BackgroundIntrafollicular melatonin maintains the DNA integrity of granulosa cells and protects them against apoptosis. This ubiquitous indoleamine compound serves as a potent free radical scavenger. It reduces oxidative stress and modulates DNA damage response, which improves oocyte's quality with a higher fertilization rate.Methods:This prospective study was designed to investigate the antioxidant property of intrafollicular melatonin and its impact on IVF outcome parameters by exploring the relative expression of five microRNAs (miR-663b, miR-320a, miR-766-3p, miR-132-3p, and miR-16-5p) and levels of cfDNA by real-time PCR in unexplained infertile patients. We collected 425 follicular fluid (ff) samples containing mature oocytes from 295 patients undergoing IVF. Results:Patients were sub-grouped based on intrafollicular melatonin concentration (Group A; ≤ 30 pg/mL, Group B; >70 to ≤110 pg/mL), Group C; >111 to ≤ 385 pg/mL). Our results showed that patients with ≤ 30 pg/mL intrafollicular melatonin levels have a significantly higher cfDNA levels and lower relative expression of miR-663b, miR-320a, miR-766-3p, miR-132-3p, and miR-16-5p compared to other subgroups (p<0.001). Similarly, they have a low fertilization rate and a reduced number of high-quality day 3 embryos. For the probability of obtaining a good quality embryo, miRNAs had AUC value of 0.89 [95% Cl; 0.71; 0.93] with 87% sensitivity and 83 % specificity (p = 0.001), whereas cfDNA have AUC value 0.76 [cut-off points, 95% Cl: 0.61, 0.67; 0.85] along with 82.4 % senstivity and 66.7 % specificity.Conclusion:Conclusively, our study provided evidence that melatonin's antioxidant capability significantly impacts cfDNA concentration and miRNAs relative expression profile in the follicular microenvironment for optimal oocyte development and embryo quality. Therefore, it may be a potent non-invasive diagnostic tool to select high-quality day 3 embryos with such promise.

During the last few decades, assisted reproductive technologies (ARTs) have flourished rapidly and accompanied a set of advanced procedures such as intracytoplasmic sperm injection (ICSI), electronic witnessing, digital monitoring through embryoscope time-lapse systems, consistent decision-making algorithms with advanced statistics modes and preimplantation genetic testing (PGT). In usual practice, manual procedures were routinely used in IVF (in vitro fertilization) laboratories worldwide, but automation and artificial intelligence (AI) systems are promising techniques for quality assurance, which reduced the burden on the working staff in the embryology laboratory. In addition, these systems are equipped with powerful mathematical tools that minimize technician variability in the IVF lab and efficiently generate data for impaired gametes and embryos. The principal challenge of single-sperm selection out of 108 gametes can be sorted out by incorporating machine learning algorithms coupled with advanced data processing capabilities. In the same line, the emergence of closed embryo culture systems (CECSs) in human embryology has enabled the accurate morphokinetic evaluation of the more rapid cell division and the identification of normal and abnormal hallmarks of embryo viability. In particular, these CECSs are guided by the latest time-lapse microscopy (TLM) facility to continuously monitor embryo development kinetics without removing them from controlled and stable incubator conditions. In conclusion, AI-driven models can reduce technical variability in sample handling and remove the burden of the most subjective, tedious and/or monotonous aspects of the IVF lab. Furthermore, these systems also highlight environmental stressors that could hamper embryo development competence. In a broader sense, AI-based approaches are more accurate, precise and rapid in predicting embryo quality noninvasively.

