International Journal of Reproductive BioMedicine
Year:2003 | Volume:1 | Issue:1|Papers Count:10

Spermatids are the earliest male germ cells with one set of haploid chromosomes. After experiments, mainly in rodents, the spermatid injection was introduced in human assisted reproduction to the treatment of men with non-obstructive azoospermia. Spermatid injection is a technique with particular difficulties that may negatively influence the outcome. The identification, isolation and the assessment of viability, especially for round spermatids, require intensive work and considerable experience. Up to date, it appears that the rates of fertilization and implantation with round spermatid injection are dramatically low and significantly less compared to the use of elongated spermatid injection. The extremely low fertilization potency of the round spermatids led to attempts for their in-vitro culture and maturation. The immaturity of round and elongated spermatids has raised concerns regarding the potential genetic risk for the offspring. Under these facts, a reconsideration of the use of spermatids in assisted human reproduction is necessary.

Background: Spinal cord injury (SCI) has a significant impact on male reproductive functions which may lead to infertility. A large number of spinal cord injured men suffer from impaired spermatogenesis. Currently, in vivo gene transfer of molecules with potential therapeutic value has been recognized as a viable method for inducing functional recovery after SCI. This study characterized the role of adenovirus-mediated gene transfer into experimentally injured spinal cords of rats on possible restoration of spermatogenic cell lines.Materials and Methods: Young adult Sprague-Dawley rats (200-250g) were assigned into one of the three different groups of control, SCI, and adenovirus transfer (Ad) (n=3/ group). Control rats received no injury, nor any surgery. For SCI rats, SCI was produced by a 10g brass rod with a tip diameter of 2 mm which was dropped from a height of 12.5 mm onto exposed spinal cord at level of T10 with NYU impactor. Animals were perfused transcardially 43 days post SCI. Both spinal cord and testicular tissues were cryo-sectioned and ultra thin-sectioned, respectively. Cellular morphology and morphometry were done for spinal cord tissues. The testicular samples were processed for both light and transmission electron microscopy (TEM). The third group of rats underwent SCI first, followed by microinjection of LacZ adenoviral vectors (5x106 p.f.u./ µl) along the T6-T10 dorsal root entry zone bilaterally. The immune system of animals were suppressed before the Ad administration. Each Ad injection was done using a glass micropipet and a Nonoject injector. Rats were killed 43 days after Ad injections, and the tissues were studied as for other groups.Results: The spinal cord lesion extents for SCI and Ad groups were 8.1±3 and 5.8±2.2 mm, respectively (p<0.05). The testicular tissue of controls revealed a normal arrangement of spermatogenesis cell types. However, impaired spermatogenesis including vacuolization of germ cells along with incomplete spermatogenesis were noted in the tubles of SCI group. Also, nuclei and cell membranes of spermatozoa were damaged. In Ad rats, relatively active spermatogenesis, ranging from reappearance of proliferating spermatogonia to the presence of mature spermatozoa were observed in some seminiferous tubles. Conclusion: Bilateral adenovirus-mediated gene transfer into experimentally injured spinal cords of rats can restore the ultrastructure of spermatogenesis including mature spermatozoa.

Background: Embryonic stem cells (ESc) are pluripotent cells which have been used as a model to study the mechanism that control the embryogenesis and early mammalian development in vitro. The aim of this study was to isolate and produce embryonic stem cells from late blastocyst stage embryos in mice.Materials and Methods: Blastocyst stage embryos from pregnant NMRI mice were obtained and cultured for 24 h in DMEM medium. 4-6 days after hatching, the inner cell masses (ICM) formed colonies which were then collected mechanically and trypsinized. Several subcultures were prepared in the medium supplemented with 0.1 mM 2 Mercaptoethanol, 1000 U/ml Leukemia Inhibitory Factor (LIF) and 10% Fetal Bovine Serum (FBS). The (ESc) were recognized by alkaline phosphates histochemistry using azo-coupling method.Results: The results demonstrated that a highly pluripotent stem cell line was derived from the blastocyst stage embryos of NMRI mice; however, the rate of colonies was as low as 10%.Conclusion: The LIF is effective to culture and maintain the isolated ICM colonies in undifferentiated condition in the absence of feeder layer.

