Asian Pacific Journal of Reproduction

252-261

Authors: Monalisa Biswas, Vijetha Shenoy Belle, Nihaal Maripini, Krishnananda Prabhu

Pregnancy associated diseases/disorders are associated with significant maternal and neonatal morbidities and mortalities. Devising/validating cost effective and easily accessible predictive, diagnostic and risk stratification markers are critical to the management and improved outcome in these diseases. Inflammation forms the backbone of most of the routinely encountered maternal complications of pregnancy. Hematological markers can be considered as a direct reflection of the systemic inflammatory milieu. Recently, the neutrophil lymphocyte ratio has been explored for its potential to assess the severity of inflammation and thus the severity of the underlying disorder. The neutrophil-lymphocyte ratio has gained scientific attention as a potential prognostic/predictive marker of acute as well as chronic inflammatory diseases including gynecological and reproductive disorders. This present study reviews the mechanistic role of neutrophils and lymphocytes in fueling or propagating the inflammatory cascades in the three most common maternal complications of pregnancy and the evidence of clinical importance of the neutrophil to lymphocyte ratio in predicting, diagnosing, and prognosticating pregnancy-associated complications.

269-273

Authors: Maryam Sadat Jalili, Samira Asadollahi, Seyed Morteza Seifati, Hamid Reza Ashrafzadeh, Nasrin Ghasemi

Objective: To investigate the frequency of -786T>C variant in endothelial nitric oxide synthase (eNOS) gene promoter in Iranian women with recurrent pregnancy loss. Methods: Blood samples were obtained from 100 unrelated women affected by recurrent pregnancy loss and 100 unaffected women as the controls. Genomic DNA was extracted and -786T>C polymorphism in eNOS gene promoter was investigated by PCR-RFLP method. Statistical analyses and Hardy-Weinberg equilibrium in the groups of patients and controls were performed by Chi-square test and SPSS standard software (Version 21). Results: The frequency of homozygous TT was 40% in cases and 46% in the control group; the frequency of CC was 7% in cases and 5% in the control group; frequency heterozygote TC was 53% in cases and 49% in the control group. Genotype frequencies between the two groups showed no significant differences (P>0.05). Conclusions: The -786T>C polymorphism is not more frequent in recurrent pregnancy loss in this population.

284-290

Authors: Seyyed Shamsadin Athari, Keivan Lorian, Haniye Kashafroodi, Saadat Ghafarzadeh, Rasool Choopani

Objective: To evaluate the protective effects of honey compound syrup on sperm count and testis tissue in rats. Methods: Thirty rats were randomly assigned to five groups. The control group received 1 mL normal saline with dimethyl sulfoxide intraperitoneally; the busulfan group received busulfan 10 mg/kg body weight at the first and twenty-first days of the experiment via intraperitoneal injection; the last three groups received busulfan 10 mg/kg body weight to induce azoospermia, and then received 1.0, 1.5, or 2.0 mg/kg honey compound syrup, respectively, after induction of azoospermia. After administration, the testis and epididymis of all rats were removed. Then, reproductive organ weight and sperm parameters (sperm concentration, epididymal sperm reserve and daily sperm production) were measured. After hematoxylin-eosin staining, seminiferous tubule cells and diameters were assessed. Results: Busulfan damaged the testis tissue and impaired spermatogenesis. Administration of honey compound syrup in three doses improved testis tissue and spermatogenesis. The protective effects of honey compound syrup may relate to the antioxidant properties of honey and other compounds in this syrup. Conclusions: Administration of honey compound syrup could be an ameliorative agent for the side effects of chemotherapy drugs such as busulfan on the male reproductive system.

203-214

Authors: Awanish Jaiswal, Anurag Pandey, Mamta Tiwari, Akhtar Ali, Rohit Sharma

According to the latest data, globally 15% of couples have infertility and male infertility contributes to 10% of all cases. Infertility can be caused by certain biological changes in the gonads and the reproductive system like azoospermia, oligospermia, asthenospermia, teratozoospermia and hypospermatogenesis. Genetic causes of azoospermia include chromosomal abnormalities, Y chromosome microdeletions and deletion or other mutations of Y-linked genes. The maximum number of the genes are located in the azoospermia factor region of the long arm (Yq) of the Y chromosome. Y chromosome microdeletion is known as the second major genetic cause of spermatogenetic failure. This article aims to review the latest updates on the involvement of Yq microdeletions in male infertility. The diagnostics, prevalence and phenotypic spectrum related to Yq gene microdeletions are discussed.

