Association of Follicular Fluid Antioxidants Activity with Aging and In Vitro Fertilization Outcome: A Cross-Sectional Study.

BACKGROUND: This research was aimed at assessing the relationship between the follicular fluid (FF) antioxidants activity, aging and in vitro fertilization (IVF) outcome. MATERIALS AND METHODS: The present cross-sectional study was carried out on 65 women undergoing IVF/intracytoplasmic sperm injection (IVF/ICSI) cycles due to unexplained infertility. Ovarian stimulation was performed using the long gonadotropin-releasing hormone (GnRH) agonist protocol. After ovum pickup, FF was collected and processed to measure the level of superoxide dismutase (SOD), catalase (CAT) activity, total antioxidant capacity (TAC) and glutathione (GSH). Day 3 after ICSI, fresh embryos were transferred and later, possible pregnancy was assessed. Patients participating in this study were divided into four groups on the basis of age and pregnancy outcome. RESULTS: SOD activity was not significantly different between the groups (P=0.218). GSH in the group whose participants were aged ≤35 years and were pregnant was higher than that in other groups. CAT activity in groups with younger participants was higher compared to the other groups. The mean TAC was higher in groups with pregnant participants compared to the non-pregnant women. Correlation analysis showed that: GSH level had a significant negative correlation with age (P<0.001, R -0.55) and a significant positive correlation with pregnancy (P=0.015, R=0.30). CAT level also had a significant negative correlation with age (P<0.001, R=-0.42) and the level of TAC had a significant positive correlation with pregnancy (P<0.001, R=0.59). CONCLUSION: According to our results, the levels of TAC, GSH and CAT in younger and pregnant women were higher compared with those undergoing ICSI cycles. Given the correlation of FF antioxidant activity with age and pregnancy, it is necessary to carry out more research on these compounds and the maintenance of pregnancy.

Negative effect of varicocele on sperm mitochondrial dysfunction: A cross-sectional study.

Background: Varicocele is an abnormal dilation and enlargement of the scrotal venous pampiniform plexus that impairs normal blood drainage and finally leads to infertility if not treated. Objective: This study aimed to figure out the impact of mitochondria status through the mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) assessment and its correlation with semen parameters to illuminate the impact of sperm mitochondria healthiness on normal sperm functionality. Materials and Methods: This analytical cross-sectional study was conducted with 100 men including 50 cases in the normozoospermic group (normal) and 50 in an infertile group with the non-varicocelectomy operation (varicocele) referring to Infertility Research and Treatment Center, ACECR Khuzestan, Iran. Routine semen analysis was performed according to World Health Organization guidelines, DNA fragmentation index, the MMP assay, ATP content, and apoptosis were carried out for all samples. Results: The results showed that the concentration, progressive motility, normal morphology, MMP, and ATP contents of sperm in varicocele were significantly lower than the normal group. In addition, the sperm DNA fragmentation index was significantly higher in the varicocele group in comparison with the normal group. Conclusion: Reduction in MMP and ATP contents, besides the loss of sperm parameters quality and increase in sperm DNA fragmentation, were seriously implicating sperm mitochondria dysfunctionality in varicocele men.

Betaine ameliorates impaired steroidogenesis and apoptosis in mice granulosa cells induced by high glucose concentration.

