Clinical use of Anti-Müllerian Hormone to Monitor Resumption of Ovarian Activity following Removal of a 4.7 mg Deslorelin Implant in Queens.

Objective: The use of deslorelin implantsto control reproduction in cats is increasing but because of its prolonged duration, cat breeders oftenrequest implant removal before the end of the treatment. Assaying Anti Mullerian Hormone (AMH) concentrations might be useful to predict time of resumption of ovarian activity in deslorelin-treated queens following implant removal. In queens a minimum of 3 weeks during increasing photoperiod after implant removal has been described for resumption of ovarian activity but no information about AMH concentrations were observed for determining ovarian activity.Animals: Sixteen queens in whichdeslorelinimplants were surgically removed after3, 6 or 9 months (n= 6, 4 and 6 queens, respectively) were used in this study. Procedures: A general and reproductive health check with a GnRH stimulation test were performed before the treatment.After implant removal queens were checked every 1-2 weeks withreproductive ultrasonography, a vaginal smear and blood collection to assay AMHconcentrations. Results: AMH concentrations decreased significantly during treatment to < 2.5+0.6 ng/ml (p<0.05) and reached a nadir at 1.9+0.9 (p<0.05) one-week post-removal.Following implant removal AMH concentrations started to rise reaching a value of 4.3+1.2 ng/ml on the third week and were not different from pre-treatment levels on week 6 post-removal (5.8 ng/ml +0.9, p>0.05).AMH values did not differ depending on duration of deslorelin treatment but were lower in adult queens (p<0.05).Clinical relevance: AMH assay can be a useful tool to follow resumption of feline ovarian function following a deslorelin treatment.

Piper longum (Linn.) restores ovarian function in Letrozole induced PCOS in Rats: Comparison with Metformin and Clomiphene citrate

This study evaluates the prospective use of an herbal plant Piper longum in letrozole induced polycystic ovary syndrome using a rat model. The study used female wistar rats, which were divided into 9 groups, each containing 6 animals. Group I (Control) daily received 1% Carboxy Methyl Cellulose (CMC)as a vehicle control. Letrozole (1mg/kg) was administered by oral route for period of 21days for induction of PCOS in group (II-IX). During experimental period, vaginal smear of all females were collected daily for the estrous cycle determination. During 28 days of letrozole administration, changes in estrous cycle of females were observed and studied. This study showed that PCOS was induced. After Letrozole treatment, 6 animals from group III-IX treated orally with, standard drugs Metformin (300mg/kg/oral route), Clomiphene citrate (100mg/kg/oral route), plant drug Piper longum L. at a concentration of 200mg/kg, 400mg/kg and 800mg/kg, drug interaction groups: Metformin + Piper longum L, 800mg/kg and Clomiphene citrate +Piper lonum L., 800mg/kg body weight and studied for consecutive estrous cycles. Vaginal smear were examined, it showed that hydro alcoholic extract of fruits of Piper longum Linn. group has potential effect on PCOS bringing estrous cycle to normalcy. Also, after Letrozole treatment ovary and reproductive weights of normal rats increased which is normalizes with plant drug treatment. Further studies of hydroalcoholic extract of fruits Piper longum Linn. need to be carried out to check other related parameters of PCOS.

Genotype distribution of cervical HPV among Caribbean women in a population-based study in Martinique: The DEPIPAPUFR study

The Caribbean ranks seventh among the world regions most affected by cervical cancer. HPV-prevalence and genotype distributions also differ from regions. Knowledge of HPV genotype profiles is important for patients care and HPV vaccination implementation. The objective of this study was to describe HPV genotype distribution and risk factors in a population-based cohort of women in Martinique. In this study, 1312 women were included and underwent cervical cancer screening with successful sample collection between 2009 and 2014. Sociodemographic and clinical variables were recorded. Cytological examination of cervical vaginal smear was performed and classified(Bethesda). Detection of HPV DNA was performed with the PapilloCheck© Kit from Greiner Bio-one. Genotypes were analyzed for18 high-risk HPV (hrHPV) and 6low-risk HPV(lrHPV) types. A total of 1075 women were included with a mean age of 49.1±10.5 years. HPV prevalence was 27.6% (297/1075) with 19.4% (209/1075) women with only hrHPV, 5.3% (57/1075) with only lrHPV. Multiple infections (hrHPV/lrHPV) were detected in 31/240 cases of hrHPV (12.9%). A total of 353 hrHPV genotypes were analyzed; the most common HPV types were HPV51 (11.0%), HPV68 (10.8%), HPV53 (9.1%) and HPV 52 (7.1%). HPV16 and HPV18 represented respectively 4.8% and 4.0% of hrHPV genotypes. Abnormal cytology was observed in 34 cases (3.2%), with 14 ASCUS (1.3%), 10 LSIL (0.9%), 5 HSIL (0.5%), 3 ASC-H (0.3%) and 2 AGC (0.2%). Fifteen (44.1%) were hrHPV and 4 (14.7%) lrHPV; 7 cases of hrPHV were in the age-group 25–34 years. Among 1041cases of normal cytology, 225 had positive hrHPV detection (21.6%). This is the first population-based study of HPV profiles in our country, and we found a high prevalence of hrHPV. The most common genotypes were HPV51, 68, 53. These results could serve for cancer vaccination strategies and HPV surveillance in Martinique.

