En 1680-6433 2008-2177 jalali 1382 8 10 gregorian 2003 11 1 1 1 online 1 fulltext

En 1680-6433 2008-2177 Is There a Place for Round and Elongated Spermatids Injection in Assisted Reproduction? - 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. - - 1 6 Byron Asimakopoulos - No Nikos Nikolettos - No Safa Al-Hasani Sf_alhasani@hotmail.com No

En 1680-6433 2008-2177 The Effect of Murine Leukemia Inhibitory Factor on In Vitro Differentiation of Mouse Embryonic Stem Cells - 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. - Embryonic stem cells, Mouse embryo, Leukemia Inhibitory Factor 12 15 Ali A Movassagh-Pour - No Mojdeh Salehnia mogdeh@dr.com No Mojdeh Salehnia mogdeh@dr.com No Ali A Pourfatollah - No Sayed M Moazzeni - No

En 1680-6433 2008-2177 Comparison between Transvaginal Sonography and Cytological results for detection of ovarian cysts - 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 - transvaginal Sonography , ovarian cyst, cytology 16 19 Abass Aflatoonian abbas_aflatoonian@yahoo.com No Abass Aflatoonian abbas_aflatoonian@yahoo.com No Reza Aflatoonian - No Zahra Khashavi - No

En 1680-6433 2008-2177 Progesterone Shifts the Pinopodes Expression of Mouse Endometrium to Pre-Implantation Time After Ovarian Hyperstimulation - 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. - Implantation Window, Pinopodes, Progesterone, Electron Microscopy 20 23 Mojdeh Salehnia mogdeh@dr.com No Mojdeh Salehnia mogdeh@dr.com No

En 1680-6433 2008-2177 Evaluation of the Predictive Value of Semen Parameters in Sperm Fertility Potential Using Intracellular Calcium Increase in Response to Progesterone - 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+]i elevation 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. - Ca2+]i increases, Progesterone, Spermatozoa] 24 28 Mir-Mehrdad Farsi m.farsi@mubabol.ac.ir No Mir-Mehrdad Farsi m.farsi@mubabol.ac.ir No

En 1680-6433 2008-2177 The Correlation Between Semen Parameters and Pregnancy Outcome after Intrauterine Insemination - 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. - IUI, pregnancy, female age, semenm parameters 28 32 Marzieh Mehrafza dr_mehrafza@yahoo.com No Marzieh Mehrafza dr_mehrafza@yahoo.com No Nadia Nobakhti - No Zahra Atrkar Roushan - No Havva Dashtdar - No Mane Oudi - No Ahmad Hosseini - No

En 1680-6433 2008-2177 The Chromosomal Abnormality of Failed Fertilized Human Oocytes in an In Vitro Fertilization Program - 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. - chromosome abnormality, cytogenetic, unfertilized oocytes, Total Fertilization Failure 33 36 Seyed M Kalnatar kalantarsm@ystp.ac.ir No Seyed M Kalnatar kalantarsm@ystp.ac.ir No Ahmad Ebrahimi - No Mehrdad Solimani - No Hossein Fazli - No

En 1680-6433 2008-2177 Restoration of Spermatogenesis by Adenoviral Gene Transfer into Injured Spinal Cords of Rats - 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. - Spinal cord injury, Gene therapy, Spermatogenesis, Rat 7 11 Mohammad A Khalili Khalili59@hotmail.com No Mohammad A Khalili Khalili59@hotmail.com No Alexander G Rabchevsky - No

En 1680-6433 2008-2177 Ultrastructural study of isolated mouse preantral follicles co-cultured with cumulus cells with leukemia inhibitory factor Background: The ultrastructural analysis of cultured follicles could direct us to understand subcellular changes during in vitro culture. Objective: This study was done to verify the ultrastructural characteristics of in vitro cultured mouse isolated preantral follicles in co-culture system in the presence and absence of leukemia inhibitory factor (LIF) Materials and Methods: Mechanically isolated preantral follicles were divided into four groups: control without LIF, control with LIF, co-cultured group with LIF, co-cultured group without LIF. In co-culture groups the follicles were cultured with cumulus cells. After 4 days the follicles were processed and sectioned for transmission electron microscopic examination. Results: The oocytes of cultured preantral follicles in all studied groups demonstrated a homogeneous cytoplasm and they had the round or ovoid shaped mitochondria with light matrix and cristae. Their endoplasmic reticulum cisternae were in association with mitochondria and Golgi complex. The cortical granules and the aggregation of mitochondria around the germinal vesicle were prominent in both co-cultured groups. The organelle distribution in granulosa cells was normal in all groups of study and no sign of cell death was observed. In both co-cultured systems the granulosa cells contained mitochondria with tubular cristae, a well developed smooth endoplasmic reticulum and several large lipid droplets, characteristics of steroid synthesis cells. Conclusion: The oocyte and granulosa cells in co-cultured system showed more remarkable maturation features than that of control. Background: The ultrastructural analysis of cultured follicles could direct us to understand subcellular changes during in vitro culture. Objective: This study was done to verify the ultrastructural characteristics of in vitro cultured mouse isolated preantral follicles in co-culture system in the presence and absence of leukemia inhibitory factor (LIF) Materials and Methods: Mechanically isolated preantral follicles were divided into four groups: control without LIF, control with LIF, co-cultured group with LIF, co-cultured group without LIF. In co-culture groups the follicles were cultured with cumulus cells. After 4 days the follicles were processed and sectioned for transmission electron microscopic examination. Results: The oocytes of cultured preantral follicles in all studied groups demonstrated a homogeneous cytoplasm and they had the round or ovoid shaped mitochondria with light matrix and cristae. Their endoplasmic reticulum cisternae were in association with mitochondria and Golgi complex. The cortical granules and the aggregation of mitochondria around the germinal vesicle were prominent in both co-cultured groups. The organelle distribution in granulosa cells was normal in all groups of study and no sign of cell death was observed. In both co-cultured systems the granulosa cells contained mitochondria with tubular cristae, a well developed smooth endoplasmic reticulum and several large lipid droplets, characteristics of steroid synthesis cells. Conclusion: The oocyte and granulosa cells in co-cultured system showed more remarkable maturation features than that of control. Co-culture, In vitro maturation, Leukemia inhibitory factor, Mouse preantral follicle, Ultrastructure. Co-culture, In vitro maturation, Leukemia inhibitory factor, Mouse preantral follicle, Ultrastructure. 187 192 Kamran Haidari Kamran Haidari _ No Mojdeh Salehnia Mojdeh Salehnia mogdeh@dr.com,salehnim@modares.ac.ir No Mojtaba Rezazadeh Valojerdi Mojtaba Rezazadeh Valojerdi _ No Shahram Pourbyranvand Shahram Pourbyranvand _ No

