Zehra Jamil

Anti-Mullerian Hormone: Above and Beyond Conventional Ovarian Reserve Markers

Management of ovarian dysfunctions requires accurate estimation of ovarian reserve (OR). Therefore, reproductive hormones and antral follicle count (AFC) are assessed to indicate OR. Serum anti-Mullerian hormone (AMH) is a unique biomarker that has a critical role in folliculogenesis as well as steroidogenesis within ovaries. Secretion from preantral and early antral follicles renders AMH as the earliest marker to show OR decline. In this review we discuss the dynamics of circulating AMH that remarkably vary with sex and age. As it emerges as a marker of gonadal development and reproductive disorders, here we summarize the role of AMH in female reproductive physiology and provide evidence of higher accuracy in predicting ovarian response to stimulation. Further, we attempt to compile potential clinical applications in children and adults. We propose that AMH evaluation has a potential role in effectively monitoring chemotherapy and pelvic radiation induced ovarian toxicity. Furthermore, AMH guided ovarian stimulation can lead to individualization of therapeutic strategies for infertility treatment. However future research on AMH levels within follicular fluid may pave the way to establish it as a marker of “quality” besides “quantity” of the growing follicles.

Polymorphism of the renalase gene in gestational diabetes mellitus

Renalase is considered as a novel candidate gene for type 2 diabetes. In this study, we aimed to investigate the relationship of serum renalase and two single nucleotide polymorphisms with gestational diabetes mellitus. One hundred and ninety-eight normotensive pregnant females (n = 99 gestational diabetes mellitus; n = 99 euglycemic pregnant controls) were classified according to the International Association of the Diabetes and Pregnancy Study criteria. Fasting and 2-h post glucose load blood levels and anthropometric assessment was performed. Serum renalase was measured using enzyme-linked immunosorbent assay, whereas DNA samples were genotyped for renalase single nucleotide polymorphisms rs2576178 and rs10887800 using Polymerase chain reaction-Restriction fragment length polymorphism method. In an age-matched case control study, no difference was observed in the serum levels of renalase (p > 0.05). The variant rs10887800 showed an association with gestational diabetes mellitus and remained significant after multiple adjustments (p < 0.05), whereas rs2576178 showed weak association (p = 0.030) that was lost after multiple adjustments (p = 0.09). We inferred a modest association of the rs10887800 polymorphism with gestational diabetes. Although gestational diabetes mellitus is self-reversible, yet presence of this minor G allele might predispose to metabolic syndrome phenotypes in near the future.

Assessment of ovarian reserve: Anti?Mullerian hormone versus follicle stimulating hormone

Background: This study aimed to evaluate the strength of anti-Mullerian hormone (AMH) and follicle stimulating hormone (FSH) in reflecting the antral follicle count (AFC) in infertile females. Materials and Methods: This cross-sectional study was conducted on 160 females, visiting infertility clinic for assisted reproduction. Serum samples collected on the 3rd day of the cycle were assayed for FSH, luteinizing hormone, and AMH while AFC was assessed via transvaginal ultrasound. The study cohort was segregated into three groups based on AFC. Results: Chronological age and FSH was significantly high in females with very low AFC (P < 0.01 and 0.009, respectively), yet they failed to discriminate patients with normal and higher follicle count (P = 0.65 and 0.84). Conversely, AMH reported highly significant difference between very low AFC and with those having either normal AFC (P = 0.002) or higher AFC (P = 0.001). Moreover, a significant difference in AMH was observed between normal and higher AFC group (P = 0.04). Conclusion: Compared to female’s age and FSH, AMH is superior in clustering study cohort on the bases of antral follicular pool, especially in setups with nonavailability of technological expertise to assess AFC. Incorporation of AMH along with other biomarkers improves estimation of baseline ovarian reserve, required to standardize dose for optimum response; avoiding the risk of failure to retrieve oocyte or inappropriate stimulation leading to ovarian hyperstimulation syndrome. Further prospective studies are required to ascertain its role in predicting the outcomes of ART in such patients.

