International Journal of Reproductive BioMedicine
Volume:11 Issue: 1, Jan 2013

The use of ovarian stimulation for infertility treatment is associated with side effects of ovarian hyperstimulation syndrome (OHSS) and potential cancer risk. This is also true in high risk women such as those polycystic with ovary (PCO) and polycystic ovarian syndrome (PCOS). In vitro maturation (IVM) of oocytes was primarily developed to make IVF safe for women with PCO and at high risk of OHSS. The application of IVM of oocytes to assist clinical infertility treatment remains poor because of the reduced developmental competence of oocytes after IVM, despite several decades of research. Reduced meiotic maturation and fertilization rates, as well as low blastocyst production reveal short-term developmental insufficiency of oocytes when compared with in vivo-matured counterparts. In this review, the structural role of human oocytes, revealed by different technical approaches, to the success of IVM technology is highlighted.

It is important to protect oocytes and embryos from oxidative stress in the culture medium. Melatonin has been shown to be a direct free radical scavenger.

Effect of melatonin during in vitro oocyte maturation, fertilization and embryo development of mouse oocytes was evaluated.

Oocytes from supper-ovulated mouse were divided to two groups: cumulus oocyte complexes (COCs, group I) and denuded COC (d-COCs, group II). The oocytes were cultured in maturation medium with different doses of melatonin (1×101-105 nM). The cumulus expansion and nuclear status were evaluated after 24 h of in-vitro maturation. The oocytes were used for in-vitro fertilization. The fertilized oocytes were cultured in medium supplemented with different doses of melatonin.

The expansion (86.79%) and maturation (80.55%) rate of COCs increased in supplemented medium with 10 nM of melatonin vs. control group (73.33%), p=0.006 and p=0.026 respectively), but oocytes without cumulus cells indicated higher maturation rate at higher melatonin doses (10 and 100 M, 84.34% and 79.5% respectively(vs. 69.33% in control group (p=0.002). Fertilization rate was higher in treated medium with 1 μM of melatonin (93.75%, p=0.007). The rate of cleavage and blastocyst formation was promoted in medium supplemented with 10 and 100 nM of melatonin (92.37% and 89.36% vs. 81.25% in control group, p=0.002). We observed a dose dependent response to melatonin treatment in this experiment.

Exogenous melatonin can promote cumulus cell expansion, in vitro oocyte maturation, and embryo development. However we investigated a dose-dependent response in different stages of maturation and development. It may reflect sensitive rate of oocytes and embryos to culture conditions.

Meiotic genes are very important candidates for genes contributing to female and male infertility. Mammalian MutL homologues have dual roles in DNA mismatch repair (MMR) after replication errors and meiotic reciprocal recombination. The MutL homologs, MLH1 and MLH3, are crucial for meiotic reciprocal recombination and human fertility. In this study the functional polymorphisms of MLH3 C2531T was investigated in Iranian women with unexplained infertility.

Investigating the association between a common SNP (single nucleotide polymorphism) C2531T in the MLH3 gene and female infertility.

In total, 105 women with unexplained infertility as case group and 100 women with at least one child and no history of infertility or abortion as controls were recruited for this association study. The MLH3 C2531T polymorphism was tested by tetra-amplification refractory mutation system-PCR (4P-ARMS-PCR) method.

The MLH3 2531C and T alleles frequencies were 43.33% and 56.67% among infertile patients, and 61.5% and 38.5% among normal controls, respectively. In the patient and control subjects the CC (Pro 844 Pro) genotype frequency of MLH3 C2531T was 4.76% and 25%, the CT (Pro 844 Leu) genotype was 77.15% and 73%, and the TT (Leu 844 Leu) genotype was 19% and 2%, respectively (p=0.0001).

The presence of the polymorphic allele T leads to an increased risk of 2.09 times (OR=2.09, 95% CI=1.38-3.16; p=0.0001) for developing infertility in relation to the control group. Therefore, our data suggest that the MLH3 C2531T polymorphism can be associated with the risk of unexplained infertility in Iranian women.

Asphyxia is a common cause of perinatal mortality in 5-10% of all births worldwide. The present parameters for determining perinatal asphyxia, e.g. preeclampsia, cannot be considered as markers per se, and require auxiliary markers, e.g. increased number of nucleated red blood (NRBC) cells, for early diagnosis of perinatal asphyxia.

In this study, we evaluated the mean NRBC count in preeclampsia and to determine the usefulness of the NRBC as independent prognostic factors of perinatal complications.

This was a cross-sectional study in order to compare the NRBC in the umbilical cord of term neonates born to 50 mothers with preeclampsia and 150 normal mothers. The exclusion criteria were mother’s affliction with complications of pregnancy and inexact last menstrual period. The variables under study were maternal and neonatal data. The count of NRBC was determined with standard laboratory procedures in the blood samples from umbilical cord of the neonates. The acquired data were fed into SPSS 16 software and analyzed using statistical tests.

