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Wednesday, April 10, 2019

Human Reproductive Sciences

From the editor's desk
Madhuri Patil

Journal of Human Reproductive Sciences 2019 12(1):1-3



Role of oxidative stress in male infertility: An updated review
Ahmed T Alahmar

Journal of Human Reproductive Sciences 2019 12(1):4-18

Current evidence links oxidative stress (OS) to male infertility, reduced sperm motility, sperm DNA damage and increased risk of recurrent abortions and genetic diseases. A review of PubMed, Medline, Google Scholar, and Cochrane review databases of published articles from years 2000–2018 was performed focusing on physiological and pathological consequences of reactive oxygen species (ROS), sperm DNA damage, OS tests, and the association between OS and male infertility, pregnancy and assisted reproductive techniques outcomes. Generation of ROS is essential for reproductive function, but OS is detrimental to fertility, pregnancy, and genetic status of the newborns. Further, there is a lack of consensus on selecting OS test, type, and duration of antioxidants treatment as well as on the target patients group. Developing advanced diagnostic and therapeutic options for OS is essential to improve fertility potential and limit genetic diseases transmitted to offspring. 


Proteins as the molecular markers of male fertility
Eswari Beeram, Bukke Suman, Bysani Divya

Journal of Human Reproductive Sciences 2019 12(1):19-23

Proteins play a key role in many functions such as metabolic activity, differentiation, as cargos, and cell fate regulators. It is necessary to know about the proteins involved in male fertility to develop remedies for the treatment of male infertility. However, the role of the proteins is not limited to particular aspect in the biological systems. Some of the proteins act as ion channels such as catsper, and protein such as Nanos is a translational repressor in germ cells and expressed in prenatal period whose role in male fertility is not clearly understood. Rbm5 is a pre-mRNA splicing factor necessary for sperm differentiation whose loss results in deficit in sperm production. DEFB114 is a beta-defensin family protein necessary for sperm motility in lipopolysaccharide-challenged mice. TEX101 is a plasma membrane specific germ cell protein whose function is not clearly identified. Gpr56 is an another adhesion protein whose null mutation leads to arrest of production of pupps. Amyloid precursor protein in Alzheimer's disease plays a role in male fertility whose function is uncertain which has to be considered while targeting them. The study on amyloid precursor protein in male fertility is a novel thing, but requires further study in correlation to Alzheimer's disease. 


Biological significance of glutathione S-transferases in human sperm cells
Roman V Fafula, Nataliya M Paranyak, Anna S Besedina, Dmytro Z Vorobets, Ulyana P Iefremova, Olena K Onufrovych, Zinoviy D Vorobets

Journal of Human Reproductive Sciences 2019 12(1):24-28

Background: Oxidative stress is considered as one of the causes of male subfertility or infertility. Among antioxidant enzymes, the crucial role belongs to glutathione S-transferases (GSTs). Data on the biological role of GSTs in the defense mechanisms of sperm cells in fertile and infertile men are limited. Aim: The aim of this study was to demonstrate the functional role of GSTs in sperm cells on the model of H2O2-induced stress on human ejaculated spermatozoa obtained from both normospermic and pathospermic patients. Subjects and Methods: We used a H2O2-induced stress on human ejaculated spermatozoa obtained from both normospermic and pathospermic patients. Results: Here, we report the effect of GST inhibitor ethacrynic acid on sperm motility and viability. Pharmacological inhibition of sperm GSTs activity leads to spermal membrane damage and rejected in the loss of motility and decrease of viability. For similar treatment conditions, thiobarbituric acid reactive substance (TBARS) levels increased significantly leading to decrease in sperm motility and viability. It is suggested that these functional impairments are related to the intensification of lipid peroxidation as expressed by TBARS levels in spermal membranes after GST inhibitor treatment. Conclusion: This study provides evidence that sperm GSTs are important in the defense mechanism against oxidative stress. Evaluation of GSTs activity in sperm cells of infertile men can be helpful in fertility assessment and for the evaluation of treatment by antioxidants. 


A study on chromosomal analysis of patients with primary amenorrhea
Asoke K Pal, Prafulla S Ambulkar, Bharat R Sontakke, Shweta S Talhar, Pradeep Bokariya, Vijay K Gujar

