International Journal of Reproductive BioMedicine
Year:2009 | Volume:7 | Issue:2|Papers Count:8

Background: Many attempts have done to improve cryopreservation of mammalian ovaries using simple, economical and efficient technique “vitrification”. Objective: The aim of the present study was to compare the mouse ovaries cryopreservation by direct cover vitrification (DCV) using different concentrations of ethylene glycol (EG) with conventional vitrification methods (CV).Materials and Methods: Ninety NMRI mice were sacrificed by cervical dislocation; their ovaries were divided into three main experimental groups: control or non-vitrified group, CV group and DCV groups with 4, 6 and 8M EG as cryoprotectant. After vitrification-warming, the viability of mechanically isolated follicles and the morphology of ovarian follicles by light and electron microscopes were studied. Results: The normality of primary and preantral follicles in non-vitrified and CV groups were higher than those achieved by DCV groups (p<0.001). The survival rates of isolated follicles in non-vitrified, CV and DCV groups with 4M, 6M and 8M ethylene glycol were 98.32, 96.26, 84.10, 85.46 and 84.56 %, respectively and in DCV groups it was lower than other groups (p<0.001). The ultrastructure of ovarian follicles was well preserved in CV technique. The follicles in DCV groups appeared to have vacuolated oocyte with nuclear shrinkage and irregular distribution of cytoplasmic organelles. Their mitochondria were located mainly in the sub cortical part of the oocyte and the granulosa cells demonstrated some signs of degeneration.Conclusion: DCV of mouse ovarian tissue using only EG has induced some alteration on the fine structure of follicles. The integrity of mouse ovarian tissue was affected by DCV technique more than CV.

Background: Leptin is an adipokine that circulates in a free form and bound to a soluble leptin receptor. Patients with polycystic ovary syndrome have increased insulin resistance and high incidence of obesity.Objective: This study was carried out to evaluate levels of leptin and free leptin in women with polycystic ovarian syndrome (PCOS) and note any relationships with insulin resistance and adiposity.Materials and Methods: We assessed the correlation of metabolic parameters with the levels of free leptin and it’s bound form in 27 PCOS women (aged 26±5.6 years) and 27 healthy women with normal menstrual cycle as controls (aged 25±4 years).Total leptinand insulin levels were measured using ELISA. Free leptin form was purified by Gel filtration chromatography and their collected fractions were measured by a sensitive ELISA-Kit. Insulin resistance was calculated by homeostasis model assessment (HOMA).Results: In PCOS patients and control group a correlation between leptin and body mass index (BMI) was found. A significant difference was found between leptin and free leptin levels in PCOS subjects and controls (p<0.05). Significant correlations were found between free and total leptin with insulin resistant in PCOS subjects (r=0.78 p=0.00, r=0.84 p=0.003) and control groups respectively (r=0.86 p=0.00, r=0.69 p=0.00). Conclusion: Total and free leptin forms are correlated significantly with BMI in patients with PCOS and in controls. Total and free leptin forms showed significant correlations with insulin resistance but no significant difference was seen in the two groups investigated.

Background: Diazinon (DZN) is an organophosphate insecticide which is used worldwide in agriculture. The exposure to this chemical might lead to damages to the living systems.Objective: The present study was done to investigate the effects of diazinon on the structure of testis and levels of sex hormones in adult male mice.Materials and Methods: For this experiment, the mature male mice divided into three groups; Control (no injection), sham (corn oil injection) and DZN (diazinon was administrated at dose of 30 mg / kg for 30 d five consecutive days per week). Animals were killed 35 days after the latest injection. Testes tissues sections were provided to investigate the histopathological changes. Serum testosterone, LH and FSH concentrations were measured by radioimmunoassay. Data were analyzed using of oneway ANOVA. Significance was set at p<0.05.Results: A significant reduction was observed in diameter and weight of testes after DZN administration. Furthermore, DZN brought about significant reduction in sperm counts and spermatogenic, Leydig and Sertoli cells and a decrease in serum testosterone concentration. Histopathological examination of testes showed degenerative changes in seminiferous tubules (p<0.001). The levels of LH and FSH were increased in DZN groups compared to the control and sham groups (p<0.05).Conclusion: DZN is a toxicant for mammals’ spermatogenic cells during the early spermatogenesis. Therefore, application of DZN should be limited to a designed program.

Background: Fertility protection is important in young patients with Hodgkin's lymphoma.Objective: The goal of this study was to determine the effects of ABVD and ChlVPP chemotherapeutic protocols for Hodgkin's disease on the spermatozoa fertility indices of male rat.Materials and Methods: After determining tolerance dose of drugs in pilot study, 24 male rats were divided to four groups: ABVD (doxorubicin, bleomycine, vinblastin, dacarbazine) group, ChlVPP (chlorambucil, vinblastin, procarbazine, prednisolone) group and two control groups one for each treatment group. One half of the lethal dose for fifty percent of population was used for treatment of animals in each protocol. Spermatozoa were used for computer- assisted sperm analysis (CASA) and morphology analyses. Heads of spermatozoa were counted.Results: Body weight, testis and epididymis weights, spermatozoa number, and live ratio in treated rats were significantly less than their control groups (p<0.05) specifically these parameters in ABVD group was less than ChlVPP group (F= 19.6, p=0.000). Spermatozoa morphology in treated groups were more abnormal than control groups (p<0.05). Evaluation of reproductive system efficacy showed that there was no pregnancy in ABVD group and in ChlVPP group there was only one pregnant female (16.6%).Conclusion: According to this study results, the ChlVPP had fewer side effects than ABVD in tolerance doses on male rats' reproductive system. More clinical trial studies are suggested on Hodgkin's patients. With equal treatment effectiveness, it will be better to use the most reliable and safe treatment especially in young patients.

