Analysis of genetic etiology and related factors in 1 065 women with spontaneous abortions / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic etiology and related factors in 1 065 women with spontaneous abortions.@*METHODS@#All patients have presented at the Center of Prenatal Diagnosis of Nanjing Drum Tower Hospital from January 2018 to December 2021. Chorionic villi and fetal skin samples were collected, and the genomic DNA was assayed by chromosomal microarray analysis (CMA). For 10 couples with recurrent spontaneous abortions but normal CMA results for abortive tissues, non-in vitro fertilization-embryo transfer (IVF-ET) pregnancies and no previous history of live births and no structural abnormalities of the uterus, peripheral venous blood samples were collected. Genomic DNA was subjected to trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing and bioinformatics analysis. Multifactorial unconditional logistic regression analysis was carried out to analyze the factors that may affect chromosomal abnormality in spontaneous abortions, such as the age of the couple, number of previous spontaneous abortions, IVF-ET pregnancy and history of live birth. The incidence of chromosomal aneuploidies in spontaneous abortions during the first trimester was compared in young or advanced-aged patients by chi-square test for liner trend.@*RESULTS@#Among the 1 065 spontaneous abortion patients, 570 cases (53.5%) of chromosomal abnormalities were detected in spontaneous abortion tissues, which included 489 cases (45.9%) of chromosomal aneuploidies and 36 cases (3.4%) of pathogenic/likely pathogenic copy number variations (CNVs). Trio-WES results have revealed one homozygote variant and one compound heterozygote variants in two pedigrees, both of which were inherited from the parents. One likely pathogenic variant was detected in the patient from two pedigrees. Multifactorial unconditional Logistic regression analysis suggested that age of patient was an independent risk factor of chromosome abnormalities (OR = 1.122, 95%CI: 1.069-1.177, P < 0.001), the number of previous abortions and IVF-ET pregnancy were independent protective factors for chromosomal abnormalities (OR = 0.791, 0.648; 95%CI: 0.682-0.916, 0.500-0.840; P = 0.002, 0.001), whilst the age of husband and history of live birth were not (P > 0.05). The incidence of aneuploidies in the abortive tissues has decreased with the number of previous spontaneous abortions in young patients (χ² = 18.051, P < 0.001), but was not significantly correlated with the number of previous spontaneous abortions in advanced-aged patients with spontaneous abortions (P > 0.05).@*CONCLUSION@#Chromosomal aneuploidy is the main genetic factor for spontaneous abortion, though CNVs and genetic variants may also underlie its genetic etiology. The age of patients, number of previous abortions and IVF-ET pregnancy are closely associated with chromosome abnormalities in abortive tissues.

Association Between PON1 (L55M and Q192R) Genetic Polymorphism and Recurrent Pregnancy Loss in North Indian Women Exposed to Pesticides / Associação entre o polimorfismo genético PON1 (L55M e Q192R) e a perda recorrente de gravidez emmulheres do norte da Índia expostas a pesticidas

Abstract Objective The aim of the present study was to examine the relation between the PON1 polymorphisms and recurrent pregnancy loss (RPL). Methods In a cross-sectional study, blood samples were collected from 100 females. DNA was extracted and PON1 genotypes were determined by polymerase chain reaction (PCR) amplification. Results Regarding PON1 L55M, the mutated allele (M) frequency was found in 70.5% in RPL and in 53.5% in controls; theMallele was significantly associated with an increased risk of RPL (adjusted odds ratio [ORadj]=2.07; 95% confidence interval [CI]; p<0.001). However, regarding PON1 Q192R, the R mutated allele frequency was found in 28.5% in RPL and in 33% in controls. The R allele did not show any risk for RPL (ORadj 0.81; 95%CI; p=0.329). Conclusion The present study suggests that there is an effect of genetic polymorphism on RPL and provides additional evidence that combines with the growing information about the ways in which certain PON1 genotypes can affect the development of the fetus in the uterus.

