Showing papers in "Iranian Journal of Reproductive Medicine in 2008"

Effect of Cnidoscolous aconitifolius (Miller) I.M. Johnston leaf extract on reproductive hormones of female rats

Abstract: Background: The increase in the rate of infertility in women has prompted the need to search for plants with antifertility potentials. Objective: The effect of administration of aqueous extract of Cnidoscolous aconitifolius leaves for seven days on reproductive hormones of female Wistar rats was investigated. Materials and Methods: Sixty female albino rats were grouped into four (A, B, C and D) of 15 each. Group A (the control) received orally 6.5 ml kg -1 body weight of distilled water on daily basis for 7 days. Groups B, C and D were treated like the control except that they received 250, 500 and 1000 mg kg -1 body weight of the extract. Results: Phytochemical screening of the extract revealed the presence of alkaloids, saponins, phenolics, tannins, flavonoids, anthraquinones, phlobatannins and triterpenes. Administration of the extract produced significant increase (p<0.05) in the serum prolactin concentration whereas those of estradiol, progesterone, follicle stimulating and luteinizing hormones were significantly reduced. Conclusion: The alterations in the female rat reproductive hormones by the extract are indications of adverse effect on the maturation and ovulation of follicles. Consequently, the extract may impair fertility and conception in female rats. Thus, the Cnidoscolous aconitifolius leaf extract may be explored as a female contraceptive.

Adverse effects of ciprofloxacin on testis apoptosis and sperm parameters in rats

Abstract: Background: Ciprofloxacin is a commonly prescribed antibiotic in the treatment of genitourinary tract infection. Objective: The aim of this study was to investigate the effects of ciprofloxacin on testis apoptosis and sperm parameters in rat. Materials and Methods: Twenty male Wistar rats were selected and randomly divided into two groups; control (n=10) and experimental (n=10). The experimental group was orally received 12.5 mg/kg ciprofloxacin daily for 60 days and the control group just received water and food. Rats were then killed and sperm removed from cauda epididymis and analyzed for sperm motility, morphology, and viability. Testis tissues were also removed and prepared for TUNEL assay to detect apoptosis. Results: Results showed that ciprofloxacin significantly decreased the sperm concentration, motility (pl0.05) and viability (pl0.001). In addition, ciprofloxacin treatment resulted in a significant decrease in the number of spermatogenic cells (spermatogonia, spermatocyte, spermatid and sperm) in the seminiferous tubules when compared with the control group. The apoptotic germ cells per seminiferous tubular cross section was significantly increased in the experimental group (15.11±3.523) as compared with the control group (7.3±0.762) (pl0.05). Conclusion: It is concluded that ciprofloxacin has the toxicological effects on reproductive system in male rats.

Correlation between sperm parameters and sperm DNA fragmentation in fertile and infertile men

Abstract: Background: It is established that sperm DNA integrity is essential in fertilization and normal embryo and fetal development. Routine semen analysis gives an approximate evaluation of the functional competence of spermatozoa, but does not always reflect the quality of sperm DNA. Therefore, the evaluation of sperm DNA integrity, in addition to routine sperm parameters, could add further information on the quality of spermatozoa and reproductive potential of males. Objective: The objective of this study was to determine the levels of sperm DNA damage in fertile and infertile males and its correlation with semen parameters. Materials and Methods: Semen samples were collected from 45 infertile men selected from couples attending the infertility clinic with a history of infertility of ≥1 years and 75 healthy volunteers of proven fertility (initiated a successful pregnancy) served as the control group. After routine sperm analysis, DNA damage was determined using single cell gel electrophoresis (comet) assay method. Results: The mean of DNA damage (comet value) in the sperms of infertile males was significantly higher than that of fertile males (12.9±7.59 vs. 48.77±24.42, p<0.001). A significant negative correlation was observed between DNA damage and sperm motility in fertile group (p<0.02, R=-0.263). In infertile males, significant negative correlations were observed between DNA damage with sperm motility (p<0.002, R=-0.45) and morphology (p<0.03, R=-0.317). There was no significant correlation between sperm concentration and sperm DNA damage in both groups. Conclusion: These results indicate that sperm DNA damages in infertile males is significantly higher than fertile males and sperms with abnormal morphology and low levels of motility has more abnormal DNA damages than motile and normal sperms.

Vitamins C, E and lipid peroxidation levels in sperm and seminal plasma of asthenoteratozoospermic and normozoospermic men

Abstract: Background: It has been shown that reactive oxygen species (ROS) can lead to deleterious effects on a range of sperm parameters. Vitamins E and C are capable of reducing ROS levels and lipid peroxidation. Objective: The aim of study was to determine the level of lipid peroxidation as indicated by Malondialdehyde (MDA) and vitamins E and C in sperm and seminal plasma of asthenoteratozoospermic and normozoospermic men and their relationships with semen parameters. Materials and Methods: Forty men with normozoospermic and 60 infertile men with asthenoteratozoospermic semen profiles were randomly selected. Semen analysis was done according to the WHO standard. Sex hormonal profiles were measured by ELISA methods. The level of vitamins (C and E) and MDA were measured by HPLC and tiobarbiotic acid, respectively. Results: MDA concentration in the spermatozoa (0.1±0.06 nmol/ml) and seminal plasma (1.9±0.35 nmol/ml) of asthenoteratozoospermic were significantly higher than in normozoospermic males (p<0.001).The level of vitamins E and C in seminal plasma of normozoospermic were significantly higher than in asthenoteratozoospermic males (p<0.01). However, the amount of vitamin C in the spermatozoa of normozoospermic was significantly less than in asthenoteratozoospermic males (p<0.01). Sensitivity, specificity, positive and negative prognostic value of MDA of seminal plasma and spermatozoa were more than vitamins C and E. The level of vitamin C of spermatozoa had more diagnostic value when compare to vitamins C and E in seminal plasma. Conclusion: The level of MDA in seminal plasma and spermatozoa and vitamin C in spermatozoa may be a diagnostic tool for etiology of infertility in the asthenoteratozoospermic patients.