Ameliorative Potential of Aqueous Extract of Broccoli Sprouts Against Triazophos Induced Ovarian Toxicity in Wistar Rats

Traditional therapeutic procedures using antioxidant-rich fruits and vegetables have been in vogue for the development of evidence-based biomarkers for assessing reproductive health. Present investigation was designed to study the antioxidative potential of broccoli sprouts aqueous extract (BE), against ovarian toxicity in female rats induced by triazophos (TZ). In the experimental setup, six groups of rats were formed; Control (group 1), BE (group 2), TZ (group 3), and also BE+TZ groups such as BE1 (group 4), BE2 (group 5) and BE3 (group 6) groups. Body weight was weekly recorded of all the rats, while vaginal smear was observed daily during 30 days experiment. After sacrifice, oxidative stress (OS) biomarkers levels viz; catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and lipid peroxidation (LPO) were determined along with histopathological and apoptotic observation. Results revealed differentially modified changes in OS biomarkers as CAT, SOD, GR, GPx, and GST, while LPO levels were significantly improved with broccoli supplementation compared to TZ group rats. Plasma progesterone and estradiol levels were also restored along with improved ovarian histoarchitecture among all BE+TZ treated rats. Reduced apoptotic granulosa cells with reduced atresia and normal ovarian surface epithelium height were also observed with BE treatment. BE exerts multi-mechanistic protective effects against TZ induced ovarian toxicity which is attributable to its antioxidant and protective actions.

Rhamnocitrin Ameliorates Ovarian Fibrosis Via PPARγ/TGF-β1/Smad Pathway to Repair Ovarian Function in Polycystic Ovary Syndrome Rats Running Title：rhamnocitrin Ameliorates Ovarian Fibrosis in Polycystic Ovary Syndrome Rats

Background: Polycystic ovary syndrome (PCOS) is one of the major endocrine disorders in women, characterized by androgen excess, chronic anovulation and ovarian fibrosis. Rhamnocitrin is an herbal bioactive flavonoid that has anti-inflammation and antioxidant effects. We intended to investigate the impacts of Rhamnocitrin on PCOS-induced ovarian fibrosis and its underlying mechanisms.Methods: Dehydroepiandrosterone (DHEA) induced-PCOS rats were treated with Rhamnocitrin. HE staining was performed to detect ovarian histological features. Ovarian fibrosis was evaluated by Sirius Red and Masson staining. Vaginal smear was examined to exhibit estrus cycle stage and vaginal cornification. The serum hormone levels of FSH, LH, E2 and T were measured with ELISA. The related mRNAs and proteins of fibrosis factors and PPARγ/TGF-β1/Smad2 signaling were detected by RT-qPCR and western blot. The weights of rat bodies and ovaries were recorded. PPARγ inhibitor T0070907 and its agonist GW1929 were employed for the mechanistic investigation.Results: The corpus luteum and follicles were increased and irregular estrous cycle was restored after Rhamnocitrin treatment in PCOS rats. Rhamnocitrin inhibited ovarian fibrosis and down-regulated the expressions of fibrotic factors. Rhamnocitrin reduced the increased levels of LH, E2, and T, and elevated the decreased FSH level in PCOS rats. Besides, Rhamnocitrin elevated the down-regulated PPARγ, and suppressed the up-regulated TGF-β1 and p-Smad2 expressions induced by PCOS. These effects of Rhamnocitrin on PCOS rats could be antagonized by T0070907, whereas GW1929 markedly mimics the functions of Rhamnocitrin. Conclusions: Rhamnocitrin ameliorates ovarian fibrosis in PCOS rats through regulation PPARγ/TGF-β1/Smad2pathway, suggesting it can be a potentially effective therapeutic candidate for PCOS treatment.

