International Journal of Reproductive BioMedicine
Volume:10 Issue: 2, Mar-Apr 2012

Sperm cells extracted from testes (TESE) have poor chromatin quality and motility. Various substances are used in the laboratory to increase sperm motility and improve the ART outcomes; however, there are few research which considered improving both sperm motility and chromatin quality. The aim of this investigation was to evaluate the improvement of the testicular sperm motility and chromatin quality exposed to L-carnitine (LC) and L-acetyl-carnitine (LAC), which are normally concentrated in testis and epididymis, compared with Pentoxifylline (PF), which used for sperm motility enhancement in IVF procedures. TESE samples from 30 male mice divided into four parts. The sperm samples were added to Ham' F10 (control) or the media contained 1.76mM of LC, LAC or PF), then, the samples were kept in the room temperature for 30, 90 and 180 min. At each time step, sperm motility and chromatin quality were assessed. Chromatin quality was evaluated by chromomycin A3 and aniline blue. Statistical analysis was performed using one way analysis of variance (ANOVA). A p-value less than 0.05 were accepted as a statistically significant difference. The results showed LC, LAC and PF significantly increased the sperm motility. However, sperm chromatin quality only improved significantly by administration of LC and LAC. Administration of LC and LAC to the testicular sperm samples can lead to improve both sperm motility and chromatin quality. It may be because they can mimic in vivo sperm condition during late spermatogenesis.

Fertilization failure is one of the most problems in assisted reproduction technology (ART). The aim of this study was the evaluation of oocytes activation by addition of calcium ionophore in unfertilized oocytes in ICSI cycles. This study was done on 15 ICSI cycles (stimulated with standard long protocol). Mature retrieved oocytes with normal morphology that had no evidence of fertilization 24 hours after ICSI were included in the study. The oocytes with fertilization and unfertilized oocytes with degeneration were excluded from the study. The unfertilized oocytes were washed with GIVF medium and were transferred to GIVF medium that contained 5 µmol of calcium ionophore and were incubated for 10 minutes. Then again oocytes were washed with GIVF medium and consequently were transferred to GIVF medium and were incubated at 37°C in 6% CO2. After 18 hours, the oocytes were examined and activated oocytes were defined with observation of at least one pronucleus or cleaved oocytes. After ovarian stimulation and oocytes retrieval, 175 mature oocytes were obtained and injection of sperm was done for all of them. 114 of 175 oocytes (66%) showed evidence of fertilization after 24 hours. A total of 61 oocytes (34%) showed no evidence of fertilization and 10 oocytes were degenerated and were excluded from the study. Only 51 unfertilized oocytes with normal morphology were selected and were exposed to calcium ionophore. 37 (72.5%) of treated oocytes were fertilized (2PN) and 32 (62.7%) of them showed evidence of cleavage. 6 (11.8%) embryos had good quality. According to our results, oocytes activation with calcium ionophore had an acceptable fertilization rate, however high quality embryos remained low. We propose future studies to evaluate embryo quality.

Postpartum period is the critically important part of obstetric care but most neglected period for majority of Pakistani women. Only life threatening complications compel them to seek for tertiary hospital care. We describe the nature of these obstetric morbidities in order to help policymakers in improving prevailing situation. To find out the frequency and causes of severe post-partum maternal morbidity requiring tertiary hospital care and to identify the demographic and obstetrical risk factors and adverse fetal outcome in women suffering from obstetric morbidities. This prospective cross-sectional study was carried out in the Department of Gynecology and Obstetrics, Liaquat University Hospital Hyderabad, between April 2008-July 2009. The subjects comprised of all those women who required admission and treatment for various obstetrical reasons during their postpartum period. Women admitted for non-obstetrical reasons were excluded. A structured proforma was used to collect data including demographics, clinical diagnosis, obstetrical history and feto-maternal outcome of index pregnancy, which was then entered and analyzed with SPSS version 11. The frequency of severe postpartum maternal morbidity requiring tertiary hospital care was 4% (125/3292 obstetrical admissions). The majority of them were young, illiterate, multiparous and half of them were referred from rural areas. Nearly two third of the study population had antenatal visits from health care providers and delivered vaginally at hospital facility by skilled birth attendants. The most common conditions responsible for life threatening complications were postpartum hemorrhage (PPH) (50%), preeclampsia and eclampsia (30%) and puerperal pyrexia 14%. Anemia was associated problem in 100% of cases. Perinatal death rate was 27.2% (34) and maternal mortality rate was 4.8%. PPH, Preeclampsia, sepsis and anemia were important causes of maternal ill health in our population. Perinatal mortality was high.