Resumo Objetivo O objetivo deste estudo foi examinar a relação entre os polimorfismos PON1 e perda recorrente de gravidez PRG. Métodos Em um estudo transversal, foramcoletadas amostras de sangue de 100 mulheres. O DNA foi extraído e os genótipos PON1 foram determinados por amplificação por PCR. Resultados Com relação ao PON1 L55M, a frequência do alelo mutado (M) foi encontrada em 70,5% no PRG e em 53,5% nos controles; o alelo M foi significativamente associado a um risco aumentado de PRG (odds radio ajustado [ORadj] =2,07; intervalo de confiança [IC] 95%; p<0,001). No entanto, em relação ao PON1 Q192R, a frequência do alelo mutado R foi encontrada em 28,5% no PRG e em 33% nos controles. O alelo R não mostrou qualquer risco para PRG (ORadj 0,81; IC 95; p=0,329). Conclusão O presente estudo sugere que há um efeito do polimorfismo genético sobre PRG e fornece evidências adicionais que se combinam com as informações crescentes sobre asmaneiras pelas quais certos genótipos PON1 podemafetar o desenvolvimento do feto no útero.

Association of angiotensin-converting enzyme insertion/deletion polymorphism with recurrent pregnancy loss: a meta-analysis of 26 case-control studies

Abstract Objective Previous studies investigating the association between angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and recurrent pregnancy loss (RPL) risk has provided inconsistent results. The aim of our study was to assess the association between the ACE I/D polymorphism and risk of RPL. Methods All studies published up to January 30, 2018 on the association of ACE I/D polymorphism with RPL were identified by searching the PubMed, Web of Knowledge, and Google scholar databases. Results A total of 26 case-control studies with 3,140 RPL cases and 3,370 controls were included in themeta-analysis. Overall, there was a significant association between ACE I/D polymorphism and RPL risk under the allele model (I versus D: odds ratio [OR] = 0.538, 95% confidence interval [CI] = 0.451-0.643, p 0.001), the homozygote model (II versus DD: OR = 0.766, 95% CI = 0.598-0.981, p = 0.035) and the recessive model (II versus ID + DD: OR = 0.809, 95% CI = 0.658-0.994, p = 0.044). Subgroup analysis by ethnicity showed that there was a significant association between ACE I/D polymorphism and increased risk of RPL in Caucasian and West-Asian populations, but not in East-Asians. When stratified by number of recurrent miscarriages (RMs), a significant association between ACE I/D polymorphism and increased risk of RPL was detected in the group of studies with ≥ 2 RMs, but not in studies with ≥ 3 RMs. Conclusion Themeta-analysis suggests that ACE I/D polymorphism is associated with increased risk of RPL. The ACE I/D polymorphism may be a risk factor for RPL in Caucasian and West-Asian populations, but not in East-Asians.

Associação de alelos HLA e aborto espontâneo recorrente em uma população de São Luís/Maranhão, na região Nordeste do Brasil / HLA alleles association and recurrent spontaneous abortion in a São Luis/Maranhão population, in Brazilian Northeastern region

Investigar a associação dos alelos HLA-A, -B e -DRB1 com a ocorrência de Aborto Espontâneo Recorrente.

Metabolism and gene polymorphisms of the folate pathway in Brazilian women with history of recurrent abortion / Metabolismo e polimorfismos gênicos da via do folato em mulheres brasileiras com história de aborto recorrente

