An interplay of Progesterone, Leukemia Inhibitor Factor and Interleukin-6 in the window of implantation; Impact on fertility.

BACKGROUND: The process of implantation is crucial for the initiation of conception and hence fertility. In addition to a number of factors, it is regulated by a cross talk of gonadotrophins [Luteinizing Hormone (LH), Follicle Stimulatory Hormone (FSH)], ovarian steroids [Estrogen (Et), Progesterone (Pt)] and cytokines [Leukemia inhibitory factor (LIF) and Interleukin 6 (IL6)]. These biomarkers are chief players of implantation. OBJECTIVE: We aimed to explore the role of gonadotrophins (LH, FSH, LH/FSH ratio), ovarian steroids (Et, Pt) and cytokines (LIF, IL6) in the implantation process. This aim was achieved by comparing these hormones and cytokines in the fertile and infertile groups [Polycystic ovaries (PCOs), endometriosis, unexplained infertility (Uex-IF)] and finding their association in all study groups. METHODS: A case control study conducted from October 2020-March 2023. A total of 135 infertile women (with PCOs, Uex-IF, and endometriosis) and 177 fertile women (matched for age and BMI) were selected. Levels of 'Et', 'Pt', 'LIF' and, 'IL6' were estimated using Enzyme Linked Immunosorbent Assay (ELISA). LH and FSH values were obtained from hospital desk records. The Independent Student'st-test was used to compare fertile and infertile groups. One-way ANOVA test was used to compare more than two groups, and Pearson's chi-square (χ2) test was employed to compare percentages of variables. Pearson correlation analysis was performed to assess the associations and correlations. A p value < 0.05 was considered statistically significant. RESULTS: Significantly higher levels of LIF and IL6 were observed in fertile women compared to infertile women. Pt levels were significantly greater in the fertile group than in the infertile group. The FSH/LH ratio was significantly higher in the fertile group. Among infertile women, PCOs (71%) and Uex-IF (91%) exhibited lower Pt levels than the fertile controls (p < 0.01), but these levels remained within the reference range (RR). Among the fertile group (81%), levels of LIF within the RR were significantly higher compared to those with Uex-IF (49%) and females with endometriosis (37%). Moreover, the highest number of participants (57%) with Uex-IF exhibited IL6 levels significantly below the RR in comparison to the fertile group and infertile groups (PCOS and endometriosis). However, lower levels of IL6 were observed in women with Uex-IF. In the control group, LIF exhibited a significant positive correlation with IL6 (r = 0.370), Pt (r = 0.496), Et (r = 0.403), and LH (r = 0.428). Among women with PCOs, LIF showed a significant positive correlation with IL6 (r = 0.443), Pt (r = 0.607), and LH (r = 0.472). In cases of Uex-IF, LIF demonstrated a significant positive correlation with IL6 (r = 0.727). Females with endometriosis displayed a significant positive correlation between LIF and IL6 (r = 0.535) as well as Pt (r = 0.605). In fertile women, a positive correlation was observed between LH and IL6 (r = 0.197, p = 0.009), LIF (r = 0.428, p = 0.000), Pt (r = 0.238, p = 0.001), and Et (r = 0.356, p = 0.000). Furthermore, a positive correlation was found between LH and LIF (r = 0.472, p = 0.000) in women with PCOs. CONCLUSION: Elevated levels of Pt were found to increase the production of LIF in fertile females. However, infertile females with PCOs and Uex-IF exhibited deficient levels of Pt, supporting its role as a biomarker for successful implantation in infertile women. These females showed decreased levels of gonadotropins as well as reduced LH/FSH ratio and diminished secretion of receptivity marker LIF, in addition to reduced Pt secretion. This suggests that reduced gonadotropin levels contribute to a lower LH/FSH ratio, resulting in decreased Pt secretion and ultimately leading to low levels of LIF, thereby causing impaired implantation in women with PCOs and Uex-IF. The exploration of low levels of LIF in patients with endometriosis requires further investigation. The significantly low levels of IL6 in the Uex-IF group elucidate the role of this cytokine in association with decreased Pt and LIF synthesis within this group.

Impacts of adiposity parameters on peak expiratory flow rate in healthy young adults of Bahria University Medical and Dental College Karachi: Cross-sectional study.

