Journal of Reproduction & Infertility
Year:2004 | Volume:5 | Issue:1 (17)|Papers Count:11

Introduction: Mammalian reproduction looks like an immunological paradox, because fetal alloantigens encoded by father genes should induce cell meditated immune responses leading to fetal loss. Maternal immune system, in addition to local modulation, undergoes systemic modulations during pregnancy. Dendritic cells (DCs), as professional antigen presenting cells, play a key role in initiation and control of immune response and it seems that functional changes in these cells during pregnancy may contribute to the systemic immune tolerance. To address this issue, in this study we isolated and purified DCs from pregnant mice and evaluated their stimulatory potential to induce proliferative response of allogeneic T cells in unidirectional mixed leukocyte reaction (MLR). Materials and Methods: Following collagenase digestion of splenic tissue, using density gradient centrifugation (13% Nycodenz) and adherence properties of DCs to the bottom of tissue culture dish, 7×105 DCs were isolated from each spleen with more than 95 percent purity. Allogeneic T cells were isolated by nylon wool column, using their non-adhesive character to nylon wool. After radiation, isolated dendritic cells from pregnant and non-pregnant Balb/ c mice were used in mixed leukocyte culture with C57BL/6 mice T lymphocytes. T lymphocyte proliferation was measured 72 hours by H-thymidine incorporation. Results: 7 ×105 dendritic cells with the purity of >95% were isolated from each spleen. Also the yield of T-lymphocyte from inguinal and Brachial lymph nodes was about 3-5 ×105 with the purity of %85-90 The results showed that there is no statistical difference between stimulatory potential of DCs from pregnant (cpm=33000) and non-pregnant (cpm=35000) mice in induction of allogeneic T-Cell proliferation. Conclusion: These findings can result from low concentration of immune suppressor factors in circulatory system of pregnant mice or due to separation of dendritic cells from pregnancy microenvironment and their maturity in vitro in the absence of the immune suppressor factors.

Introduction: In recent years there have been great advances in vitrification of embryos. However, there is no reliable vitrification protocol to ensure a high embryo survival rate, because the mechanisms of embryo injury have not been discovered precisely. The aim of the present study was to determine the effects of vitrification on apoptosis in mouse blastocysts. Materials and Methods: Ninety five mouse blastocysts were obtained by flushing from Swiss Albino mouse and randomly divided into control and experimental groups. Blastocysts in the control group (52) were cultured in M 16 media for 2 hours and then the apoptotic index were obtained after staining by TUNEL technique with PI. Blastocysts in the experimental group (43) were vitrified just after flushing in EFS40 solution and kept in LN2 for one month. After thawing and culture in M16 for 2 hours, the apoptotic indices were obtained by TUNEL staining. Results: The results showed that the mean number of blastomeres in the vitrified blastocysts group (44.91±2.47) was not significantly different (P=0.176) from those that seen in the control group (50.23±2.9), while the mean number of apoptotic blastomeres in vitrified blastocysts group (4.08±0.28) was significantly higher (P=0.02) as compared to the control group (4.93±0.22). The mean apoptosis Index in vitrified blastocysts (11.87±0.63) was significantly higher (P<0.004) than the control group (9.12±0.67). Conclusion: We can conclude that the vitrification can increase apoptotic cell death in mouse blastocysts.

Introduction: The role of antisperm antibodies with a prevalence of 6-26% is well known in immunological infertility. Thus, there is clinical importance to determine ASA levels in both male and female. Nowadays, one of the most important discussed controversies in the field of immunological infertility is establishing a standard method to determine ASA. It seems that ELISA method will be more sensitive, specific and more diagnostic in determination of ASA if sperm surface antigens could be used as coated antigens, with least contamination to sperm intracellular antigens and nonspermic antigens. So, the aim of this study is designing an ELISA method by using the best method of sperm antigens extraction with at least contamination. Materials and Methods: In this study we designed an ELISA method with three different extraction methods of sperm antigens including sonication method, using SDS detergent, and application of LIS detergent, then we compared ELISA method based on the three extraction methods as well as two similar commercial ELISA kit (IBL Co, and Bioserv Co) with SpermMar test. Results: Comparing designed method with commercial kit indicated that among 28 sera which had 16 positive sera and 12 negative sera by SpermMar, 14 sera were true positive by LIS method and only 2 cases were false negative without any false positive results, whereas there were 5 true positive results and 11 cases false negative by the sonication method. The SDS method also had 13 true positive results with false negative and 4 false positive results. In addition, two commercial kit had in turn 7 and 4 cases true positive and both of them had I case false positive and in turn 9 and 12 cases with false negative result. ELISA method designed by LIS detergent has adequate sensitivity (87.5%) with higher specificity (100%) and efficacy (92.8%) than other extraction methods. There is a significant correlation between this designed method and Sperm Mar test (r=0.572).Conclusion: The results of this study indicated that ELISA method by LIS antigens has at least contamination with nonspermic antigens and it is better than other extraction methods and commercial ELISA kits for detection of antisperm antibody.