Effect of Nelumbo nucifera seeds on the reproductive organs of female rats

Abstract: Background: Nelumbo nucifera has been used as antifertility agent in females by the local tribals of Rajasthan especially of Udaipur district India. Objective: The present study was conducted to investigate the effect of Nelumbo nucifera on the fertility and general physiology in female rats. Materials and Methods: 20 female albino rats were taken and divided into two groups. Group 1 served as control and group 2 received ethanolic extract at dose of 800mg/kg b.wt. for investigating the nature of the drug and antifertility effect. Vaginal smear was monitored everyday in the morning to study the estrous cycle. On day 41 st all the animals were sacrificed and various haematological and biochemical parameters were estimated. Results: Data revealed that oral administration of Nelumbo nucifera extract brought about a significant decline in the weight of Ovary; Control (43±4.75mg), Nelumbo extract treated (25±3.86mg), Uterus; Control (236±0.004mg), Nelumbo extract treated (214±0.007mg) and Vagina; Control (221±0.002mg), Nelumbo extract treated (178±0.003mg) as well as protein and glycogen level, however cholesterol level increased significantly. In addition, the diestrous phase of the estrous cycle was found to be prolonged; Control (1.81±0.21) days, Nelumbo extract treated (3.62±0.42) days. Conclusion: These results suggest that Nelumbo nucifera has the anti-estrogenic nature without altering the general physiology of the female rats.

The in-vitro effects of nicotine, cotinine and leptin on sperm parameters analyzed by CASA system

Abstract: Backgrand: Cotinine (COT) is a major degradation product of nicotine (NIC). The participation of leptin in female reproduction is well-established, but any role in male reproductive function is at the best tenuous. Objective: The aim of this study was to evaluate the in-vitro effects of nicotine, cotinine and leptin on sperm parameters in normal semen of non-smokers fertile men. Materials and Methods: Ten healthy nonsmokers aged 25-40 years old were devided into 7 groups, Thier semens were divided into 7 aliquots. (A) was layered with basal solution 70 ng/ml NIC, (B) 35 µg/ml NIC, (C) 300 ng/ml COT, (D) 200 µg/ml COT, (E) 30 ng/ml leptin, (F) 300 ng/ml leptin respectively and (G) was layered with mHTF. After migration, the samples were examined at time 0, +1, +2, +4, +8, and + 24 h of incubation. Results: These findings were obtained: sperm count: 75.66±66.25x10 6 /ml, forward motility: 75.55±14.80%, progress: 33.66±13.01, VSL: 51.58±6.99 µm/s, VCL: 103.33±14.52 µm/s, ALH: 4.33±0.77 µm, BCF: 25.60±2.97 HZ, STR: 79.33±8.04 %, LIN: 52.55±10.52 %, ELO: 74.22 ± 12.76 % and ARE: 3.04 ± 1.50 u/sq. The parameters were similar before 8 hr and were being decreased after that. Conclusion: According to the results. nicotin and cotinin have negative effects on the sperm parameters but despite the positive effect of leptin, there is no correlation between leptin concentration in semen and its physical characteristics.

Neonatal mortality of low birth weight infants in Yazd, Iran

Abstract: Background: Low birth weight (LBW) is one of the major determinants of neonatal survival as well as postnatal morbidity. Objective: The main objective of the present study was to determine neonatal mortality rate (NMR) in LBW infants in Yazd, Iran. Materials and Methods: In a prospective-cohort study, all births in the maternity hospitals of Yazd, Iran in 2004 were evaluated and mortality rate in LBW population over the course of the first month of extra uterine life was determined. Results: In total, 8.4% (507 of 6016 births) of all newborns were LBW and 18.7% (95/507) of all LBW neonates died. Neonatal mortality rate in Yazd was 24/1000 live births .Two- third (95 /143) of all neonatal deaths occurred in LBW. Neonatal mortality rate (NMR) in LBW, Moderately low birth weight (MLBW), Very low birth weight (VLBW) and Extremely low birth weight (ELBW) were 23, 11.5 , 62.5 and 117 times more than that of normal weight newborns, respectively. Nearly 65% of all LBW neonatal deaths occurred in first 24 hours after birth. Overall NMR, Early Neonatal mortality rate (ENMR) and Late Neonatal mortality rate (LNMR) in LBW were 187, 118 and 9.8 in 1000 live births, respectively. The main causes of mortality among LBW in order of prevalence were respiratory distress syndrome (RDS) (59%), asphyxia (20%), septicemia (12%) and congenital malformation (9%). Conclusion: Neonatal mortality rate in Yazd is high and LBW accounted for two-third of neonatal deaths. Therefore, effort should be intensified to implement effective strategies for the reduction of LBW births and improving the care of these vulnerable neonates.