Improvements in culture media formulations have led to an increase in the ability of sheep embryo in culture throughout the preimplantation period. This study was carried out to evaluate the effects of various concentrations of MEM vitamins during in vitro maturation of sheep oocytes and subsequent embryo development. Sheep ovaries were collected from a slaughterhouse and transported to the laboratory. Oocytes were matured in SOF medium supplemented with, eCG, hCG and EGF in various concentrations of MEM vitamins (control, 0.5, 1 and 1.5) for 24h. The cumulus oocyte compelex (COCs) were co-incubated with epididymal spermatozoa of post mortem rams in synthetic oviduct fluid fertilization (SOFF) medium with 10% heat inactivated estrous sheep serum for 18h. Embryos were cultured in synthetic oviduct fluid culture 1 (SOFC1) medium for 48h followed by cultured in synthetic oviduct fluid culture 2 (SOFC2) medium for six days. Addition of 0.5 and 1 MEM vitamins significantly increased (P< 0.05) overall blastocyst development (21.62% and 22.33%; respectively) compared with 1.5 MEM vitamins (15.59%), but there was no difference between control, 0.5 and 1 MEM vitamins in the percentage of embryos successfully developing to the blastocyst stage (19.50%, 21.62% and 22.33% respectively). It seems that addition of 1.5 of MEM vitamins has detrimental effect on blastocyst rate.

Sperm parameters and motion kinetics are affected by cryopreservation. The main purpose of the current study was to determine the effect of different concentrations of Trolox as an antioxidant to freezing-thawing procedure on human sperm kinematic parameter. Semen was collected from 20 normal donors and divided into five aliquots prior to cryopreservation. The first aliquot was analyzed by computer-assisted sperm analysis (CASA). Other aliquots were mixed with cryo-protective agent containing 0, 20, 40, and 80 µmol Trolox and treated samples were cryopreserved in liquid nitrogen. After two weeks samples were thawed and sperm motion kinematics was measured by CASA. Percent motility (Mot), curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), linearity (LIN), and amplitude of lateral head displacement (ALH) were compared before and after freeze. Addition of 40µmol Trolox resulted in significantly higher (p<0.05) post thaw VCL, VSL and VAP compared to other groups. Therefore the percentage of post thaw motile spermatozoa were significantly higher (p<0.01). The supplementation of Trolox significantly improved the post-thawed human semen quality, especially progressive motility and average path velocity.

Obesity is an independent risk factor of preeclampsia with unknown mechanism and hyperlipidemia might be a probable case of it. The objective of this study was to determine the role of hyper-triglyceridemi in association with high prepregnancy body mass index and the risk of preeclampsia. The authors conducted this case-control study of 42 preeclamptic and 41 normotensive overweight pregnant women. The two groups were comparable with respect to age, gestational age, and body mass index. Blood samples were collected at the time of diagnosis of preeclampsia, after 14 hour fasting to determine plasma lipid concentrations. Enzymatic photometric tests were used to determine lipid profile. Data was analyzed with independent “t-test”, Chi-square and one-way ANOVA and post HOC Tukey HSD test. The statistical significance was set at 0.05 levels. In the subjects with preeclampsia, serum triglyceride and total cholesterol levels were significantly increased and plasma HDL-cholesterol concentrations were decreased compared with the controls, (p<0.05), but plasma LDL cholesterol levels didn’t differ between the two groups. Women who developed severe preeclampsia had higher concentrations of TG and cholesterol and lower levels of HDL compared to noromotensive group. Mean TG: 375.16 vs. 202.85, p<0.001, Mean cholesterol: 245.64 vs. 214.32, p=0.04, Mean HDL: 40.80 vs. 48.95, p=0.03). We noted that dyslipidemia, particularly hypertriglyceridemia was highly correlated with prepregnancy high BMI in preeclamptic women. These findings continue to support a role for dyslipidemia in BMI related preeclampsia.

