https://www.jri.ir/ "Journal of Reproduction & Infertility" is owned, published, and copyrighted by ©2009 Avicenna Research Institute. No parts of this journal may be reproduced in any form or by any means unless properly referenced or sent a notification letter through www.jri.ir en The effects of Matrigel on the developmental processes of mouse blastocysts Introduction: Culturing embryos on Matrigel™, is one of the most suitable methods for study-ing in vitro embryonic developments. As Matrigel has not been used extensively in different species for embryonic development studies, this study was undertaken to determine the effects of Matrigel on the developmental processes of mouse blastocysts. Materials & Methods: To a number of female NMRI mice, hMG and HCG injections were made for ovulatory stimulation and then they mated with males from the same strain. Later on, blastocysts were obtained and randomly divided into 2 groups: 150 case blastocysts and 134 control blastocysts. Blastocysts were cultured for 48 hours in M16 medium, supplemented with 4mg/ml of bovine serum albumin (BSA). Later, the blastocysts were compared with the blastocysts cultured in Matrigel plus the same medium. Developmental studies were carried out every 24 hours for 2 days. The data was analyzed by SPSS software and the results were tested by chi-squared. Results: After 24 hours, a significantly higher ratio of embryos reached the hatched blastocysts stage Ι in the case group (74%), compared with that of the control group (52.2%), (p<0.05). At the same time the percentage of fragmented blastocysts in the control group was 11.9% which was significantly higher than the case group (2%), (p<0.05). After 48 hours, 41% of blastocysts cultured in the control medium, developed to stage I, the value being significantly more than the blastocysts in the case group (p<0.05). Moreover, after the same period of time (48 hours), the percentage of stage II hatched blastocysts in the case group (79%) was higher than the control groups (59%), (p<0.05). Conclusion: Matrigel use in enriched culture media can increase development and growth of mouse blastocysts. It also seems that ultrastructural studies of cultured embryos or immunocyto-chemical studies from this regard would be beneficial in understanding the processes involved. https://www.jri.ir/article/219 Evaluation of acid phosphatase activity alterations in mouse ovary after ovarian hyperstimulation in early stages of pseudo and normal pregnancies until implantation time Introduction: Acid phosphatase (ACP) is a lysosomal enzyme which contributes in ovarian metabolic functions such as oocyte maturation, resumption of mitotic divisions, germinal vesicle breakdown and ovulation. It digestes the corpus lutetium and helps the atresia of follicles by autophagia and hetrophagia activities. Considering the hormonal control of this enzyme, the present study was designed to deterimine the ovarian ACP activity after ovulation stimulation by the administration fo PMSG and hCG during preimplantation period. Materials & Methods: For this purpose a number of 6 to 10-week old female NMRI mice were selected and randomly divided into control and hyperstimulated groups after the administeration of PMSG and hCG, and later to pregnant and pseudopregnant groups. The mice were rendered pseudopregnant by mechanical vaginal stimulatin. Five mice in each group were sacrificed by cervical dislocation at the first to the sixth day of pregnancy for biochemical assays. The ovarian samples were obtained and were hemo-genated and centrifuged at 14000 g. The activity of the enzyme was determined using paranitrophenyl phosphate as substrate and later the specific activity of the enzyme was calculalated according to the amount of total protein. The data were analysed by Mann Whitheny test. Statistical significance was indicated by a P value less than 0.05. For hisotchemical evaluations, the sampels were obtained from one of the ovaries in each mouse and then 5 m thick cryosections were prepared. Cryosections were stained by Goumory method. Results: The ACP activity of ovarian tissues in the first day of pregnancy in the normal pregnant and pseudopregnant control groups, hyperstimulated normal pregnant and pseudopregnant groups were 0.340.04 IU/mg, 0.390.04 IU/mg, 0.40.08 IU/mg, 0.450.01 IU/mg respectively and in the fourth day were 0.690.1 IU/mg, 0.61 0.06 IU/mg, 1.090.10 IU/mg and 0.79 0.05 IU/mg. The results showed that biochemical findings correlated with histochemical observations. The ACP reaction changes were seen