Reproduction

Review on canine pyometra, oxidative stress and current trends in diagnostics Rupali Rautela, Rahul Katiyar Asian Pacific Journal of Reproduction 2019 8(2):45-55 Pyometra is one of the most important and common disorders in canine. The disease results in life threatening condition associated with septicemia and toxemia. The condition commonly occurs during luteal phase of estrous cycle, generally 4 weeks to 4 months of estrous cycle. The age, parity, breeds, administration of hormones are some predisposing factors of the condition. However, interaction between potentially pathogenic bacteria and hormonally primed uterus is believed to result in pyometra. The disease is initiated in the form of cystic endometrial hyperplasia, which later progresses into purulent uterine content following bacterial infection resulting in pyometra. The disturbance in antioxidant and pro-oxidant balance succeeding bacterial infection results in oxidative stress. The resultant stress further induces endometrial degeneration, immunosuppression and additionally aggravates the condition. An important clinical sign is malodorous, sanguineous or mucopurulent vaginal discharge with general symptoms such as vomiting, polyuria, polydipsia, depression, anorexia and occasionally fever. Till date, ultrasonography is the best diagnostic method while radiology and serological test are also helpful. Recently, the estimation of inflammatory mediators (acute phase proteins and prostaglandin metabolites) has appeared as marker for diagnosis of the pyometra. Determination of levels of inflammatory mediators has emerged as an important diagnostic method because it can be helpful in prognosis of the condition. The pyometra can be best treated by surgical manipulation (ovariohysterectomy); however, hormonal treatment can be given in less severe cases which aimed at reducing the effect of progesterone on the reproductive tract. Together, provision of antioxidants could be helpful in disease recovery along with the determined treatment. In this review, incidence, predisposing factors, etio-pathogenesis, oxidative stress associated with condition, diagnostic methods and treatment are discussed.

Effect of combination of Gynura procumbens aqueous extract and Trigona spp. honey on fertility and libido of streptozotocin-induced hyperglycaemic male rats Khaidatul Akmar, Mahanem Mat Noor Asian Pacific Journal of Reproduction 2019 8(2):56-62 Objective: To study the effects of co-administration of Gynura procumbens (GP) and kelulut honey (KH) on male fertility and libido in diabetes-induced rats. Methods: A total 42 males Sprague Dawley rats aged 8 weeks were randomly and equally divided into six different groups. All groups except a normal control group were induced with 50 mg/kg of streptozotocin (STZ) intravenously to induce diabetes. A positive control group was treated with an antidiabetic drug, metformin (500 mg/kg) whereas a negative control group remained untreated throughout the experiment. Meanwhile, another three treatments on diabetic rat groups were performed and categorised as Group 1 (450 mg/kg GP + 300 mg/kg KH), Group 2 (450 mg/kg GP + 600 mg/kg KH) and Group 3 (450 mg/kg GP + 1 200 mg/kg KH). Treatments were given for seven consecutive days through oral gavage and all rats were euthanized on day 8th for fasting blood glucose analysis, sperm quality, spermatogenesis, sexual behaviour and implantation sites analysis. Results: Fasting blood glucose levels were significantly reduced after treatment of GP and KH, compared to negative and positive controls. The treated groups showed significant increment in sperm quality compared to all control groups. Testes histology illustrated significant damages on leydig and sertoli cells for both negative and positive controls. On the contrary, co-administration of GP and KH displayed regeneration of leydig and sertoli cells in the testes. Additionally, the number of implantation sites significantly increased in females copulated with treated groups, compared to controls. Besides, the libido analysis displayed improvement of libido in treated groups, compared to all controls. Throughout the study, insignificant variances were recorded between the treated groups, indicating that treatment in Group 1 was sufficient to trigger significant improvement on fasting blood glucose level, fertility, and libido in diabetic male rats. Conclusions: Co-administration of GP and KH has great potential to serve as a pro-fertility agent amongst diabetic patients.