“Polycystic ovary syndrome (PCOS) is a complex endocrine and metabolic disorder associated with ovulatory dysfunction”. “Autonomic and central nervous systems play important roles in the regulation of ovarian physiology”. The noradrenergic nucleus locus coeruleus (LC) plays a central role in the regulation of the sympathetic nervous system and synaptically connected to the preganglionic cell bodies of the ovarian sympathetic pathway and its activation is essential to trigger spontaneous or induced LH surges. This study evaluates sympathetic outflow in central and peripheral pathways in PCO rats. Our objectives in this study were (1) to estimate LC activity in rats with estradiol valerate (EV)-induced PCO; (2) to antagonized alpha2a adrenoceptor in systemic conditions with yohimbine. Forty two rats were divided into two groups: 1) LC and yohimbine and 2) control. Every group subdivided in two groups: eighteen rats were treated with estradiol valerate for induction of follicular cysts and the remainders were sesame oil groups. Estradiol concentration was significantly augmented by the LC lesion in PCO rats (p<0.001), while LC lesion could not alter serum concentrations of LH and FSH, like yohimbine. The morphological observations of ovaries of LC lesion rats showed follicles with hyperthecosis, but yohimbine reduced the number of cysts, increased corpus lutea and developed follicles. Rats with EV-induced PCO increased sympathetic activity. LC lesion and yohimbine decreased the number of cysts and yohimbine increased corpus lutea and developed follicles in PCO rats.

Nitric oxide (NO) is a molecule that incorporates in many physiological processes of female reproductive system. Recent studies suggested the possible role of endothelial isoform of nitric oxide synthase (eNOS) enzyme in female infertility. The aim of this study is to evaluate the expression of endothelial nitric oxide synthase in endometrial tissue of women with unexplained infertility. In this case-control study a total of 18 endometrial tissues obtained from 10 women with unexplained infertility and 8 normal and fertile women by endometrial biopsy, 6 to 10 days after LH surge. Specimens were fixed in 4% paraformaldhyde fixative and frozen sectioned for semi-quantitative immunohistochemical evaluation using monoclonal anti-human eNOS antibody. Hematoxilin and Eosin was used for Histological dating. Localization of endothelial nitric oxide synthase was seen in glandular and luminal epithelium, vascular endothelium and stroma in both fertile women and women with unexplained infertility. Although there were differences in immunoreactivity of glandular epithelium (p=0.44), vascular endothelium (p=0.60) and stroma (p=0.63) but only over-expression of eNOS in luminal epithelium (p=0.045) of women with unexplained infertility compared to fertile women was statistically significant (p<0.05). This study suggests that changes in luminal expression of eNOS may influence receptivity of endometrium.

Polycystic ovarian syndrome (PCOS) is a condition associated with chronic anovulation, insulin resistance and androgen excess. Women with this syndrome are at increased risk of metabolic syndrome. The aim of the present study was to determine the prevalence of metabolic syndrome (MBS) in women with PCOS referred to Arash Hospital in different ages and body mass index (BMI). A cross-sectional study was conducted in Gynecologic Clinic at Arash Hospital affiliated with Tehran University. A total of 282 women with PCOS ages between 15-40 years were included. The prevalence of Metabolic Syndrome and its components in this population were the main outcomes. Height, weight, waist circumference, blood pressure and laboratory tests (FBS, TSH, HDL-C, serum prolactin, triglycerides and total cholesterol) were measured in this population. The prevalence of MBS in PCOS women was 22.7% (64 cases). The rate of central obesity, FBS more than 110 mg/dl, triglycerides more than 150 mg/dl, high-density lipoprotein cholesterol levels (HDL-C) less than 50 mg/dl, and blood pressure? 130/85 mmHg in PCOS women was 31% (87), 3.2% (9), 33% (93), 68.8% (194), and 10.6% (30), respectively. The risk of MBS was increased in older and the obese women (BMI? 30 kg/m2). The present sample showed women with PCOS have a high prevalence of MBS and its individual components, particularly decreased HDL-C.