Antispasmodic effect of Physalis alkekengi fruit extract on rat uterus

Abstract: Background: Studies have shown that Physalis alkekengi reduces implantation and induces antifertility in rat. In Iranian traditional medicine it is believed that this plant has abortifacient and antifertility activities. Objective: The goal of this study was to evaluate the effect of Physalis alkekengi ripe fruit hydroalcoholic extract (PFE) on uterine contractility and its possible mechanism(s). Materials and Methods: Extraction of Physalis alkekengi fruit was carried out by maceration method (70% alcohol). Uterus was dissected out from adult non-pregnant rat (Wistar) and contracted by KCl (60mM) or oxytocin (10mU/ml) in an organ bath containing De Jalon solution and the effect of PFE on the uterine contractions was investigated. Furthermore, the role of α- and β-adrenoceptors, opioid receptors, nitric oxide and cyclic guanosine monophosphate synthesis inhibitors on the extract effects were evaluated. Results: KCl- and oxytocin-induced uterine contractions were inhibited (p<0.001) by the cumulative concentrations of the extract in a concentration dependent manner. Incubation of uterus with propranolol (1μM) and L-NAME (100μM) attenuated the PFE antispasmodic effect (p<0.05). But the PFE effect was unaffected by phentolamine (1μM), naloxone (1μM) or methylene blue (10μM). In Ca 2+ -free with high potassium (60mM) De Jalon solution, cumulative concentrations of CaCl2 (0.1-0.5mM) induced uterine contraction concentration-dependently (p<0.001). Uterus incubation (5min) with PFE (0.25-1.75mg/ml) attenuated the CaCl2–induced contractions (p<0.05). Conclusion: It seems that the extract induced antispasmodic effect mainly via calcium influx blockade and partially through blocking β-adrenoceptors and nitric oxide (NO) synthesis. However, neither α-adrenoceptors nor opioid receptors or cGMP synthesis were involved.

Pre and post natal exposure of 50 Hz electromagnetic fields on prostate glands of rats: an electron microscopy study

Abstract: Background: Men are unavoidably exposed to ambient electromagnetic fields (EMF) generated from various electrical gadgets and from power transmission lines. Prostate gland plays an important role in secretion of semen as largest accessory gland of male reproductive system. It seems that protection of this gland against EMF is important in spermatogenesis process. Objective: The goal of this study was to investigate the effect of non ionizing radiation on ultra structure of prostate gland. Materials and Methods: In total 50 male and 50 female rats, aged 15 weeks, were mated in animal house of Tabriz University of Medical Sciences. Then among born rats, 20 randomly were chosen as control and 30 were randomly chosen for exposure to EMF. They were exposed to 50 Hz EMF (8 M.T.) during in utero development (approximately 3 weeks) and postnatal life (5 weeks). Samples of prostate gland were processed and observed under light and transmission electron microscope. Results: In the experimental group, the secretory epithelial cells were generally inactive and cuboidal and their nuclei were dense with more corpus amylace compared to the control. Smooth muscle fibers spread out in different directions with heterochromatic nuclei. Mitochondria seemed without cristae and electron opaque. Conclusion: The results indicate that EMF had a deleterious effect on ultra structure of prostate gland in rat.

Pregnancy complications of grandmultiparity at a rural setting of South Africa

Abstract: Background: Intrapartam complications such as foetal malpresentation, placental abruption, dysfunctional labour, and postpartum haemorrhage are associated with grandmultiparity. In developing countries and many parts of Africa and sub-Saharan Africa, there is limited access to medical care. There is a need in these areas to identify women whose pregnancies are at risk of complication and it is an important part of antenatal screening and care during delivery to reduce adverse outcomes. Objective: The objectives of this study were to evaluate the complications during pregnancy and delivery of grandmultiparity and to compare it with other parity groups. Materials and Methods: A retrospective case control study was conducted targeting women delivered at Empangeni Hospital during April to December 2004. Among all women who delivered at the hospital, 352 grandmultimaras, 3326 nulliparas and 3772 who had parity 1 to 5, were taken as cases and controls respectively for the study. Results: Significantly higher rates of ante partum (1%) and post partum (2%) haemorrhages, post term delivery (3%), and intrauterine foetal deaths (4.5%) were observed in grandpultiparas compared to nulliparous women but there was no difference with parity group 1-5. Assisted vaginal delivery rates were significantly lower in grandpultiparas women (2%) compared to nulliparous (5%) group (p < 0.05) but there was no difference with parity group 1-5. There were no significant differences in anaemia (14%), hypertension (10%), eclampsia (0.5%) and diabetes (3%), elective and emergency caesarean delivery (14.2%) and low-birth-weight delivery rates (13.2%) among different parity groups. Conclusion: Grandmultiparity was not safer compared to other lower parity groups. Thus strategies are needed to guide women to seek proper care during pregnancy and if possible to avoid pregnancy if they had higher parity.

Assessment of testis histological changes and sperm parameters in experimentally–induced unilateral and bilateral cryptorchid mouse model

Abstract: Background: In most mammals, the testis is always maintained at a lower temperature than that in the abdomen, and exposure of the testis to body temperature causes degeneration of germ cells. Objective: In this research, the long effect of heat exposure on sperm parameters and microstructure of mouse testis were investigated. Cryptorchid mouse were induced by exposure to abdominal heat. Materials and Methods: Immature mice were anesthetized and a small incision was made in the abdominal skin, then fat pad at the upper end of testis was sutured to peritoneum. Weight of testis, spermatogenic cell numbers, tubular ectasis (rate of tubular lumen comparing to the thickness of germinal epithelium) as well as epididymal sperm parameters were measured. Results: The results showed that spermatogenesis was arrested and testicular weights, seminiferouse tubular diameters and epididymal sperm parameters were significantly reduced in the bilateral undescended testis compared with unilateral undescended testis and the control mice. However, complete depletion of seminiferous tubules and absence of germ cells was not found in the animals. Conclusion: In general, high temperature caused a decreased in all analyzed parameters except spermatogonial cell number probably due to the apoptosis and these changes significantly increase in bilateral groups compared with unilateral groups. We believe that the present model is a suitable tool for enrichment of spermatogonial stem cells, also it is useful for treatment of cryptorchidism and further biological research on spermatogenesis.