Introduction: Antisperm antibodies (ASA) are present in % 8-21 of infertile men. In vitro Fertilization (IVF) has been recommended as an effective procedure in couples with immunological male factor. Although this procedure has been found to bypass the inhibitory effect of antisperm antibodies on fertilizing ability of spermatozoa but the fertilization rate is reduced about %40 for ASA positive samples. The goal of present study was to investigate the correlation between anti-sperm antibodies measured by indirect flow cytometry and fertilization rate in infertile couples undergoing In vitro Fertilization (IVF).Materials and Methods: Semen samples were collected from 80 infertile men undergoing IVF cycle in Isfahan fertility and infertility center. Couples were classified based on fertilization rate into high and low groups. 52 couples had high (>50%) and 28 couples had low fertilization 8rate (<50%). Seminal plasma samples were incubated with normal motile spermatozoa from donor. Sperm bounded antibody was detected with FITC- labeled immunoglubin against human IgA and IgG by flow cytometer. The statistical analysis performed using c#, t-test, and Pearson's correlation. Results: There was significant difference between the mean levels of antisperm antibodies in high and low fertilization rate groups (p<0.001). However there was a significant inverse relationship between IgA antisperm antibody level and fertilization rate (r=-0.47 and p<0.001). Inverse relationship between IgG antisperm antibody level and fertilization rate was not significant (r=-0.2 and p=0.08).Conclusion: The results of this study clearly show that high level of IgA antisperm antibody decreases the fertilization rate. Therefore, it can be suggested that patients with high level of IgA antisperm antibody should become candidate for intracytoplasmic sperm injection (ICSI).

Introduction: Nowadays the wide spread use of GnRH agonists in ART protocols has emerged the need for luteal phase support by progesterone. However the time of starting progesterone administration is still obscure, some investigators recommend the day of oocyte retrieval and the others later. The present study was designed to investigate the effect of the progesterone administration timing before or after embryo transfer on the outcome of ART.Materials and Methods: A randomized clinical trial was designed to study a total of 575 women referred to Shahid Sadooghi University of Medical Sciences and Yazd Madar Hospital, undergone ART treatment. Using long GnRH-a protocol, the patients were randomly divided into two groups. Progesterone administration was started on oocyte retrieval day in the first group (n=307), versus after embryo transfer in the second group (n=268). Pregnancy rate were analyzed using statistical trials including X2 and T.Results: Regarding the age of couples, the duration and etiology of infertility, the number of follicles, retrieved oocytes and zygotes (3.0±1.4 in first group versus 2.8±1.4 in second group) no statistically significant difference was observed. The outcome, defined as a positive pregnancy test was not different in two groups.Conclusion: Considering the results and the disturbances following intramuscular injection of progesterone including the risk of bleeding, the administration of progesterone after embryo transfer is recommended.

Introduction: The third and forth stages of labor which involve the separation and expulsion of the placenta and an hour immediately following delivery are the two critical phases. Many maternal deaths in the developing countries result from complications of the third stage of labor and postpartum hemorrhage. Active management of labor and use of oxytocic drugs can reduce postpartum hemorrhage. This study compares the effects of oxytocin and syntometrine in preventing postpartum hemorrhage which is an important problem causing maternal morbidity and mortality after delivery.Materials and Methods: A clinical trial was designed to study 618 patients admitted to Zahedan Ghods Birth Center in 2001 for normal delivery. The patients were ran randomly divided into two groups. All Pregnancies were singleton, normotensive (BP<140/90) and free from medical disease. After exiting the anterior shoulder of fetus, 5 IU of oxytocin in the first group and 0.5 mg ergometrine plus 5 IU of oxytocin in the second group was injected intramuscularly. The abnormal postpartum hemorrhage by obstetrician's estimation, the need for repeated oxytocic or other interventions and less than 100 mm Hg fall in systolic blood pressure was determined. The length of third stage was determined for all of the patients.Results: Among the first group there were 20 cases (%6.47) with abnormal hemorrhage, compared to 8 cases (%2.58) in the second group. There was a significant difference between the two groups in postpartum hemorrhage (p<0.05) (using Chi-square test, CI=%95). There was no significant difference between two groups concerning the duration of third stage and the need for manual removal of placenta. The side effects were uncommon and the incidence of hypertention (Bp>140/90) was not different between two groups. Conclusion: some studies do not recommend the use of syntometrine, believing that there is no clinical difference in the effectiveness of oxytocin and syntometrine, whereas syntometrine can case hypertention and retained placenta. Our results similar to other researches show that syntometrine is more effective than oxytocin in preventing postpartum hemorrhage.

