Implication of Novel BMP15 and GDF9 Variants in Unexpected Poor Ovarian Response.

Unexpected poor ovarian response (UPOR) occurs when nine or fewer oocytes are retrieved from a young patient with normal ovarian reserve. Bone morphogenetic protein15 (BMP15) and growth differentiation factor 9 (GDF9) are two oocyte-specific factors with pivotal role in folliculogenesis. The aim of this study was to assess the relation between BMP15 and GDF9 variants with UPOR. Hundred women aged ≤ 39 with AMH ≥ 1.27 IU/ml participated as UPOR and normal ovarian responders (NOR) based on their oocyte number. Each group consisted of 50 patients. After genomic DNA extraction, the entire exonic regions of BMP15 and GDF9 were amplified and examined by direct sequencing. Western blotting was performed to determine the expression levels of BMP15 and GDF9 in follicular fluid. Additionally, in silico analysis was applied to predict the effect of discovered mutations. From four novel variants of BMP15 and GDF9 genes, silent mutations (c.744 T > C) and (c.99G > A) occurred in both groups, whereas missense variants: c.967-968insA and c.296A > G were found exclusively in UPORs. The latter variants caused reduction in protein expression. Moreover, the mutant allele (T) in a GDF9 polymorphism (C447T) found to be more in NOR individuals (58% NOR vs. 37% UPOR (OR = 2.3, CI 1.32-4.11, p = 0.004).The novel missense mutations which were predicted as damaging, along with other mutations that happened in UPORs might result in ovarian resistance to stimulation. The mutant allele (T) in C447T polymorphism has a protective effect. It can be concluded that there is an association between BMP15 and GDF9 variants and follicular development and ovarian response.

Improving the knowledge of pregnant women using a pre-eclampsia app: A controlled before and after study.

OBJECTIVE: The objective of this study was to investigate the effect of a pre-eclampsia mobile application on the knowledge of pregnant women. METHODS: Following development of a pre-eclampsia mobile application, we conducted a controlled before and after study during a three-month period in 2018. The study population consisted of pregnant women attended to obstetrician clinics and offices in Kerman, Iran of whom, 110 sample participants were divided into two intervention and control groups. The participants completed a questionnaire of pre-eclampsia knowledge at baseline and 1-month follow up. Data were analyzed using inferential statistics including chi-square, independent sample t-test, paired t-test and linear regression. RESULTS: A total of 108 pregnant women with an average age of 28 years participated in this study. There was no significant difference between the scores of the two groups before the intervention (p=0.94). Their difference after the intervention was highly significant (p<0.001). The difference between the knowledge of the participants before and after the intervention was significant in the both groups (p<0.05). The results showed that the knowledge score of the participants after the intervention was significantly associated with their group and assessment score before the intervention (p<0.001). CONCLUSION: The results showed that the use of a mobile-based educational application improves the knowledge of pregnant women about pre-eclampsia. Increasing women's knowledge about pre-eclampsia may enables them to identify its signs and symptoms, resulting in the early detection and management of this condition, and likely reduction of its adverse consequences. TRIAL REGISTRATION: IRCT2017050633837N1.

Association of SNP rs.2414096 CYP19 gene with polycystic ovarian syndrome in Iranian women.

BACKGROUND: Genetic factors are believed to play an important role in the etiology of polycystic ovarian syndrome (PCOS) which is the most common endocrinological disorder of women in their reproductive age. Androgen metabolism is impaired in PCOS and, thus, CYP19 gene which is involved in this pathway can be a candidate gene. Previous studies have shown a relationship between single nucleotide polymorphism (SNP) of CYP19 in hyperandrogenism and PCOS in some racial groups. OBJECTIVE: This study was designed to elucidate the role of CYP19 gene in PCOS in Iran. MATERIALS AND METHODS: In this case-control study, 70 PCOS women and 70 non-PCOS women as normal control were selected. Following the informed consent, 5 ml blood was taken from individuals and subsequently, genomic DNA was extracted by salting out method. Furthermore, a set of polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) was carried out using specific primers for SNP rs.2414096 followed by enzyme digestion, with HSP92II. RESULTS: Genotype frequencies of SNP rs. 2414096 in PCOS women were as follows: AA (14.4%), AG (44.3%), and GG (41.4%) while in normal group, genotypes were 24.3%, 52.8%, and 22.9%, respectively. Allele frequencies in PCOS group were 49.3% for A and 50.7% for G, whereas normal group had a different percentage of A (36.4%) and G (63.6%). The calculations for both genotypic and allelic frequencies showed statistical significance difference. CONCLUSION: Variants of SNP rs. 2414096 in CYP19 could play a role in the development of PCOS in Iranian women.

Mutation analysis of exon1 of bone morphogenetic protein-15 gene in Iranian patients with polycystic ovarian syndrome.