Effect of anxiety and depression on ART outcome

Abstract: Background: There is increasing evidence that psychological factors like anxiety and depression can affect IVF/ICSI treatment results. Objective: This study aimed to clarify the role of women’s anxiety and depression on the outcome of ART cycles using Intra Cytoplasmic Sperm Injection (ICSI). Materials and Methods: This was a prospective pilot study. One hundred six (106) consecutive women undergoing ICSI cycles were enrolled between January 2006 and 2007. Age, duration and cause of infertility, number and score of transferred embryos were recorded for each patient. Data regarding the state of anxiety and depression of each volunteer were collected using the translated and validated Iranian Cattle Anxiety and Beck Depression Inventories. Results: Among 106 women enrolled in the study, 25 cases (23.5%) of clinical pregnancies occurred. In univariate analysis, there was no significant difference regarding age and cause and duration of infertility between groups. Number of transferred embryos was significantly associated with higher pregnancy rates (3.4± 1.15 vs. 2.5±1.38 in pregnant and nonpregnant group respectively). Among the 106 participants, 73.58% had anxiety and 30.18% showed various degrees of depression. Out of 28 patients with no anxiety, 21(75%) and out of 74 patients with no depression, 24(32%) became pregnant. There was significant association between depression/anxiety and pregnancy rate (p=0.034 and p=0.00 respectively). Logistic regression model showed that anxiety/depression affect the outcome of ART significantly. Conclusion: It is crucial to identify infertile patients at greater demand for psychological support before starting ART cycles.

Comparison of coasting with Cabergoline administration for prevention of early severe OHSS in ART cycles

Abstract: Background: One of the major and life-threatening side effects of Assisted Reproductive Technique (ART) is ovarian hyperstimulation syndrome (OHSS). The available data however, have been showed that both Cabergoline (anti VEGF) and coasting reduce the severity of OHSS. Objective: We aimed to compare coasting and Cabergoline administration in prevention of severe OHSS. Materials and Methods: A total of 60 IVF/ICSI cycles were selected. Patients at risk of developing OHSS were divided into two groups as patient's convenience. For 30 patients in coasting group, exogenous gonadotropins were withheld to allow E2 to decrease while GnRH-a was maintained. Then 10,000 unit hCG was administrated and oocyte retrieval was performed 36 hours later. In Cabergoline group, 30 patients were administered with 0.5mg Cabergoline tablet on day of hCG injection, continued for 8 days. Results: The mean number of retrieved, good quality, mature oocytes and the mean number of embryos were significantly different in two groups (p 0.05). The incidence of severe OHSS was similar in two groups. Conclusion: The Cabergoline was as effective as coasting in the prevention of early severe OHSS in high risk patients, but yielded more retrieved oocytes.

Reactive oxygen species (ROS) production in seminal fluid correlate with the severity of varicocele in infertile men

Abstract: Background: Reactive oxygen species (ROS) are a group of free radicals that in excessive amounts have negative influence on sperm quality and function. Objective: To study the effect of varicocele and its severity on the level of ROS in infertile men with clinical varicocele. Materials and Methods: In this controlled prospective study, 42 men with clinically diagnosed left varicocele and 30 fertile men were studied. Patients were asked about history of urogenital infection, using any antioxidant medication and smoking. Grade of varicocele was determined by physical examination. Levels of ROS in seminal plasma were measured in each group by a chemiluminiscence assay. The sperm parameters were also determined and compared in different groups. Results: The ROS levels were significantly higher in patients with varicocele than normal men (mean: 1575.42 RLU (Radio Luminescence Unit) vs. 53.79 RLU, p=0.005). In total 20 patients had grade I, 20 patients grade II and 2 patients had grade III varicocele. The mean ROS levels were 669.12 RLU, 2406.87 RLU and 2324 RLU in patients with grade I, II and III varicocele respectively (p=0.144). In case group, 15 patients were smoker and 27 were non-smokers. The mean ROS levels in patients with the history of smoking was 1367.71 RLU while in non-smokers it was 897.672 RLU (p=0.729). Conclusion: Our study showed that increased levels of ROS production in the seminal fluid can be an important factor in the etiology of male infertility in patients with varicocele, and this effect is more prominent with higher grade of varicocele.

