SIRT1 and antioxidants in infertile females: Exploration of the role of vitamin D.

BACKGROUND: Deficiency of silent information regulator 1 (SIRT1) can trigger inflammation, mitochondrial malfunctioning, and apoptosis through the hypothalamic-pituitary-ovarian axis, producing poor quality oocytes, leading to infertility. Normal vitamin D (VD) levels promote SIRT1 activity required for optimal fertility, and low levels of either may result in fertility problems owing to cell-membrane de-stabilization, increased autophagy, DNA damage leading to increased reactive oxygen species and mitochondrial dysfunction. Therefore, in this study, we want to estimate the levels of VD, SIRT1 and antioxidants (MnSOD; manganese superoxide dismutase, GR; glutathione reductase, visfatin) and oxidants (adrenaline & cortisol) in individuals living with infertility and explore the association of VD with SIRT1 expression (levels), antioxidants, and oxidants contributing to infertility in women. The significance of this study is that it highlights the importance of maintaining optimal levels of VD for reproductive health in females. METHODS: This cross-sectional study included 342 (135 infertile and 207 fertile) female subjects. Serum levels of MnSOD, SIRT1, visfatin, GR, VD, adrenaline, and cortisol were analyzed by ELISA and were compared in fertile and infertile samples using the Mann Whitney U test. RESULTS: There were significantly high levels of VD, SIRT1, GR, MnSOD and visfatin in fertile female participants. However, mean adrenaline and cortisol levels were higher in infertile samples with a significant negative correlation with VD. A significant negative correlation of VD with MnSOD, SIRT1, visfatin and GR was observed (p <0.01). In VD subset groups, MnSOD levels were significantly high in VD sufficient groups however, adrenaline and cortisol levels were significantly high in groups suffering from VD deficiency. CONCLUSIONS: Deficiency of VD is associated with a decrease in SIRT1 and other antioxidants, which may deter natural reproductive functions leading to infertility. Further studies are required to determine the cause-effect relationship of VD deficiency on conception and interpretation of the involved mechanism.

Multiple mini-interviews is a predictor of students' academic achievements in early undergraduate medical years: a retrospective study.

BACKGROUND: Our study determined Multiple Mini-Interview (MMI) effectiveness in evaluating specific skill sets based on medical students' performances during the undergraduate years and compared the academic performances of medical students who appeared for onsite/online MMI. METHODS: A retrospective study of 140 undergraduate medical students between 2016 and 2020 included data on age, gender, pre-university results, MMI scores, and examination results. Appropriate non-parametric tests were applied to compare the students' MMI and academic performances. RESULTS: Ninety-eight students from cohorts 12 to 15 had an overall MMI score of 69.0(IQR: 65.0-73.2)/100 and an overall Cumulative Grade Point Average(GPA) of 3.64 (3.42-3.78)/5.0. Spearman's correlation revealed a significantly positive relationship between MMI and cGPA (rho = 0.23) and GPA from the first 2 semesters (GPA1 rho = 0.25, GPA2 rho = 0.27). This observation was similar to that for station A in the first year (cGPA rho = 0.28, GPA1 rho = 0.34, GPA2 rho = 0.24), and in station B (GPA4 rho = 0.25) and D (GPA3 rho = 0.28, GPA4 rho = 0.24) in the second year. Of twenty-nine cohort16 students, 17(58.6%) underwent online and 12(41.4%) offline modes of MMI assessment, respectively. The overall median MMI score was 66.6(IQR: 58.6-71.6)/100, and the overall median cGPA was 3.45 (3.23-3.58)/5.0. When comparing the median marks of cohort16 groups, the online group scored significantly higher marks for station D than the offline group (p = 0.040). CONCLUSION: Correspondence between MMI scores and cGPA predicted MMI scoring during student selection and entry process might ensure the success of their academic performance in medical school.

Interplay between oxidative stress, SIRT1, reproductive and metabolic functions.