Pengaruh ekstrak daun binahong (Anredera cordifolia ) terhadap pemulihan siklus estrus mencit (Mus musculus)

This study aimed to determine the effect of binahong (Anredera cordifolia) leaf extract on the recovery of the estrous cycle of mice (Mus musculus) which has been induced for the extension of the estrous cycle using angsana (Pterocarpus indicus) bark extract. In this study vaginal smear was performed daily at 10.00 WIB to determine the phase of the estrous cycle for 26 days. Mice were divided into 5 groups, each with 6 replicates. The treatments were T1 (positive control), T2 (negative control), T3 (0.8 mg/head/day), T4 (2.4 mg/head/day) and T5 (4 mg/head/day). The data obtained were processed using Analysis of Variance (ANOVA) and followed by the Duncan's New Multiple Range Test (DNMRT) at a 95% level of significance. The results showed that the angsana (Pterocarpus indicus) bark extract adminsitration (T2) lengthen (p <0.05) the estrous cycle of mice compared to those of normal mice (T1). The administration of binahong (Anredera cordifolia) leaf extract on T3 and T4 groups mice were followed by shorter (p <0.05) estrous cycle than those of T2, and it were not significantly different (p >0.05) compared to T1 group. However, the higher dose of binahong (Anredera cordifolia) leaf extract (T5) revealed the length of estrous cycle was not significantly different (p >0.05) compared to T2 group. It can be concluded that the administration of binahong leaf extract was able to recover the normal estrous cycle of mice.

Loss of ASPM Disrupts Female Folliculogenesis in Mice

Background: The abnormal spindle-like, microcephaly-associated (ASPM) gene is a causative gene of autosomal recessive primary microcephaly (MCPH) 5 in humans, which is characterized by a reduction in brain volume. It was previously reported that truncated ASPM proteins in transgenic mice caused major defects in the germline, a severe reduction in ovary weight and the number of follicles accompanied by reduced fertility. However; it remains unknown whether a loss of ASPM induces abnormal ovarian function, resulting in female infertility. Methods: In order to assess the ovary function, we examined vaginal smear cytology from the age of 7 weeks to 100 weeks in CAG-mediated Cre-loxP conditional Aspm-/- knockout mice and control female mice. In addition, we evaluated the ovarian size, fibrosis ratio and the number of follicles (primordial, primary, secondary, antral and atretic follicles) in mice from 15 weeks to 100 weeks old by image analyses. Mann-Whitney U-test was used for statistical analysis. Results: The size of the ovary was significantly reduced in Aspm knockout mice at 15-20 weeks, 40-50 weeks and 70-80 weeks old. compared with the control mice. Furthermore, at all stages, we found a severe decrease in the number of developing follicles at 10-15 weeks, 40-50 weeks and 70-80 weeks old, accompanied by disrupted cyclic changes of vaginal cytology. Conclusion: The results showing that folliculogenesis was significantly decreased and associated with abnormal vaginal cytology in Aspm knockout female ovaries suggested that ASPM might play an important role in the folliculogenesis and estrous cyclicity of the postnatal ovary.

The Effect of Different Concentration of Aqueous Extracts of Labisia pumila in Preventing Osteoporosis and Improvement of Dermal Elasticity in Polycystic Ovary Syndrome Rats

Background: Labisia pumila (LP) a Malaysian herb thought to have phytoestrogenic effects in rats with PCOS. In this study we investigate the effects of LP aqueous extracts on bone composition, bone biomarkers and metabolic disorder in female rats fed continuously with high fat diet to induce PCOS. Hypothesis: The administration of Labisia pumila treatment in PCOS rats for 90 days will ameliorate the adverse effect of osteoporosis by reducing the inflammatory cytokine and improving the dermal elasticity of PCOS rats. Study design: On the 90th day of the development of PCOS model rats fed with high fat diet and after the vaginal smear analysis indicating a prolonged estrus cycle of more than 2 weeks, all PCOS rats were divided into 4 groups which consisted of control, placebo (water), LP25 (LP 25mg/kg) and LP50 (LP 50 mg/kg) respectively. All PCOS continue to received the formulated high fat diet and control animals continued to received normal chow and water ad libitum. Method: Vehicle and treatments, which were given orally by using stomach gavaging needle size 16 gauge straight and curve retrolingual administration. The rats were sacrificed at the end of the trial phase and organs, tissues and blood samples were harvested for multiple assays and analysis listed below.Results: The level of estradiol was significantly increased in LP25 and LP50 as compared with placebo. The inflammatory cytokine of C-reactive protein and TNF-ɑ were significantly decreased in LP25 and LP50.Conclusion: Treatment of LP extract might reduce the inflammatory cytokine related to the formation of osteoporosis and loss of bone mass, increase the insulin sensitivity and reduce of osteoporosis in PCOS patients. The phytoestrogenic of LP all of the above significant positive results are proven in lowering osteoporosis and metabolic disorder in PCOS rats.