BACKGROUND: With the prevalence of 6-10%, polycystic ovarian syndrome (PCOS) is considered the most common endocrinological disorder affecting women in their reproductive age. It has been suggested that genetic factors participate in the development of PCOS. Follicular development has been considered as one of the impaired processes in PCOS. Bone morphogenetic protein-15 (BMP-15) gene is a candidate gene in follicular development and its variants may play role in pathogenesis of PCOS. OBJECTIVE: To investigate whether BMP-15 gene mutations are present in Iranian women with PCOS. MATERIALS AND METHODS: In this cross-sectional study 5 ml venous blood samples was taken from 70 PCOS women referring to Afzalipour Hospital, Kerman University of Medical Sciences, Kerman, Iran, between January to December 2014. Genomic DNA was extracted from the blood sample by salting out method. Then a set of PCR reactions for exon1 of BMP-15 gene was performed using specific primers followed by genotyping with direct sequencing. RESULTS: Two different polymorphisms were found in the gene under study. In total 20 patients (28.6%) were heterozygote (C/G), and 2 patients (2.86%) were homozygous (G/G) for c.-9C>G in 5´UTR promoter region of BMP-15 gene (rs3810682). In addition, in the coding region of exon1, three patients (4.3%) were heterozygote (G/A) for c.A308G (rs41308602). Two PCOS patients (2.86%) appeared to have both c.-9C>G (C/G) and c.A308G (G/A) variants simultaneously. CONCLUSION: Our research detected two polymorphisms of BMP-15 gene among PCOS patients, indicating that even though it cannot be concluded that variants of BMP-15 gene are the principal cause of polycystic ovarian syndrome; they could be involved in pathogenic process in development of PCOS.

The effectiveness of luteal phase support with cyclogest in ovarian stimulated intra uterine insemination cycles: A randomized controlled trial.

BACKGROUND: Controlled ovarian stimulation combined with intra uterine insemination (IUI) is a convenient treatment of infertility with a success rate of 11%. The clinical observation and pattern of progesterone secretion in this method is suggestive of luteal phase defect and postulated as an implicating factor of treatment failure. OBJECTIVE: To investigate the efficacy of luteal phase support with intravaginal cyclogest in women undergoing controlled ovarian stimulation combined with intrauterine insemination. MATERIALS AND METHODS: In this single-blinded clinical trial, 196 consecutively seen women eligible for the study protocol, were randomized to receive either intravaginal progesterone (cyclogest pessary, Actavis) or no medication in luteal phase. Blood samples were collected and serum progesterone level in 7th and 11th day of the cycle, biochemical and clinical pregnancy and luteal phase duration were compared in case and control groups. RESULTS: The mean age in case and control group was 28 and 27.9 years, respectively and the most frequent cause of infertility was unexplained. Additionally, ovulatory dysfunction was the most common cause of female infertility in both groups. Based on these variables, there was no statistically significant difference between the two groups. Mean serum progesterone level in the case group were 48.34 and 34.24nmol/day on day 7 and 11 after insemination, respectively and both values were significantly higher than the control group. There was no difference between the two groups in terms of biochemical and clinical pregnancy. Luteal phase duration in the case group was significantly longer than the control group. CONCLUSION: Luteal phase support by Cyclogest pessary increases progesterone level and prolongs the luteal phase, but does not affect success rate of IUI cycles in terms of achieving pregnancy.

Comparison of maternal and cord blood nucleated red blood cell count between pre-eclamptic and healthy women.

AIM: The aim of this study was to evaluate the influence of pre-eclampsia on the cord and maternal nucleated red blood cell (NRBC) count. METHODS: Immediately after delivery, 1 mL of maternal venous blood and 1 mL of cord blood from 50 pre-eclamptic and 150 healthy pregnant women were collected separately in tubes containing 1.5 mg ethylene diamine tetra-acetic acid. Blood smears were prepared and stained using the Giemsa method. The number of NRBC per 100 leukocytes in maternal and cord blood was counted and compared between the two groups using SPSS software package for Windows. Any correlation of the NRBC count in maternal and umbilical cord blood was also evaluated. P-values < 0.05 were considered significant. RESULTS: The mean (+/-SD) NRBC per 100 white blood cell (WBC) level in cord blood of newborns in the pre-eclamptic group (18.2 +/- 31.8, range 0-142) was significantly greater than in the control group (6.2 +/- 8.1, range 0-36). Low birth weight and intrauterine growth restriction showed a statistically significant relationship with abnormal NRBC count in pre-eclamptic patients. A significant correlation was found between the maternal and cord blood NRBC count in the pre-eclamptic group. CONCLUSION: Fetal response to utero-placental insufficiency in pre-eclampsia leads to elevated NRBC in the cord blood, particularly in the presence of low birth weight and intrauterine growth restriction. The positive correlation between maternal and cord blood NRBC counts in pre-eclamptic patients indicates that maybe the hypoperfused placenta plays a role in the correlated alteration of the maternal and fetal NRBC count.

