International Journal of Reproductive BioMedicine
Year:2012 | Volume:10 | Issue:3|Papers Count:15

Background: In this study we aimed to evaluate the impact of chronic exposure to the Gly-phosate (GP) on rat’s testicular tissue and sperm parameters. Objective: Testicular tissue, morphology of sperms and testosterone level in serum of mature male rats were analyzed.Materials and Methods: Animals were divided into two test and control-sham groups. The test group was subdivided into 4 groups (10, 20, 30 and 40 days GP administrated). Each test group (n=8) received the compound at dose of 125 mg/kg, once a day, orally for 40 days while control-sham group (n=16) received the corn oil (0.2 ml/day).Results: Microscopic analyses revealed increased thickness of tunica albuginea, obvious edema in sub-capsular and interstitial connective tissue, atrophied seminiferous tubules, arrested spermatogenesis, negative tubular differentiation and repopulation indexes, decreased Leydig cells/mm2 of interstitial tissue, hypertrophy and cytoplasmic granulation of Leydig cells, elevated death, immature sperm and increased immotile and abnormal sperm percentage. The carbohydrate ratio was reduced in first three layers of the germinal epithelium (GE) cytoplasm. The upper layers of the GE series were manifested with low rate of lipid accumulation in cytoplasm, while the cells which were located in first layers were revealed with higher amount of lipid foci. Hematological investigations showed significant (p<0.05) decreasing of testosterone level in serum.Conclusion: The current data provide inclusive histological feature of chronic exposure against GP with emphasizing on reproductive disorders including histological adverse effect on the testicular tissue, spermatogenesis, sperm viability and abnormality which potentially can cause infertility.

Background: Epigenetic reprogramming of differentiated cells can modify somatic cells into pluripotential state. Pluripotency can be induced in somatic cells by several approches. One of the easy ways to induce pluripotency is the exposure of the somatic cells to the embryonic stem cell (ESC) extract. Objective: The objective of this study was to increase the efficiency of reprogramming of granulosa cell as a differentiated cell into pluripotential state by using epigenetic modifier agents and extract.Materials and Methods: The human granulosa cells were cultured in the medium containing 5-Aza-Deoxycytidine and trichostatin A. Then, the cells were exposed to mouse ESCs extract and co-cultured with mouse embryonic fibroblast in the presence of leukemia inhibitory factor (LIF). Alkaline phosphatase test and also immonohistochemistery staining for Oct4, Sox2 and Nanog were performed after 24 and 72 hours and 1 week.Results: The granulosa cells showed the alkaline phosphatase activity after 24 hours and the enzyme activity maintained for 72 hours. They also expressed Oct4 after 24 hours. The cells also expressed Sox2 and Nanog, 72 hours after exposure to the ESCs extract. The expression of the pluripotency markers decreased after 1 week. It seems that the extract can induce dedifferentiation in granulosa cells and they can express the stem cell markers.Conclusion: It seems that the inhibitors of the methyl transferase (5-Aza-Deoxycytidine) and histone deacetylase (trichostatin A) could delete the epigenetic markers and prepare the cells for reprogramming by administration of the extract.

Background: Detection of best predictor of ovarian reserve in patients with temporarily or consistently elevated early follicular phase serum levels of FSH is one of the most important goals in assisted reproductive technique (ART).Objective: To evaluate whether high level of anti-mullerian hormone level is related to success of ART in patients with temporarily or consistently elevated early follicular phase serum levels of FSH.Materials and Methods: Sixty three women underwent intracytoplasmic sperm injection (ICSI) with GnRH-agonist long protocol or intrauterine insemination (IUI) in a prospective cohort study. FSH, inhibin B and anti-Mullerian hormone (AMH) levels were measured in these women whom were divided to three groups (persistently elevated FSH, variably elevated FSH and, normal FSH level). Basal characteristics, stimulation parameters, and pregnancy occurrence were evaluated.Results: AMH was significantly higher in women with persistently elevated early follicular phase FSH achieving pregnancy. Women with normal FSH did not have significant difference in AMH level between conceived and non conceived cycles. Women with only one elevated early follicular phase FSH achieving pregnancy did not have significant difference in AMH level with non pregnant women. Response to gonadotropin stimulation, recommendation to oocyte donation significantly differed between the groups.Conclusion: This study has demonstrated that relatively young women with persistently or intermittently elevated day 3 FSH levels have diminished ovarian reserve and lower ART success. However, in women whose FSH levels were constantly elevated, AMH (not inhibin B) concentrations were significantly higher in ART cycles resulting in pregnancy. Therefore, AMH level is a good predictor of ART outcome in patients with elevated early follicular phase serum levels of FSH.