Anti mullerian hormone: Ovarian response indicator in young patients receiving long GnRH agonist protocol for ovarian stimulation

Objective: Anti Mullerian hormone (AMH) is gaining place as ovarian marker, chiefly in infertility assistance. We explored its correlation with oocytes retrieval after long GnRH agonist protocol for stimulation, in younger and older infertile population. Methods: This retrospective analysis compiled data of 166 females, receiving ICSI treatment from June 2014 to March 2015. Serum FSH, LH, Estadiol, AMH and antral follicle count were assessed. Outcomes were measured as good (5 to 19 oocytes) and bad responders. Results: Higher discriminatory power of AMH (AUROC; 0.771; p < 0.05) was seen in comparison to FSH (0.692; p < 0.05) and AFC (0.690; p < 0.01). AMH reported strongest association with oocyte retrieved (odds ratio of 15.06). Subgroup analysis reported 68.6 % risk of bad response with AMH levels of less than 1.37ng/ml. This association was observed more significant in young infertile patients <35 year of age (r=0.245; p=0.012) versus older population >35 year (r=0.169; p>0.05). Conclusion: Our study reaffirms that serum AMH correlates well with oocytes retrieved, particularly in females younger than 35 years. We suggest incorporation of AMH in baseline assessment of infertile females, who are falsely advised to postpone interventions based on their age and normal FSH levels.

Cross talk between serum Kisspeptin-Leptin during assisted reproduction techniques

Background & objective: Leptin facilitates onset of puberty by impact on hypothalamic Kisspeptin, gonadotropin releasing hormone, follicle stimulating and luteinizing hormone. The link of peripheral Leptin-Kisspeptin in regulating the ovarian and endometrial tissue in relation to adiposity is unknown. Therefore, we wanted to identify Kisspeptin-Leptin association with body mass index (BMI) and success of assisted reproductive treatments (ART) in infertile females. Methods: A cross sectional study was carried from August 2014 till May 2016 after receiving ethical approval at Australian Concept Infertility Medical Centre, and Aga Khan University. The study group comprised of females with an age range of 25-37 year who had duration of unexplained infertility for more than two years. They were grouped as; underweight (<18 kg/m2), normal weight (18-22.9 kg/m2), overweight 23-24.99 kg/m2 and obese (>25 kg/m2). Kisspeptin and Leptin levels were measured by enzyme linked immune sorbent assay before down regulation of ovaries and initiation of treatment protocol of ART. Failure of procedure was detected by beta human chorionic gonadotropin <25mIU/ml (non-pregnant) whereas females with levels >25mIU/ml and cardiac activity on trans-vaginal scan were declared pregnant. Results: Highest Kisspeptin and Leptin levels were seen in normal weight group (374.80 ± 185.08ng/L; 12.78 ± 6.8 pg/ml) respectively, yet the highest number of clinical pregnancy was observed in overweight group (42%).A strong correlation of Kisspeptin with Leptin (r=0.794, p=0.001) was observed in the overweight females. Conclusion: Leptin-Kisspeptin-fertility link is expressed by maximum number of clinical pregnancies in the female group that showed strongest relationship between serum Leptin and Kisspeptin levels, irrespective of their BMI.

Flipping the classroom: training the flippers

are taught through reflection-based exercises during virtual and classroom-based learning. A robust departmental faculty development exercise was conducted for curriculum development, active teaching and learning, VL master training and the use of the flipped classroom. What lessons were learned? A majority of students (78.6%) reported that they found the blended curriculum more engaging both during and between classes. They provided positive feedback on both the curricular elements and the use of VLE. One challenge was that faculty members found it hard to change their T/L roles from the traditional expert-led teaching culture; they were not comfortable using Moodle and had limited engagement with VL. Faculty members’ comfort was increased by a non-clinical VL team who provided supervision and facilitation. Another challenge identified by the core team was aligning outcomebased formative assessment to the institution-based summative assessment system. Overall, the use of VL was helpful in creating an integrated and blended curriculum for a medical student psychiatry clerkship. This comprehensive curriculum can be offered to other Pakistani medical colleges in the future that face challenges of limited faculty members with minimal VL expertise, as long as it is accompanied by faculty development in T/L methods.