Background: Sperm selection for ICSI based on morphology and motility might not be relevant to chromatin integrity. Thus sperm selection based on sperm characteristics has been suggested.Objective: The aim of this study was to compare the efficiency of Zeta method with routine Density Gradient Centrifugation method (DGC) for the selection of sperm with higher DNA integrity.Materials and Methods: Semen samples were obtained from 63 individuals referring to Andrology Unit of Isfahan Fertility and Infertility Center. Semen analysis was carried out according to WHO criteria. Each semen sample was divided into three equal portions. One portion was used as control, the second portion was used for Zeta method and the third portion underwent DGC method. Each portion was evaluated to DNA integrity by TUNEL assay. Student t-test was carried out using SPSS and p-value lower than 0.05 was considered significant.Results: The mean number of sperm DNA fragmentation in Zeta and DGC methods were significantly decreased compare to the control group (p<0.001). In addition, Zeta method was more efficient than the DGC method in the selection of sperm with intact DNA (p<0.001).Conclusion: The Zeta method appears to be a suitable procedure to recover sperm with normal DNA integrity.

Background: The aim of this study was to determine the incidence of AZF (Azoospermia Factor) microdeletions of the Y chromosome in infertile Turkish male patients and intracytoplasmic sperm injection (ICSI) outcome of these patients. Objective: This study was undertaken in order to evaluate the outcome of intracytoplasmic sperm injection (ICSI) in infertil man with AZF microdeletions.Materials and Methods: We evaluated 348 azoospermic and oligozoospermic patients retrospectively. Fourty of these patients had various types of AZF microdeletions. These patients had non-obstructive severe oligoasthenospermia or azoospermia with normal karyotype. Azoospermic patients underwent testicular sperm extraction and aspiration (TESE, TESA). Then ICSI was performed to patients who had testicular sperm or ejeculat.Results: Fourty patients with AZF microdeletion were evaluated in this study. No spermium could be found in 27 patients. Three of these patients had only AZFa microdeletion, three had AZFb microdeletion, three had AZF (b+c), six had AZF (a+b+c) and 12 patients had AZFc microdeletion. Only two of all patients achieved a pregnancy and both had only AZFc microdeletion.Conclusion: AZFc microdeletions have a better prognosis for achieving spermium in ejaculate or TESE, TESA materials.

Background: Considerable attention is focused on effects of electromagnetic field (EMF) and its increasing use in everyday life. Appliances and various equipments are sources of electromagnetic fields with a wide-range of technical characteristics. Objective: In this study we investigated the effect of EMF (50 Hz, 0.5 mT) on epididymis and deferens duct in mice.Materials and Methods: 30 BALB/C mice were selected and divided into three groups (control, sham and experimental). While control and sham groups were not exposed to EMF, the experimental group was exposed to EMF (50 Hz, 0.5 mT) 4 hours a day, 6 days per week and for 2 months. At the end of 2 months, the mice were sacrificed, dissected and samples from epididymis and vas deferens in all groups were taken and processed for light microscopic studies. 40 microscopic fields from each group were randomly selected. The diameters and the height of epithelial cells of epididymis and deferens duct in 3 groups were measured and compared using statistical methods. Results: The data showed that the mean diameter of epididymis and deferens duct in EMF group was significantly decreased compared to the control group (p=0.001). The height of epithelial cells in epididymis and deferens duct in EMF group was considerably reduced compared to the control and sham groups (p=0.001). In addition, the weight of testes in EMF group was significantly decreased compared to the control and sham groups (p<0.007).Conclusion: It could be concluded that the exposure to EMF leads to detrimental effects on male reproductive system in mice as seen by a decrease in diameter of reproductive ducts, the height of epithelial cells and weight of testes.

Background: Induction of ovulation in ART is necessary for superovulation and the side effects of superovulatory drugs are debated. Oxytocin as a natural hormone, have receptors and is synthesized by several reproductive organs. Preovulatory presence of oxytocin receptor mRNAs in granulosa cells indicating a role for oxytocin in follicular development.Objective: The aim of the present study was to investigate the effect of exogenous oxytocin injection on folliculogenesis, ovulation and endometrial growth in mice. Materials and Methods: Forty adult female mice were divided into two groups as control and experimental. The mice at their sterous cycle received 1 IU/gr oxytocin, in experimental, and the same volume of solvent in control groups. Half of the mice in each group are sacrificed at 24 hours post injection and the other half, 48 hours after the injection. Ovarian samples fixed in 10% formalin, embedded in paraffin and sections were stained with H and E and studied using stereological techniques. The data were analyzed with Man – Whitney test.Results: Microscopic examination revealed that the number and morphological features of follicles at different stages were similar at 24 and 48 hours post injection in both groups. The volumes of the ovaries were similar in both groups at 24 hour. However, at 48 hour, the volume of ovaries, corpora lutea and endometrial thickness, in experimental group, were significantly higher than those in control group (p< 0.05). Conclusion: According to the increased volume of corpus luteum in the experimental group, it is concluded that oxytocin injection has a stimulatory effect on induction of ovulation.