Safety and efficacy of misoprostol in induction of labour in prelabour rupture of fetal membrane in nigerian women: a multicenter study

Abstract: Background: Misoprostol, a prostaglandin E1 analogue compared to prostaglandin E2, has the advantage of being inexpensive and stable at room temperature, with its proven efficacy and safety. However studies on the effect of pH on the efficacy of misoprostol have yielded conflicting results. Thus its use in the induction of labour in patients with premature rupture of membrane requires further investigation. Objective: To evaluate the safety and efficacy of misoprostol in induction of labour in Nigerian women with prelabour rupture of membrane after 34 weeks of gestation. Materials and Methods: Three hundred and forty six Nigerian women with prelabour rupture of membrane who consented to participate in the trial were randomised into two arms of misoprostol and oxytocin. Labour was managed with WHO partograph. The primary outcome was the caesarean section rate and induction vaginal delivery interval. Results: The mean induction to vaginal delivery interval was significantly shorter in the misoprostol arm (504 mins) compared to 627 mins in the oxytocin arm (t=3.97; p=0.005). The caesarean section rate of 18.1% among the misoprostol arm was also significantly lower than the 41.4% recorded in the oxytocin arm (p=0.002). Among patients with Bishop score greater than 6 there were no statistically significant differences between the two groups in the outcomes measured. Conclusion: Misoprostol is not only effective but also safe when compared with titrated oxytocin in Nigerian parturients with prelabour rupture of membrane after 34 weeks.

Cytogenetic and molecular genetic analysis of dicentric Y chromosome and its relation to male azoospermia

Abstract: Background: Cytogenetic analysis, Y-chromosome microdeletion screening, FISH techniques and other genetic methods have helped in finding the causes of infertility in azoospermic or severe oligoazoospermic cases in the last decade. Objective: In the present study, we characterized an abnormal Y-chromosome, detected as a mosaic in an azoospermic male ascertained for infertility. Materials and Methods: Chromosome analysis, using G, Q and C banding techniques and FISH analyses with several different DNA probes specific for Y and X chromosome sequences [XY centromeric α-satellite, Y non-α-satellite III, LSI-probes of the Y chromosome, WCP of Chromosome Y, SRY gene, subtelomeric Xp and Yp, which cover the SHOX (short stature-homeobox containing) gene, and subtelomeric Xq and Yq probes] were performed. A total of 20 sequence tagged sites were analyzed using primer sets specific for the Y-chromosome microdeletion loci. The primers were chosen to cover AZFa, AZFb, and AZFc regions as well as the SRY gene. Results: Chromosome analysis revealed a gonosomal mosaicism of monosomy X (51%) and a pseudodicentric Y (49%) chromosome: mos 45, X/46,X psu dic (Y) (qter→p11.32 :: p11.2→qter). Molecular genetic studies did not show deletions in the AZFabc regions, but a deletion was found in the short arms of the dicentric Y chromosome. One of the SRY genes was also missing. Conclusion: The azoospermia in this patient could be explained by either the presence of an abnormal Y-chromosome that cannot form a sex vesicle (which appears to be necessary for the completion of meiosis process and the formation of sperm), or the presence of the 45, X cell line.

The changes in morphology and morphometrical indices of endometrium of ovariectomized mice in response to exogenous ovarian hormones

Abstract: Background: The preparation of endometrium for embryo reception and implantation are controlled by ovarian hormones. These hormones have distinct cyclical changes during estrus cycle. Objective: The aim of this study was to evaluate the changes in morphology and morphometrical indices of endometrium by daily injections of estrogen and progesterone in ovariectomized mouse. Materials and Methods: In total 60 adult NMRI female mice were ovariectomized and after two weeks, they were randomly divided into five groups: control, sham group, estrogen treated mice (which received daily dosage of 0.5 ml/mouse of hormone for five days), progesterone treated mice (which received daily dosage of 0.2 ml/mouse of progesterone hormone for five days) and estrogen-progesterone treated mice (they received 0.5 ml/mouse estrogen on the first day and 0.2 ml/mouse progesterone injections from the second day to the fifth day of treatment). The mice were sacrificed in every day (n=5) up to five days after treatment and their uterine horns were obtained and processed for morphological and morphometrical studies. Results: On the second day of treatment, the diameter of glands was observed to be more in the progesterone group (53.75±6.32μ) than this in the estrogen (45.13±7.78 μ) and estrogen-progesterone treated groups (48.17±13.58 μ). While, the number of glands (76.25±17.37) and thickness of endometrium (39.58±3.37 μ) were observed to be more in the estrogen treated group (p=0.01). Conclusion: Progesterone had effect on the gland whereas estrogen caused increased in height of surface epithelium of endometrium. Overall, the day 2 after treatment (in all experimental groups) is suitable day for sampling for further studies.

GnRHa stop protocol versus long protocol in poor responder IVF patients

Abstract: Background: Recently different studies suggested that discontinuation of gonadotrophin releasing hormone analogue (GnRHa) at beginning of ovarian stimulation (improvement of ovarian response to gonadotrophins) may have some benefit to poor responder patients in invitro fertilization (IVF) cycles. Objective:The efficacy of GnRHa stop protocol in poor responder patients in IVF cycles was assessed. Materials and Methods: This study was a prospective, randomized controlled trial that 40 poor responder patients (less than three mature follicles in a previous cycle) with normal basal follicle stimulating hormone (FSH) were randomly allocated into two protocols: 1) Non-stop protocol: long GnRHa suppression, and start gonadotrophins from day 3 of mense. 2) Stop-protocol: GnRHa is stopped with the onset of menses, and gonadotrophin doses remained similar to group 1. Results: A significantly higher number of follicles, oocytes, embryos and fertilization rate also shorter stimulation days and lower human menopausal gonadotropins (HMG) ampoules were recorded in the stop protocol compared to the control group. Both protocols resulted in a similar cancellation rate, pregnancy rate, estradiol level and LH level. Conclusion: Early follicular cessation of GnRHa permitted the retrieval of a significantly higher number of follicles, oocytes and embryos, and can reduce the number of HMG and stimulation days.