Interleukin-18 polymorphism as an inflammatory index in metabolic syndrome: A preliminary study

AIM To assess circulatory levels of interleukin-18 (IL-18) and determine whether the presence of IL-18 promoter polymorphism influences metabolic syndrome phenotypes. METHODS This study recruited one hundred and eighty individuals divided into three groups with sixty subjects each as: Normal weight (18.0-22.9 kg/m2), overweight (23.0-25.9 kg/m2) and obese (> 26.0 kg/m2) according to South Asian criteria of BMI. Fasting blood glucose (FBG), Lipid profile, insulin, IL-18 and tumor necrosis factor (TNF)α were measured using ELISA kits, whereas low density lipoprotein (LDL)-cholesterol, insulin resistance (HOMA-IR) and insulin sensitivity (QUICKI) were calculated. The body fat percentage (BF) was measured through bioelectrical impedance analysis; waist and hip circumference were measured. Genotyping of IL-18 -607 C/A polymorphism was performed by using tetra-primer amplification refractory mutation system. Student t test, One-way analysis of variance, Hardy-Weinberg equilibrium, Pearson’s χ2 test and Pearson’s correlation were used, where a P value < 0.05 was considered significant. RESULTS In an aged matched study, obese subjects showed higher levels of FBG, cholesterol, triglycerides and LDL levels as compared to normal weight (P < 0.001). Highest levels of IL-18 and TNF levels were also seen in obese subjects (IL-18: 58.87 ± 8.59 ng/L) (TNF: 4581.93 ± 2132.05 pg/mL). The percentage of IL-18 -607 A/A polymorphism was higher in overweight and obese subjects vs normal weight subjects (P < 0.001). Moreover, subjects with AA genotype had a higher BF, insulin resistance, TNFα and IL-18 levels when compared with subjects with AC (heterozygous) or CC (wild type) genotypes. However, we did not find any difference in the lipid profile between three subgroups. CONCLUSION This preliminary data suggests that IL-18 polymorphism affects IL-18 levels that might cause low grade inflammation, further exacerbated by increased TNFα. All these increase the susceptibility to develop MetS. Further studies are required to validate our findings.

Kisspeptin and embryo implantation after ICSI

A cross-sectional study was carried out at the Australian Concept Infertility Medical Centre from June 2014 to June 2015 to relate serum kisspeptin levels on human chorionic gonadotrophin (HCG) day with pregnancy outcome after intracytoplasmic sperm injection (ICSI). A total of 176 women aged 20 to 42 years, with regular menstrual cycles, normal thyroid function and prolactin levels selected for ICSI were included in the study. Patients with uterine fibroids, metabolic disorders, short agonist and antagonist protocol were excluded. Long protocol for down-regulation of ovaries was observed and kisspeptin levels were estimated on HCG day. Results were categorized into groups: Group A, non-pregnant with β-HCG <25 mIU/ml; and Group B, clinical pregnancy with β-HCG >25 mIU/ml and cardiac activity on transvaginal scan. Kisspeptin levels were significantly higher in Group B versus Group A (P < 0.001), independently associated with positive pregnancy (r = 0.388; P < 0.001), but just borderline with endometrial thickness (r = 0.294; P = 0.05). The area under the curve was highest for kisspeptin, 0.784 (95% CI, 0.681 to 0.886) for positive pregnancy, which indicated that kisspeptin measured on HCG day can be used as a marker for success of treatment in women after ICSI.