Semen is a pale whitish fluid secreted by male during ejaculation and containsspermatozoa which are male gametes essential of fertilizing the oocytes which are femalegametes. In a quest to evaluate male’s fertility potential semen is analyzed to look into some ofits characteristics and of the sperms contained within the semen analyzed. Method of collectioninfluences the results of Semen analysis as does the technique of analysis. Spermatozoa areexamined for number (count), shape (morphology) and movement (motility) in order to assesstheir quality. Non sperm cells, volume, Fructose level, pH, liquefaction are also checked asa part of routine analysis. Objectives: To describe the pattern of semen parameters in subfertilemales. To look into frequency and distribution of abnormal semen parameters in a groupof Pakistani males in Lahore. Methods: In this Retrospective, cross sectional, observationalstudy all males undergoing for evaluation and treatment for sub-fertility at a private AssistedReproductive Technology clinic in Lahore, Pakistan were included. Approval of the IRB wassought and data collection instrument was a specially designed Performa which was validatedby the biostatistician of LIFE research cell. Data was extracted from the files of LIFE (LahoreInstitute of Fertility and Endocrinology) and entered in SPSS version 15. Sampling techniquewas non-probability, consecutive. Semen analysis was done by methods defined by the WHO(World Health Organization). Results: Of total patient (n=679) 92.2% (626) males passed sampleat LIFE (Lahore institute of fertility and endocrinology) and (7.8%) 53 brought sample fromhome. Of the males who passed sample at LIFE (78.8%) 535 collected semen by masturbation,(11.9%) 81 by coitus; the source of sample of (9.3%) 63 males was not known. As 2-6 ml semenwas consider to be normal by WHO criteria, (80.6%) 547 males were in normal range (14.1%)96 found to be less than 2-6 ml and (5.3%) 36 found to be more than normal range. Accordingto WHO criteria 15 million/ml count is said to be normal, in our research (82.0%) 557 were foundto be normal, in (2.9%) 20 count was found to be less than 15 million/ml and in (5.9%) 40 countwas less than 1 million/ml. In (9.1%) 62 counts was found to be abnormally low. In this research(66.1%) 449 had normal sperm motility, (21.8%) 148 had less than 40% and abnormally lowsperm motility was found in (12.1%) 82 males. Conclusion: The results of the single semenanalysis are of limited utility and no decision should be taken on the bases of these results interm of diagnosis and treatment strategies.

Various ovarian reserve tests were developed to estimate the ovarian reserve andpredict about the outcome in subfertile females undergoing evaluation for assisted reproduction.FSH and AMH levels are considered to be good ovarian reserve indicators along with antralfollicle count. Objectives: To explore relationship of AMH and FSH in patients undergoing IVFwith respect to ovarian reserve and outcome of the treatment. Study Design: Prospective cohort.Study Period: 1st January 2015 to 31st December 2015. Place of study: Lahore Institute ofFertility and Endocrinology, Hameed Latif Hospital, Lahore Material and Methods: In 346 IVF/ICSI patients after anthropometric measurements and transvaginal ultrasound antral folliclecount were assessed in each ovary. For the hormone measurements blood samples were takenduring the early follicular phase of menstrual cycle. Clinical pregnancy was also visualizedthrough transvaginal ultrasound. Results: From the 346 IVF/ICSI patients 89 (25.79%) clinicalpregnancies resulted. The mean age in pregnant group was 32.89 ± 2.99 years and in nonpregnantgroup was 33.62 ±4.36. Mean FSH and AMH in pregnant group was 6.38 ±2.38,3.27 ±1.86 and in non- pregnant group was 7.54±3.76, 2.72 ± 1.82 respectively. Age andFSH are significantly associated with each other (p-vale = 0.000) and mostly patients had FSHbelow 9(mIU/mL). Age and AMH are significantly associated with each other (p-vale = 0.000)and mostly patients had AMH above 1.5 (ng/mL). Conclusions: Better pregnancy rate wasassociated with FSH below than 9 (mIU/mL) and AMH above 1.5 (ng/mL).

The premature ejaculation diagnostic tool (PEDT) assesses premature ejaculation (PE). However, there is insufficient evidence about its validity in evidence-based‑ defined PE. The main aim of this study was to assess the validity of PEDT and its association with an international index of erectile function (IIEF-15) in Acquired premature ejaculation (APE) patients. A total of 50 men complaining of APE from Lahore institute of fertility and endocrinology, Hameed Latif Hospital, and 50 healthy control males without PE from a medical center were enrolled in this study. All individuals were asked to complete questionnaires including demographics, sexual history, and PEDT and IIEF‑ 15. The results of this research indicated men with APE showed higher PEDT scores (13.51 ± 3.04) and lower IIEF‑ 15 (39.45 ± 6.54) than men without PE (PEDT: 4.89 ± 2.14, IIEF‑ 15: 51.36 ± 2.36, P < 0.001 for both). Results also reported that a score of ≥ 8 indicated PE in Acquired premature ejaculation (APE) individuals by sensitivity and specificity analyses (sensitivity: 0.985; Specificity: 0.891). In APE men IIEF was negatively correlated to PEDT (adjust r = -0.311 P < 0.001) after adjusting for age. It was concluded that evidence-based‑ defined PE was diagnosed by PEDT. Moreover, PEDT was negatively related to IIEF-15 in APE men.