International Journal of Reproductive BioMedicine
Year:2007 | Volume:5 | Issue:4|Papers Count:10

Fertilization involves direct interaction of the sperm and oocyte, fusion of the cell membranes and union of male and female gamete genomes. The completion of this process and subsequent embryo development depend in part on the inherent integrity of the sperm DNA. Sperm genome quality has been emphasized for several years as playing a major role in early embryogenesis. There is clinical evidence showing that human sperm DNA damage may adversely affect reproductive outcomes and that spermatozoa of infertile men possess substantially more DNA damage than do spermatozoa of fertile men. Testing DNA integrity may help selecting spermatozoa with intact DNA or with the least amount of DNA damage for use in assisted reproductive techniques (ARTs). This review will focus on how sperm DNA is organized, what causes sperm DNA damage and what impact this damage may have on reproductive outcome.

Background: Determination of transgenic embryos from non transgenic embryos sibling is an important step in producing homozygous transgenic mice. These steps need by PCR or southern blotting followed extraction of DNA, but both techniques require skill and consume time. Objective: The aim of this study was simulation of high accuracy method using novel enhanced green fluorescent protein (EGFP) gene cassette to eliminate some consume time in livestock industry to assay high quality embryos and morphological plasticity.Materials and Methods: We modified pQE-Tri systemic vector with EGFP and IRES sequence to trace out coming planning of molecular farming transgene using coinjection method.Results: The combination of these sequences successfully showed the faint and normal expression of transgene in mouse pre-implantation stage embryos. The low rate of surviving green positive embryos as compare as only medium and physical treatments could be partly from the physical damage caused by microinjection and gene integration.Furthermore, application of the enhanced GFP marker facilitated subtle detection of asymmetrical division appeared in some of transgenic embryos.Conclusion: The results of current mouse simulation model imply that an efficient production and propagation of transgenic livestock can be done by co-injection of every economical gene with this novel transgene and also it can be suitable gene cassette for numerous experiments and study of protein behaviors in living cells.

Background: Preparation of oocytes is one of the critical factors that determine the developmental competence of embryos produced by in vitro fertilization (IVF).Objective: In this study, the effect of cysteamine, type of media and glutathione (GSH) level on blastocysts development after in vitro maturation of mouse oocytes were investigated.Materials and Methods: Premature female mice were primed with pregnant mare stimulating gonadotrophin (PMSG), and germinal vesicle (GV) stage oocytes were obtained 45 hr later. GV oocytes were cultured in presence of 0, 50, 100, 200 and 500 mm cysteamine in TCM199 and MEME media. After IVM, MII oocytes were in vitro fertilized (IVF) and in vitro cultured (IVC) in order to observe embryo development. A group of In Vivo Ovulated (IVO) oocytes after priming with PMSG and HCG also were included in this study. 5,5-Dithio-bis (2nitrobenzoic acid) DTNB-recycling protocol was used for GSH assay.Results: Rate of IVM and IVF were improved in all oocytes treated with cysteamine in the two medium except 500 mm (81% MII rate in TCM and 64% MII in MEME). Rate of blastocyst in 100 mm cysteamine in TCM1199 and 200 mm in MEME was higher compared to control groups (In TCM 45% and in MEME 35%). In vivo MII and GV oocytes represented the highest and lowest GSH level respectively.Conclusion: Our results revealed that the media and concentration of cysteamine can affects on IVM, IVF and rate of blastocysts development on dose dependant manner.

Background: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimizing.Objective: The aim of this study was to improve the single step and step-wise vitrification effects on maturing mouse GVBD oocytes by ethylene glycol (EG) in conventional straws.Materials and Methods: Oocytes with compact cumulus cells were cultured for 3hr in TCM199 supplemented with 10% fetal bovine serum (FBS) in 5% CO2 in air. GVBD oocytes were randomly allocated into three groups. (1) Control (non-vitrified group), (2) exposed to single-step vitrification (contained of EG 20%+0.5M sucrose), (3) exposed to step-wise vitrification (2%, 5%, 10%, 20%EG +0.5M sucrose). In vitrification groups,oocytes were thawed and underwent additional 21 hr maturation. Viability of oocytes and maturation to MII stage were analyzed using inverted microscope and additionally by staining of propidium iodide and Hoechst 33342.Results: All non-vitrified oocytes were viable after 24 hr; however, viability of vitrified samples in single-step group was significantly lower than that of the step-wise and controls Groups. Also, the maturation rate in the step-wise group was significantly higher (p<0.05) compared to single-step.Conclusion: These results suggest that step-wise vitrification of GVBD oocytes as compared to single step vitrification was better in the rate of survival and in vitro maturation of oocytes.