Background: Ovarian functional cyst is one of the most common pelvic mass in reproductive age which mostly resolves spontaneously. Sonography is a valuable tool for diagnosis of benign cyst with high accuracy. The objective of this cross sectional study was to evaluate the accuracy of transvaginal sonography in detecting type of ovarian cyst and compare the results wieh cytological results.Materials and Methods: 82 women in reproductive age who have had simple ovarian cysts with benign criteria which unresolved after taking contraceptive pills for 3 months were considered for this clinical study. Transvaginal ultrasound-guided aspiration of cysts were done and were then sent to the pathological evaluation. Also, all data regarding the size of the cysts and aspirated fliud were recorded in charts for further statistical analysis.Results: The accuracy of transvaginal ultrasound comparing with cytology on diagnosis for functional cysts was 94.9%, for epithelial ovarian cyst was 97.5% and for endometrioma was 97.5% (P= 0.0001).The size of cysts with diameter of <10cm was not related to the quality of cysts. Conclusion: The results showed that sonography is a valuable and reliable tool for diagnosis of benign ovarian cyst. It seems that if a mass appears benign by ultrasound morphologic criteria, probability of it being malignant is near to zero, which can be aspirated by transvaginal route without any fear from missing of malignancy or complication.

Background: The aim of this study was to determine the correlation between ultrastructural studies for pinopodes expression after ovarian hyperstimulation and progesterone injection in mice. Materials and Methods: Adult NMRI mice were superovulated using human menopasual gonadotropic (hMG) and human chorionic gonadotropic (hCG) hormones; after that, daily injection of progesterone (1 mg/mouse) was performed. Animals were sacrificed by cervical dislocation 3.5 and 4.5 days after hCG injection. Tissues of uterine horns were obtained and processed for scanning (SEM) and transmission (TEM) electron microscopy studies. The pseudopregnant control samples were studied same as experimental groups. Results: The SEM and TEM observations showed that in control groups on 3.5 days of pregnancy, there were some pinopodes. All apical cell surfaces expressed these projections on the forth day. In progestrone-injected group, well developed pinopods were expressed 3.5 days after hCG injection and they were transformed to small projections on the fourth day following hCG injection. Also, the life span of pinopods was limited to a short time. At the TEM levels, the pinopods were seen as swelling process on the apical surface, which were more pronounced on day 3.5 of hCG injection in hyperstimulated and progestrone injection.Conclusion: The progestrone may cause premature expression of pinopodes and the implantation failure after ovarian induction may be due to these timing changes.    

Background: While traditional semen parameters are of significant clinical value, total fertilization failure in IVF cycles is not uncommon. Sperm function testing such as; Hamster egg penetration test has severed limitations as a clinical test. The aim of this study was to evaluate the predictive value of semen parameters by using of intracellular calcium [Ca2+]I increase in response to progesterone. Materials and Methods: The [Ca2+]i response to progesterone was measured in spermatozoa of 86 patients referring to the Assisted Conception Unit for semen analysis. The patients were divided into 3 groups; according to their semen parameters and measured intracellular [Ca2+]i increasing in response to progesterone . Results: There was no significant correlation between each individual semen parameter and[Ca2+]Ielevation in response to the progesterone, but most of the patients in each group had [Ca2+]I increasing as expected based on sperm parameters. However, there were cases in groups 1 and 2 (Normal and IVF) that demonstrated [Ca2+]i increases which were poor or lower than expected. Out of the 22 patients in the normal category, 8 cases had poor response to [Ca2+]I increase and out of the 47 patients in the IVF group, 9 patients were as well. In addition we measured [Ca2+]I increases in 6 fertile donor samples for comparison purposes. Conclusion: [Ca2+]i increase in response to progesterone is related to predicting value of sperm parameters in most cases. However, the response of sperm to progesterone could be different in some cases that are expected in normal or IVF category based on our semen analysis criteria. We suggest that the [Ca2+]i measurements may perfect the sperm fertility potential

Backgroud: Intrauterine insemination (IUI) is generally attempted before proceeding to more expensive and invasive assisted reproductive techniques such as invitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). This procedure is most commonly performed as a therapeutic method for couples with a wide variety of subfertility etiologies, such as low count or low motility of sperm, or an incompatibility between the sperm and the cervical mucus. The objective of this clinical trial study was to compare the correlation between the semen parameters and pregnancy rates in patients undergoing hyperstimulation and IUI.Materials and Methods: 336 infertile couples that underwent 336 cycles of IUI with washed husband’s semen were included in this study. All patients’ charts were reviewed for age, etiology and duration of infertility, semen characteristics and pregnancy rates. The SPSS 9 software and Chi-square tests were applied for statistical analysis. P<0.05 was determined as statistical significance.Results: Total pregnancy rates were18.2% (61 out of 336 cycles). Postwash semen parameters including: sperm count ≥10´ 106 ,motility ≥50% (grade III and IV≥20%) had significant effect on pregnancy rates after IUI. The Outcome of this procedure was not significantly affected by female age, duration or etiology of infertility.Conclusion: Postwash semen quality was the most important factor for predication of successful pregnancy in this study.