225-231

Authors: Anjaneyababu Naik Banavath, Sridevi Nangali Srinivasa

Objective: To study the effect of mobile emitted radio frequency electromagnetic radiation (RF-EMR) on sperm parameters (sperm count, viability, motility, progressivity, and morphology) and ameliorative effect of pomegranate juice in rats. Methods: Thirty male Sprague Dawley rats were divided into five groups (n=6 in each group). Group I was neither exposed to RF-EMR nor given pomegranate juice, group II, III and IV were exposed to mobile emitted RF-EMR for 60 min/day for 90 days. After 90-day exposure to RF-EMR, group III was supplemented with pomegranate juice for 90 days (1 mL/day) and group IV was allowed to recover for 90 days without supplementation of pomegranate juice. Group V was supplemented with pomegranate juice for 90 days without exposure to RF-EMR. At the end of intervention, epididymal sperm parameters (sperm count, viability, motility, progressivity, and morphology) were measured. Results: The microscopic examination of sperm parameters such as sperm count, sperm viability, sperm motility, progressivity were significantly decreased in group II (the mobile RF-EMR group) compared to group I (the control group) (P<0.05). In addition, sperm morphology was also significantly altered (abnormal) in group II compared to group I (P<0.05). However, the sperm parameters including sperm morphology were significantly altered in group III (mobile RF-EMR + pomegranate juice) compared to group II (P<0.05). The sperm parameters including sperm morphology were not significantly altered in group IV (the mobile RF-EMR recovery group) compared to group II (P>0.05). The sperm parameters were non-significantly increased in group V (the pomegranate juice group) compared to group I (P>0.05). Conclusions: Mobile RF-EMR exposure reduces the sperm parameters, which, however, can be reversed by treatment with pomegranate juice, indicating that pomegranate juice can be used as a nutritional supplement to improve sperm quality.

239-240

Authors: Mário Sousa, Jan Tesarik

The terms syngamy, pronucleus, pronuclear breakdown and zygote are common for experts dealing with reproductive cell morphology. However, these terms are often not correctly used in scientific and didactic literature.

102-112

Authors: Arijit Chakraborty

To prevent iodine deficiency disorders, the universal salt iodization programme has been introduced all over the globe, including environmentally iodine sufficient regions irrespective of their iodine status. As a result, iodine-induced thyroid dysfunctions namely hyperthyroidism, hypothyroidism, autoimmune thyroid diseases, endemic goiter and even thyroid cancer including infertility, still births, abortions and embryo toxicity have emerged as a major public health problem. In other words, the consequence of iodine deficiency and excess is almost ‘U’-shaped. Hypothyroidism caused by iodine deficiency affects reproductive functions of organisms; however, such undesirable effects of iodine overload on male gonadal physiology together with hormonal profiles are yet to be adequately explored. The discovery of iodide transporter in the testis justifies an independent role of iodine in male reproductive function, which is not entirely known. Recent studies on human subjects and animal models are now revealing further perceptions into the effect of excess iodine on male infertility with euthyroid status. Excess iodine exposure has been linked with deterioration of structural and functional changes of testis leading to compromised spermatogenesis by affecting various cellular and molecular signaling pathways culminating into disrupted the blood-testis barrier and cytoskeleton. This review provides an update and summarizes various novel insights of excess iodine exposure on reproduction by establishing the independent role of iodine on male reproductive endocrinology, which might help in formulating future strategies to prevent iodine-induced male infertility, an emerging global concern, especially in the post-salt iodization era.