Repeated IVF/ICSI-ETs failures and impact of hysteroscopy

Abstract: Background: Despite numerous developments in the field of assisted reproduction, the implantation rate remains low. Among the various reasons of implantation failure, endometrial regularity has an important role. Hysteroscopy is an accurate method for evaluating the endometrial characteristics, with the ability to treat uterine pathology. Objective: The aim of the present study was to evaluate the findings on hysteroscopy and thereafter the result of subsequent IVF/ICSI in infertile women with the history of frequent unexplained and unsuccessful IVF/ICSI attempts. Materials and Methods: In this observational study, the hysteroscopy findings and the outcomes of subsequent IVF/ICSI were evaluated in 89 infertile women admitted in Avicenna Infertility Clinic, with previous repeated (more than two) failed IVF/ICSIETs, including the patients with normal Hysterosalpinography (HSG) and excluding severe male factor infertility and also thrombophilia, genetic and immunologic problems. The data were analyzed with SPSS software and Fisher exact, chi-square, and MC-Nemar tests. Results: In 53 (59.5%) cases, hysteroscopy revealed abnormal intrauterine findings including adhesions 7 (13.7%), single polyp 11 (20.7%), endometrial polyposis 10 (18.8%), endometrial hyperplasia 10 (18.8%), uterine cavity hypoplasia 4 (7.8%) and myoma 5 (9.8%). These abnormalities were significantly higher in women with more than 8 years of infertility (chi-square=4.7, p-value=0.03). After hysteroscopy and subsequent IVF/ICSI-ET attempt using standard long protocol, pregnancy rate were significantly higher compared with the previous repeated IVF/ICSI attempts (35.8% versus 0%). Conclusion: According to this study, we strongly suggest evaluation of endometrial integrity by hysteroscopy in patients with repeated IVF/ICSI-ETs failure, before entering any other fertilization procedures.

Morphological and morphometrical changes of endometrim after application of estrogen and progesterone during luteal phase in the superovulated mice

Abstract: Background: The application of luteal phase supplementation hormones to increase the implantation rate is on debate among researchers. Objective: In this study, the morphological and morphometrical assessment of superovulated mice endometrium were investigated at window implantation period superovulated mice. Materials and Methods: Female mice were superovualated then were mated with vasectomized mice; the mice were divided in experimental and control groups. Experimental group included five groups which in them pseudopregnant mice were given a four consecutive daily injection of 1-progesterone (P), 2-estrogen (E), 3-estrogen + progesterone, 4-antiprogesterone+estrogen and 5-sham group. The uterine of all groups were collected after 4.5 day of pseudopregnancy and were prepared for histological and morphometrical studies. Results: Morphological studies of endometrial tissue showed that the luminal epithelium in group P appeared cuboidal shape. Endometrial folding was very high in group E+P. The luminal epithelium in groups E, E+P and RU 486 + E were seen in different morphology in comparison to control group. Morphometrical evaluation also showed height of luminal epithelium in group E (32.7±0.67) and E+P (33.6±1.3) were higher than those were seen in control (22.5±1.5) and group P (15.3±1.2). Conclusion: Progesterone caused the lowest endometrial development compare to other groups. It is concluded that the adding of E to P may improve endometrial condition to implant at luteal phase.

Fusion and development of 2-cell bovine embryos to tetraploid blastocyst with different voltages and durations

Abstract: Background: The values of embryonic stem cell and cloning are evident. Production of clone from embryonic stem cells can be achieved by introduction of stem cell into a tetraploid blastocyst. Tetraploid blastocyst can be produced in vitro by electrofusion of 2-cell embryos. Objective: The aim of this study was to assess the effect of different voltages and durations on fusion rate of bovine 2-cell embryos and their subsequent development in vitro. Material and Methods: The in vitro produced bovine 2-cell embryos were categorized into 3 groups: (1) fused group (FG); 2-cell embryos fused by exposure to different voltages (0.5, 0.75, 1, 1.25 and 1.5 kV/cm) and durations (20, 40, 60, 80 and 100 μs), (2) exposed control group (ECG); 2-cell embryos exposed to different voltages and durations but remained unfused and (3) unexposed control group (UCG); embryos cultured without exposure to any voltage. The embryos from each group were cultured and fusion, cleavage and developmental rates were compared in each group. Results: The results show that increased voltage, increases the fusion rate up to 88% for 1.5 kV/cm; however, the rate of cleavage and blastocyst formation decreases significantly to 18% and 10% respectively (p<0.05). Increased duration does not significantly increase fusion rate, however, in high voltage, increased duration decreases cleavage rate and blastocyst formation rate. Blastocyst formation rate in UCG showed a better development (32%) compared to FG (20%) or ECG (22.5%) (p<0.05). Conclusion: It can be concluded that for optimal fusion, cleavage and development, one pulse of 0.75 kV/cm for 60μs should be applied.

