradiol to progesterone in H follicles were significantly higher than those in A follicles. On the other hand, the level of progesterone was not significantly different among these follicle types. We also found that the level of IGF-Ⅰ in H follicles was higher than in SA and A follicles, whereas the amount of IGF-Ⅱ did not vary significantly. The expression of IGF receptor also decreased in A follicles as compared to that in H and SA follicles. These results suggested that estradiol and IGF-Ⅰ might be involved in controlling apoptosis in granulosa cells during follicular atresia.

Ultrastructural and enzymatic alterations in the ovary of Rhodnius prolixus infected with Trypanosoma rangeli

Objective:To investigate the morphological structure of ovarian follicular cells and biochemical parameters of both ovaries and fat bodies(sites of vitellogenesis)from Rhodnius(R.)prolixus infected with Trypanosoma(T.)rangeli.Methods:Adult virgin females of R.prolixus were fed upon a membrane apparatus containing heat-inactivated citrated rabbit blood and a suspension of T.rangeli epimastigotes(Macias strain).Females from the control group and all the males received parasite-free blood.Transmission electron microscopy was used to reveal the morphological aspects of ovarian follicle cells in both control and parasite-infected groups.Protein profile,proteolytic activities and Western blotting analyses were performed in either ovary or fat body samples of control and parasite-infected groups.Results:According to the ultrastructural data,T.rangeli infection elicited a degeneration process in the ovarian follicular cells of R.prolixus.Proteolytic assays indicated a reduction in the activity of aspartic peptidases in the ovary and fat body from parasite-infected group,while a significant increase in the cysteine peptidase activity was measured in both insect organs.Additionally,immunoblotting revealed that vitellogenin was overexpressed in the ovary of parasite-infected insects.Conclusions:T.rangeli infection seems to elicit an early programmed cell death in the ovarian follicle cells as well as induces the modulation on the activities of different peptidase classes in either ovaries or fat bodies and the overexpression of the vitellogenin in the ovary of R.prolixus.

Localization of cystathionine β synthase in mice ovaries and its expression profile during follicular development

Background In vitro fertilization （IVF） researches have suggested that cystathionine β synthase （CBS） is involved in oocyte development. However, little is known about the regional and cellular expression patterns of CBS in the ovary. The purpose of this study was to analyze the localization of CBS in mice ovaries and to investigate the expression profile during follicular development. Methods We used in situ hybridization and immunohistochemical analysis to determine CBS expression in the ovaries of female Balb/c mice. Then the follicles were collected from F1 （C57BL×Balb/c） mice and cultured in vitro. With the method of semi-quantitative RT-PCR, we also investigated the expression profile of CBS during follicular development. Results CBS was absent in the oocytes, although it was ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, CBS expression was markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte. The semi-quantitative RT-PCR showed that CBS mRNA was detected in follicles at all stages in vitro. In cumulus-oocyte complexes superovulated, CBS expression also increased rapidly. Conclusions CBS was located mainly in the follicular cells in the ovaries. The level of CBS expression is high in follicles during folliculogenesis in mice. Differences in the CBS expression profile between oocyte and follicular cells suggest a role for CBS as a mediator in interactions between oocyte and granulosa cells.

Involvement of Quebracho tannins in diet alters productive and reproductive efficiency of postpartum buffalo cows

This study was conducted to investigate the effects of 10 weeks supplementation of Quebracho tannins(QT; 0 [control], 100 [QT100] or 200 g/[cow$d] [QT_(200)]) to 30 multiparous postpartum buffalo cows(10 cows per group) on milk yield and composition, blood metabolites and reproductive performance.Supplementation of QT100 had no significant effect on milk yield, whereas QT_(200) decreased(P < 0.05) this trait. Compared with the control group, both QT levels decreased(P < 0.05) fat-corrected milk(FCM)yield, but no significant effects were found on percentages of milk fat and protein. Contrariwise, yields of milk fat, lactose and milk protein were decreased(P < 0.05) when QT_(200) was supplemented. The solids nonfat(SNF) percentage and yield were decreased(P < 0.05) with QT100 supplementation. Moreover, QT tended to numerically reduce total number of ovarian follicles, number of small follicles, peripheral progesterone concentration and conception rate. Supplementation of QT_(200) numerically increased number of large follicles, mean diameter of large follicle, number and diameters of corpora lutea. The inclusion of QT_(200) shortened days open(DO) and decreased number of services per conception.Contrariwise, QT did not show significant effects on serum total protein, albumin, globulin, glucose,cholesterol and triglycerides concentrations. Supplementation of QT100 caused an increase(P < 0.05) of serum urea compared with that in control and QT_(200) groups. Generally, QT decreased(P < 0.05) serum creatinine concentration. Therefore, the supplementation of a commercial QT to early lactating Egyptian buffalo cows displayed negative consequences on their productive and reproductive performances.

Changes of Serum Follicular Stimulating Hormone,Luteinizing Hormone and Follicular Sizes duringElectroacupuncture for Induction of Ovulation

Changes in follicular stimulating hormone (FSH) . luteinizing hormone (LH) and follicularsizes were observed in 10 patients with chronic anovulation during electroacupuncture treatment. Sevencases were diagnosed as suffering from polycystic ovary syndrome, 2 from dysfunctional uterine bleeding,and 1 from hypogonadotropic amenorrhea. Among them 8 cases complained of infertility for an average of2. 7years. Ovulation was confirmed by pregnancy or the combination of biphasic basal body temperatureand ultrasonographic signs. During one course of 3 consecutive days of electroacupuncture treatment onacupoints Guanyuan (Ren 4), Zhongji (Ren 3), Zigong (Extra 16) and Sanyingjiao (SP 6). ovulation re-sulted in 5 patients (ovulating group) and 3 of the 4 infertile women became pregnant. Five cases failed toovulate (non-ovulating group) , 3 of them reached a biphasic basal body temperature without ovulatorysigns on ultrasonograph. Serum FSH, LH values and FSH pulse frequency increased significantly during-electroacupuncture treatment in the ovulating group (from 2. 10±0. 42 pulses/4h to 3. 70±1. 64 pulses/4h), but not in the non-ovulating group. No apparent change was found in LH pulse frequency, or in pulseamplitudes for FSH and LH. In the ovulating group, diameters of ovarian follicules markedly increased,but their growth was limited in the non-ovulation group. The results suggest ovulation may be induced byelectroacupuncture via a regulation on the hypothalamic-pituitary function, leading to normal secretion ofFSH and LH.