PURPOSE: To investigate the association between polymorphisms in genes that encode enzymes involved in folate- and vitamin B12-dependent homocysteine metabolism and recurrent spontaneous abortion (RSA). METHODS: We investigated the C677T and A1298C polymorphisms of the methylenetetrahydrofalate reductase gene (MTHFR), the A2756G polymorphism of the methionine synthase gene (MS) and the 844ins68 insertion of the cystathionine beta synthetase gene (CBS). The PCR technique followed by RFLP was used to assess the polymorphisms; the serum levels of homocysteine, vitamin B12 and folate were investigated by chemiluminescence. The EPI Info Software version 6.04 was used for statistical analysis. Parametric variables were compared by Student's t-test and nonparametric variables by the Wilcoxon rank sum test. RESULTS: The frequencies of gene polymorphisms in 89 women with a history of idiopathic recurrent miscarriage and 150 controls were 19.1 and 19.6% for the C677T, insertion, 20.8 and 26% for the A1298C insertion, 14.2 and 21.9% for the A2756G insertion, and 16.4 and 18% for the 844ins68 insertion, respectively. There were no significant differences between case and control groups in any of the gene polymorphisms investigated. However, the frequency of the 844ins68 insertion in the CBS gene was higher among women with a history of loss during the third trimester of pregnancy (p=0.003). Serum homocysteine, vitamin B12 and folate levels id not differ between the polymorphisms studied in the case and control groups. However, linear regression analysis showed a dependence of serum folate levels on the maintenance of tHcy levels. CONCLUSION: The investigated gene polymorphisms and serum homocysteine, vitamin B12 and folate levels were not associated with idiopathic recurrent miscarriage in the present study. Further investigations are needed in order to confirm the role of the CBS 844ins68 insertion in recurrent miscarriage. .

OBJETIVO: Investigar a associação entre polimorfismos nos genes que codificam enzimas envolvidas no metabolismo da homocisteína dependente de folato e vitamina B12 e aborto espontâneo recorrente. MÉTODOS: Investigamos os polimorfismos C677T e A1298C no gene methilenotetrahidrofalato redutase (MTHFR); o polimorfismo A2756G no gene metionina sintase (MS) e a inserção 844ins68 no gene da cistationina beta-sintetase (CBS). A técnica de PCR seguido por RFLP foi utilizada para investigar os polimorfismos. Os níveis séricos de homocisteína, vitamina B12 e de folato foram investigados pela técnica de quimioluminescência. O Software Epi Info versão 6.04 foi utilizado para realizar a análise estatística. As variáveis paramétricas foram comparadas pelo teste t de Student e as variáveis não paramétricas pelo teste de Wilcoxon rank sum. RESULTADOS: As frequências dos polimorfismos gênicos em 89 mulheres com história de aborto recorrente idiopático e 150 controles foram de 19,1 e 19,6% para o C677T; 20,8 e 26% para o A1298C; 14,2 e 21,9% para o A2756G e 16,4 e 18% para a inserção 844ins68, respectivamente. Não houve diferenças significantes entre os grupos caso e controle em todos os polimorfismos dos genes investigados. No entanto, a frequência da inserção 844ins68 no gene CBS foi maior entre mulher com histórico de perdas no terceiro trimestre da gravidez p=0.003). Os níveis de homocisteína, vitamina B12 e folato séricos não foram diferentes entre os polimorfismos estudados nos grupos casos e controles. No entanto, a análise de regressão linear mostrou dependência dos níveis séricos de folato na manutenção dos níveis de homocisteína. CONCLUSÃO: Os polimorfismos gênicos investigados, assim como homocisteína, vitamina B12 e os níveis séricos de folato não foram associados com abortos recorrentes idiopático no presente estudo. Novas investigações devem ser realizados a fim de confirmar o papel da inserção 844ins68-CBS nos abortos recorrentes. .

Chromosomal abnormalities in couples with recurrent first trimester abortions / Alterações cromossômicas em casais com aborto recorrente de primeiro trimestre

PURPOSE: To investigate the prevalence of chromosomal abnormalities in couples with two or more recurrent first trimester miscarriages of unknown cause. METHODS: The study was conducted on 151 women and 94 partners who had an obstetrical history of two or more consecutive first trimester abortions (1-12 weeks of gestation). The controls were 100 healthy women without a history of pregnancy loss. Chromosomal analysis was performed on peripheral blood lymphocytes cultured for 72 hours, using Trypsin-Giemsa (GTG) banding. In all cases, at least 30 metaphases were analyzed and 2 karyotypes were prepared, using light microscopy. The statistical analysis was performed using the Student t-test for normally distributed data and the Mann-Whitney test for non-parametric data. The Kruskal-Wallis test or Analysis of Variance was used to compare the mean values between three or more groups. The software used was Statistical Package for the Social Sciences (SPSS), version 17.0. RESULTS: The frequency of chromosomal abnormalities in women with recurrent miscarriages was 7.3%, including 4.7% with X-chromosome mosaicism, 2% with reciprocal translocations and 0.6% with Robertsonian translocations. A total of 2.1% of the partners of women with recurrent miscarriages had chromosomal abnormalities, including 1% with X-chromosome mosaicism and 1% with inversions. Among the controls, 1% had mosaicism. CONCLUSION: An association between chromosomal abnormalities and recurrent miscarriage in the first trimester of pregnancy (OR=7.7; 95%CI 1.2--170.5) was observed in the present study. Etiologic identification of genetic factors represents important clinical information for genetic counseling and orientation of the couple about the risk for future pregnancies and decreases the number of investigations needed to elucidate the possible causes of miscarriages. .