Tubal patency after ultrasound guided local injection of KCL to tubal ectopic pregnancy with alive fetus

Abstract: Background: Ectopic pregnancy is one of the pathologic entities that it destroys the fallopian tube and impairs the future pregnancy. There are different medical and surgical therapies in order the treatment and reserve of fertility. The aim of this report is discuss a case of successful intrauterine after management of ectopic pregnancy with local injection of KCL in an infertile PCOD patient. Case: The patient was a PCOD woman with gestational age of 8.5 weeks. One gestational sac and alive fetus with normal heart beat was seen in the right adnexa. Hemodynamic situation was stable. 0.5cc KCL 10% was injected to the fetal heart and the extracted tissue sent for pathology. We evaluated the effects of this treatment by measuring serial human chorionic gonadotropin (hCG) titers. Moreover, we used hysterosalpingogrphy to diagnose and to evaluate the patency of the tubes. Induction ovulation and intrauterine insemination was done and pregnancy occurred. Patient passed the pregnancy very good and she delivered by cesarean section due to breech presentation at 39 weeks of gestational age. There was no sign of pathologic finding in the tubes or adhesion or sequel of ectopic pregnancy. Conclusion: The beneficial outcomes of this kind of treatment suggest that local injection of KCL as a low invasive treatment can be the choice treatment for alive and progressed ectopic pregnancy or heterotopic pregnancy. Because of rarity of this management and successful intrauterine we reported the usefulness of local KCL injection for the successful conservative treatment of alive ectopic pregnancy in Iran.

The effect of active immunization with gonadotropin releasing hormone conjugate (gnrh-bsa) on gonadosomatic indices (gsi) and sperm parameters in mice

Abstract: Background: Gonadotropin releasing hormone conjugate (GnRH-BSA) raises antibodies against biologically active gonadotropin releasing hormone (GnRH) which affects body weight, gonadosomatic indices (GSI) and sperm quality in male mice. Objective: The objective of this experimental investigation is to develop an effective and reliable hormonal immunocontraceptive vaccine to suppress spermatogenesis by using GnRH-BSA conjugate. Materials and Methods: Forty sexually mature mice, Mus musculus were divided into two groups of twenty each. Group 1, served as control, while group 2 were immunized at monthly intervals for four times against GnRH with a GnRH-BSA conjugate (50 µg) with aqueous adjuvant (Freund's adjuvant). After 30 days of each immunization, body weight, GSI and sperm quality were observed in the immunized groups and compared with the control group. Results: Body weight showed alterations in immunized animals as compared to control. However, GSI, sperm motility, sperm count and sperm morphology were significantly decreased in immunized animals throughout the experimental investigation and these effects were more prominent and significant in the later part of the experiment. Conclusion: These results suggested that the active immunization against GnRH produced bioeffective antibodies as indicated by significant reduction in GSI level and sperm quality and induced infertility in male mice.

Fatty acid composition in normozoospermic, asthenozoopermic,asthenoteratozoospermic and oligoasthenoteratozoospermic ejaculates

Abstract: Background: The lipids of the spermatozoa membrane are important for the fluidity and flexibility of spermatozoa. However, spermatozoa’s lipids are the main substrates for peroxidation, which may provoke severe functional disorder of sperm. Objective: The aim of this study was to investigate the fatty acids composition of spermatozoa in men with asthenozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia compared with normozoospermic males. Materials and Methods: A cross-sectional study was designed. The patients were 51 men with seminal parameters abnormalities undergoing infertility screening. The patients were grouped into asthenozoospermic (n=15), asthenoteratozoospermic (n=21) and oligoasthenoteratozoospermic (n=15). The patients were compared with 21 males with normozoospermia. Sperm fatty acid analysis was performed using capillary gas chromatography. Results: Levels of stearic acid and oleic acid were significantly higher in oligoasthenoteratozoospermic subjects compared with these levels in normozoospermic males. Levels of arachidonic acid and DHA were significantly lower in the sperms of oligoasthenoteratozoospermic males than normozoospermic men. Sperm motility and morphology were correlated positively with levels of arachidonic acid and DHA while a negative correlation was observed with levels of stearic acid and oleic acid. Conclusion: In conclusion, impaired sperm function can originate from the disorder of sperm lipid metabolism. Low levels of DHA and arachidonic acid in spermatozoa of oligoasthenoteratozoospermic subjects may be the result of breakdown of them.

Expression of c-Jun in human granulosa cells from patients participating in in vitro fertilization programs

Abstract: Background: The mitogen-activated protein kinase (MAPK) pathway is one of the major signaling pathways that transmit intracellular signals initiated by extracellular stimuli to the nucleus. The stress-activated protein kinase (SAPK)/c-Jun NH2-terminal kinase is a subfamily of MAP kinases implicated in cytokine and stress responses. Objective: In this study, we have examined total and phosphorylated c-Jun in the mural and cumulus granulosa cells, and investigated also whether c-Jun can be responsible for the difference in the expression of apoptosis between mural and cumulus regions. Materials and Methods: A total of 14 consecutive couples participating in IVF program were investigated. Aspirated follicular fluid was transferred into tissue culture dishes and oocyte-cumulus cells complexes were isolated. The cells were centrifuge and fixed with Bouin’s solution and then were put on a glass slide. After fixation, the slides were stained by immunocytochemistry method. The incidence of apoptotic granulosa cells was examined by a fluorescence microscope. Results: The incidence of apoptotic granulosa cells was 1.27 ± 0.12 in the mural region and 0.38 ± 0.07 in the cumulus regions. All mural and cumulus cells expressed total c-Jun in 7 patients while phosphorylated c-Jun was also expressed in all cells of the other 7 patients. There was no difference between apoptotic and nonapoptotic cells in the expression of total and phosphorylated c-Jun. Conclusion: C-Jun may not be responsible for apoptotic effect on mural and cumulus cells.