Background: Spinal cord injury (SCI) occurs most often to young men at the peak of their reproductive health. Only 10% of SCI men can father children without medical assistance due to potential impairments in ejaculation and sperm quality.Objective: The main objective of this experimental study was to evaluate the epididymal necrospermia- sperm death, after chronic SCI in rat.Materials and Methods: Forty-five adult Wistar rats were divided into 3 groups of SCI, sham, and control. Following laminectomy, SCI was induced onto exposed dura matter (T10) of anesthetized rats. Sham group underwent laminectomy of T10 only; while, control rats were not exposed to any type of injury or medication. The spermatozoa from cauda epididymis were aspirated after 50 days for analysis of necrospermia with three assays of Eosin-Y staining, Hypo-osmotic swelling (HOS), and Hoechst 33258 fluorescent dye.Results: The rate of necrospermia in SCI rats was significantly increased when compared with other groups (p<0.05). Also, the rates of necrspermia in SCI samples were similar with application of 3 assays (Eosin-Y: 46.11±9.41; HOS: 45.88±8.89; Hoechst: 46.76±9.31). Total necrospermia was not observed in any of the epididymal samples.Conclusion: The results showed that chronic SCI is associated with high rate of epididymal necrospermia in mammals such as rats. It is, therefore, recommended that an effective laboratory technique, such as Hoechst 33258 should be used for separation of live and motile sperms from necrospermic ones for assisted reproduction program.

Background: During the detorsion of a torsioned ovary, oxidant agents are released and melatonin as an antioxidant can reduce ischemia. We studied the histopathological changes after using melatonin on experimental torsioned ovary in cat.Objective: The aim of this experimental study was to investigate the effects of Melatonin on histopathological changes in torsion – detorsion injury in cat ovary.Materials and Methods: An adnexal torsion – detorsion model was created by using 20 adult cats randomly divided equally in to 2 groups of Saline and Melatonin. Ischemia was induced by iathrogenic 360° clockwise torsion of the cat adnex for 3 hr. Reperfusion was achieved for 3 hr. Melatonin or saline were injected intra peritoneally (10mg/kg) 30 min before ovarian detorsion in both groups. After 3 hr of ovarian detorsion, ovarian tissue was removed and fixed in 10% formalin solution, embedded in paraffin and evaluated for ischemic indices.Results: Histological examination showed a significant improvement in ovarian morphology in the melatonin treated cats. Edema and vasoconstriction in saline group were more severe than Melatonin group (p-value = 0.009). Hemorrhage and leukocyte infiltration were also more obvious in saline group (p-value 0.0018).Conclusion: Our results demonstrated that Melatonin administration reduced ovarian histopathological damage due to oxidative injury associated with torsion.

Background: Severe preeclampsia is a quite well-known entity with high incidence of both maternal and fetal morbidity and mortality. Although little is known about its etiology, inherited disorders of hemostasis and antiphospholipid syndrome have been postulated as common causes. The present study was conducted to evaluate the association of these two entities with preeclampsia in a group of Iranian patients. Materials and Methods: A case-control study was performed on 26 parturients with severe preeclampsia and 26 healthy pregnant women who were matched according to the age, parity, gestational age and previous history of abortion. A 10cc blood sample was obtained and the following factors were measured: factor V Leiden, protein S, protein C, antithrombin III, anticardiolipin antibodies (IgM and IgG) and the presence of the lupus anticoagulant antibody.Results: We have not found any significant difference in the values of factor V Leiden, antithrombin III, protein C, protein S, and anticardiolipin-IgG between preeclamptic (case) and non-preeclamptic (control) parturients. Meanwhile, lupus anticoagulant antibody was detected in one case and one control. However, anticardiolipin IgM was shown to be significantly higher in the preeclamptic patients. Severe preeclamptic parturients were 4.4 times more likely to develop elevated levels of IgM (OR=4.4, 95% CI=1.9-10, p<0.05).Conclusion: Our results failed to reveal any significant association between preeclampsia and indices of inherited disorders of hemostasis, except for anticardiolipin IgM. Thus, routine screening of these indices are not recommended due to high expenses and shortness of reliability.