Effect of Vitex agnus-castus plant extract on polycystic ovary syndrome complications in experimental rat model Amal H Hamza, Widad M AlBishri, Mona H Alfaris Asian Pacific Journal of Reproduction 2019 8(2):63-69 Objective: To investigate ameliorative effects of Vitex agnus-castus (VAC) and VAC containing pharmaceutical supplement (VPS) against polycystic ovary syndrome (PCOS). Methods: PCOS in the rats was induced by daily administration of letrozole at 1 mg/kg body weight concentration for 21 d. PCOS rats were then treated daily either with metformin, VAC plant extract or VPS at 70, 8 or 8 mg/kg body weight concentration for 15 d. Rats that received none of these treatments were considered as control. Blood and ovaries were collected from all the rats. Serum glucose, cholesterol, triglyceride and high density lipoprotein-cholesterol were measured spectrophotometrically. Serum insulin, estrogen, progesterone, testosterone, luteinizing hormone, follicle-stimulation hormone, catalase, superoxide dismutase, malondialdehide and reduced glutathione were measured using enzyme-linked immunosorbent assay. Results: Rats treated with letrozole demonstrated a significant increase in serum testosterone, estrogen, cholesterol, luteinizing hormone, triglycerides, glucose, insulin, and malondialdehide levels, and a significant decline in progesterone, follicle-stimulating hormone, high density lipoprotein-cholesterol, catalase and reduced glutathione levels compared to control. Contrarily, no significant change in superoxide dismutase was noted in response to letrozole treatment. Rats treated with metformin, VAC or VPS showed a remarkable reversal in the levels of parameters affected by letrozole treatment. Conclusions: Data indicate that VAC and VPS exert potential ameliorative effects against PCOS through the modulation of hormonal and lipid profile as well as oxidative stress. Moreover, the favorable effects of these compounds are comparable to that of metformin.

Improvement of Phaseolus vulgaris on breastfeeding in female rats Shiva Roshankhah, Cyrus Jalili, Mohammad Reza Salahshoor Asian Pacific Journal of Reproduction 2019 8(2):70-74 Objective: To evaluate effect of Phaseolus vulgaris (P. vulgaris) on the breastfeeding in female rats. Methods: This experimental study was done from May 2018 to December 2018 in the Anatomical Department of Medical School in Kermanshah University of Medical Sciences in Iran. In this study, after one-week adaptation and fertilization by male, 40 female rats within 20 days of pregnancy (on average, every mother had 10 newborns) were equally separated into four groups (animals were administrated after delivery of offspring). Group 1 was control group receiving normal saline interaperitoneally, and groups 2, 3, 4 were treatment groups receiving the dose of 20, 50, 100 mg/kg of P. vulgaris interaperitoneally respectively once a day for 60 days. The prolactin hormone was measured by radio immune assay, number and diameter of alveoli via histological and morphometrical examinations, and receptor prolactin gene expression by real-time polymerase chain reaction method. Results: P. vulgaris significantly improved alveoli’s number and diameter, prolactin hormone and receptor prolactin expression when compared to the control group (P<0.001). Conclusions: P. vulgaris is helpful to improve the breastfeeding parameters of rats’ mammary glands.

Oestrous cycle of Wistar rats altered by sterol and triterpenes rich fraction of Adansonia digitata (Linn) root bark - A scientific rationale for contraceptive use Chibuogwu Ijeoma Chika, Nwannenna Agnes Ifeyinwa, Ubah Simon Azubuike, Ogwu David Asian Pacific Journal of Reproduction 2019 8(2):75-82 Objective: To investigate the effect of different fractions from the aqueous methanolic root bark of Adansonia digitata (Linn) on the estrous cycle of female Wistar rats and to elucidate their phytochemical content. Methods: Crude aqueous methanol extract of Adansonia digitata (Linn) root bark was fractionated by column chromatography using various ratios of hexane, acetyl acetate, methanol and distilled water. Fractions with similar trails on a thin layer chromatography were pooled into four fractions. Each fraction was then screened for its phytochemical content. Forty mature Wistar female rats weighing (135±28) g with regular estrous cycles were given each fraction at two dose levels while a control group received distilled water of equal volume. Treatments were given per os for 15 days starting from the day of estrus. Vaginal smears were observed daily for the duration of each cycle, frequency of occurrence of each stage of the estrous cycle and number of cycles for 15 days before and during treatment. At the end of treatment, rats were humanely sacrificed. Ovaries and uteri were weighed and processed for histopathology. Results: Hexane acetyl acetate and acetyl acetate methanol fractions at 150 mg/kg body weight and 300 mg/kg body weight prolonged the occurrence of proestrus and diestrus and produced atretic cyst-like ovaries at 300 mg/kg body weight. Acetyl acetate methanol treated rats also had low relative ovarian weights when compared to the control. Conclusions: Sterols and triterpenes in hexane acetyl acetate and acetyl acetate methanol fractions of the root bark may be responsible for the disruption of estrous cycle in female Wistar rats and may have a contraceptive effect.