Silent information Regulators (SIRT1) gene stimulates antioxidants' expression, repairs cells damaged by oxidative stress (OS), and prevents the cells' dysfunction. In particular, the role of different Sirtuins, particularly SIRT1 in reproduction, has been widely studied over the past decade. Decreased SIRT 1 causes mitochondrial dysfunction by increasing Reactive Oxygen Species (ROS), lipid peroxidation, and DNA damage in both male and female gametes (Sperms and Oocytes), leading to infertility. In the female reproductive system, SIRT1 regulates proliferation and apoptosis in granulosa cells (GCs), and its down-regulation is associated with a reduced ovarian reserve. SIRT1 also modulates the stress response to OS in GCs by targeting a transcription factor vital for ovarian functions and maintenance. ROS-mediated damage to spermatozoa's motility and morphology is responsible for 30-80% of men's infertility cases. High levels of ROS can cause damage to deoxyribo nucleic acid (DNA) in the nucleus and mitochondria, lipid peroxidation, apoptosis, inactivation of enzymes, and oxidation of proteins in spermatozoa. SIRT 1 is a cardioprotective molecule that prevents atherosclerosis by modulating various mechanisms such as endothelial injury due to impaired nitric oxide (NO) production, inflammation, OS, and regulation of autophagy. SIRT 1 is abundantly expressed in tubular cells and podocytes. It is also found to be highly expressed in aquaporin 2 positive cells in the distal nephron suggesting its involvement in sodium and water handling. SIRT1 improves insulin resistance by reducing OS and regulating mitochondrial biogenesis and function. It also decreases adiposity and lipogenesis and increases fatty acid oxidation. So, its involvement in the multiple pathways ensures its unique role in reproductive and metabolic derangement mechanisms.

Myeloperoxidase: Indicator of cardiovascular disease in chronic kidney disease patients of tertiary care hospital of Karachi.

OBJECTIVE: To estimate the levels of myeloperoxidase in various stages of chronic kidney disease, and to correlate them with an inflammatory marker and lipid profile. METHOD: The cross-sectional study was conducted at the Biochemistry Department, Basic Medical Sciences Institute, in collaboration with the Nephrology Department, Jinnah Post Graduate Medical Centre, Karachi, from January 2013 to September 2014, and comprised chronic kidney disease patients and healthy controls. Serum cholesterol, triglycerides, high-density lipoprotein, C-reactive protein and myeloperoxidase levels were noted. Data was subjected to statistical analysis. RESULTS: Of the 150 subjects, 84(56%) were cases and 66(44%) were controls. Weight, body mass index, triglycerides, very low-density lipoprotein, C-reactive protein and myeloperoxidase levels were significantly higher among the cases compared to the controls (p<0.05). Serum myeloperoxidase had a significantly positive association with C-reactive protein (p<0.01), cholesterol (p<0.01), triglycerides (p<0.01), low-density lipoprotein (p<0.01) and very low-density lipoprotein (p<0.01), and had a negative correlation with high-density lipoprotein (p<0.01). CONCLUSIONS: Myeloperoxidase concentration had association with lipid profile and C-reactive protein.

Suggested role of silent information regulator 1 (SIRT1) gene in female infertility: A cross-sectional study in Pakistan.

AIM & OBJECTIVE: Silent information regulator 1 (SIRT1) gene stimulates the expression of antioxidants and repairs damaged cells. It affects the mitochondrial activity within the oocytes to overcome the oxidant stress. We aimed to assess an association of SIRT1 polymorphism (Tag SNPs rs10509291 and rs12778366) with fertility, and assess serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), oestradiol, progesterone, manganese superoxide (MnSOD) and SIRT1. MATERIAL AND METHODS: In this cross-sectional study, 207 fertile and 135 infertile subjects between the ages of 18-45 years were recruited. Polymerase chain reaction (PCR) was performed; products were electrophoresed in a 2% agarose gel. Descriptive analysis of continuous variables was expressed as mean ± standard deviation. Mann-Whitney test was performed for comparison of groups, P value <.001 was considered significant. Single Nucleotide Polymorphism (SNP) data were analysed by applying chi-squared statistics. RESULTS: All subjects were age matched (P = .896). SIRT1 levels were significantly lower in infertile females when compared with fertile subjects (P < .001). AA (rs10509291) and CC (rs12778366) variant frequency was higher in the infertile than fertile subjects (P < .01). Similarly, the frequency of A allele (rs10509291) and C allele (rs12778366) was higher in infertile subjects (P < .001). Infertile females (29%) showed existence of SNP rs10509291 while 49% demonstrated genetic variation of rs12778366. MnSOD and SIRT1 levels were found to be lower in these subjects. CONCLUSION: The presence of SIRT1 genetic variants (rs10509291 and rs12778366) apparently disturbs the expression of SIRT1 deteriorating mitochondrial antioxidant function within the oocytes, instigating oxidative stress within. Their probable effect on modulating oocyte maturation may be the cause of infertility in females.

Peer-Assisted Learning (PAL): An innovation aimed at engaged learning for undergraduate medical students.