Introduction: Despite the success of family planning programs in Iran during the recent years, surveys indicate that a significant proportion of pregnancies are unwanted. The objective of the present study was to examine the rate of unwanted pregnancies in Iran and to analyze the effect of relevant socio-economic and demographic variables on unwanted pregnancies.Materials and Methods: The data and information for this study were obtained from the Iran Demographic and Health Survey (IDHS) undertaken by the Ministry of Health and Medical Education in 2000. The study was performed on 5420 married pregnant woman.Results: Results showed that around 35 percent of pregnancies were unintended, of which 19 percent were mistimed and 16 percent were unwanted. There was a meaningful relation between unintended pregnancy rate and pregnancy turn. Besides, around 42 percent of women who experienced unintended pregnancy were using a contraceptive method. Traditional methods and pills were the most prevalent methods used prior to unintended pregnancies in rural and urban areas respectively. Conclusion: Based on the results of the present study we suggest that the improvement of quality of family planning services is likely to decrease the level of unintended pregnancies in the future. The programs should target the couples who rely on traditional methods and pill in urban and rural areas respectively.

Introduction: Iran is one of the Middle East countries involved in AIDS crisis. Around 17% of cases are between 20-29 years old and 35% between 30-39. Considering the long incubation period (around 10 years), obviously a great percentage of cases have become involved in their adolescent ages. To address this issue, school based AIDS educational program for adolescents is being considered as an important strategy of the ministry of education and the ministry of health in Islamic Republic of Iran. Accordingly, studying and recognition of the most effective method of education which leads to increasing knowledge, improvement of attitude and enhancement of life skills related to prevention of AIDS among adolescents seems to be a priority. This study was designed to compare the effects of two educational interventions for prevention of AIDS, conducted by peers and adults (school counselors), on knowledge, attitude and self efficacy of female students in high schools of Tehran, studying in human science field, in 4th region of ministry of education in 2002-3. The objective was to found new initiatives in school based AIDS educational program. Materials and Methods: The mode of education used in this survey was Bendora Socio - Cognitive theory. A total of 441 students from 7 high schools in 4th region of ministry of education, were recruited using random cluster sampling and were allocated based on the educator, to 2 intervention groups (peer-led and adult-led group) and one control group. The education subjects included AIDS prevention and correct attitudes towards AIDS (vulnerability and rejecting patients) and self-efficacy skills (decision making, problem solving and assertiveness). Anonymous questionnaires were filled by the students before and 30 days after the intervention. Data analysis was done using SPSS-11, and statistical test including Analysis of variance, and Chi-Square were used.Results: The mean score of knowledge significantly i2n both interventional compared to control group after intervention (p<0.0001), but no significant difference was detected between peer-led and adult-led groups in this regard. Despite the equality between two interventional groups in the rise in knowledge, the type of information imparted by them was significantly different. Concerning the unique mode of education used in both groups, it seems that peer-led group more likely imparted those information which were a mater of concern among their peers and friends (e.g transmission mode) while adults (school counselors) imparted some other informations (e.g treatment, vaccination). There was no significant improvement in the attitude of vulnerability to wards AIDS among interventional groups from pre-intervention to post-intervention. Whereas the attitude of rejecting AIDS patients in both interventional groups has improved compared to control group (p<0.0001), no difference has been detected between the two interventional group in this regard. The educational intervention only in adult-led group has led to enhancement of self-efficacy skills significantly compared to control group, from pretest to posttest, however, there was no significant difference between peer-led and adult-led group in this issue.Conclusion: we recommend both peer-led and adult-led programs to be employed in school-based AIDS education, particularly peers in imparting information according to their peers need and adults in training self efficacy skills. However further studies with lager sample size and longer educational periods are recommended to confirm the current results.