Background: The value of serial measurement of serum b subunit of human chorionic gonadotropin (bHCG) and ultrasonography in the early diagnosis of ectopic pregnancy has well established.Objective: The objective of this study was to explore the diagnostic value of raising level of serum bHCG, single measurement of progesterone (P) and estradiol (E2) in early diagnosis of ectopic pregnancy.Materials and Methods: Serum levels of bHCG and estradiol were measured by Radio Immuno Sorbent Assay (RIA) and progesterone level was measured by Enzyme Linked Immuno Sorbent Assay (ELISA) techniques in 43 symptomatic women with ectopic pregnancy and 42 women with normal intrauterine pregnancy in Alzahra Hospital, Tabriz, Iran. These values were compared by T-test. By determining cut-off levels of these parameters the efficiency and sensitivity of them in prediction of ectopic pregnancy was estimated.Results: The mean serum levels of bHCG, estradiol and progesterone in patients with ectopic pregnancies (940±552 mlu/ml, 593±237 pg/ml, 5.83±3.41 ng/ml, respectively) were significantly lower than these levels in normal intrauterine pregnancies (4620±2030 mlu/ml, 1627±435 pg/ml, 24.8±6.08 ng/ml, respectively). The average rate of bHCG rising was 8.2% for 24 hours in patients with ectopic pregnancy (EP) and 32.8% in normal intrauterine pregnancies (NIUP).Conclusions In this study single measurement of serum progesterone level has the greatest sensitivity (100%) and specificity (98%) in the diagnosis of ectopic pregnancy.

Background: Group B streptococcus is regarded as a potential factor for adverse outcomes of pregnancy such as preterm birth.Objective: To study the association of maternal vaginal colonization with group B streptococcus (GBS) and preterm labor.Materials and Methods: From April 2005 to May 2006, vaginal culture for GBS were conducted in 101 laboring women with a gestational age of 24-37 weeks and 105 women admitted for term delivery at maternity center of Afzalipour Hospital in Kerman, Iran. Student`s t test and Chi square test were used to compare continuous and categorical data between the groups. Using multivariate logistic regression the association between GBS colonization and preterm labor was analyzed. P-values<0.05 were considered as significant.Results: Colonization was detected in 9.2% of all mothers. Although GBS colonization was found more frequently in preterm than term patients (12 v/s 7 cases), the difference was not statistically significant. However, GBS positivity was roughly associated with preterm labor. Age was also a risk factor for GBS colonization. No case of perinatal sepsis occurred during the study period.Conclusion: Maternal colonization for GBS is relatively low in our center. Increasing age enhances the risk of colonization. Vaginal colonization of GBS is relatively associated with preterm labor.

Background: Smoking has negative effects on reproductive process. Exposing to cigarette smoking (passive smoking) may exert some effects as the direct smoking.Objective: The aim of this study was to evaluate the correlation between ovarian response and passive smoking in women who underwent ART cycles.Materials and Methods: One hundred-sixty patients who underwent ICSI between 2000 and 2001 were studied in a prospective cohort study. The case group included women whose husbands smoked at least 5 cigarettes daily for 1 year or more. The control group included women with nonsmoking husbands. Women with high FSH level (>12 IU/ml) were excluded. Long standard protocol with GnRH agonist and HMG were used in all patients. In vitro fertilization and embryo transfer was carried out in a standard fashion.Results: Eighty one women were in case group and 82 in control group. Ovarian response variables were not significantly different between two groups but there was a significant relation between passive smoking and fertilization (RR= 1.18, 95% CI: 1.07- 1.31). However pregnancy rate was not significantly different between two groups. Moreover there were no significant differences between heavy and light smokers in ovarian response outcomes.Conclusion: This study showed no correlation between ovarian response parameters and passive smoking in women underwent ART cycles, whereas fertilization rate is significantly lower in this group compared to control group. It may be related to sperm quality than oocytes. Assessment of nicotin in follicular fluid and cytogenetic evaluation of embryo before transfer are recommended for more information and confirmation.