OBJETIVO: Determinar a prevalência de alterações cromossômicas em casais com dois ou mais abortos recorrentes do primeiro trimestre, sem causa definida. MÉTODOS: Foram incluídas 151 mulheres e 94 parceiros com história obstétrica de 2 ou mais abortos consecutivos de 1º trimestre (1-12 semanas de gestação). Os controles foram 100 mulheres saudáveis, sem histórico de perda da gravidez. A análise cromossômica foi realizada em linfócitos de sangue periférico, cultivados 72 horas e tratados com a técnica Tripsina-Giemsa (GTG banda). Em todos os casos, foram analisadas 30 metáfases e montados 2 cariótipos, sendo utilizada microscopia de luz. A análise estatística foi realizada por meio do teste t de Student para dados com distribuição normal e o teste Mann-Whitney para os dados não paramétricos. Foi usado o teste de Kruskal-Wallis ou Análise de Variância para comparação dos valores médios entre três ou mais grupos. O software utilizado foi o Statistical Package for the Social Sciences (SPSS), versão 17.0. RESULTADOS: A frequência de alterações cromossômicas das mulheres com aborto recorrente foi de 7,3%, incluindo 4,7% com mosaicismo do cromossomo X, 2% com translocações recíprocas e 0,6% com translocações Robertsonianas. No total, 2,1% dos parceiros das mulheres com abortos recorrentes tinham anormalidades cromossômicas, sendo 1% com mosaicismo do cromossomo X e 1% com inversões. Entre os controles, 1% apresentou mosaicismo. CONCLUSÃO: No presente estudo, foi observada associação entre alterações cromossômicas e aborto recorrente no primeiro trimestre da gestação (OR=7,7; IC95% 1,2-170,5). A identificação etiológica de fatores genéticos é uma informação ...

Análisis de asociación de los polimorfismos -725C>G (rs1233334), -201G>A (rs1233333) y 14 bp deleción/inserción (14-pb del/ins) (rs66554220) del gen HLA-G en mujeres mexicanas con pérdida gestacional recurrente / Analysis of association of polymorphisms -725C >G ( rs1233334 ), -201G >A ( rs1233333 ) and 14 bp deletion / insert (14 - bp / ins ) ( rs66554220 ) of HLA -G gene mexican women with recurrent pregnancy loss