The mean value of NRBC count was significantly higher in preeclamptic women (p<0.0001). The average 1st and 5th minute Apgar scores were significantly higher in normal mothers (p<0.001).

Increase of NRBC in neonates born to mothers with preeclampsia may be due to chronic hypoxia; this group of neonates has increased risk and requires more precise and extensive care during delivery and after birth in order to have reduced mortality and complications during the neonatal period.

Research in reproductive health (RH) has been located in the core of women’s health research. Providing accurate information through conducting scientific and controlled research is essential, but increased number of research in the world especially in developing countries in RH area in order to introduce advanced technologies has been resulted in much unethical, illegal and abusive research on women, which needs particular attention to ethical issues by the practitioners who are involved in RH research.

This study was conducted to develop a practical ethical framework for RH research.

45 expert academics and clinicians in various disciplines included in a three rounds Delphi study through purposeful sampling method. In round 1 Delphi data were gathered using open-ended questions by e-mail and answers were analyzed by conventional content analysis and the findings merged and validated with the results of a thorough literature review. Face and content validity index were determined in round 2 Delphi and consensuses were attained in round 3.

Emerged categories were 1) management of the research process 2) protection of participants’ rights 3) third party consent 4) gender sensitive research and 5) conflict of interest.

This study has provided a practical ethical framework according to the socio-cultural context of Iran for all practitioners who are involved in research on women. Adherence to this framework may protect practitioners against unethical and illegal lawsuits and help them to respect their clients’ reproductive rights.

Methamphetamine (MA) is a potent psychomotor stimulant with high abuse and addictive potential. MA is a neurotoxic drug which is widely abused by females of childbearing age, raising serious public health concerns in terms of exposure of the fetus to the drug. Neurotoxic effects of MA on adult are well known, such as dopaminergic nerve terminal degeneration and cell death in regions of brain in some doses.

In the present study, we examined effect of prenatal MA exposure on mouse fetuses.

In this study, forty 8-12 week-old NMRI female mice were used which were mated with male mice in serial days. When sperm plug was observed it was designated as gestational day (GD) 0. Pregnant mice were individually housed in plastic cages. Pregnant mice were divided into four groups: in first group 10 mg/kg /day MA, in second group 5 mg/kg /day MA and in third group saline were injected subcutaneously from GD 6 to GD 14, corresponding to organogenesis period, while fourth or control group were without injection. On GD 14 fetuses were removed and accomplished chromosome preparation from fetal liver. Then fetal were fixed in formalin for brain hematoxilin and eosine staining and TUNEL assay.

We observed morphological abnormality including exencephal fetus in 5mg/kg MA group and premature fetuses in 10 mg/kg MA group. Also brain histological study showed subarachnoid hemorrhage in fetal brain in both experimental groups. Fetal liver karyotyping analysis was normal in fetuses of all groups and TUNEL assay in fetal striatum did not show significant difference in number of apoptotic cells between groups.

From our results, it could be concluded that chronic abuse of MA by pregnant females during organogenesis period can cause teratogenic effect and brain hemorrage in fetus.

Fetal color Doppler is important for evaluation of hypoxia in intrauterine growth restriction (IUGR) fetus.

In this study we compare fetal and maternal color Doppler with blood gas analysis to detect fetal acidosis.

In this cross-sectional study we evaluated 100 hospitalized patients with IUGR for comparison of color Doppler results with arterial blood gas analysis. Results of Doppler sonography of fetus middle cerebral arteries, umbilical and uterine artery and umbilical artery ABG were studied in these neonates.

Mean maternal age was 28±7 years, mean gestational age was 31.79±2.59 weeks and mean growth restriction was 3±2 weeks. Resistance increasing was observed in right uterine arteries of 37 mothers. It was normal in 60 mothers. Resistance increasing was observed in left uterine arteries of 36 mothers and nuch was seen in four cases. PCO2, PO2, and pH mean were 48.41±9.50 mmHg, 26.00±12.34 mmHg, and 7.28±0.10 in the neonates respectively.

In this study abnormal color Doppler in IUGR fetuses have no significant correlation with umbilical cord blood gas.

Diabetes mellitus (DM), primary or idiopathic is a chronic disorder of the carbohydrate, lipid and protein metabolism. DM may impact male reproductive function at several levels. It is shown that DM has detrimental effects on sperm parameters in human and experimental animals.

The aim of this study was to observe the effects of diabetes on sperm parameters (viability, count, morphology and motility) and evaluation of sperm chromatin quality in mice.

Totally twenty adult male Syrian mice were divided randomly into 2 groups (n=10). The animals of group A were considered as controls while group B mice were diabetic that received a single dose (200 mg/kg) streptozotocin (STZ) intra peritoneally. After 35 days, the cauda epididymis of each diabetic mouse was dissected and placed in culture medium for 30 min. The swim-out spermatozoa were analyzed for count, motility, morphology and viability. The sperm chromatin quality and DNA integrity, was evaluated with Aniline Blue (AB), Toluidine blue (TB), Acridine orange (AO) and Chromomycin A3 (CMA3) staining.