OBJECTIVE: To evaluate the effectiveness of Peer Assisted Learning in teaching at undergraduate level and to assess its effects on Peer Leaders and Peer Learners. METHODS: The cross-sectional study was conducted at the Aga Khan University, Karachi, from May to October 2017, and comprised Peer Learners who were trained by faculty members in workshops and pre-run of experiments. Students were divided into two groups; Group A had Peer Learners taught by Peer Leaders, and Group B had those taught by trained lab technologists. Knowledge of the groups was assessed by a quiz using Kahoot. Post-session feedback questionnaires were also filled by the participants. Data was analysed using SPSS 23. RESULTS: There were 10 Peer Leaders with a mean age of 19.5±0.85 years, and 62 Peer Learners with a mean age of 19.08±0.81 years. Among the learners, there were 35(56.5%) males and 27(43.5%) females. Post-session assessment showed a significant difference in the test performance by the two groups (p<0.05). Feedback indicated that the learners found Peer Leaders more accessible than lab staff, leading to enhanced understanding of the subject. CONCLUSIONS: Peer-Assisted Learning was found to promote learning by creating an informal student-friendly learning environment.

Changes in vitamin E levels as a marker of female infertility.

OBJECTIVE: To study the impact of Vitamin E (VE) levels of follicular fluid (FF) on oocyte competence, embryo development and pregnancy outcome in patients after intra cytoplasmic sperm injection (ICSI). METHODS: It was a cross-sectional study conducted in Islamabad Clinic Serving Infertile Couples in which follicular fluid of 137 females booked for ICSI, was obtained during oocyte retrieval, centrifuged and stored for analysis. VE levels in FF were analyzed by enzyme linked immune sorbent assay. Receiver Operating Curve (ROC)was used to demarcate VE levels required for acquiring pregnancy. Generalized linear model using log binomial regression was applied to see the effect of VE on pregnancy, the effect of VE on oocyte and embryo parameters was assessed by linear regression; all p-values less than 0.05 were considered statistically significant. RESULTS: ROC suggested 5.49 (unit) as the cutoff value of VE in the pregnancy group, with 72.9% area under the curve. Ninety-one females comprised Group I with VE > 5.49, whereas forty six females formed Group II with VE < 5.49. Follicular fluid VE levels were significantly high in 39 (28.5%) females who compromised pregnancy group. Chances of pregnancy increased to 4% with an increase in VE levels (p-value 0.01). VE gave significant positive relationship with all oocyte (retrieved, mature and fertilized) parameters, cleavage of embryo till its differentiation to blastocysts (p<0.01). CONCLUSIONS: Adequate amount of VE in follicular fluid enhances the possibility of maturation of oocytes which resulted in better reproductive outcome after ICSI.

Impact of serum and follicular fluid kisspeptin and estradiol on oocyte maturity and endometrial thickness among unexplained infertile females during ICSI.

OBJECTIVE: To relate serum and follicular fluid (FF) kisspeptin and estradiol levels in different stages of stimulation during Intracytoplasmic Sperm Injection (ICSI) with oocyte maturity and endometrial thickness among unexplained infertile females. METHODS: This cross-sectional study was carried out at the Australian Concept Infertility Medical Centre from March 2017 till March 2018. Fifty unexplained infertile females, booked for ICSI, were included in the study. Serum kisspeptin and estradiol were estimated by Enzyme-Linked Immunosorbent Assay in all four stages; 1: follicular stimulation, 2: ovulation induction, 3: oocyte pickup, and 4: embryo transfer. FF was aspirated during oocyte retrieval (stage 3) for the analysis of KP and estradiol. Pregnancy outcomes were categorized as non-pregnant, preclinical abortion, and clinical pregnancy. RESULTS: The age of the study subjects was 32.04 ± 2.29 (Mean±SD) years, with mean BMI of 28.51 ± 4.15 (Mean±SD) kg/m2. Mean serum kisspeptin and estradiol levels increased in all subjects as the stimulation proceeded stages 1-3; however, the mean dropped after retrieval of the oocytes (stage 4). Out of 27 female subjects who completed the cycle, 17 remained non-pregnant, 4 had preclinical abortion, and 6 acquired clinical pregnancy. The FF kisspeptin concentration was significantly higher than serum concentrations and positively correlated with serum and FF estradiol concentrations. FF-kisspeptin correlated with serum kisspeptin in Stage 3 (r = 0.930, p<0.001), maturity of oocyte (r = 0.511, p = 0.006) and endometrial thickness (r = 0.522, p = 0.005). Kisspeptin in stage 3 was also found to correlate with endometrial thickness (r = 0.527, p = 0.005) and with estradiol (r = 0.624, p = 0.001) independently. CONCLUSION: Increase in serum and FF-kisspeptin and estradiol levels from stages 1 to 3, resulted in an optimum endometrial thickness, probability of fertilization of oocytes and chances of clinical pregnancy in Assisted Reproductive Techniques /ICSI cycles of unexplained infertile females.