Antecedentes: El antígeno leucocitario humano (HLA)-G es una molécula inmunomoduladora que contribuye a la aceptación del feto semialogénico. Algunos polimorfismos de un solo nucleótido (SNP) en las regiones no codificantes del gen HLA-G inducen a la disminución de moléculas HLA-G, lo cual contribuye a complicaciones en el embarazo, tales como la preeclampsia o pérdida gestacional recurrente. Objetivo: Analizar la asociación de los polimorfismos -725C>G (rs1233334), -201G>A (rs1233333) y 14 bp deleción/inserción (14-pb del/ins) (rs66554220) del gen HLA-G en mujeres mexicanas con PGR. Métodos: Los polimorfismos -725C>G (rs1233334), -201G>A (rs1233333) y 14-pb del/ins (rs66554220) se identificaron por medio de PCR-SSOP (Polymerase Chain Reaction-sequence-specific oligonucleotide probe) y PCR (Polymerase Chain Reaction), respectivamente, en 58 mujeres con pérdida gestacional recurrente (> 2 abortos), sin factores de riesgo identificables y 56 mujeres fértiles no relacionadas (> 2 nacidos vivos). Resultados: El polimorfismo -725C>G (rs1233334) presentó diferencias significativas entre los grupos de estudio pero no se asoció con PGR (p=0,02601; OR=11,484; IC95 por ciento =0,617-213,659). Los polimorfismos -201G>A (rs1233333) y 14-pb del/ins (rs66554220) no se distribuyeron de manera diferente entre los grupos de estudio ni se asociaron con pérdida gestacional recurrente. Los polimorfismos analizados se encontraron en equilibrio de ligamiento (D'>0,3563; r²<0,1140). Conclusión. Este estudio sugiere que los polimorfismos -725C>G (rs1233334), -201G>A (rs1233333) y 14-pb del/ins (rs66554220) del gen HLA-G están en equilibrio de ligamiento y no influyen en el riesgo de pérdida gestacional recurrente en mujeres mexicanas.

Background: The human leukocyte antigen (HLA)-G is an important immunomodulatory molecule that contributes to the acceptance of the semi-allogeneic fetus. Some single nucleotide polymorphisms (SNP) in the noncoding regions of the HLA-G gene may influence the cellular levels of HLA-G, contributing to pregnancy complications such as preeclampsia or recurrent pregnancy loss. Objective: To analyze the association of -725C>G (rs1233334),-201G>A (rs1233333) and 14 bp deletion/insertion (14-bp del/ins) (rs66554220) polymorphisms in the HLA-G gene in Mexican women with RPL. Methods: -725C>G (rs1233334), -201G>A (rs1233333) and 14-bp del/ins (rs66554220) polymorphisms in the HLA-G gene were identified by PCR-SSOP (polymerase chain reaction-sequence-specific oligonucleotide probe) and PCR (polymerase chain reaction), respectively, in 58 women with recurrent pregnancy loss (> 2 miscarriages) without identifiable risk factors and 56 unrelated fertile women (> 2 live births). Results: -725C>G (rs1233334) polymorphism showed significant differences between the study groups but it was not associated with recurrent pregnancy loss (p=0.02601, OR=11.484; 95 percent CI=0.617-213.659). -201G>A (rs1233333) and 14-bp del/ins (rs66554220) polymorphisms were not distributed differently in study groups and not associated with RPL. Analyzed polymorphisms were in linkage disequilibrium (D' > 0.3563, r² < 0.1140). Conclusion: This study suggests that -725C>G (rs1233334), -201G>A (rs1233333) and 14-pb del/ins (rs66554220) in the HLA-G gene are in linkage equilibrium and do not influence the risk of recurrent pregnancy loss in Mexican women.

Anexinas y pérdidas recurrentes de embarazo / Annexins and recurrent pregnancy loss

Las pérdidas de embarazo recurrentes (PER), afectan a la salud pública y comprometen en forma directa la calidad de vida de cientos de mujeres, con detrimento de su salud física y psíquica. Aproximadamente un 50% de las PER no se asocian a alguna de las etiologías conocidas, y por lo tanto se consideran idiopáticas. Recientemente se ha demostrado que la expresión de la anexina 5 (ANXA5), una proteína ubicada en la superficie trofoblástica, juega un papel fundamental en el mantenimiento del embarazo ya que cumple un rol como inmunomodulador y anticoagulante a nivel de la placenta. Algunos haplotipos genéticos de la ANXA5 se asocian a alteraciones en la expresión de este gen, como el haplotipo M2 que se vincula a una reducción en la expresión de la ANXA5. La presencia de dicho haplotipo se relaciona con los siguientes eventos del embarazo: PER, restricción del crecimiento fetal intrauterino, bajo peso al nacer, preclampsia y tromboembolismo pulmonar materno. Esta revisión describe la estructura, función y expresión genética de la ANXA5, así como también su posible implicancia en la PER.