Journal of Human Reproductive Sciences 2019 12(1):29-34

Background: Primary amenorrhea is one of the most common disorders seen as gynecological problems in adolescent girls. It refers to the participants who did not attain menarche by the age of 11–15 years. Chromosome abnormalities contribute as one of the etiological factors in patients with primary amenorrhea. Aims: The aim of this study was to evaluate the frequency of chromosomal abnormalities and to investigate the abnormal karyotypes in patients referred with the symptom of primary amenorrhea for better management and counseling. Setting and Design: One hundred and seventy-four cases of primary amenorrhea were referred from the obstetrics and gynecology department to our cytogenetic laboratory for chromosomal analysis. G-banded chromosomes were karyotyped, and chromosomal analysis of all patients was done. Results: Out of 174 patients, we observed 23 (13.22%) participants with abnormal karyotype. In 23 cases of chromosomal abnormalities, 10 cases were sex reversal female (46,XY) and Turner karyotype (45,X) in 6 females. Other numerical and structural abnormalities were also seen such as 47,XXX; 45,X/47,XXX; 45,X/46, X,dic(X); 46,XX, inv (9); 45,X/46,X,i(Xq); 46,X,mar(X); and 45,X/46,XY in the primary amenorrhea cases. Conclusion: This study definitely attests the importance of chromosomal analysis in the etiologic diagnosis of primary amenorrhea patients. Karyotyping will help to counsel and manage the cases of primary amenorrhea in a better way. This study reveals the frequencies and different types of chromosomal abnormalities found in primary amenorrhea individuals and that might help to make the national database on primary amenorrhea in relation to chromosomal aberrations. 


Menstrual blood versus endometrial biopsy in detection of genital tuberculosis by using nested polymerase chain reaction in an endemic region
Lavina Chaubey, Deepak Kumar, Vidyut Prakash, Gopal Nath

Journal of Human Reproductive Sciences 2019 12(1):35-39

Aim: The aim of this study is to compare the results of nested polymerase chain reaction (PCR) for early detection of genital tuberculosis (GTB) using menstrual blood (MB) and endometrial tissue (ET) as samples in females presenting as infertility. Methods: The ET and MB samples were collected from a total of 194 females, enrolled in this study. DNA isolation from samples was done using standard, phenol–chloroform method. Heat shock protein gene (hsp65/groEL2) of Mycobacterium tuberculosis was targeted and amplified, and the final products were analyzed. Results: Overall, 126 (65%) cases of infertility were positive for M. tuberculosis complex by nested PCR. The detection rates in the two samples were statistically insignificant. The combined positivity rate of ET and MB, when compared with positivity rate in MB showed a positive predictive value, negative predictive value, and accuracy of 100%, 68.7%, and 84%, respectively. The results of nested PCR using MB as sample alone showed good agreement with the nested PCR results of the combined samples. Conclusions: The hsp65 Nested PCR of MB can be used as a noninvasive screening test for early diagnosis of GTB. 


Evaluation of oxidative stress and severity of endometriosis
Shaiesta Amreen, Pratap Kumar, Priyanka Gupta, Pragna Rao

Journal of Human Reproductive Sciences 2019 12(1):40-46

Aims: The aim was to evaluate the association between oxidative stress with the severity of endometriosis in blood and peritoneal fluid. Settings and Design: This study was a prospective observational study conducted in the department of obstetrics and gynecology in a tertiary center over 2 years in patients with endometriosis requiring laparoscopy. Methodology: Patients diagnosed with endometriosis and undergoing laparoscopy for infertility and/or chronic pelvic pain were included in the study. Out of the 64 patients recruited, only 55 patients formed the part of our study. Blood and peritoneal fluid was evaluated for oxidative stress parameters such as glutathione peroxidase, superoxide dismutase (SOD), and lipid peroxides. Results: The severity of endometriosis was assessed intraoperatively by the revised American Society for Reproductive Medicine scoring in all 55 patients who underwent laparoscopy into minimal (n = 3), mild (n = 7), moderate (n = 32), and severe (n = 17). Median activity of SOD and glutathione peroxidase was lowest in the severe stage of endometriosis, whereas the median activity of lipid peroxide was highest in the severe stage of endometriosis in both blood and peritoneal fluid samples. There was a statistically significant increase in oxidative stress with the severity of endometriosis. Conclusions: Oxidative stress might play a role in the disease process of endometriosis. Control of oxidative stress can be used as the latest treatment option for the management of endometriosis. 


Anxiety and stress at different stages of treatment in women undergoing In vitro fertilization–intracytoplasmic sperm injection
Manisha Awtani, Gitanjali Kaur Kapoor, Parveen Kaur, Jayeeta Saha, Diana Crasta, Manish Banker