121-126

Authors: Zhila Khodabandeh, Vahid Nejati, Ali Shalizar-Jalali, Gholamreza Najafi, Fatemeh Rahmani

Objective: To scrutinize the protective role of royal jelly as an anti- oxidant on nicotine-induced changes in malondialdehyde (MDA) level, p53 expression, in vitro fertilization (IVF) rate, and early embryo development in adult female rats. Methods: A total of 56 adult female Wistar rats were divided into 8 groups (n=7 in each group). Group 1 served as an untreated control group, group 2, 3 and 4 received nicotine at a dose of 0.50, 1.00 and 2.00 mg/kg respectively, group 5 received royal jelly at a dose of 100.00 mg/kg, and group 6, 7 and 8 received 0.50, 1.00 and 2.00 mg/kg nicotine, respectively, with 100.00 mg/kg body weight royal jelly. Nicotine and royal jelly were administered daily for 49 days in the experimental groups intra-peritoneally and orally, respectively. At the end of the experimental period, p53 expression, IVF rate and early embryo development as well as MDA concentration were measured. Results: The IVF rate, number of cumulus oocytes, two-cell embryos and blastocysts decreased in the nicotine-treated groups in a dose-dependent manner. In addition, p53 mRNA expression and MDA levels increased in the nicotine-treated groups. Royal jelly co-administration led to partial improvement in the aforementioned parameters. Conclusions: Royal jelly may have a repro-protective effect in nicotine-administered female rats in terms of its anti-oxidant and anti-apoptotic properties.

137-144

Authors: Mimi-Sophia Sarbandi, Nor-Ashikin Mohamed Noor Khan, Nor-Shahida Abdul Rahman, Zolkapli Eshak, Fathiah Abdullah, Mastura Abd Malek, Aqila-Akmal Mohammad Kamal

Objective: To observe the effects of vitamin E on post-vitrification preimplantation development, gross morphology as well as mitochondrial distribution and ultrastructure. Methods: Twenty-four female C57BL/6NTac mice, aged 12-16 weeks, were randomly divided into four groups. Group A did not receive any treatment and served as the control group. Group B was treated with corn oil stripped of tocopherols and served as the vehicle group. Group C was treated with 60 mg/kg body weight of tocotrienol-rich-fraction with corn oil stripped of tocopherols. Group D was treated with 60 mg/kg body weight of alpha-tocopherol with corn oil stripped of tocopherols. All treatments were administered orally for 7 consecutive days. After superovulation and mating with fertile males, 2-cell stage embryos were harvested for vitrification. Post vitrification development in vitro, gross morphology and ultrastructure were compared between groups. Results: The number of 2 and 8-cell embryo, and blastocysts in the treatment and control groups were not significantly different (P>0.05). Following vitrification, all 2-cell embryos had equal-sized blastomeres and intact zona pellucida. Mitochondrial aggregation toward the perinuclear region was seen in all of the treatment groups. Both groups C and D had vacuolated mitochondria, which was reflected in the trend of preimplantation development reduction. Conclusions: Vitamin E supplementation of 60 mg/kg body weight does not improve the viability of healthy embryos according to this study. As a result, the most effective dose of vitamin E supplementation may be determined by the initial quality of the embryos.

11-20

Authors: Temidayo S. Omolaoye, Shantal Windvogel, Stefan S. du Plessis

Objective: To investigate the testicular oxidative stress and apoptosis status, as well as the sperm functional parameters in streptozotocin (STZ) induced diabetic rats following treatment with rooibos (Aspalathus linearis), honeybush (Cyclopia intermedia) and sutherlandia (Lessertia frutescens) infusions. Methods: Diabetes was induced by injecting fourteen-week-old adult male Wistar rats (250-300 g) with a single intraperitoneal injection of STZ (45 mg/kg body weight). Fifty rats were randomly divided into five groups: the vehicle group received 0.1 M citrate buffer, the diabetic control group received 45 mg/kg STZ, the diabetic+rooibos group received 45 mg/kg STZ + 2.0% rooibos, the diabetic+honeybush group received 45 mg/kg STZ + 4.0% honeybush, and the diabetic+sutherlandia group received 45 mg/kg STZ + 0.2% sutherlandia. Rats were sacrificed 7 weeks after induction of diabetes mellitus. The testes and epididymides were harvested and weighed after induction. Spermatozoa were retrieved from the cauda epididymis for motility, concentration, and morphology analysis, and the testis was used for all biochemical assays. Oxidative stress was determined by measuring malondialdehyde levels, catalase, and superoxide dismutase activities, while apoptotic biomarkers were evaluated by Western blotting assays. Results: After induction of diabetes, rats in the diabetic control group, diabetic+rooibos group, diabetic+honeybush group, and diabetic+sutherlandia group presented with significantly elevated blood glucose levels as compared with the vehicle group (P<0.001). Rats in the diabetic control group had a reduction in sperm progressive motility, while rats in the diabetic+rooibos group and the diabetic+sutherlandia group displayed an increase in progressive motility as compared with the diabetic control group. The diabetic control animals showed a 40.0% decrease in sperm concentration when compared to the vehicle group, and there were no significant differences in sperm kinematic and speed parameters between the groups. In addition, the percentage of morphologically normal spermatozoa was increased by 13.0%, 16.0%, and 15.0% after treatment with rooibos, honeybush, and sutherlandia, respectively and the rats in the diabetic+infusion groups also displayed an increase in superoxide dismutase activity when compared to the diabetic control group. Conclusions: Rooibos, honeybush and sutherlandia infusions may partly alleviate diabetes-induced sperm function impairment by reducing oxidative stress.