mainly in granulosa cells with a minimum enzyme activity in the first day (zero activity) and a maximum activity in the fourth day of pregnancy (+3). Conclusion: The increased ACP activity on the 3rd- 4th days of pregnancy, may be due to the este-roidogenic activity of granulosa cells. Also, the results showed that ovarian hyperstimulation could not change the pattern of ovarian ACP activity during early stages of pregnancy. More research is required in this area for a better understanding of the processes. https://www.jri.ir/article/220 A morphologic and morphometric study of adult mouse testis following different doses of busulfan administration Introduction: Anti-cancer drugs have adverse effects on spermatogenesis. Therefore, informa-tion on their role for the prevention of germinal epithelium destruction is necessary. The aim of this study was morphologic and morphometric evaluations of testes, measurement of volume and volume density of testes parameters, measurement of tubular diameters, germ and somatic cell counts following administration of different doses of busulfan in adult mice. Materials & Methods: In the present study, 42 male NMRI mice aged 6-8 weeks were used. The animals were divided into 5 groups. Case groups received a single dose of busulfan by intraperitoneal injection as 5, 10, 20 and 40 mg/kg in the first, second, third and forth groups respectively. The control group received only the solvent for busulfan. All the animals were killed 35 days after treatment and their testes were dissected out and processed for light microscope studies. Then morphometric studies were performed on testicular parameters. The data were analyzed using ANOVA and Tukey test and p values less than 0.05 were considered significant. Results: Busulfan administration in 5, 10, 20 and 40 mg/kg doses significantly reduced most morphometric parameters in testes with a maximum effect in the 40 mg/kg group. Volumes of testes, tubules and germinal epithelia were decreased significantly in the experiment groups (p<0.05) however, the volume of interstitial tissue increased (p<0.05). Tubular diameters and thickness of epithelia were also decreased in the experiment groups. Number of germ cells was reduced, but number of sertoli cells was not affected. The number of leydig cells were not affected in 10 and 20 mg/kg busulfan treated groups, however in the 40 mg/kg treated group they were increased significantly (p<0.003). In 5 mg/kg treated group there were no significant differences in morphologic and morphometric studies. Conclusion: Busulfan could reduce testicular parameters and disrupt spermatogenesis through affecting both germ and somatic cells in a dose dependent manner. Therefore, the side effects of busulfan on spermatogenesis should be considered during cancer therapies. https://www.jri.ir/article/221 Influence of cryoprotectants on DNA fragmentation of in vitro produced porcine blastocysts Introduction: The aim of the present study was to investigate the effects of different cryoprotectants (CPs) on DNA fragmentation of in vitro produced blastocysts to determine an appropriate cryoprotectant for embryo cryopreservation. Therefore, the precise aims of the study were to assess the toxic effects of different cryoprotectants in terms of survival rate and to evaluate the effects of different CPs on DNA fragmentation in in vitro produced porcine blastocysts. Ethylene glycol, 1,2 propanediol and glycerol are common cryoprotectants widely used for embryo cryopreservation in different animals as well as humans. Materials & Methods: 197 porcine blastocysts were produced in vitro and 160 blastocysts were random-ly selected and divided into 4 groups. 40 blastocysts were placed in phosphate buffer solution (PBS) with-out any cryoprotectants for 1 hour in room temperature (23-25C) as the control group. The rest of the blastocysts were exposed to 3 different cryoprotectants (10% solutions) ethylene glycol (EG), 1, 2 pro-panediol (PG) and glycerol (Gly) for 1 hour in a 3- step method in room temperature. The survival rate was assessed after culture in NCSU-37 medium for 24 hours as the proportion of recovered embryos with the reformation of blastocele observed by stereomicroscopy at 40 magnifications. The apoptotic indices were evaluated after staining by TUNEL technique to label apoptotic nuclei and later were counter-stained by propidium iodide (PI) to label all nuclei and were analyzed by fluorescent microscopy. Then, the sur-vival rate was compared with the data obtained from the control group. Through ANOVA and Fishers