OBJECTIVE: To explore the association of various adiposity parameters in young adults. METHODS: The cross-sectional study was conducted at Bahria University Medical and Dental College, Karachi, from January to February 2021, and comprised healthy medical students aged 19-21 years. Height, weight, body mass index, waist circumference, waist-hip ratio and percentage body fat were recorded. Peak expiratory flow rate was determined using Wright's meter, and its association with adiposity markers was determined using regression analysis. Data was analysed using SPSS 22. RESULTS: Of the 162 subjects, 96(59.3%) were females and 66(40.7%) were males. The overall mean age was 20.18±1.01 years. Peak expiratory flow rate was significant positively associated with height, body mass index, waist circumference, and percentage body fat (p<0.05). It had weak negative association with waist-hip ratio(p>0.05). CONCLUSIONS: Peak expiratory flow rate increased with height, body mass index, waist circumference, and percentage body fat in healthy subjects.

Association of novel stop-gained leukaemia inhibitory factor receptor gene (rs121912501) variant, leukaemia inhibitory factor and ovarian steroids with unexplained infertility among Pakistani women.

AIMS AND OBJECTIVES: Embryo implantation is a complex process that requires sequential steps at the interface of embryo interaction with decidual endometrium. Many women after experiencing multiple attempts of assisted reproductive techniques fail to get implantation because of instability of leukaemia inhibitory factor and leukaemia inhibitory factor receptor-signal transducer and activator of transcription factor 3 (LIF-LIFR STAT3) signalling cascade. Therefore, this study explores the association of ovarian steroids, LIF and LIFR stop-gained variant using the tetra primer amplification refractory mutation system-polymerase chain reaction (TARMS-PCR) with unexplained infertility (UEX-IF) among Pakistani women. MATERIALS AND METHODS: This is a case-control study, a total of 81 unexplained infertile women and 162 fertile controls (with age and BMI matched) were inducted. Serum estradiol, progesterone and LIF were determined using enzyme-linked immunosorbent assay (ELISA). T-ARMS-PCR was designed using Primer 1 software. Genomic DNA was extracted from peripheral blood and amplified using T-ARMS-PCR followed by sequencing for validation and comprehensive concordance. RESULTS: This study established differences in LIF levels (χ2  = 9.857, P < .05) between patients and controls as well as explored the decreased LIF significantly raised the risk of UEX-IF (OR = 2.316; 95% CI = 1.214, 4.416). Progesterone (P) was significantly associated with UEX-IF between fertile and infertile counterparts (χ2  = 20.347, P < .05). It was also observed that increased Progesterone reduced the risk of UEX-IF (OR = 0.306; 95% CI = 0.166, 0.567). A rapid and inexpensive method for genotyping novel LIFR gene polymorphism through T- ARMS-PCR was successfully developed. LIFR gene SNP (rs121912501) had significant association (χ2  = 200.681, P < .05) with UEX-IF. LIFR rs121912501 "TT" genotype (OR = 5.417; 95% CI = 1.868, 15.709) and "CT" genotype (OR = 3.104, 95% CI = 1.586,6.076) were at increased risk of infertility. CONCLUSION: UEX-IF can be caused by LIFR gene variation irrespective of increased P. It may open the doors for the discovery of new management plans for infertile women.

Embryo quality in intracytoplasmic sperm injection: A quasi experimental design in Pakistan.

OBJECTIVE: To assess the role of good-quality embryos in acquiring pregnancy after intracytoplasmic sperm injection and to predict factors required for development of embryos. METHODS: The quasi-experimental study was conducted from June 2010 to August 2012 at Islamabad Clinic Serving Infertile Couples and comprised infertile females who received long-term gonadotropin agonist protocol for intracytoplasmic sperm injection. Basal estradiol and antral follicle count was measured at baseline, while estradiol, progesterone and interleukin levels with pre-ovulatory follicle count were evaluated on the ovulation induction day. Follicular output rate was measured as ratio of pre-ovulatory follicle count to antral follicle count. The quality of embryos was graded as good, fair and poor on the third day after the injection. Linear regression was employed to determine unadjusted and adjusted estimate of effect of different factors on good-quality embryo. RESULTS: There were282 subjects with a mean age of 32.1±4.7 years (range: 23-41 years). There was an inverse correlation of body mass index with good-quality embryo (r=-0.131). In adjusted analyses, basal estradiol and progesterone on the ovulation induction day showed negative effect (95% confidence interval: -1.42 - -1.01), while estradiol and interleukin measured positive effect on quality of embryo (p<0.05). Follicular output rate was highly positively correlated with good quality embryo (p<0.05).. CONCLUSIONS: Females with low body mass index produced good-quality embryos. A high follicular output rate with production of estradiol and interleukin complemented good-quality embryos.

Self-reported academic performance in relation to health behaviours among Bahria University students.