P–589 The effects of vanillic acid on production of testosterone and function of insulin in adipose and ovarian tissues in rat model of polycyctic ovary syndrome

Study question What is the possible therapeutic effects of vanillic acid (VA) on the treatment of Polycystic Ovary Syndrome (PCOS)? Summary answer Vanillic acid successfully alleviated metabolic and endocrine abnormalities in adipose and ovarian tissues in a rat model of PCOS. What is known already Polycystic Ovary Syndrome (PCOS), as a common endocrine disorder, is accompanied by hyperandrogenism, insulin resistance, ovulation problems, and infertility. In this study, the effects of vanillic acid (VA) on metabolic and endocrine abnormalities were evaluated in the adipose and ovarian tissues of a letrozole-induced rat model of PCOS. Study design, size, duration In this study, a letrozole-induced rat model was established and then the experimental groups were treated by VA and MET through oral gavage for one month. Before and after the treatment with drugs, the vaginal smear was carried out . Ovarian and adipose tissues were collected and frozen at − 80 for further analysis following euthanasia of the rats. Participants/materials, setting, methods Thirty Six -week- old Wistar rats were divided into four groups (six rats per each group). Twenty-four rats were received letrozole for five weeks. Then, two experimental groups were treated by metformin (MET), and VA . Serum lipid profile, fasting glucose levels, and hormonal assays were done by corresponding commercial kits. The phosphorylation of AMP-activated protein kinase (AMPK) was detected by western blotting and the expression of studied genes were measured by Real-time PCR. Main results and the role of chance Our results showed that both VA and MET successfully reversed the abnormal estrous cycles of PCOS rats and reduced the serum testosterone levels and Steroid 17-alpha-hydroxylase/17,20 lyase (CYP17A1) gene expression. Furthermore, they improved Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) and decreased the serum glucose and triglyceride levels, and gene expression of resisitin. The phosphorylation of AMPK was significantly decreased in adipose and ovarian tissues in PCOS group and both therapeutic factors successfully activated the AMPK in these tissues. However, VA had not such a significant effect in adipose tissue. Limitations, reasons for caution The main limitation of the current study was its design as a rat model, which may have limitation in the translation of knowledge to the clinics. Wider implications of the findings: In summary, to the best of our knowledge, this study is the first study reported beneficial effects of VA on the treatment of PCOS. The findings of the current study shed light on an urgent need for discovering novel therapeutic pharmaceuticals regarding the treatment of the PCOS. Trial registration number 0

Expression of Vascular Endothelial Growth Factor, Matrix Metalloproteinase-2 and Matrix Metalloproteinase-9 in Polycystic Ovarian Syndrome Rats and Its Implication

Polycystic ovary syndrome (PCOS) is a female endocrine disorder and frequently leads to infertility. Vascular endothelial growth factor (VEGF) has crucial roles and matrix metalloproteinase (MMPs) is correlated with cell migration. Both of them are involved in the occurrence and progression of PCOS. This study established a rat PCOS model using letrozole to measure the expression of VEGF, MMP-2 and MMP-9 (MMP-2/9), to analyze its correlation with PCOS. Letrozole was applied by gavage to establish rat PCOS model. General condition and ovarian tissue morphology were observed under a light field microscope. ELISA and immunohistochemistry (IHC) were used to detect serum or tissue expression of VEGF, MMP-2/9. Estrous cycle of rats was disrupted after 12 d for using letrozole. Vaginal smear showed abundant leukocytes with sparse keratinocytes. Ovary showed whitening and increased volume, with early phase small follicles plus lower granular cells or corpus luteum. Compared to control group, experimental group had significantly higher VEGF, MMP-2/9 (P < 0.05), which were higher in antral follicles than those in preantral follicle with higher expressions than primordial follicle (P < 0.05). In conclusion, VEGF, MMP-2/9 are abundantly expressed in both serum and tissues of PCOS rats.