exact test the data were analyzed while employing StatView software and the level of significance was considered as 0.05%. Results: Exposing porcine blastocysts to different cryoprotectants results in an increase in DNA fragmentation, although the apoptotic index in blastocysts with blastocele compared to those without them were lower in the study, disregard of the kind of cryoprotectant. Conclusion: It is concluded that CPs can decrease the survival rate of porcine blastocysts by increasing the percentage of DNA fragmentation but EG has the least effect on DNA fragmentation. https://www.jri.ir/article/222 The teratogenic effects of Lamotrigine on mouse fetus Introduction: One of the most important complications of antiepileptic drugs (AEDs) is increased congenital malformations. The risk of malformations in such fetuses is 7-10% compared to 2-4% in the general population. Lamotrigine is a recently introduced AED, with fewer teratogenic effects than that of the older ones and without reported teratogenic effects in most cases. Recently some malformations have been reported related to Lamotrigine use in human beings but the reports are scanty and contradictory and no particular patterns of malformations have been presented. This study has been done to detect the teratogenic effects of this drug in mouse fetus. Materials & Methods: In this study, NRMI mice were divided into four groups: I) control group 1, II) control group 2, with intraperitoneal administration of ethanol solution, III) case group 1, with intraperitoneal administration of three 25 mg/kg doses of Lamotrigine and IV) case group 2, with intraperitoneal administration of three 75 mg/kg doses of Lamotrigine. Injections were made during the 9th to the 18th days of gestation. On the 18th day of gestation, the fetuses were harvested. The body weight and height were measured and malformations in vertebral column, limbs and cranium were looked for and recorded. Malformations were compared in the four groups by SPSS software. Results: In the groups with 25 & 75 mg/kg Lamotrigine administrations, reduction of body weight and height and increased malformations of vertebral column and limbs were noticeable in a dose dependent fashion compared to the control groups number 1 & 2_ cranial malformations were insignificant. Conclusion: Based on the results, Lamotrigine can be considered as a risk factor for the increase of malformations in the treated animals. As a number of researchers believe that decrease of serum folate and methionine are effective in the appearance of malformations and they may implicate the situation, further studies on the mechanisms of Lamotrigine from this point of view are recom-mended. https://www.jri.ir/article/223 The comparison of serum Follicle-Stimulating Hormone (FSH) and vaginal pH for the diagnosis of menopause Introduction: Because of improvements in the measurement methods of FSH levels in serum, there is less interest in evaluating vaginal pH for the diagnosis of menopause nowadays. The traditional test to diagnose menopause has been FSH, so far. In recent studies it has been shown that diagnosis of menopause by testing for vaginal pH is a simple, non-invasive and inexpensive method .This study intended to compare diagnostic value of vaginal pH and serum FSH in the diagnosis of menopause. Materials & Methods: This article is a descriptive cross-sectional study taking place from June to July 2005. 120 menopausal women, aged 31 to 95 years that were being referred to a meno-pausal clinic in Qaem Hospital during the study, were chosen by a target-based nonrandom sampling technique for the study. A questionnaire was used for collecting data from the patients. In addition, pH test strips were used for vaginal pH measurements and the patients' serum FSH were evaluated in a definite laboratory and their clinical signs and symptoms were recorded. The data was analyzed by SPSS software and the statistical significant of the results were tested by 2 in comparison with the kappa coefficient of agreement. P0.05 was consi-dered as the significance level of the study. Results: According to the gathered data in the study, the average vaginal pH in the patients, in the absence of vaginal infections, was 5.330.53 (with a minimum of 4.1 and a maximum of 5) With attention to the cut of point of 4.5 for vaginal pH and the baseline FSH >20 mIU/ml that characterizes menopause, the sensitivity of pH-meter in diagnosing menopause was determined to be 97%. The mean average and the standard deviation of FSH levels in this study was 80.79 