OBJECTIVE: To find an association between self-reported academic performance with different socio-demographic factors, health behaviours and mental health amongst university students. METHODS: This cross-sectional study was conducted at Bahria University, Karachi, from January 2012 to December 2013, and comprised university students of different disciplines. An anonymous, self-reported questionnaire was distributed among the subjects. Convenient sampling technique was used. Demographic information, including age, gender and field of study, were obtained. Depresion was evaluated via Centre for Epidemiological Studies Depression Scale. SPSS 22 was used to analyse data. RESULTS: Of the 813 respondents, 334(41.1%) were males and 479(58.9%) females. The mean age was 19.9±1.8 years. Overall, 126(15.5%) subjects reported excellent, 242(29.8%) very good, 310(38.1%) good, 100(12.3%) satisfactory and 35(4.3%) not satisfactory academic performance. Residential status of students played a significant role on their academic performance (p=0.011). Breakfast eating behaviour depicted a significant association with the academic performance (p=0.04).The proportion of unsatisfactory academic performances among students having severe sleep disorder was the highest, followed by mild/moderate (p=0.01). The depression scale's item 'troubling in mind' was highly associated with academic performance (p<0.05). CONCLUSIONS: A constructive association existed among healthy behaviours and academic performance. .

Utilization Focused Evaluation at Bahria University Medical & Dental College.

OBJECTIVE: To evaluate integrated learning program of neurosciences for continuation of integrated learning in the forthcoming teaching and learning modules of undergraduate medical curriculum at Bahria University Medical & Dental College (BUMDC). METHODS: A mixed method design was conducted from August 2016to February2017 after ethical approval from BUMDC. The quantitative aspect was evaluated retrospectively by desk records ofmarks obtained in integrated module and nonintegrated module. Focused group discussionwere conducted with primary intended users (chair of integration committee, faculty and students of first and second year MBBS)to share their expectations and concerns and get responses on key evaluation questions for implementationand outcome evaluation of integrated learning program. RESULTS: The desk record revealed a positive perception of students and faculty at the time of implementation with improvement in results after integration in subjects of basic sciences. The discussions highlighted reasons which resulted in failure of its continuation and affirmedreadiness for re-induction and continuation of integration with clinicalsciences. CONCLUSION: Evaluators considered approval and re-application of integrated curriculum at BUMDC after utilization focused evaluation.

Interplay of "leukemia inhibitory factor receptor gene" (rs3099124) polymorphism, leukemia inhibitory factor and ovarian steroids with unexplained infertility.

OBJECTIVE OF THE STUDY: To explore the association of leukemia inhibitory factor receptor (LIFR) gene variant rs3099124, ovarian steroids, and leukemia inhibitory factor with unexplained infertility in Pakistani females. METHODOLOGY: A case-control investigation in which eighty-one (81) females with unexplained infertility and one hundred and sixty-two (162) fertile counterparts (age and body mass index compared) were recruited between October 2016 and 2018. Ten milliliters of venous blood was collected from all participants. "Genomic DNA" was taken out from lymphocytes in peripheral blood samples. "Tetra Amplification Refractory Mutation System Polymerase Chain Reaction (T-ARMS-PCR)" was constructed through software "Primer-I". Amplification was carried out by "T-ARMS-PCR" followed by subsequent sequencing for confirmation and extensive consonance. Estradiol, Progesterone and Leukemia Inhibitory Factor (LIF) were measured in serum by ELISA. RESULTS: Statistically significant difference was noticed in genotype frequency in "LIFR-gene variant; rs3099124" (χ2 = 28.222, P value < 0.01) between research participants. Although, rs "3099124" "AA" (OR = 0.000; 95%CI = 0-0) and "GA" genotypes (OR = 0.525; 95%CI = 0.226-1.22) showed non-significant safety/protection against unexplained infertility yet minor/risk allele "A" frequency was greater in women with unexplained infertility suggesting a possible explanation of implantation failure. LIF concentration varied between fertile and infertile groups (χ2 = 9.857, P < 0.05) revealing significant threat of unexplained infertility in women with decreased LIF concentration (OR = 2.316, 95%CI = 1.214-4.416). Progesterone was significantly related to unexplained infertility in both study groups (χ2 = 20.347, P < 0.05). High progesterone reduced the possibility of unexplained infertility (OR = 0.306; 95% CI = 0.166-0.567). CONCLUSION: LIFR gene variation (rs3099124) and reduced LIF secretion may cause implantation failure in women with unexplained infertility.

General Public Perception of Social Media, Impact of COVID-19 Pandemic, and Related Misconceptions.