29-35

Authors: Jonah Sydney Aprioku, Benjamin Toochukwu Okpe, Doupere Ben

Objective: To investigate the effects of 30-day treatment with therapeutic dose equivalent levels of tramadol on serum testosterone level, sperm parameters, and testicular histology in rats. Methods: Thirty-five Wistar rats were equally divided into seven groups. Group 1 (the control group) received distilled water (0.5 mL) daily for 30 days. Groups 2-4 were gavaged with therapeutic dose equivalent levels of tramadol (1.25, 2.50 and 5.00 mg/kg/day body weight, respectively) in two equal divided doses for 30 consecutive days, and sacrificed on day 31. Groups 5-7 received similar tramadol treatments as above but they were allowed for another 30 days to recover after receiving the last dose and sacrificed on day 61 for reversibility study. Serum testosterone level and epididymal sperm were analyzed, and histopathological examination of the testis was carried out. Results: Tramadol treatment significantly decreased serum testosterone levels compared with the control group. Furthermore, tramadol treatment inhibited sperm motility and significantly and dose-dependently decreased sperm count and viability compared with the control group. In addition, tramadol significantly increased morphological abnormalities in sperm (P<0.05). The above effects of tramadol were reduced in the reversible groups. Testis histopathological examination revealed disintegrated cell architecture, eroded and atrophied seminiferous tubules, and a marked decrease in the number of spermatogenic cells in the tramadol treated groups. The histopathological changes were restored in the reversible groups, but improvement was not complete in the 5.00 mg/kg tramadol treated reversible group. Conclusions: Long term treatment with tramadol at clinical dose levels may adversely affect testosterone level, sperm parameters, and testicular histology, but they are reversible at lower doses.

43-48

Authors: Kambulu Lukusa, Abubeker Hassen, Khoboso C. Lehloenya

Objective: To evaluate the effect of cooling rates, selenium supplementation, and three semen extenders on cooled and frozenthawed Saanen buck sperm. Methods: Twenty Saanen bucks were divided into two groups: the selenium supplemented group and the control group. Ejaculates were collected once weekly by artificial vagina. The first experiment examined the effects of cooling rates and selenium supplementation on semen characteristics of Saanen bucks. Pooled semen was diluted with triladyl extender and split into two aliquots for slow and fast cooling. The second experiment explored the effect of selenium supplementation and semen extenders on post-cryopreserved sperm quality. Ejaculates from each group were divided into three aliquots and diluted with three extenders (i.e. clarified egg-yolk, whole egg yolk, Tris without egg-yolk extenders). All samples were cooled for 2 h at 4 曟 and frozen at −196 曟 for 24 h. The sperm characteristics such as sperm motility, acrosome integrity, normal morphology, and viability were evaluated by using a phase-contrast microscope. Results: In the first experiment, all sperm characteristics were significantly increased in selenium supplemented samples when slow cooling was used (P<0.05). In the second experiment, in cooled semen, sperm motility and viability were significantly increased in both clarified egg yolk and Tris without egg yolk extenders in selenium supplemented samples as compared with whole egg yolk extender, respectively (P<0.05). After freezing-thawing, all sperm parameters of selenium supplemented samples in clarified egg yolk extender were significantly greater than those in Tris without egg yolk extender (P<0.05). However, normal morphology and acrosome integrity of selenium supplemented samples in whole egg yolk extender were similar to those of selenium supplemented samples in clarified egg yolk extender. Conclusions: The characteristics of cooled and post-cryopreserved sperm are greater when clarified egg yolk extender is used in semen from selenium supplemented bucks.