Role of oxidative stress and altered thyroid hormones in unexplained infertility.

OBJECTIVE: To explore the link between altered thyroid profile and oxidative stress marker in females with unexplained infertility. METHODS: The cross-sectional case-control study was carried out at the Islamabad Clinic Serving Infertile Couples, Islamabad, Pakistan, from June 2016 to August 2017, and comprised women aged 18-40 years regardless of ethnic background who were divided into two groups; those with unexplained infertility were the cases, while fertile women acted as the controls. Serum was analysed for triiodothyronine, thyroxine and thyroid stimulating hormone as well as for oxidative stress markers including manganese superoxide dismutase, glutathione reductase and adrenaline using enzymelinked immunosorbent assay. Data was analysed using SPSS 19. RESULTS: Of the 88 subjects, there were 44(50%) in each of the two groups. There was no significant difference in terms of thyroids markers except thyroxine and thyroid stimulating hormone (p<0.05). There were significant differences in terms of oxidative stress markers between the groups (p<0.05). A significant positive correlation of thyroid stimulating hormone was observed with manganese superoxide dismutase and adrenaline (p<0.05) with a weak non significant association of glutathione reductase (p>0.05). CONCLUSIONS: Increased thyroxine levels in females with unexplained infertility was associated with decrease in the serum levels of antioxidants.

Association of circulatory Kisspeptin levels and hormonal interplay with male infertility: A cross-sectional study in Karachi, Pakistan.

OBJECTIVES: To determine the impact of Kisspeptin on male reproductive axis in a selected urban population. METHODS: The cross-sectional study was conducted at the Sindh Institute of Reproductive Medicine in collaboration with Aga Khan University, Karachi, from July, 2017, to February, 2018, and comprised infertile males with abnormal sperm parameters who were placed in Group A and fertile males with normal sperm parameters placed in Group B. Serum samples were estimated for Kisspeptin, follicle stimulating hormone, luteinizing hormone, testosterone and sex hormone-binding globulin using enzyme-linked immunosorbent assay. Data was analysed using SPSS 22. RESULTS: Of the 313 male subjects, 178(57%) were in Group A and 135(43%) in Group B. Median Kisspeptin levels were higher among fertile males compared to infertile males (p<0.001). Mean follicle stimulating hormone, luteinizing hormone and testosterone values were higher among the fertile males (p<0.001). There was significant interaction between follicle stimulating hormone and testosterone (p<0.1). CONCLUSIONS: Fertility in males depended on optimal secretion of Kisspeptin which exert edits effect on hypothalamic pituitary gonadal axis to increase male reproductive hormone production.

SIRTI and cortisol in unexplained infertile females; a cross sectional study, in Karachi Pakistan.

OBJECTIVE: To explore a disparity in anti-oxidants (SIRT1) and pro-oxidants (cortisol) levels as a plausible cause of unexplained infertility in females. METHODOLOGY: It was a cross sectional study in which 342 females with unexplained infertility were recruited from ACIMC outpatient clinic: 135 infertile cases and 207 fertile controls. Biochemical estimation of serum cortisol and SIRT1 was performed using Enzyme Linked Immuno Sorbent Assay. Statistical comparisons were performed using Student-t test and Mann-Whitney U test. Associations between circulating hormone levels and infertility were determined using Spearman's rank correlation. Associations were considered significant where value of p was less than 0.05. RESULTS: The stress hormones profile of case and control demonstrated that the antioxidant SIRT1 was significantly lower in infertile females when compared with the fertile (p =< 0.001) while (the oxidant) Cortisol showed a contrast of results with higher values of in infertile females when compared with fertile counterparts (p =< 0.01). There was a strong negative association observed between SIRT1 and cortisol serum level (r = 0.244, p < 0.001). CONCLUSION: Chronic stress causes oxidative stress that is depicted by a decrease in antioxidant levels in infertile females.

Association of fetuin-A with dyslipidemia and insulin resistance in type-II Diabetics of Pakistani population.

