Évelyn Traina

Polymorphisms in VEGF, progesterone receptor and IL-1 receptor genes in women with recurrent spontaneous abortion

Hormonal, inflammatory and vascular alterations during pregnancy are thought to be involved in pregnancy loss. The role of progesterone, the actions of which involve cytokines and vascular endothelial growth factor (VEGF), has been thoroughly studied. Genetic factors are involved in modulation of these molecules, and several genes have been associated with recurrent spontaneous abortion (RSA), as well other obstetric conditions. Here we investigate the relationship between RSA and gene polymorphisms of the progesterone receptor (PROGINS), interleukin-1 receptor 1 (PstI) and vascular endothelial growth factor (VEGF) (-634, 936). This case-controlled study comprised a total of 89 women with idiopathic RSA and a control group of 191 women with at least two successful pregnancies and no miscarriages. Genomic DNA was extracted from whole blood, and polymorphism genotyping was conducted by digesting PCR products with specific restriction endonucleases. No correlations were found in any of the investigated polymorphisms, even among dominant, co-dominant and additive inheritance models and alleles. Our results support the hypothesis that these specific gene polymorphisms are not the major determinant of pregnancy success. Although we did not find any correlations, the investigation of these and other polymorphisms remains a challenge in the evaluation of RSA.

Acurácia diagnóstica da histerossalpingografia e da ultra-sonografia para avaliação de doenças da cavidade uterina em pacientes com abortamento recorrente

OBJETIVO: avaliar a acuracia da histerossalpingografia e da ultra-sonografia transvaginal bidimensional para o diagnostico de defeitos uterinos em pacientes com aborto recorrente. METODOS: oitenta pacientes com historia de dois ou mais abortos espontâneos consecutivos foram submetidas a histerossalpingografia, ultra-sonografia bidimensional pelvica e transvaginal e histeroscopia ambulatorial. Os achados foram divididos em tres grupos: alteracoes da forma, sinequias e lesoes polipoides. A histeroscopia foi considerada padrao-ouro. A concordância entre os diferentes metodos foi avaliada pelo coeficiente kappa e sua significância foi testada. O nivel de significância adotado foi de 0,05 (a=5%). Sensibilidade, especificidade, valores preditivos positivo e negativo, com intervalos de confianca de 95%, foram calculados. RESULTADOS: defeitos cavitarios foram diagnosticados em 29 (36,3%) pacientes, sendo 11 (13,7%) alteracoes da forma, 17 (21,3%) sinequias e uma (1,3%) lesao polipoide. A concordância global da histerossalpingografia com a histeroscopia foi de 85,0%, ao passo que da ultra-sonografia foi de apenas 78,7%. Os melhores resultados da histerossalpingografia foram obtidos para os diagnosticos de alteracoes da forma e sinequias (acuracia de, respectivamente, 97,5 e 95%). Para lesoes polipoides, a histerossalpingografia teve acuracia de apenas 92,5%, devido ao baixo valor preditivo positivo (14,3%). A ultra-sonografia teve acuracia inferior para todos os diagnosticos, 93,7% para alteracoes da forma e 85% para deteccao de sinequias, as custas principalmente de baixa sensibilidade. CONCLUSOES: a histerossalpingografia revelou boa acuracia diagnostica para a pesquisa da cavidade uterina em pacientes com aborto recorrente. A ultra-sonografia mostrou especificidade alta, porem com baixa sensibilidade.

Thrombophilic Mutations and Polymorphisms, Alone or in Combination, and Recurrent Spontaneous Abortion

Generally, recurrent spontaneous abortions (RSAs) have no identifiable cause; yet, vascular alterations during pregnancy may be associated with pregnancy loss. Therefore, we evaluated the association between thrombophilic mutations and RSAs. This case–control study was conducted in 112 patients who had RSAs and 98 health control women. Genomic DNA was extracted from whole blood, and polymorphism genotyping was conducted using polymerase chain reaction. The following 6 genetic variants were analyzed: factor V Leiden, prothrombin mutation, methylenetetrahydrofolate reductase C677T and A1298C, plasminogen activator inhibitor type 1 (4G>5G), and factor XIII G103T (V34L). No correlations were found in any of the investigated polymorphisms. Moreover, 35.0% of cases and 25.5% of controls had at least 2 mutations in combination, and 4.8% of cases and 5.1% of controls had 3, but these combinations were not associated with additional risk. In conclusion, we found no association between the polymorphisms studied and the occurrence of RSAs.

FAS and FAS-L genotype and expression in patients with recurrent pregnancy loss.