En 1680-6433 2008-2177 - - 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 co-injection 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. - Enhanced green fluorescent protein (EGFP), Transgenic animals, Embryo 151 158 - - Abdolhossein Rezaeian - No - - Seyed Mehdi Kalantar kalantarsm@ystp.ac.ir No - - Safar Farajnia - No - - Mehrdad Soleimani - No - - Abbas Baghi - No - - Abbas Aflatoonian - No - - Sirous Zeinali - No

En 1680-6433 2008-2177 In vitro application of Matrigel enhances human blastocyst formation and hatching Background: Matrigel (extracellular matrix) can improve the growth of many cell types in vitro. Objective: The aim of the present study was to determine the effect of Matrigel on the development of 2-4 cells human embryos in culture. Material and Methods: Surplus 2-4 cells human embryos, resulting from ICSI, were divided into two groups (control and test). Quality of embryos in both groups was morphologically similar. The test group (n=140) was cultured in Hams’ F10 supplemented with 10% human serum albumin and 150 µl liquid Matrigel. The control group (n=140) was cultured in the same medium devoid of Matrigel. Embryos were cultured for an additional 4 days and their morphology was assessed every 24 hours. Both groups were then statistically compared. Results: The percentage of the human embryos that reached the morula stage in the control and test groups were 79.2% and 80%, respectively (p>0.05). However, 36.4% of embryos reached the blastocyst stage in the test group as compared to 5.7% in the control group after 144 hours in culture. This difference was statically significant (p <0.01). In addition, culture of embryos on Matrigel and medium versus medium alone significantly improved in vitro hatching (25.7% versus 3.5%; p <0.01). Conclusion: Matrigel at low concentration enhances human blastocyst formation and hatching in vitro. Background: Matrigel (extracellular matrix) can improve the growth of many cell types in vitro. Objective: The aim of the present study was to determine the effect of Matrigel on the development of 2-4 cells human embryos in culture. Material and Methods: Surplus 2-4 cells human embryos, resulting from ICSI, were divided into two groups (control and test). Quality of embryos in both groups was morphologically similar. The test group (n=140) was cultured in Hams’ F10 supplemented with 10% human serum albumin and 150 µl liquid Matrigel. The control group (n=140) was cultured in the same medium devoid of Matrigel. Embryos were cultured for an additional 4 days and their morphology was assessed every 24 hours. Both groups were then statistically compared. Results: The percentage of the human embryos that reached the morula stage in the control and test groups were 79.2% and 80%, respectively (p>0.05). However, 36.4% of embryos reached the blastocyst stage in the test group as compared to 5.7% in the control group after 144 hours in culture. This difference was statically significant (p <0.01). In addition, culture of embryos on Matrigel and medium versus medium alone significantly improved in vitro hatching (25.7% versus 3.5%; p <0.01). Conclusion: Matrigel at low concentration enhances human blastocyst formation and hatching in vitro. Matrigel, Extracellular matrix, Human embryos, Blastocyst, Hatching Matrigel, Extracellular matrix, Human embryos, Blastocyst, Hatching 103 107 - - - - - No

En 1680-6433 2008-2177 Non-puerperal uterine inversion in a virgin woman Background: Inversion of the uterus is very uncommon. Patients may present with pelvic pain, vaginal discharge, or hemodynamic shock. Case: We report a case of 35 years old women (virgin) who was admitted with profuse vaginal bleeding and cramps of uterus. In the vaginal examination at litothomy position a mass of 5ď‚´8 cm in size was protruded from the vagina. At first myomectomy was performed and after that laparotomy with total abdominal hysterectomy was done. Conclusion: Early diagnosis, immediate treatment of shock, and replacement are essential in uterine inversion. Background: Inversion of the uterus is very uncommon. Patients may present with pelvic pain, vaginal discharge, or hemodynamic shock. Case: We report a case of 35 years old women (virgin) who was admitted with profuse vaginal bleeding and cramps of uterus. In the vaginal examination at litothomy position a mass of 5ď‚´8 cm in size was protruded from the vagina. At first myomectomy was performed and after that laparotomy with total abdominal hysterectomy was done. Conclusion: Early diagnosis, immediate treatment of shock, and replacement are essential in uterine inversion. Non-puerperal, Inversion, Uterine, Myoma Non-puerperal, Inversion, Uterine, Myoma 135 136 - - - - - No