Vascular endothelial growth factor (VEGFA) gene variation in polycystic ovary syndrome in a Tunisian women population.

BACKGROUND: Polycystic ovary syndrome (PCOS) is characterized by the growth of a number of small cysts on the ovaries which leads to sex hormonal imbalance. Women who are affected by this syndrome suffer from irregular menstrual cycles, decline in their fertility, excessive hair growth, obesity, acne and most importantly cardiac function problems. The vascular endothelial growth factor (VEGF) plays a pivotal role in tissue vascularization in general and in the pathogenesis of many diseases. The PCOS was found to be associated with high expression levels of VEGF. In women who undergo assisted reproductive procedures (ART), VEGF was found to be a key mediator of other factors to control ovary angiogenesis. Here, we set out to examine the association of VEGFA gene polymorphism with PCOS and its components in a population of Tunisia women to enhance our understanding of the genetic background leading angiogenesis and vascularization abnormalities in PCOS. METHODS: The association of VEGFA gene with PCOS and its components was examined in a cohort of 268 women from Tunisia involving 118 PCOS patients and 150 controls. VEGFA gene variations were assessed through the analysis of the following SNPs rs699947 (A/C), rs833061 (C/T), rs1570360 (G/A), rs833068 (G/A), rs3025020 (C/T), and rs3025039 (C/T). The linkage disequilibrium between SNPs was assessed using HAPLOVIEW software while combination of SNPs into haplotypes in the population and the reconstruction of the cladogram were carried-out by PHASE and ARLEQUIN programs, respectively. Genetic association and genotype-phenotype correlations were calculated by logistic regression and non-parametric tests (Kruskall-Wallis and Mann-Whitney tests), respectively, using StatView program. RESULTS: We observed 10 haplotypes in our studied cohort whereH1 (ACGG), H2 (ACAG), H7 (CTGG) and H8 (CTGA) were the most frequent. We observed the association of the genotype CT of the SNP rs30225039 with PCOS phenotype (P = 0.03; OR 95 % CI = 2.05 [1.07-3.90]) and a trend for correlation of the pair of haplotypes H2/H2 with prolactin levels in plasma (P = 0.077; 193.5 ± 94.3 vs 45.7 ± 7.2). These data are consistent with literature and highlight one more time the role of vascularization in the pathogeny of PCOS. CONCLUSIONS: LD pattern in VEGF locus showed a similar LD pattern between the Tunisian population and the CEU. More haplotypes in the Tunisian population than in CEU was observed (22 haplotypes vs 16 haplotypes) suggesting higher recombination rate in Tunisians. The study showed that there was any advantage of using haplotypes compared with SNPs taken alone.

Chediak-higashi syndrome presented as accelerated phase: case report and review of the literature.

Chediak-Higashi syndrome (CHS) is a rare autosomal recessive disease, characterized by partial oculocutaneous albinism, recurrent pyogenic infections (skin, mucosa and respiratory system), and neurologic deficit. The hallmark of this syndrome is the presence of abnormal intracytoplasmic giant granules in all granule containing cells including leukocytes in blood and bone marrow. A majority (85 %) of patients with CHS develop an accelerated phase consisting of a lymphoproliferative syndrome with hemophagocytosis and infiltration of most tissues. This phase is characterized by fever, jaundice, hepatosplenomegaly, lymphadenopathy, pancytopenia and neurological abnormalities. In this paper, we report a case of CHS presented as accelerated phase in a 9-month-old girl child.

Polymorphisms in genes coding for the NK-cell receptor NKG2D and its ligand MICA in recurrent miscarriage.

PROBLEM: To investigate the possible association of Natural Killer Group (NKG) receptors gene polymorphisms and MHC class I chain-related protein A (MICA) gene polymorphism with recurrent miscarriage (RM). METHODS: Seven SNPs in NKG2D gene (rs1049174, rs2255336, rs2617160, rs2617161, rs2246809, rs2617169, and rs2617170), one SNP in NKG2A gene (rs1983526), and one SNP in MICA gene (MICA129) were assessed by allelic discrimination (real-time PCR) in both patients and control women. RESULTS: The rs2617170 T/T genotype significantly protected against RM [OR (95%) = 0.63 (0.40-0.98)]. The NKG2D haplotypes analysis on the basis of pairwise LD revealed two haplotype blocks. In block1, we found an increased frequency of CAT (Pc = 0.007; OR = 2.13; 95% CI = 1.24-3.68) and GGA haplotypes (Pc = 0.041; OR = 2.02; 95%CI = 1.03-3.96) and reduced frequency of CAA haplotype (Pc = 0.027; OR = 0.72; 95% CI = 0.54-0.96) in patients. In block2, increased frequency of GATG haplotype (Pc = 10(-4) ; OR = 9.25; 95% CI = 3.04-28.12) and reduced frequency of ATTC haplotype (Pc = 0.035; OR = 0.69; 95%CI = 0.50-0.97) were seen in patients. CONCLUSION: The NKG2D gene polymorphisms may influence the success of pregnancy in Tunisian women.