Conventional IVF and ICSI are two common techniques to achieve fertilization. IVF has long been used for treatment of infertility, although it is not an effective treatment in severe male infertility. The use of ICSI has been expanded in severe male factor and fertilization failure after IVF cycle. In spite of the widespread use of ICSI in patients with non-male factor infertility, there is still little evidence to confirm its effectiveness in this population. To evaluate assisted reproductive technology outcomes between IVF and ICSI cycles in non-male factor, normoresponder patients. A total of 220 non-male factors, normoresponder patients who were indicated for ART were enrolled in this study. The patients received standard long GnRH agonist or GnRH antagonist protocols for ovarian stimulation and after oocytes retrieval, the patients were divided into two groups (IVF and ICSI groups). In IVF group (n=112), all of retrieved oocytes were treated by conventional IVF and in ICSI group (n=88), microinjection (ICSI) was done on all of retrieved oocytes. In IVF group, fertilization and implantation rates were significantly higher than ICSI group (66.22% and 16.67% in IVF group versus 57.46% and 11.17% in ICSI group, respectively). Chemical and clinical pregnancy rates were statistically higher in IVF group as compared with the ICSI group (42.9% vs. 27.3% and 35.7% vs. 21.5%, respectively). According to our study, the routine use of ICSI is not improved fertilization, implantation and chemical pregnancy rates and is not recommended in non-male factor, normozoospermic patients.

Asphyxia is a medical condition in which placental or pulmonary gas exchange is impaired or they cease all together, typically producing a combination of progressive hypoxemia and hypercapnea. In addition to regional differences in its etiology; it is important to know its risk factors. This is a case-control study, all neonates born from May 2002 to September 2005 in Vali-e-Asr Hospital were studied. 9488 newborns were born of which 6091 of the live patients were hospitalized in NICU. 546 newborns were studied as case and control group. 260 neonates (48%) were female and 286 neonates (52%) were male. Among the neonates who were admitted, 182 of them were diagnosed with asphyxia and twice of them (364 newborns) were selected as a control group. The variables consist of; gestational age, type of delivery, birth weight, prenatal care, pregnancy and peripartum complications and neonatal disorders. Our studies showed that 35 (19.2%) patients had mild asphyxia, 107 (58.8%) had moderate asphyxia and 40 (22%) were diagnosed as severe asphyxia. Mean maternal age was 34.23±4.29yr; (range: 23-38 yr); and mean of parity was 2±1.2; (range: 1-8). Risk factors in our study included emergent Caesarian Section, preterm labor (<37w), low birth weight (<2500g), 5 minute Apgar (less than 6), need for resuscitation, nuchal cord, impaired Biophysical Profile, neonatal anemia, and maternal infertility. All risk factors listed above play a role in asphyxia. The majority of these factors are avoidable by means of good perinatal care.