Background: The exact pathophysiology of testicular degeneration, following varicocele has not been completely understood yet. Objective: The current study was designed to determine the effect of varicocele on germinal epithelium (GE) cytoplasmic biohistochmical alterations.Materials and Methods: To follow-up this study, left varicocele was induced in test groups. Non-varicocelized rats were served as control-sham (n=6). Following 4, 6 and 8 months, right and left testes were dissected out and the blood serum sample was taken. The GE cytoplasmic carbohydrate, lipid accumulation, lipase and alkaline-phosphates (ALP) ratios were analyzed. Serum levels of LH, FSH and testosterone were measured.Results: Observations demonstrated that in varicocele-induced rats, the spermatogenesis cell lineage exhibited lower number of cells with periodic acid shift positive cytoplasm, higher number of cells with lipid and ALP positive stained cytoplasm in comparison to control animals. Lipase enzyme decreased by the time in the test animals. In varicocelized groups the number of Leydig cells decreased in to 2.25±0.41 and 1.16±0.75 per one mm2 in left and right testicles respectively after 8 months, and these cells demonstrated an ALP positive feature. In test groups, the serum levels of LH and FSH reduced into 1.12±0.01 and 2.03±0.05 ng/ml respectively after 8 months. Although testosterone level diminished by the time in the test animals, and this decreasing was significant (p=0.031) after 8 months (3.08±0.10 ng/ml).Conclusion: Our results suggest that following varicocele induction major alterations occur in GE, which may lead to loss of GE cells physiological function and ultimately result in fertility problems.

Background: About 75% of the symptomatic patients who involved with endometriosis have pelvic pain and dysmenorrhea. Pentoxifyllin is one of the drugs that according to its mechanism could be effective for pain relief of endometriosis which has been used for endometriosis treatment recently.Objective: We conducted a comparative study for detecting the effect of pentoxifylin (as an immonomodelator) in preventing recurrence endometriotic pain with pentoxifylin plus a combined contraceptive pill with low dose estrogen (LD) and also the LD pill alone.Materials and Methods: This was a comparative clinical trial on 83 patients with the chief complaint (CC) of pain (dysmenorrheal /or pelvic pain) and with the end diagnosis of endometriosis, in an operative laparoscopy. Patients, dividing to 3 groups, were treated with pentoxifylin, pentoxifylin+LD and LD alone for 10 months. The severity of pain (dismenorhea and/or pelvic pain) was detected by visual analogue scale (VAS) before and after the treatment. The severity of endometriosis in the patients was: I in class I and II in class II and III in class III. The groups were matched for the pain. The number of the patients in group 1, 2 and 3 were 28, 28 and 27 respectively.Results: The pain was reduced in the groups of pentoxifylin+LD (p<0.001) and LD alone (p=0.00). The pain relief was not significant in the group of pentoxifylin alone (p=0.136). After treatment, the severity of pain was not significantly different between the LD group and the LD+penthoxyfillin group, but there was difference between these two groups and the group of penthoxyfillin alone.Conclusion: This study showed that penthoxyfillin actually could not have any effect on the pain relief of endometriosis. It also made it clear that penthoxyfillin could not increase the efficacy of LD when used with this medication.