Recurrent Pregnancy Loss (RPL) affects public health and directly compromises the quality of life of hundreds of women, with a detrimental effect on their physical and mental health. Approximately 50% of RPL are not associated to any of the currently known etiology and will be considered idiopathic. Recently, it has been demonstrated that the expression of annexin 5 (ANXA5), a protein found on the trophoblastic surface, plays a fundamental role in the development of pregnancy due to its immunomodulator and anticoagulant function at the placentary level. Some genetic haplotypes of ANXA5 are associated to alterations in the expression of this gene, such as haplotype M2 which is associated to a decrease in the expression of ANXA5. The presence of this haplotype is related to the following conditions occurring during pregnancy: RPL, foetal intrauterine growth restriction, low child weight at birth, preeclampsia and maternal pulmonary thromboembolism. This review describes the structure, function and genetic expression of ANXA5, as well as its possible implication in RPL.

study of chromosomal abnormalities and heteromorphism in couples with 2 or 3 recurrent abortions in Shahid Beheshti hospital of Hamedan

Different studies show that chromosomal balance translocation in the parents can cause recurrent spontaneous abortions. Incidence of chromosomal translocation abnormalities in couples with repeated abortions is from 0% to 31%. The purpose of this research was studying the presence or absence of chromosomal abnormalities and heteromorphism in couples with recurrent abortions and also the role of this anomaly in the abortions. This study is a cross sectional descriptive study which have investigated 75 couples who had three abortions or more, and 65 couples who had two abortions that referred by gynecologist to the lab of Beheshti Hospital in Hamedan for cytogenetical investigation. Also 40 healthy individuals without history of abortion investigated as control group.GTG bonding technique [staining banding with gymsa and trypsin] is used in this study. Frequency of chromosomal abnormalities and heteromorphism among couples with three or more abortions were reported respectively 5.3% and 9.3%. This frequency in couples with two abortions was respectively 3.07%and 6.15%. The frequency of chromosomal heteromorphism in control group was 7.5% and no chromosomal abnormalities were observed in them. This study shows that chromosomal abnormality can be one reason of recurrent spontaneous abortions and more abortion increases the probability of this anomaly. Also, existence of chromosomal heteromorphism in the general population without clinical abortion symptoms shows that chromosomal heteromorphism cannot be the reason of these spontaneous abortions

relationship between cytokine gene polymorphism and unexplained recurrent spontaneous abortion in Saudi females

To investigate the relationships between unexplained recurrent spontaneous abortion [RSA] and single nucleotide polymorphisms tumor necrosis factor-alpha [TNF-alpha] [-238 G/A, -308 G/A], interleukin [IL]-6 [-634 G/C] and IL-10 [-592 C/A] in the promoter region of 3 different interleukin [TNF- alpha, IL-6, and IL-10] genes. The study group comprised 65 women [mean age: 34.1 +/- 6. 2; range: 15-45 years] with unexplained RSA, consecutively referred to the Recurrent Abortion Clinic, King Khaled University Hospital, Riyadh, Kingdom of Saudi Arabia from January 2010 to January 2011. The control group consisted of 65 females with at least 2 successful pregnancies and no history of abortion. Blood samples were drawn and deoxyribonucleic acid [DNA] was extracted using Puregene DNA purification kit. Utilizing polymerase chain reaction, the promoter region was amplified and sequenced on an Applied Biosystems Integrated sequencer to study the polymorphic sites of interest. All polymorphisms were identified in the case and control samples. A significant association was identified only between the -308 G/A polymorphism in the TNF- alpha gene promoter and the occurrence of unexplained RSA, and there was no significant association with other positions. The TNF- alpha gene polymorphism at position -308 could be a genetic predisposing factor for unexplained RSA

Genetic counseling in carriers of reciprocal translocations involving two autosomes.