We assessed FAS and FAS-L gene polymorphisms and messenger RNA (mRNA) levels in patients with recurrent pregnancy loss (RPL). This case–control study compared 129 women with RPL with 235 healthy multiparous women (control group). Genomic DNA and total mRNA were extracted from whole blood, and polymorphisms genotyping was performed by polymerase chain reaction (PCR). Messenger RNA expression levels were analyzed by real-time PCR. Data were analyzed by chi-square and Fisher exact tests; P < .05 was considered significant. There were no significant differences in the FAS (670 A/G) genotype or allelic frequencies between the RPL and control groups. We found significant differences in the FAS-L (844 C/T) genotype and allelic frequencies between women with RPL and controls. Patients with RPL had significantly higher FAS-L expression. Our data suggest that FAS-L gene polymorphism is associated with increased susceptibility to RPL. Moreover, women with RPL seem to abnormally express FAS-FAS-L molecules.

Polimorfismo do receptor de progesterona como fator de risco para o parto prematuro

PURPOSE to investigate the association between gene polymorphism of the progesterone receptor (PROGINS) and the risk of premature birth. METHODS In this case-control study, 57 women with previous premature delivery (Case Group) and 57 patients with delivery at term in the current pregnancy and no history of preterm delivery (Control Group) were selected. A 10 mL amount of peripheral blood was collected by venipuncture and genomic DNA was extracted followed by the polymerase chain reaction (PCR) under specific conditions for this polymorphism and 2% agarose gel electrophoresis. The bands were visualized with an ultraviolet light transilluminator. Genotype and allele PROGINS frequencies were compared between the two groups by the χ2 test, with the level of significance set at value p < 0.05. The Odds Ratio (OR) was also used, with 95% confidence intervals. RESULTS PROGINS genotypic frequencies were 75.4% T1/T1, 22.8% T1/T2 and 1.8% T2/T2 in the Group with Preterm Delivery and 80.7% T1/T1, 19.3% T1/T2 and 0% T2/T2 in the term Delivery Group. There were no differences between groups when genotype and allele frequencies were analyzed: p = 0.4 (OR = 0.7) and p = 0.4 (OR = 0.7). CONCLUSIONS the present study suggests that the presence of PROGINS polymorphism in our population does not constitute a risk factor for premature birth.

Progesterone receptor gene polymorphism and recurrent spontaneous abortion

PURPOSE To assess a possible association between polymorphism of the progesterone receptor gene (PROGINS) and recurrent spontaneous abortion (RSA). METHODS In this case-control study, 85 women with at least three previous spontaneous abortions without an identifiable cause (RSA Group) and 157 women with at least two previous term pregnancies without pathologies and no previous miscarriage (Control Group) were selected. An amount of 10 mL of peripheral blood was collected by venipuncture and genomic DNA was extracted by the DTAB/CTAB method, followed by the polymerase chain reaction (PCR) under specific conditions for this polymorphism and by amplification by 2% agarose gel electrophoresis. The bands were visualized with an ultraviolet light transilluminator and the gels were photographed. Differences in the PROGINS genotype and allele frequencies between groups were analyzed by the χ² test, with the level of significance set at p<0.05. The Odds Ratio (OR) was also used, with 95% confidence intervals 95%CI. RESULTS PROGINS genotypic frequencies were 72.3% T1T1 and 27.7% T1T2 for the RSA group and 76.4% T1T1, 22.3% T1T2 and 1.3% T2T2 for the control group. There were no differences between groups when the genotype and allele frequencies were analyzed: respectively p=0.48 (OR: 0.8) and p=0.65 (OR: 0.9). CONCLUSIONS Our results suggest that PROGINS polymorphism is not associated with RSA.

Porque usamos imunoglobulina anti-D em excesso no abortamento precoce?

OBJECTIVE: evaluation of fetomaternal hemorrhage (FMH) in patients who would need Rh alloimmunization with anti-D immunoglobulin (300 mg) prophylaxis after early miscarriage. METHOD: we included in the study Rh (D) negative blood group patients with positive or unknown Rh (D) partners, who had had a miscarriage up to 12 weeks of gestation, and had been admitted to hospital for uterine curettage. After this procedure 5 ml of venous blood was collected from the patients and the rosette test was applied to screen which patients would need quantitative determination of fetal blood transferred to the maternal circulation, by the Kleihauer-Betke test (K-B). RESULTS: out of 26 patients evaluated the rosette test was positive in one , who showed an FMH of 1.5 ml in the K-B test. CONCLUSIONS: the dose of anti-D immunoglobulin used in cases of miscarriage up to 12 weeks of gestation should be substantially reduced. The availability of preparations of 50 mg is recommended, for a more inexpensive and rational treatment.