In sperm analysis, the diabetic mice had poor parameters in comparison with control animals (p=0.000). Regarding sperm chromatin quality, the results of TB and AO tests showed statically significant differences between two groups, but in AB and CMA3 staining, we didn’t see any differences between them.

The results showed that STZ-induced diabetes mellitus may influence the male fertility potential via affecting sperm parameters and DNA integrity in mice. However, according to our data, the diabetes doesn’t have any detrimental effects on histone-protamines replacement during the testicular phase of sperm chromatin packaging.

Recurrent pregnancy loss (RPL) caused by various genetic and non-genetic factors. After chromosome abnormality, thrombophilia is one of the most important genetic factors that could cause RPL. Factor V Leiden and factor II G20210A mutation were the most common mutations cause thrombophilia in the world.

The purpose of this study was to determine the frequency of factor V Leiden and prothrombine gene mutations in women with RPL compared with women who had uneventful pregnancies.

This case control study evaluates the frequency of factor V-Leiden and factor II G20210 genotypes in 80 women with two or more pregnancy losses, compared with 80 women without adverse pregnancy outcome. The mutations were assessed by PCR-RFLP.

Frequency of the factor V Leiden among cases was 2.5%, which was higher than controls (1.25%), but the difference was not significant. No factor II G20210 mutation was found among cases and controls.

These data did not confirm that factor V Leiden and factor II G20210 mutation might play a role in recurrent pregnancy loss in Iranian women.

The up-regulation of galectin-3, galectin-9, and galectin-15 expression in the luminal and glandular epithelium was reported in preparation of the endometrium for embryo implantation at the midlutheal phase. However, no data was available regarding the expression and the distribution pattern of galectin-8 in the human endometrium during a regular menstrual cycle.

The current study designed to investigate the expression and the distribution pattern of galectin-8, a beta-galactoside-binding lectin in the human endometrium during both proliferative and luteal phases of a regular menstrual cycle.

Endometrial biopsies were obtained from the anterior wall of the uterine cavity of 16 women (proliferative phase: n=4, lutheal phase: n=12). All female patients with mean age of 37.5 years were fertile (range 25-45). Each biopsy was divided into three pieces; one piece was fixed in formaldehyde for light microscopy and immunohistochemistry. The second portion fixed in glutaraldehyde for scanning electron microscopy and the third portion was prepared for western blot analysis.

Data of immunoblotting revealed a molecular weight of 34 kD band with high intensity in the lutheal phase samples. The immunohistochemistry staining demonstrated that galectin-8 expressed at a very low concentration during the proliferative phase, but showed a high expression throughout the lutheal phase. The expression of galectin-8 observed in luminal surface epithelium, glandular epithelium and stroma.

The up-regulation of the expression of galectin-8 during lutheal phase may suggest galectin-8 as one of the potential molecular marker of the endometrial receptivity. These data propose that galectin-8 may play an important role during the initial events of human embryo implantation.

Polycystic ovary syndrome (PCOS) causes an increased risk of metabolic cardiovascular syndrome. Also, cystatin C serum levels are associated with the risk of cardiovascular events in metabolic syndrome patients.

To investigate the relationship between cystatin C in PCOS patients.

35 women with PCOS were compared to 35 women with healthy matched age and body mass index. They all underwent tests to determine plasma levels of C-reactive protein (CRP), cystatin C, lipid profile and apo-lipoprotein. Blood pressure and demographic variables of each subject were obtained.

Systolic and diastolic blood pressure were higher in PCOS patients compared to control group. Triglyceride and low-density lipoprotein cholesterol levels were higher in PCOS; contrariwise, high-density lipoprotein was lower from that of healthy volunteers. Cystatin and CRP levels were significantly higher in patients with PCOS in comparison with healthy subjects (p<0.0001). Among measured determinants, only PCOS status was independently associated with cystatin C.

Cystatin C was positively correlated with PCOS status concentrations but not with systolic and diastolic blood pressure, or any of the lipid profile variables or demographic characteristics. Indeed, no correlation was found between cystatin C and CRP levels. Therefore, cystatin C might be related to PCOS beyond its use as a marker of the renal function.

Repeated implantation failure (RIF) is defined as pregnancy failure after two to six times with at least ten high grade embryo transfer to uterus. A variety of causes have been anticipated for RIF, including anatomical, autoimmune, genetics, endocrine and thrombotic anomalies. Factors responsible for RIF have important implication regarding treatment however in many couples a perfect cause cannot be found.

Cases:

 In these case series, we reported nine couples with RIF that after investigation no definitive etiology was found for RIF and empirical therapy by heparin, aspirin and or immunotherapy was not effective. In these cases we recommended transfer of embryos to surrogate uterus. Nine patients were studied and six of them developed a normal pregnancy (pregnancy rate=66.66%).

This study showed that surrogacy is a good option for treatment of RIF.