Contribution of JAK2 and STAT3 variants to the genetic susceptibility of recurrent spontaneous miscarriage in a Tunisian population.

AIMS: Th1 and Th2 balance is crucial for maintenance of pregnancy, and intracellular JAK and STAT proteins significantly contribute to it. In view of evidence linking JAK2 and STAT3 variants with recurrent spontaneous miscarriage (RSM), here we investigated the association of JAK2 (rs2230724) and STAT3 (rs1053023 and rs1053004) to RSM susceptibility in Tunisians. SUBJECTS AND METHODS: A retrospective case-control study. Subjects comprised 235 RSM cases and 235 control subjects. JAK2 and STAT3 were genotyped by the allelic discrimination method. RESULTS: STAT3 rs1053023 and, to a lower extent, rs1053004 were significantly associated with RSM under the additive and dominant, but not recessive models. This remained significant after adjustment for the covariates age, smoking, and gravida. In contrast to STAT3 variants, JAK2 rs2230724 was not associated with RSM under any of the genetic models tested. Two-locus STAT3 (rs1053023/rs1053004) haplotype analysis revealed increased frequency of the C/G haplotype in patients with RSM. Multivariate regression analysis confirmed the association of C/G haplotype with RSM (p=0.001; odds ratio=2.01; 95% confidence interval=1.32-3.07), thus conferring RSM susceptibility nature. These differences remained significant after applying the Bonferroni correction for multiple testing (Pc=0.004). CONCLUSIONS: STAT3 rs1053023, more so than the STAT3 rs1053004 or JAK2 rs2230724 polymorphisms, is associated with RSM risk.

Interleukin-18 promoter polymorphisms and risk of idiopathic recurrent pregnancy loss in a Tunisian population.

IL-18 is a pro-inflammatory cytokine that regulates the differentiation and effector functions of CD4+ (Th1) and CD8+ (CTL) T cells, which are implicated in the pathogenesis of recurrent pregnancy loss (RPL). We investigated the association of the IL-18 gene promoter single nucleotide polymorphisms (SNPs) -656C/A (rs1946519), -137G/C (rs187238), -119A/C (rs360718), and -105G/A (rs360717), by TaqMan assays in analysis in 470 Tunisian women comprising 235 RPL cases and 235 multi-parous controls. The association of IL-18 alleles, genotypes, and haplotypes with RPL was evaluated by Fisher's exact test and regression analysis. The frequency of minor alleles -105G/A (P<0.001) and -656C/A (P<0.001), but not -119A/C (P=0.93) or -137G/C (P=0.32), were higher in RPL cases. Significant differences were also noted in the genotype distribution of -105G/A (P<0.001) and -656C/A (P<0.001) between cases and controls. Four-locus (-656C/A, -137G/C, -119A/C, -105G/A) IL-18 haplotype analysis identified AGAA (corrected P<0.001), and CGAA (corrected P<0.001) haplotypes to be associated with increased RPL risk, after adjusting for age and BMI. These results demonstrate that -105G/A and -656C/A IL-18 variants are significantly associated with RPL.

Endothelial protein C receptor 1651C/G polymorphism and soluble endothelial protein C receptor levels in women with idiopathic recurrent miscarriage.

High levels of soluble endothelial protein C receptor (EPCR) induce coagulation dysfunction by inhibiting protein C activation, and activated protein C (APC) activity. We tested whether EPCR 1651C/G promoter variant and changes in plasma soluble EPCR levels are risk factors for idiopathic recurrent spontaneous miscarriage (RSM). A case-control study involving 283 RSM cases and 380 age and BMI-matched control women. EPCR 1651C/G genotyping was performed by PCR-RFLP method. Plasma-soluble EPCR levels were measured with ELISA. The 1651G allele frequency and C/G genotype were significantly higher in RSM cases than controls; none of the cases or control participants was a 1651G/G homozygote. Lower soluble EPCR levels were seen in RSM cases compared to controls, and higher soluble EPCR levels were seen in 1651C/G compared to 1651C/C carriers in cases and controls. Lower soluble EPCR levels were seen in cases, both in 1651C/C (P = 0.0046) and 1651C/G (P = 0.0032) genotype carriers. Multivariate analysis demonstrated strong association of EPCR 1651C/G [P = 0.011; adjusted odds ratio (aOR) (95% confidence interval [CI] = 3.13 (1.31-7.60)], but not soluble EPCR plasma levels [P = 0.067; aOR (95% CI) = 1.01 (1.00-1.10)], with increased RSM risk. In addition, smoking was independently associated with increased RSM risk [P = 0.002; aOR (95% CI) = 2.86 (1.48-5.52)]. EPCR 1651C/G polymorphism and elevated soluble EPCR levels but low soluble EPCR levels increase the risk of idiopathic RSM. Replication studies on other racial groups, and other EPCR gene variants, are warranted.