Chromosome abnormality (CA) including Sex chromosomes abnormality (SCAs) is one of the most important causes of disordered sexual development and infertility. SCAs formed by numerical or structural alteration in X and Y chromosomes, are the most frequently CA encountered at both prenatal diagnosis and at birth. This study describes cytogenetic findings of cases suspected with CA referred for cytogenetic study. Blood samples of 4151 patients referred for cytogenetic analysis were cultured for chromosome preparation. Karyotypes were prepared for all samples and G-Banded chromosomes were analyzed using x100 objective lens. Sex chromosome aneuploidy cases were analyzed and categorized in two groups of Turners and Klinefelter’s syndrome (KFS). Out of 230 (5.54%) cases with chromosomally abnormal karyotype, 122 (30%) cases suspected of sexual disorder showed SCA including 46% Turner’s syndrome, 46% KFS and the remaining other sex chromosome abnormalities. The frequency of classic and mosaic form of Turner’s syndrome was 33% and 67%, this was 55% and 45% for KFS, respectively. This study shows a relatively high sex chromosome abnormality in this region and provides cytogenetic data to assist clinicians and genetic counselors to determine the priority of requesting cytogenetic study. Differences between results from various reports can be due to different genetic background or ethnicity.

Peroxisome proliferative-activated receptors (PPARs) are nuclear receptors that involved in cellular lipid metabolism and differentiation. The subtype? of the PPAR family (PPAR?) plays important roles in physiologic functions of ovaries. To determine correlation between PPAR? protein level in granulosa cells and pregnancy rate in women undergoing in-vitro fertilization (IVF) treatment. In this cross-sectional study, twenty-five samples of granulosa cells were collected from women referred to an IVF treatment center. PPAR? protein expression level in granulosa cells was determined in comparison with? -actin level as control gene with Western blot test. Laboratory pregnancy was determined by a rise in blood? -hCG level fourteen days after embryo transfer. Correlation analyses were used to test for associations between the oocytes and pregnancy occurrence as outcome variables and PPAR? protein expression level. Correlation analysis indicated that there was no significant relationship between granulosa cells PPAR? protein level with IVF parameters including number of matured oocytes and the ratio of fertilized to matured oocytes. Comparison of granulosa cells PPAR? protein level with positive and negative laboratory pregnancy revealed also no significant relationship. According to the results of this study, PPAR? protein level in granulosa cells could not be directly correlated to the success rate of IVF.

Prolonged GnRH-a administration in IVF cycles may have some advantages related to the treatment outcomes. In this study, we aimed to analyse the effect of prolonged gonadotropin releasing hormone agonist (GnRH-a) administration on controlled ovarian hyperstimulation outcomes of in vitro fertilization (IVF) patients. In this retrospective study, 55 patients with a GnRH-a administration period more than 10 days were compared with 55 patients whose same period was? 10 days with respect to the demographic characteristics, metaphase II (MII) oocyte ratio, grade I (GI) embryo ratio, blastocyst ratio, fertilization, implantation, and the clinical pregnancy rates. The mean hospital visit count of the prolonged GnRH-a patients was 2.6±0.4. As we expected, total GnRH-a doses used during hypophyseal down regulation were significantly different between the groups (p<0.0001). MII oocyte, G1 embryo and the blastocyst ratios were also significantly different between the groups (p<0.0001; p<0.01 and p<0.05). All the other parameters were insignificant. Prolonged GnRH-a administration during ovarian suppression in IVF patients may have positive impacts on the oocytes and the embryos, but this affect may not be observed in the overall pregnancy rates.

Rabies is a fatal infectious disease and rabies post-exposure prophylaxis is the method of choice for prevention of human rabies. Case series: We report rabies antibody levels in cord blood and also in serum of pregnant women who were bitten by suspected animals to rabies and were immunized by purified Vero cell rabies vaccine (PVRV) and Human Rabies immunoglobulin (HRIG) serum. During the years of 2007-2010, six pregnant women by the age range of 22-35 years were admitted in treatment and prevention of rabies center in Pasture institute of Iran, in Tehran. Among them two cases were at first trimester, one at second trimester and three at third trimester of conception. The interval between biting with delivery was 5-265 days (mean 121 days). Results of immunoglobulin illustrate that levels of rabies antibody in maternal sera with the fetus are not equal and uniform but it is proved that baby will find efficient immunity as well with minimum protective level of 0.5 IU/ml in all cases except a newborn whom had been born just 5 days after the mother’s immunization and in a shorter time than the appropriate immunization of the mother who had received her second vaccination courses.