Betaine is a bioactive peptide whose beneficial effects on diabetes complications have been considered, previously. The present study aimed to investigate the possible protective effects of betaine against hyperglycemia-induced steroidogenesis impairment and apoptosis in mice granulosa cells. Ovarian granulosa cells were isolated from C57/BL6 mice and cultured in steroidogenesis medium (SM) containing 30 ng/ml FSH and 0.5 µM testosterone. The cells were cultured in SM containing low (5 mM) or high (30 mM) glucose concentrations for 24 h in the presence or absence of betaine (5 mM). At the end of the experiment, estradiol and progesterone were measured by ELISA in the culture medium. Expression of apoptosis and steroidogenesis associated genes and caspase-3 activity were determined by qRT-PCR and colorimetric assays, respectively. Exposure of mice granulosa cells to high glucose concentration inhibited the steroidogenesis by decreasing estradiol and progesterone secretion and downregulation of steroidogenesis-related genes including 3ßHSD, Cyp11a1, Cyp19a1, and StAR. Betaine treatment could ameliorate the steroidogenesis impairment at molecular and biochemical levels. High glucose concentration also enhanced apoptosis in mice granulosa cells that were characterized by elevation of caspase-3 activity, upregulation of bax gene and downregulation of bcl2 gene. Betaine treatment could attenuate the apoptotic-related changes induced by high glucose concentration in granulosa cells. According to the results of the present study, betaine could ameliorate the adverse effects of hyperglycemia on the physiological function of ovarian granulosa cells. The results highlight the potential role of betaine for the intervention of ovarian dysfunction in diabetic patients. Abbreviations: AABA: Betaine-α-aminobutyric acid; AGEs: Advanced glycation end products; bax: bcl2 Associated X; bcl2: B-cell lymphoma 2; AMPK: AMP-activated protein kinase; BHMT: Betaine homocysteine methyltransferase; C/EBP: CCAAT-enhancer-binding proteins; Cyp11a1: Cholesterol side-chain cleavage cytochrome P450; Cyp19a1: Cytochrome P450 aromatase; DM: Diabetes mellitus; E2: Estradiol; ERS: Endoplasmic reticulum stress; GCs: Granulosa cells; GLUT: Glucose transporter; FSH: Follicle-stimulating hormone; 3ßHSD: 3ß-hydroxysteroid dehydrogenase; IL-1ß: interleukin-1ß; LH: Luteinizing hormone; MDCK: Madin-Darby Canine Kidney cell; MT: Methionine synthase, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NLRP3: NLR Family Pyrin Domain Containing 3; NF-κB: Nuclear factor κB; P4: Progesterone; ROS: Reactive oxygen species; SGLT: Sodium dependent glucose transporter; SLC7A6: Solute Carrier Family 7 Member 6; StAR: Steroidogenic acute regulatory protein; STZ: Streptozotocin; Tumor necrosis factor α: TNF-α; TXNIP: Thioredoxin interacting protein.

A successful pregnancy following recurrent implantation failure with clinical laboratory strategy.

In treatment cycles of in vitro fertilization (IVF), 15% of the oocytes are immature and in the germinal vesicle (GV) phase. In rare occasions, more than 50% of the oocytes of a patient in a cycle are immature. Selecting fertility treatment for patients in this situation can be very challenging. The patient described in this report was a 35-year-old woman with primary infertility for 10 years. She had undergone fertility treatment six times, with no success. In her cycles, more than 50% of the oocytes were immature and in the GV phase. To address the situation, we developed a coordinated protocol involving medical and embryology procedures, analyzed the patient's medical records, and looked into the reasons of prior treatment failure. The development of a special and coordinated effort - instead of having medical and embryology teams working in isolation - proved efficient at providing better outcomes to the patient.

Effects of Tyrode's solution osmolarities and milk on bull sperm storage above zero temperatures.

BACKGROUND: Control of the medium osmolarity and temperature during long or short time sperm manipulation is essential. OBJECTIVE: The objectives of the present study were to find the effects of different osmolarities of modified Tyrode's solution and milk on the bull sperm during incubation at above zero temperatures. MATERIALS AND METHODS: Semen samples were collected twice from five Najdi bull. Centrifuged and most parts of seminal plasma were removed. First experiment: The concentrated semen were splited into nine aliquots to incubate in three different osmolarities (200, 300 and 400 mOsm) at three temperatures (5, 25 and 39°C) for 15 and 60 min of incubation. Second experiment: The semen samples were splited, mixed with the same volume of whole cow milk (5 and 25°C) and milk with 7% glycerol (5°C) and incubated for 15 and 60 min. RESULTS: Sperm motility severely affected (p<0.05) by incubation at low ionic tension (200 mOsm/l) especially at low temperature (5°C). The impact of low osmolarity on sperm viability can reduce by increasing the incubation temperature to 39°C. The decreased sperm motility, which was induced by lowering osmolarity, was not improved (p>0.05) by increasing temperature during 1 h of incubation. Milk can protect the sperm viability and motility at cool conditions and there is no beneficial effect of glycerol in combination of milk on sperm incubation at above zero temperatures (p<0.05). CONCLUSION: Iso- and hyper-osmotic solutions protect bull sperm motility and viability at 25 and 39°C, while milk can be used for protecting sperm at 5°C.