Common polymorphisms in the P-selectin gene in women with recurrent spontaneous abortions.

BACKGROUND: To investigate possible associations of P-selectin polymorphisms with idiopathic recurrent pregnancy loss (RPL). METHODS: Study subjects comprised 270 consecutive RPL cases attending outpatient maternity services, and 322 multi-parous control women. P-selectin genotyping was done by PCR-RFLP and PCR-ASA methods. RESULTS: The P-selectin variants rs1800807, rs1800805, and rs6127, were in Hardy Weinberg equilibrium, and low linkage disequilibrium was noted between the three studied SNPs. The frequency of rs6127 A allele (P<0.001I), but not rs1800807 C allele (P=0.957) or rs1800805 A allele (P=0.760), was higher in RPL cases than in control women. Significant differences in the distribution of rs6127 (P<0.001), but not rs1800807 (P=0.444) or rs1800805 (P=0.391) genotypes were seen between cases and controls, and only rs6127 showed a significant association with RPL, with increments of 2.65 and 4.96 in disease risk seen for heterozygous and homozygous carriers, respectively. Among the 8 three-locus Pselectin haplotypes constructed (rs1800807/rs1800805/rs6127), increased frequency of GGG (Pc=0.0249), CGG (Pc=0.0256), and CAG (Pc=0.0174) haplotypes, and lower frequency of CGA haplotype (Pc=0.0091) were seen in RPL cases, thus conferring disease susceptibility and protective nature to these haplotypes, respectively. CONCLUSIONS: P-selectin gene polymorphisms and haplotypes contribute to RPL development.

Association between adverse pregnancy outcomes and maternal factor V G1691A (Leiden) and prothrombin G20210A genotypes in women with a history of recurrent idiopathic miscarriages.

Thrombophilia was implicated in the development of pregnancy complications, including recurrent idiopathic pregnancy loss, and is aggravated in women who are carriers of factor V G1691A (FV Leiden) and prothrombin (PRT) G20210A single-nucleotide polymorphisms (SNPs). Previous studies examined the role of FV-Leiden and PRT G20210A in recurrent pregnancy loss with conflicting results. Here we examined the prevalence of FV Leiden and PRT G20210A SNPs, in 200 women with 3 or more consecutive early (n = 87), late (n = 41), or early-late (n = 72) recurrent pregnancy losses, and 200 age-matched fertile parous control women. APC resistance (APCR) was detected functionally (measuring the activated clotting time triggered by activated factor X in presence of a fixed amount of purified APC), and FV-Leiden and PRT G20210A genotypes were assessed by PCR. The frequency of the mutant FV (0.1400 vs. 0.0276; P < 0.001) but not PRT 20210 (0.0100 vs. 0.0225; P = 0.159) allele was higher in patients than controls, respectively. APC resistance with factor V Leiden was seen in 27% of patients compared to 11.5% of controls, while APC resistance without factor V Leiden was seen in 12.5% of patients compared to 9.5% of controls. Regression analysis demonstrated that the significant predictors for early abortion was FV Leiden; those for late abortion were oral contraceptive, APCR, and FV Leiden; and predictors for early-late abortions were oral contraceptives, obesity, FV Leiden, and smoking. APC resistance and FV Leiden, as well as combination of both, are common thrombotic defects seen in women with idiopathic recurrent pregnancy loss, thus testing for these is recommended in women who have experienced recurrent miscarriages.

Prevalence of antiphospholipid antibodies, factor V G1691A (Leiden) and prothrombin G20210A mutations in early and late recurrent pregnancy loss.

OBJECTIVE: We assessed the prevalence of inherited (FV-Leiden and PRT G20210A), and acquired (anti-PL antibodies) risk factors among habitual aborters in Tunisia. STUDY DESIGN: We studied prospectively 146 patients with > or =3 consecutive early, late, or early-late recurrent pregnancy losses, together with 99 age-matched controls. Anticardiolipin antibodies (ACL), lupus anticoagulant (LA), and APC resistance (APCR) were detected by ELISA, dilute Russell Viper Venom Time (dRVVT), and coagulation tests, respectively, and FV-Leiden and PRT G20210A genotypes were assessed by PCR. RESULTS: Anti-PL antibody frequencies were 45 and 9% among patients and controls, respectively (P < 0.001), with positive LA only (P = 0.004), or combined elevated ACL-positive LA being consistently higher (P < 0.001) among patients than controls. FV-Leiden (20.54% versus 6.06%), but not PRT G20210A (2.74% versus 4.04%) was significantly higher in patients versus controls. Among LA-positive cases higher prevalence of G/A (14/146 versus 1/99) and A/A genotypes (4/146 versus 0/99) were seen, and among ACL-positive cases higher prevalence of G/A (10/146 versus 0/99) and A/A genotypes (2/146 versus 0/99) were recorded. CONCLUSIONS: Anti-PL antibodies and FV-Leiden, but not PRT G20210A, are associated with recurrent idiopathic pregnancy losses in Tunisian women.