BACKGROUND & OBJECTIVE: Fetuin-A, hepatokine is responsible for instigating insulin resistance by inhibiting tyrosine kinase receptors. Our objective was to investigate the relationship of fetuin-A with dyslipidemia and insulin resistance in type-II diabetics of Pakistani population. METHODS: In this cross sectional study which was conducted at Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre, Karachi between October 2013 to March 2014, a total of 330 participants were selected and divided into two groups. Group-A (n = 165) normal healthy individual and Group-B (n = 165) Type-II diabetes mellitus mellitus with no comorbidities. Serum fetuin-A and insulin levels were determined by commercially prepared ELISA kits while fasting blood glucose (FBG) and lipid profile were performed by enzymatic kit method. Employing independent t-test, comparison of groups was done and correlation was achieved by using spearman correlation. RESULTS: The results demonstrate a significant difference in mean values of fetuin-A, lipid profile, glucose, insulin and Homoeostasis Model Assessment of Insulin resistance (HOMA-IR) in type-II diabetics when compared to normal healthy individuals (p<0.01). A positive correlation was found between serum fetuin-A levels and FBG(r= 0.495, p< 0.001), insulin(r= 0.227, p< 0.001), HOMA-IR(r= 0.336, p<0.001, triglycerides(r= 0.197, p< 0.001) and LDL-cholesterol(r= 0.170, p= 0.002), while negative correlation with HDL-cholesterol(r= -0.251, p< 0.001). CONCLUSION: The study concludes that fetuin-A might be accountable for dyslipidemia and insulin resistance in type-II diabetes mellitus mellitus. So the high levels of Fetuin-A responsible for insulin resistance might alters endothelium and causes inflammation, vasoconstriction and thrombosis and ultimately atherosclerosis.

Single nucleotide polymorphisms in Renalase and KCNQ1 genes and female infertility: A cross-sectional study in Pakistan.

A global increase in the incidence of subfertility is observed, and research suggests strong genetic influences that might restrict fertility directly or indirectly. It therefore becomes important to rule out the existence of genetic causes and counsel infertile couples before offering "Advanced Infertility Treatment Techniques." This cross-sectional study aimed to explore the association of KCNQ1 (rs2237895) and Renalase (rs2576178 and rs10887800) single nucleotide polymorphisms with different causes of infertility by analysing 508 fertile and 164 infertile women. Gene variant (AC/CC) of KCNQ1 rs2237895 showed a slight difference in the endometriosis group compared to the fertile group (p = .049), with the C allele showing a significant association with infertility overall (OR = 1.42 [1.100-1.833]; p < .0069). The variant AG/GG of Renalase rs2576178 was significantly associated with overall infertility (OR = 2.266; p < .001), with a strong G allele association with unexplained infertility OR = 2.796 (p = .002) that remained significant after adjusting for age and body mass index. Similarly, Renalase rs10887800 AG/GG and G allele showed significant association with both infertility due to polycystic ovarian syndrome and unexplained infertility. Expression of single nucleotide polymorphism rs2237895 and rs2576178 in both KCNQ1 and Renalase genes might be responsible for altering reproductive potential, hence leading to infertility in women.

Kisspeptin and attributes of infertile males and females: A cross-sectional study in a subset of Pakistani population.

Kisspeptin, a peptide hormone, plays a pivotal role in fertility and neuroendocrine regulation of hypothalamo-pituitary-gonadal axis. Increased kisspeptin and reproductive hormones are responsible for fertility in male and females. This study aimed to explore the role of kisspeptin on hypothalamo-pituitary-gonadal axis by comparing the levels of kisspeptin in fertile and infertile subjects and identifying single nucleotide polymorphisms (SNPs) of KISS1 gene in exon 2 and exon 3 of infertile male and female cohorts. A cross-sectional study was carried out on 80 males (44 infertile and 36 fertile) and 88 females (44 in each group). Significantly high levels of kisspeptin (KP), follicle-stimulating hormone (FSH), luteinizing hormone and testosterone were observed in fertile male and female subjects except low FSH levels in comparison with infertile female subjects. One polymorphism in exon 2 (E1225K [G/A 3673]) and three in exon 3 (P1945A [C/G 5833]; Insertion of T at 6075; G2026G [C/G 6078]) in infertile group were detected, with low KP and hormonal levels. Male subjects had abnormal sperm parameters and unsuccessful attempt of intracytoplasmic sperm injection in females. Expression of SNP in exon 2 and exon 3 of KISS1 could be responsible for alteration in release of reproductive hormones and gonadal functions, hence causing infertility.