Background: Main function of corpus luteum is progesterone synthesis that is significantly accompanied with an increase in levels of mRNA encoding of steroidogenic enzymes known as luteal markers.Objective: This study was designed to evaluate effects of lithium chloride on the release of steroid hormones and steroidogenic enzymes in gonadotropin-stimulated rats.Materials and Methods: Immature 23 days old Wistar rats were divided into 10 groups; each group comprised of 8 rats, and induced with single injection of pregnant mare’s serum gonadotrophin (PMSG) and followed by single injection of human chorionic gonadotropin (hCG). Then, rats were given lithium chloride (LiCl) or saline at 12 hours post-hCG injection. Ovaries were collected in 4-hour interval from 8-24 hour post-hCG injection. Expression pattern of steroidogenic acute regulatory protein (StAR), side-chain cleavage cytochrome P450 (P450scc) and 3b-hydroxysteroid dehydrogenase (3b-HSD) genes were determined by semi-quantitative RT-PCR. In addition, serum levels of progesterone and 17b-estradiol were measured by ELISA.Results: Our results showed that hCG stimulation of progesterone was markedly diminished and transcript levels of key steroidogenic enzymes were altered in the hormone-stimulated rats following LiCl treatment.Conclusion: These results suggest that critical steps in the function of corpus luteum are disrupted by lithium. It is concluded that LiCl is an effective factor for suppressing of steroid genes expression.

Background: Polycystic ovary syndrome (PCOS) is a common cause of ovulatory disorders and infertility with high LH to FSH ratio. In order to prevent further increase of LH and follicle atresia, different regimens for ovulation induction have been recommended using FSH alone. Objective: This study was performed in PCOS patients to compare ART outcomes in cycles induced by FSH alone, using either recombinant or urinary products.Materials and Methods: In a randomized trial, from 623 patients who underwent down regulation with GnRH analogue in a long protocol, 160 PCOS patients were randomly divided into two groups of 80. Group A received 150 IU/d recombinant FSH (Gonal-F) and group B 150 IU/d urinary FSH (Fostimon).Results: 33 cases (41.2%) in group A and 36 (45%) in group B achieved clinical pregnancy, which was not significantly different (p=0.67). Total number of oocytes retrieved (13.03±5.56 vs. 14.17±4.89, p=0.17), quality and number of embryos (7.42±3.35 vs. 7.63±3.28, p=0.68) and OHSS rate were similar in group A compared to group B. Endometrial thickness which was 9.66±1.67 mm in group A and 10.36±1.35 mm in group B, showed a significant difference (p=0.004).Conclusion: It seems that in PCOS patients, both pure FSH products used for controlled ovarian hyperstimulation have similar effects on ART outcome and can be used according to availability and patient acceptance without significant difference.

Background: Gestational diabetes mellitus (GDM) is common problem during pregnancy. Diagnostic criteria of this problem are based on foreign population. Because of differences in racial, cultural, and nutritional characteristics, we need to determine these criteria are suitable for Iranian population.Objective: To determine whether different diagnostic criteria of gestational diabetes mellitus (GDM) are suitable for Iranian population.Materials and Methods: Prospective study was performed on 617 pregnant women. 1804 subjects referred for 50 g glucose challenge test (GCT) between 24th and 28th weeks of gestation. 617 women with abnormal GCT (blood glucose ³130 mg/dl) underwent 100-g 3-h oral glucose tolerance test (OGTT). The results were classified by three diagnostic criteria: new "Iranian" diagnostic criteria based on the results from the 100-g 3-h OGTT performed in healthy participating women; the Carpenter and Coustan (CC) criteria; and the National Diabetes Data Group (NDDG) criteria. Obstetric and neonatal outcomes were recorded.Results: With 89% as the statistical cutoff value for the 100-g 3-h OGTT, the new diagnostic criteria were 92, 179, 153, and 121 mg/dL at 0, 60, 120, and 180 min. The K value was 0.945 for the new criteria vs. the CC criteria and 0.657 for the new criteria vs. the NDDG criteria (p<0.001). In women with GDM, the incidence rates of adverse outcomes by the new and CC criteria were similar, but higher than NDDG criteria (p<0.05).Conclusion: Carpenter and Coustan criteria are applicable to Iranian pregnant women for diagnosis of GDM.