Ionized and total magnesium concentration in patients with severe preeclampsia-eclampsia undergoing magnesium sulfate therapy.

AIM: As ionized magnesium is the active form of magnesium and exerts a therapeutic effect, the present study was performed to determine the levels and correlations between ionized and total magnesium under baseline and therapeutic conditions in patients with severe preeclampsia and eclampsia receiving magnesium sulfate. METHODS: Fifty singleton patients with severe preeclampsia received a loading dose of 4 g of magnesium sulfate, followed by 2 g per hour as maintenance dose until 24 h after delivery, or 24 h after the last seizure in case of postpartum convulsions. Serial blood samples were taken before magnesium sulfate infusion, 30 min and 240 min after the initiation of the infusion and 4 h after the discontinuation of the drug. Data were analyzed by repeated measure ANOVA and paired t-test. RESULTS: Baseline levels of total and ionized magnesium were 2.4+/-0.6 mEq/L and 1.3+/-0.5 mEq/L (mean+/-SD), respectively. Putative level of 4 mEq/L of total magnesium was not obtained in up to 42% of patients during the treatment. There was not any significant correlation between the two forms of magnesium under baseline and therapeutic conditions. CONCLUSION: Despite the effectiveness of the standard regimen of magnesium sulfate in the treatment and prevention of eclamptic seizures, it can not provide the proposed therapeutic level of magnesium in all patients. With respect to the lack of correlation between ionized and total magnesium, further studies are necessary to investigate the superiority of measurement of ionized, rather than total magnesium, for titration of therapeutic magnesium sulfate infusion.

Severe preeclampsia and eclampsia in Kerman, Iran: complications and outcomes.

BACKGROUND: Our objective was to assess the frequency, epidemiological factors, complications, and outcome of severe preeclampsia and eclampsia and compare them according to severity of the condition in Kerman, southeastern Iran. MATERIAL/METHODS: A prospective study was performed on 200 consecutive cases of severe preeclampsia and eclampsia referred to the Kerman University Maternity Center from June 2001 to December 2002. Demographic data and complications were described and severe preeclampsia and eclampsia compared using t-student's and chi2 tests. RESULTS: Severe preeclampsia occurred in 3% and eclampsia in 0.6% of deliveries during the period of study. The majority of patients were young and primigravida, especially in the eclampsia group. 72.7% of eclamptic patients suffered their first convulsion at home and not in the presence of medical help. HELLP syndrome, disseminated intravascular coagulation, acute renal failure, neurological complications, and acute respiratory distress syndrome were the main maternal complications, all occurring more frequently in eclamptic patients. The frequency of pulmonary edema, placental abruption, postpartum hemorrhage, and aspiration pneumonia was not significantly different between the two groups. More than half of patients underwent cesarean delivery. Early neonatal mortality rate was 13% in the entire cohort. All six maternal deaths during the period of the study occurred in eclamptic cases (mortality rate 18%), four relating to patients who had not received prenatal care. CONCLUSIONS: Severe preeclampsia carries a high risk of maternal and neonatal morbidities and mortality in Kerman. This risk increases in the presence of eclampsia, particularly in neglected cases who have not sought prenatal care.

Nifedipine or hydralazine as a first-line agent to control hypertension in severe preeclampsia.

BACKGROUND: Pre-eclampsia is one of the most serious and common complications of pregnancy. Nifedipine, a calcium channel blocker, and the vasodilator hydralazine have both been used as antihypertensive agents in this condition. The aim of this study was to determine which of these two agents is the most appropriate antihypertensive in the management of severe pre-eclampsia. METHODS: One hundred and twenty-six pre-eclamptic patients with a gestational age of more than 20 weeks were randomized to receive either 8 mg nifedipine sublingually or 5-10 mg intravenous hydralazine. Women with a history of heart failure and women receiving antihypertensive treatment during the course of the current pregnancy were excluded. For each patient the following data were recorded; the number of drug administrations, the time needed to control blood pressure, mean urinary output, the time interval between effective control and a new hypertensive crisis after each drug administration and relevant adverse effects in mother or fetus. RESULTS: Effective control of blood pressure was achieved in both treatment arms. Data analysis indicated significantly fewer drug administrations in the nifedipine arm of the study. The time interval before a new hypertensive crisis following initial effective control of blood pressure was significantly longer in the nifedipine group when compared with hydralazine. Effective control of blood pressure was achieved more rapidly in multiparous patients receiving nifedipine (p=0.026). Mean urinary output before and after delivery was greater in the nifedipine arm of the study. There were no significant differences between the two groups in other variables. In addition, in neither group were there any serious adverse effects in mother or fetus. CONCLUSION: Nifedipine is safe and more effective than hydralazine in controlling blood pressure in severe pre-eclampsia. It has the added advantage of being cheaper and more widely available than the latter and is easily administered.