Oocyte activation: The impact of calcium signals on fertilization

Abstract: Fertilization is triggered by changes in intracellular calcium concentration. In mammals, these transients in ooplasmic calcium concentration take the form of repetitive spikes, so called calcium oscillations (Ca2+-oscillations). These oscillations are important for relieve of meiotic arrest and to induce all the other events of oocyte activation. Although a surface mediated way of oocyte activation has been proposed, there is now substantial evidence to suggest that the sperm cell induces these Ca2+-oscillations by introducing a sperm specific phospholipase C, PLC., in the ooplasm. Ca2+-oscillations are also observed after intracytoplasmic sperm injection ( ICSI), a successful technique in human assisted reproduction. In the rare cases that no fertilization is observed following ICSI, this may be due to a deficiency in PLC.. However, artificial activating the oocytes after ICSI by increasing the calcium concentration can restore fertilization rates in these cases and support further development, as evidenced by successful pregnancies. Further evaluation of the current protocols for assisted oocyte activation is appropriate and investigation of the future application of PLC. is warranted.

The study of detecting sperm in testis biopsy in men with severe oligospermia and azoospermia by two methods of wet prep cytologic and classic histopathologic

Abstract: Background: Many azoospermic patients with non obstructive azoospermia (NOA) are candidate for testicular sperm extraction (TESE) and in vitro fertilization. Because sperm might be present in some but not all parts of the testes of such men, multiple sampling of testicular tissue are usually necessary to increase the probability of sperm finding. Sperm finding can be done by two methods: 1) classic histopathology and 2) wet smear. Objective: Comparative study of pathology and wet smear methods for discovering sperm in testis biopsy of azoospermic men. Materials and Methods: We prospectively studied 67 consecutive infertile men who referred to Fatemieh Hospital, Hamedan, Iran between April 2002 and September 2004. All patients were either azoospermic or severely oligozoospermic. They underwent intraoperative wet prep cytological examinations of testis biopsy material and then specimens were permanently fixed for pathologic examination too. Results: Among the 67 testes that underwent wet prep cytological examination, 44 (65.7%) were positive and 23 (34.3%) had no sperm in their wet smear. On the permanent pathologic sections, 19 (28.4%) were positive and 48 (71.6%) cases were with no sperm in their sections. Among all the individuals 18 (26.8%) were negative in both studies, while 14 (20.8%) had minimum 1 sperm in their smears in both examinations. The positive cases in wet prep cytological examination were significantly more than the cases in the permanent histopathologic sections (p-value=0.000). Conclusion: It seems that wet prep cytological examination is more reliable than permanent histopathologic sections in detecting sperm in testis biopsy of azoospermic men.

Conventional versus 3D ultrasound for the investigation of infertile women

Abstract: Background: With the technical development, ultrasound tends to overpass its limits and to become the main investigation tool even for situations when it was traditionally considered “second best”. It is more convenient both for patient and doctor, less expensive and has real diagnostic value. Objective: The present paper, and its graphic back-up, are intended as a visual testimony of the fact that the newer three dimensional (3D) ultrasound technique is much more than a simple generator of “nice”, commercial, images. Moreover, its use may be extended from obstetrics to gynecological pathology, especially in cases related to infertility. Materials and Methods: We performed a review of the main local infertility causes and their perception by the 3D ultrasound, as compared to the conventional methods. General literature citations were completed with personal cases and images. In our research, we used a Kretz – Voluson Expert 730 ultrasound machine with a full logistic equipment. Our database was completed over a period of 18 months and included various pathology, such as uterine anomalies, fibroids, polyps, ovarian cysts and many others. Results: Three dimensional ultrasound represents the best tool in evaluating the uterine cavity, the endometrium, assessing its volume and vascularity pattern. It also offers a very good image of the uterine structure, the adnexal morphology and their relationship. It performs a thorough pelvic assessment by a single examination. Moreover, it may realize a histerosalpingography, which is as efficient as the radiological method, but easier and with fewer side effects. Conclusion: Even though it is technically more difficult and time consuming, a good practice and high quality ultrasound equipment offer a series of benefits over any other kind of investigation.

Conservative management in young patients with borderline and malignant ovarian tumors: outcomes and pregnancies

Abstract: Background: Recent studies have showed conservative management in selective patients with borderline and malignant ovarian tumors is safe; therefore this management is considered in patients with ovarian tumor who desire to preserve fertility. Objective: This study has been performed to evaluate the clinical outcome and fertility in patients with ovarian tumors who were treated conservatively. Materials and Methods: All patients who were treated conservatively (preservation of uterus and at least one ovary) or were on follow-up and had recurrence were evaluated in Vali-e-Asr Hospital during 2000-2004. Results: Among 410 patients with ovarian tumors, 60 were treated conservatively. Age range was 13-34 years. Twenty-six of patients (43.3%) were desired pregnancy and 34 (56%) patients did not. Three (5%) patients had history of infertility. Histological types of tumors were as follows; 15(25%) borderline tumors, 10(16.7%) epithelial tumors, 26(43.3%) germ cell tumors, and 9(15%) sex cord tumors. Range of follow-up time was 12-48 months. Seven term pregnancies in 6 patients had been occurred, 1 in epithelial group, 2 in germ cell group, 1 in sex cord group and 3 in borderline group. Nine patients had recurrence and 2 patients expired, including one patient with serous cyst carcinoma (Stage IIIC).This patient had refused radical surgery and referred to our center with recurrence. Another patient had immature teratoma (Stage IIIC). Conclusion: Conservative surgical management in young patients with stage I (grade 1, 2) of epithelial ovarian tumor and sex cord-stromal tumor and in patients with borderline and germ cell ovarian tumors could be performed in order to preserve fertility.