Background: In the recent years, the use of laboratory blood factors such as FSH and inhibin-B for the assessment of spermatogenesis in different studies has increased; of course, the conflicting results have also been achieved.Objective: To investigate if the measurement of inhibin-B can help surgeon to reduce unnecessary diagnostic testicular biopsies in males with azoospermia.Materials and Methods: This cross-sectional study was done during July 2006 to September 2007 on 41 patients with azoospermia. FSH and inhibin-B were measured and bilateral open testicular biopsy was performed for all patients.Results: Sperm was seen in 29% of biopsies that in 100% of these samples inhibin-B was more than 100 pg/mL and FSH was less than twice the normal (p=0.001). Inhibin-B had significant correlation inversely with testicular fibrosis and Sertoli cell only syndrome (p=0.043 and p=0.011, respectively) and directly with incomplete spermatocytic maturation arrest and obstructive azoospermia (p=0.027 and p=0.013, respectively). FSH was only correlated with obstructive azoospermia (p=0.001).Conclusion: We suggest that if FSH is less than twice the normal, inhibin-B should be measured and if its level is less than 100 pg/mL, we can cancel about the half of unnecessary diagnostic testicular biopsies.

Background: Arsenic as an environmental toxicant is able to exert malformations in male reproductive system by inducing oxidative stress. Vitamin E (Vit.E) is known as antioxidant vitamin.Objective: The aim of this study was to investigate the harmful effects of sodium arsenite on sperm parameters and the antioxidant effects of Vit.E on sperm anomalies in sodium arsenite treated rats.Materials and Methods: Adult male rats were divided into 4 groups: control, sodium arsenite (8 mg/kg/day), Vit.E (100 mg/kg/day) and sodium arsenite+Vit.E. Oral treatments were performed till 8 weeks. Body and left testis weight were recorded and then left caudal epididymis was cut in Ham's F10. Released spermatozoa were used to analyze number, motility, viability and abnormalities of the sperm. Sperm chromatin quality was assessed by nuclear staining using acridine orange and aniline blue.Results: Body and testis weight showed no significant change in 4 groups (p>0.05). A significant decrease in the number, motility, viability and normal sperm morphology was found in sodium arsenite-treated rats compared to the control (p<0.001). Sodium arsenite had no effect on sperm DNA integrity and histon-protamine replacement (p>0.05). In sodium arsenite+Vit.E group, Vit.E could significantly compensate the harmful effects of sodium arsenite on sperm number, motility, viability and morphology compared to sodium arsenite group. In addition, sperm viability and motility was significantly increased in rats treated with Vit.E alone compared to the control and sodium arsenite+Vit.E group.Conclusion: Vitamin E could compensate the adverse effects of sodium arsenite on sperm parameters in adult rats.

Background: Cryopreservation has limited successes and in-vitro maturation is used to improve its results. Hydrostatic pressure (HP) plays an important role in follicular development. Objective: This study was designed to examine the effects of HP on in-vitro maturation of oocytes and cell death in cumulus cells derived from vitrified-warmed mouse ovaries.Materials and Methods: Preovulatory follicles were harvested from non-vitrified and vitrified-warmed 6-8 week-old female NMRI mouse ovaries and randomly assigned to following groups: non-vitrified (control), non-vitrified with HP exposure (treatment I), vitrified-warmed (treatment II) and vitrified-warmed with HP exposure (treatment III). The follicles of treatments I and III were subjected to HP (20 mmHg) for 30 min and after that all groups were cultured for 24h and assessed for in-vitro maturation of oocytes. The viability and apoptosis of cumulus cells and oocytes were assessed using supravital nuclear staining and TUNEL assay, respectively.Results: Oocytes harvested follicles in both control and treatment II had a significantly lower percentage of metaphase II oocytes (MII) than the treatment I and III (23.5±3.1, 15.03±4.6 and 32.7±3.2, 25.5±4.6; respectively) (p<0.05). Viability of the cumulus cells reduced in treatment I, II and III (83.4, 83.3 and 77.7%) compared to control (86.9%), (p<0.05). The apoptotic index in cumulus and oocyte complexes in treatments I and III (10.7±0.8 and 15.3±0.8) was higher than in control and treatment II (6.7±0.5 and 9.7±0.5) (p<0.05).Conclusion: These results demonstrate that HP had a mild effect on cell death incidence in cumulus cells without any effect on oocyte. However, it can be used as a mechanical force to improve in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries.