Journal of Human Reproductive Sciences 2019 12(1):47-52

Aim: The aim of the present study is to evaluate the state anxiety (the present state of mind), trait anxiety (general anxiety), as well as perceived stress in women undergoing in vitro fertilization (IVF) treatment at three stages: T1 (on the day of start of stimulation), T2 (on the day of embryo transfer), and T3 (10 days after embryo transfer). The data at T3 level were collected telephonically. Methodology: The present study was carried out on 137 women undergoing IVF intracytoplasmic sperm injection cycle at four different clinics of four cities from October to April 2016. State-trait anxiety inventory (Spielberger) and perceived stress scale (Okun, et al.) were used as the tools. Statistical Analysis: The analysis was done at two levels; descriptive and inferential (analysis of variance [ANOVA], Student's t-test, Levene's test) using SPSS v16. Results: The state anxiety was higher at all the three levels than trait anxiety. The overt anxiety was highest at T3 level (mean = 45.77) followed by T1 level (mean = 44.23) and T2 level (mean = 43.04). Perceived stress was elevated at T1 level (mean = 17.93) followed by T3 level (mean = 17.28) and T2 level (mean = 16.72). The results of ANOVA showed a significant difference in anxiety among all the three levels (P = 0.036), but no significant difference was found for perceived stress (P = 0.169). t-test revealed that there was a significant difference between state and trait anxiety at T1, T2, and T3 levels (P = 0.01, P = 0.21, P = 0.00, respectively). A significant difference was only seen between the T1 and T2 levels in perceived stress (P = 0.052). In state anxiety, a significant difference was observed only between T2 and T3 levels (P = 0.23). Conclusion: It was observed that anxiety and stress are present in women throughout the treatment. The waiting period (T3) is the most anxious for them and their level of state anxiety is higher compared to their trait anxiety. Perceived stress is observed to be more on the day of start of stimulation followed by the waiting period. 


A multicenter, randomized, equivalence trial of a new recombinant human chorionic gonadotropin preparation versus ovitrelle® for ovulation in women undergoing intrauterine insemination following ovarian stimulation
Abha Majumdar, Rekha Sachan, Yogeshwar S Nandanwar, Rahul V Mayekar, Neelu Soni, Manish R Banker, Rajan S Vaidya, Manjeet Arora, Girish P Godbole, Gautam V Daftary, Ganesh H Divekar, James John

Journal of Human Reproductive Sciences 2019 12(1):53-58

Context: A new indigenous recombinant human chorionic gonadotropin (r-hCG) has been developed in India with a comparable pharmacological profile to that of Ovitrelle® (Merck Serono). Aims: This study aims to compare the efficacy and safety of the new r-hCG with that of Ovitrelle for induction of ovulation in women undergoing intrauterine insemination (IUI). Settings and Design: Randomized (2:1), multicenter, open-label, equivalence clinical trial conducted in India. Subjects and Methods: A total of 217 women, aged 20–37 years, undergoing IUI were administered the new r-hCG (test) 250 mcg or Ovitrelle 250 mcg (comparator) after ovarian stimulation with gonadotropins. The ovulation rate was compared as the primary outcome. In addition, pregnancy rates, incidence of adverse events (AEs), and development of immunogenicity were assessed. Statistical Analysis Used: The ovulation and pregnancy rates were compared using Chi-squared test with statistical significance at P < 0.05. Results: With 144 women in the test group and 73 in the comparator group, the ovulation rate (85.4% vs. 78.1%; P = 0.17) and pregnancy rate (serum β hCG test) (11.8% vs. 12.3%; P = 0.91) were similar in both groups. A total of 15 AEs were reported (11 in the test r-hCG group and 4 in the comparator group) in 11 women; none of these were serious, and all were judged to be unrelated to the study drug. No subject developed immunogenic reaction to the test drug. Conclusions: The new preparation of r-hCG was equivalent to the conventional preparation of r-HCG in the induction of ovulation in patients undergoing IUI. 


Personalized embryo transfer helps in improving In vitro fertilization/ICSI outcomes in patients with recurrent implantation failure
Jayesh A Patel, Azadeh J Patel, Jwal M Banker, Sandeep I Shah, Manish R Banker

Journal of Human Reproductive Sciences 2019 12(1):59-66

Aims: This study aims to compare clinical outcomes in patients of recurrent implantation failure (RIF), who had embryo transfer (ET) following a receptive (R) endometrial receptivity array (ERA) and a personalized embryo transfer (pET) after a nonreceptive (NR) ERA. Settings and Design: This was a retrospective observational study. Study Period: July 2013–September 2017. Subjects and Methods: Two hundred and forty-eight patients having unexplained RIF who underwent ERA test were included in the study. Clinical outcomes were compared between patients having a receptive (R) ERA and those having a NR ERA who underwent a pET-based on ERA. Statistical Analysis Used: Chi-square and t-test. Results: ERA predicted receptive (R) endometrium at P + 5 in 82.3% (204/248) patients and NR in 17.7% (44/248) patients. Average failed previous in vitro fertilization cycles were 3.67 ± 1.67 among receptive ERA patients and 4.09 ± 1.68 among NR ERA patients. Pregnancy rate (PR), clinical PR, implantation rate (IR), abortion rate (AR), ongoing pregnancy rate (OPR), and cumulative PR were comparable between patients having receptive ERA who had a routine Embryo Transfer (ET) and those with an NR ERA who underwent a pET. Conclusions: ERA is helpful in identifying the window of implantation (WOI) through genetic expressions of the endometrium to pinpoint embryo transfer timing. pET guided by ERA in patients of RIF with displaced WOI improves IRs and OPRs. 


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