155-161

Authors: Maria Ruth B. Pineda-Cortel, Karlo Baybayan, Peter Louie Bello, Latiffa Lois Camenforte, Stefany Jane Ching, Kathleen Conti, Jeremiah Jose Ignacio, Jiovanni Diaz, Adrian Villavieja, Jefferyl Kae Pandac, Emilyn U. Alejandro

Objective: To investigate the possible association between rs7754840 and rs7756992 polymorphisms of CDKAL1 gene and susceptibility to gestational diabetes mellitus (GDM) in a Filipino pregnant population. Methods: A total of 101 patients with GDM and 99 women without GDM were included. Two CDKAL1 gene single nucleotide polymorphisms (SNPs), namely rs7754840 and rs7756992, were genotyped by using TaqMan allelic discrimination assays. Mann￾ Whitney U test, median and interquartile range were used to describe physical and biochemical characteristics. The differences in the genotype and allele distribution of the target genetic variants among the two groups of participants were assessed by using Chi￾ square test. Conformity to Hardy-Weinberg equilibrium was tested prior to conducting further analysis. Multiple logistic regression model was used to investigate the effects of the genotype models on GDM development. Results: There was no observed correlation between the genotypes of the rs7754840 SNP and oral glucose tolerance test parameters. Consequently, there was no significant association between genetic models of the rs7754840 SNP and GDM risk (additive OR 1.43, 95% CI 0.82-2.50, P=0.21; dominant OR 1.21, 95% CI 0.57-2.59, P=0.62; recessive OR 1.63, 95% CI 0.86-3.09, P=0.13). Conclusions:The results of this study suggest no association between CDKAL1 gene variant rs7754840 and GDM development in Filipino pregnant women. Further studies with a larger population should be performed to validate our findings.

176-186

Authors: Temidayo S. Omolaoye , Stefan S. du Plessis

Objective: To evaluate the testis and the cauda epididymis after treating both healthy and diabetic rats with rooibos (Aspalathus linearis), honeybush (Cyclopia intermedia), and sutherlandia(Lessertia frutescens), respectively. Methods:Ninety male Wistar rats were randomly divided into nine groups (n=10 per group). Fifty rats were randomized to receive tap water, 0.1 M citrate buffer, 2.0% fermented rooibos, 4.0% fermented honeybush, or 0.2% unfermented sutherlandia. Diabetes was induced via a single intraperitoneal injection of streptozotocin (STZ) at 45 mg/kg in 40 rats. The diabetic rats were randomized to receive tap water, 2.0% fermented rooibos, 4.0% fermented honeybush, or 0.2% unfermented sutherlandia. Rats were sacrificed after seven weeks of diabetic induction. Blood was collected for hormonal assay, while the testes and caudal epididymis were retrieved for histomorphological evaluations. Results: Diabetic rats treated with 2.0% fermented rooibos exhibited a significant increase in fasting plasma glucose (FPG) versus the vehicle group (P=0.002), and a 3.6% decrease compared to the diabetic control group (P>0.05). Diabetic rats treated with honeybush or sutherlandia showed 23.6% and 15.4% decrease in FPG when compared to the diabetic control group, respectively (both P>0.05). The diabetic control rats had a significant decrease in the percentage of histologically normal seminiferous tubules compared to the control group (P=0.049). There was 21.0%, 14.0%, and 5.4% rise in the percentage of normal seminiferous tubules in diabetic rats receiving rooibos, honeybush or sutherlandia, respectively, compared to the diabetic control group. The infusion control groups (rooibos, honeybush and sutherlandia) showed normal seminiferous tubules, presence of spermatozoa in the epididymal lumen, and had normal overall architecture. Both testicular and epididymal morphology were altered in the diabetic control group, but these disruptions were mildly ameliorated by rooibos, honeybush, and sutherlandia. Conclusions: The detrimental effects of diabetes on the histomorphological architecture of the testis and epididymis are mildly ameliorated by the infusions (2.0% fermented rooibos, 4.0% fermented honeybush, and 0.2% unfermented sutherlandia).