Background: Women with polycystic ovary syndrome (PCOS) often are infertile and even if they become pregnant, there are complications with some adverse outcomes. It has been reported that aspirin and metformin improve uteroplacental circulation and reduce pregnancy complications.Objective: To determine and compare uteroplacental circulation and obstetrics complications in pregnant women with PCOS treated with metformin, aspirin and control group.Materials and Methods: 105 pregnant women with PCOS were enrolled in this study after assessing uterine artery pulsatility index (PI) with Doppler ultrasonography at 12 weeks of gestation. The patients were divided into three groups and received metformin 2000 mg or aspirin 80 mg daily, or no intervention until the end of pregnancy. PI was assessed for the patients at 20 week of gestation and groups were followed up till delivery. PI and obstetrics complications such as gestational diabetes, preterm labor, preeclampsia and IUGR were compared among groups.Results: All groups had significant reduction in the mean uterine artery PI at 20 weeks measurement (p<0.05), but this reduction was more in metformin and aspirin groups than control group (p=0.002). There was a significant difference in mean uterine artery PI 20 week of gestation in three groups (p=0.005). Adverse outcomes have seen 4 out of 35 in metformin group, 7 out of 35 in aspirin group and 11 out of 35 in control group. There weren’t significant differences among groups (p=0.12).Conclusion: Metformin and low dose aspirin reduced uterine artery impedance but there was not associated with reduced obstetrics complication in women with PCOS.

Background: Abnormal placental invasion has increased parallel with persistent rise in Caesarean delivery. Management relies on accurate diagnosis and delivery should be planned at an institution with appropriate expertise and resources for managing this condition.Case: We present a case of a placenta invasion anomaly which is the major risk factors of peripartum deaths. In this case we try to explain our approach which reduces unnecessary hysterectomy rates.Conclusion: In order to avoid postpartum hemorrhage and hysterectomy protocols, our approach which consists bilateral hypogastric arterial ligation, Bakri balloon tamponade and If necessary methotrexate therapy can be applied succesfully.

Background: Acute appendicitis with appendicial endometriosis is a very infrequently encountered condition during pregnancy. Decidualization is the hypertrophy of endometrial stromal cells by the effect of progesterone. Similarly, in pregnancy, ectopic stromal endometrial cells in endometriosis can also be transformed by the same mechanism and ectopic decidua (deciduosis) may occur.Case: Here we report a 30 year old pregnant woman presenting twice with acute abdominal symptoms requiring surgery for appendicial and ovarian endometriosis and deciduosis. We emphasize that deciudualized endometriosis may first present during pregnancy with acute abdomen necessitating emergency laparotomy and complicating the course of gestation.Conclusion: To our knowledge only 9 cases in which decidualized endometriotic tissue causing acute abdomen necessitating surgery during pregnancy were reported in the literature. What makes our case special is that the patient needed two laparotomies during the pregnancy period which was a very stressful situation for both the patient and the physicians.

The big public health concern in this year is the nuclear crisis in Japan that causes contamination of nuclides around the world. The effect of the exposure to leaked nuclides from the present nuclear crisis on reproductive health is a topic to be concerned. In my previous publication, the evidence on the relationship between nuclear exposure and infertility can be confirmed. However, there are also other adverse effects on the reproductive system of the exposed subjects.