Background: The high fertilization failure after IVF treatment cycles could be related to chromosomal abnormalities. This study was carried out to assess the frequency of chromosomal abnormality on human oocytes lacking signs of fertilization 18-20 h after insemination.Materials and Methods: On day one, 18-20 h after insemination (IVF), fertilization was confirmed when two pronuclei (normal IVF) or more pronuclei (poly pronucleus FR) were present. Chromosomal analysis of unfertilized oocytes was carried out within 20-24 h of collection. All oocyte did not sign of pronuclei were collected from total fertilization failure, TFF (FR=0) or partial fertilization failure, PFF (FR=10-90%). Chromosomal preparation was carried out as described by Tarkowski’s techniques. The average of finding between two groups was compared by X2 test. Results: Chromosome spreading permitted adequate analyzing in 348 unfertilized oocytes. In 33.6% chromosomal aneulpoidy was observed with the following frequencies; hypo-hyploidy, 22/348 (6.4%), hyper-hyploidy, 42/348 (12.2%) and diploidy, 52/348 (15%). The frequency of aneuoplidy was significantly higher in TFF group 33/80 (41%) than PFF group 83/268 (31%), p<0.01, X2. The most frequent numerical aberration was observed in chromosome group, G of the human karyotyped. Conclusion: Since cytogentic analysis of failed fertilized oocytes and sperm function tests are very helpful for direct information on low success rate of fertilization, further studies analyzing on both gametes function in TFF cycles will be needed.    

Introduction: Unwanted pregnancies are one of the important problems of countries. In our country despite the numerous efforts of authorities and cover a wide family planning, unwanted pregnancies and the consequences of that such as infectious abortions are common problems. Unwanted pregnancies are often due to lack of use or uncorrected use of common methods of contraception due to low knowledge and information. Therefore, its researchers have done to assess knowledge of young couples about contraceptive methods. Materials and Methods: This study was an interventional study that was conducted in 1389. Environmental research was center health and temperament of Khoy city and the study populations were young couples in the health center that comprised within two months.Simple sampling done and available sample included 40 persons. Data collection tool was (teenage knowledge of contraception and sexual heath). The scientific validity has done by five faculty members and content validity determined by using Test-Retest (a=0.79). Results analysis was investigated by using SPSS software before and after training.Results: The results showed that participating couples in all areas of contraception had moderate and poor information. The least information was about emergency contraception, and most information was about menstrual period. T test showed the significant difference before and after training (p=0.001), so that couples in all areas of knowledge after the training had reached the desired or medium level.Conclusion: Considering the average knowledge about the emergency methods and poor knowledge of some cooperation about useful items, including the proper time to use the emergency contraceptive. This point shows that strengthening the quantitative and qualitative aspects of this centers can play a vital role in giving information, establish correct beliefs about contraceptive methods and all of this should be lead to efficient application of these methods and reduction in failure contraceptive methods.

Introduction: This study investigates the relationship between daily intakes of protein in pregnant women and BUN, Cr levels in the first trimester of pregnancy.Materials and Methods: This cross sectional study was conducted on 436 18-35 year-old pregnant women, whose gestational age was equal or less than 20 weeks and singleton pregnancy who attended prenatal clinics in the hospitals of Shahid Beheshti, Iran and Tehran Universities in 2010. The prenatal clinics were selected by stratified cluster sampling. All subjects completed two questionnaires that included demographic characteristics and food frequency questionnaire (FFQ). Received food and calories of each person was determined by using nutritional software. To descriptive tests, Pearson's correlation coefficient and linear regression analysis, we used SPSS Version16.Results: In this study, no relation between daily protein intake of dietary (p>0.05) with BUN, Cr first trimester in pregnant women has been seen. To more accurately evaluate, we run linear regression and no significant relationship among intake proteins, BUN, Cr first trimester (p>0.05) and parity, chronic diseases, smoking and maternal age was revealed.Conclusion: According to the results, there wasn’t any relationship between the daily protein intake and BUN, Cr first trimester that probably was due to comparatively low samples. It is suggested that more studies with larger samples be performed to establish these relationships.