Comparison of p38 MAPK, soluble endoglin and endothelin-1 level in severe preeclampsia and HELLP syndrome patients Efendi Lukas, Maisuri T Chalid, Upik A Miskad, Syakib Bakri Asian Pacific Journal of Reproduction 2019 8(2):83-87 Objective: To compare the levels of p38 mitogen-activated protein kinase (MAPK), soluble endoglin and endothelin-1 in the serum of patients with severe preeclampsia, hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome and normal pregnancies. Methods: This study was an observational analytic cross-sectional study performed at Wahidin Sudirohusodo Hospital, Makassar, Indonesia, in the period of 5th February 2016 to 20th January 2017. P38 MAPK, soluble endoglin and endothelin-1 levels of patients with normal pregnancies, severe preeclampsia and HELLP syndrome were measured by enzyme-linked immunoabsorbentassay technique, using kits of human soluble endoglin, endothelin-1 and p38 MAPK, Quantikine immunoassay: R&D System Inc. Results: Level of serum p38 MAPK in HELLP syndrome group was higher than in severe preeclampsia and normal pregnancy groups. Soluble endoglin and endothelin-1 levels in pregnancies with severe preeclampsia and HELLP syndrome were higher than normal pregnancy but there was no significant difference between these two groups (P>0.05). Levels of p38 MAPK, soluble endoglin and endothelin-1 also had a positive linear correlation with systolic and diastolic blood pressures (P<0.05). Conclusions: P38 MAPK in serum may be a marker for evidence of the severe hypoxia and its application may be considered for the diagnosis of HELLP syndrome.

Secondary sex ratio of assisted reproductive technology babies Nathira Abdul Majeed, Charley Zheng, Alex Polyakov, Megan Pucci, Mohamed Hatta Tarmizi, Mie Mie Cho Win Asian Pacific Journal of Reproduction 2019 8(2):88-92 Objective: To assess the secondary sex ratio (SSR) of assisted reproduction technology babies within a fertility clinic in Melbourne, Australia. Methods: A retrospective cohort study was conducted on 3 369 babies who were born following single embryo transfer between 2011 and 2016. Variables examined included embryo creation date, maternal and paternal ages, maternal body mass index (BMI), type of infertility, sperm parameters, fertilization method, type of embryo and stage of transfer. Multivariate regression analysis was performed on the data set using STRATA V9.2. Results: More males were found to be born to embryos created in summer (adjusted odds ratio=1.46, 95% confidence interval: 1.08-1.95; P=0.01). Lower BMI of mother (≤30 kg/m2) and younger age of parents (≤35 years) were associated with an increase in SSR (50.1% vs. 47.1%; 50.6% vs. 48.8%; 50.0% vs. 48.9%) respectively. Decreased SSR value was associated with primary subfertility, using in-vitro fertilization procedure and fresh embryo compared to those with secondary subfertility, using intracytoplasmic sperm injection procedure and thawed embryo (49.3% vs. 50.6%; 47.1% vs. 50.8%; 49.4% vs. 50.1%) respectively. Conclusions: The fertilization methods, type of embryo, stage of embryo transfer, parameters of the sperm and status of subfertility do not significantly affect the SSR. SSR is affected by the seasonality, maternal age, and BMI.