6.42. Conclusion: Vaginal pH sensitivity in the diagnosis of menopause is similar to serum FSH and could be used as a suitable alternative. https://www.jri.ir/article/224 Prevalence of leg cramps during pregnancy and effects of supplemental therapy Introduction: Leg cramps are common painful spasms especially in the lower extremities during pregnancy. The purpose of this study was to find a suitable alternative in the treatment of the disease. Materials & Methods: 217 patients reported leg cramps with different degrees and frequencies among 401 pregnant women in the second half of their pregnancies who visited the Prenatal Clinic of Imam Khomeini Hospital from July to December 2002. After preliminary evaluations for ruling out electrolyte imbalances or insufficient supply of dietary micronutrients, patients were randomly allocated into supplemental therapy (Calcium, magnesium or vitamin B intake for 2 weeks) and control groups. The groups were compared after 4 weeks according to their complete relief from leg cramps. The clinical findings and the data collected from the patients were analyzed by chi-square and t-student tests and a logistic regression model using the SPSS (V.10) software. Results: There was a significant improvement in patients receiving vitamin B with 71% complete and 19% relative relief from leg cramps, compared to 9% complete relief in the control group- 29% and 52% in the groups receiving magnesium and calcium respectively (p<0.0001). Conclusion: Although the high prevalence of leg cramps (55%) in the patients was not necessary-ly related to dietary habits, but it seemed that supplementing symptomatic patients with vitamin B could be beneficial. https://www.jri.ir/article/225 The reasons for the termination of pregnancies in the third trimester in Shahr-e-kord’s Hadjar hospital during 2005 Introduction: Observations show that the majority of pregnant women hospitalized in the third trimester of pregnancy, their pregnancies are terminated in the absence of any labor pain. Because of the importance of mother and baby health, this study was designed to assess the reasons for these terminations in Hadjar’s University Hospital in Shahr-e-Kord. Materials & Methods: In this descriptive and analytical study, 750 women who were hospi-talized for the termination of pregnancy, were selected randomly and assessed in morning, evening and night shifts. A questionnaire and a check list were used for data collection. Hospital records, interviews with mothers, a vaginal exam and physical exam of infants were used to complete the form and the check list. Data were analized by SPSS software and t, Chi square tests were used and p<0.05 was considered significant. Results: Based on the results, 298 (39.7%) of pregnancies were terminated for labor pain and 452 (60.3%) were terminated according to physicians’ advice or other health care providers’ without presence of labor pain. In the latter group, 23.2% of pregnancies were terminated for a history of previous cesarean section, elective cesarean sections, reduction of fetal movements, post-term pregnancies, maternal hypertention, placenta previa, placenta abruption or oligohydro-amnious.The mean Bishop score for induction was greater in women whose pregnancies were terminated for labor pain than those terminated according to physicians’ or any other health care providers’ advice (df=541, p<0.001). The correlation between Bishop score and mode of delivery was significant and in women whose Bishop score was less than 5, cesarean section was higher (df=20, p<0.001). After the elimination of previous and elective cesarean sections, the correlation between the causes of terminations and mode of delivery was significant and most hospitalized women terminated their pregnancieis, by cesarean section without presence of labor pain (df=16, p<0.001), although in this group the mean gestational age based on LMP and sonography was greater than those with labor pain (t= 3.7, df= 311, p<0.001). There were no significant differences in the weight of infants in the two groups. Conclusion: Taking the exact information in prenatal cares about the gestational age, carrying out a sonography in the first 26 weeks of pregnancy, educating pregnant women about the time of hospitalization for the termination of pregnancy and complications of early hospitalization, forming a specialty committee to decide on the termination of pregnancies in hospitals and setting practice guidelines in this regard,are efforts to prevent early termination of pregnancies. https://www.jri.ir/article/226