OBJECTIVES: This research aimed at investigating the general public perception of social media (SM), impact of COVID-19 (SARS-CoV-2) pandemic, and related misconceptions among the Pakistani population. METHODOLOGY: Cross-sectional study conducted during the peak of COVID-19 in Pakistan between May and June, 2020 comprised of 2307 Pakistani male and female participants. Subjects under 18 years of age and nationality other than Pakistani were excluded. An online questionnaire was administered via the Internet using various kinds of social media. RESULTS: The study was comprised of 2307 male and female participants; 2074 (89.90%) used SM for seeking COVID-19 information, 450 (20%) used both Facebook (FB) and WhatsApp (WA), and 267 (11.6%) used FB, WA, Twitter, and Instagram. Respondents' perceptions showed that: 529 (23%) believed in SM information and 1564 (67.8%) stated that COVID-19 affected their social and mental wellbeing. Respondents' knowledge revealed that: 1509 (65.40%) had poor knowledge (≤ 50% score), and 798 (34.6%) had good knowledge (> 50% score) (P < 0.001) about COVID-19. Binary logistic regression analysis showed that higher-earning positively correlated, while private jobs were negatively associated, with good knowledge. CONCLUSION: FB and WA were the 2 common social media used by study participants (a third had good knowledge). COVID-19 affected the social, mental, and psychological well-being of individuals. Good knowledge was greater in individuals with higher earning and less with private job involvements.

COVID-19 Pandemic Awareness, Attitudes, and Practices Among the Pakistani General Public.

Background: Outbreak of COVID-19, in many countries, has imposed a lockdown on their residents. The usefulness of extenuative actions is extremely reliant on society's knowledge, attitudes, and practices (KAP) toward pandemic control. Objective: This study aimed to explore the awareness, attitudes, and practices of the general Pakistani population to COVID-19. Methods: From June 13, 2020, until June 30, 2020, a cross-sectional online KAP survey was conducted among the Pakistani public. For data collection, a validated self-administered questionnaire was used. The survey instrument consisted of six demographic characteristics, 14 items on knowledge, four on attitudes, and six items on practices, modified from a previously published questionnaire on COVID-19. Results: The present study included 2,307 participants, 58.3% males and 41.7% of females. The majority (86.7%) sought information from social media (SM) and television, 95% had good practices, 89.9% had positive attitudes, and two-thirds (67.4%) of the respondents had adequate knowledge. The students and people from younger age groups had more positive attitudes compared with others. Highly educated w with other groups (p < 0.001). In logistic regression analysis, the odds ratio indicated that the private job was negatively associated, and high monthly income was positively associated with adequate knowledge (OR = 0.595). Old age was the predictor of negative attitude, and high school degrees and master's degrees were associated with good practice scores. Conclusion: The Pakistani general population has an overall positive attitude and proactive practices against COVID-19, but their knowledge is inadequate. The most important source of information was SM, followed by television. These are playing a crucial role in educating the Pakistani public.

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.

Influence of body mass index and polycystic ovarian syndrome on ICSI/IVF treatment outcomes: A study conducted in Pakistani women.

BACKGROUND: Obesity may establish a crucial barrier for effective fertility treatment in polycystic ovary syndrome (PCOS) females. OBJECTIVE: To compare results of intra-cytoplasmic sperm injection (ICSI) in females with and without polycystic ovarian syndrome and further appraise the effect of obesity in PCOS females. MATERIALS AND METHODS: A cross-sectional study from June 2015 to July 2016 included non-PCOS and PCOS (recognized by Rotterdam criteria) females who underwent ICSI. The PCOS were further stratified into non-obese and Obese according to the South Asian criteria for body mass index. Results were categorized on the basis of beta-human chorionic gonadotropin (ß-hCG) and transvaginal scan into non-pregnant (ß-hCG <25 mIU/ml), preclinical abortion (ß-hCG >25 mIU/ml with no fetal cardiac activity) and clinical pregnancy (ß-hCG >25 mIU/ml with fetal cardiac activity on transvaginal scan). In addition, reproductive outcomes; implantation rate, clinical pregnancy rate and miscarriage rate among obese and non-obese PCOS and non-PCOS patients were compared. RESULTS: Our results revealed 38.5% clinical pregnancy rate in non-PCOs females, 23.8% in non-obese PCOS females whereas 26.4% in obese PCOS. Preclinical abortions were found to be highest (31.5%) in non-obese PCOS females and were the lowest (26.2%) in non-PCOS females. In non-PCOS group and non-obese PCOS females 35.4% and 44.6%, respectively, failed to become pregnant. CONCLUSION: The success after ICSI in terms of number of clinical pregnancies was more in non-PCOS patients as compared to PCOS. Increase in body mass index reflected a negative impact on the reproductive outcome in PCOS patients.