One of the main genetic causes involve in the pathogenesis of recurrent abortion is parental chromosomal abnormalities. The central concept in genetic counseling with such families is to estimate the probability of recurrence of unfavorable pregnancy outcomes. The main questions that consultants usually ask are: Why did this happen? What is the risk to be done again? Our cases were two families with repeated miscarriage. The pedigrees were drawn, the chromosomes of couples were studied, and estimation for recurrent risk was done. We tried to answer those two main questions and clear the results for them. Parental chromosome abnormalities were founded after karyotyping with GTG technique at 450 band resolution, revealing 46 chromosomes with balanced translocation of autosomes in one of the partner in both families. Recurrent risk was estimated as "high" for their future pregnancies in each family. Couples in which one partner is the carrier of such balanced translocation have increased risks of infertility, recurrent abortion, and delivery of chromosomally abnormal offspring. Genetic counseling of such couples, therefore, presents a unique challenge and should be considered in dealing with such families.

The ADA*2 allele of the adenosine deaminase gene (20q13.11) and recurrent spontaneous abortions: an age-dependent association

OBJECTIVE: Adenosine deaminase acts on adenosine and deoxyadenosine metabolism and modulates the immune response. The adenosine deaminase G22A polymorphism (20q.11.33) influences the level of adenosine deaminase enzyme expression, which seems to play a key role in maintaining pregnancy. The adenosine deaminase 2 phenotype has been associated with a protective effect against recurrent spontaneous abortions in European Caucasian women. The aim of this study was to investigate whether the G22A polymorphism of the adenosine deaminase gene is associated with recurrent spontaneous abortions in Brazilian women. METHODS: A total of 311 women were recruited to form two groups: G1, with a history of recurrent spontaneous abortions (N = 129), and G2, without a history of abortions (N = 182). Genomic DNA was extracted from peripheral blood with a commercial kit and PCR-RFLP analysis was used to identify the G22A genetic polymorphism. Fisher's exact test and odds ratio values were used to compare the proportions of adenosine deaminase genotypes and alleles between women with and without a history of recurrent spontaneous abortion (p<0.05). The differences between mean values for categorical data were calculated using unpaired t tests. The Hardy-Weinberg equilibrium was assessed with a chi-square test. RESULTS: Statistically significant differences were identified for the frequencies of adenosine deaminase genotypes and alleles between the G1 and G2 groups when adjusted for maternal age. CONCLUSIONS:The results suggest that the adenosine deaminase *2 allele is associated with a low risk for recurrent spontaneous abortions, but this association is dependent on older age.

Polimorfismo do gene dos receptores de progesterona e o aborto espontâneo de repetição / Progesterone receptor gene polymorphism and recurrent spontaneous abortion

OBJETIVO: investigar se polimorfismos dos genes que codificam o receptor de progesterona (PROGINS) estão relacionados à ocorrência de aborto espontâneo de repetição (AER). MÉTODOS: em estudo caso-controle, foram selecionados 85 pacientes com antecedente de pelo menos três abortos precoces sem etiologia definida (Grupo Caso) e 157 mulheres com história de pelo menos duas gestações de termo sem intercorrências e sem passado de abortamento (Grupo Controle). Realizada coleta de 10 mL de sangue por punção venosa periférica e extração de DNA pela técnica DTAB/CTAB. As genotipagens foram feitas por reação em cadeia de polimerase (PCR), nas condições de ciclagem específica para o polimorfismo em estudo, seguida de amplificação em gel de agarose a 2 por cento. A visualização das bandas foi feita sob luz ultravioleta e os géis foram fotografados. As diferenças genotípicas e alélicas entre os dois grupos para o polimorfismo PROGINS foram calculadas pelo teste de χ2, adotando-se como nível de significância valores de p<0,05. Calculou-se ainda o Odds Ratio (OR, razão de chances), com intervalos de confiança de 95 por cento (IC95 por cento). RESULTADOS: As frequências genotípicas encontradas para o polimorfismo PROGINS foram de 72,3 por cento T1/T1 e 27,7 por cento T1/T2 no grupo com AER e 76,4 por cento T1/T1, 22,3 por cento T1/T2 e 1,3 por cento T2/T2 no Grupo Controle. Não houve diferenças entre os grupos, analisando-se as frequências genotípicas e alélicas: respectivamente p=0,4 (OR: 0,8) e p=0,6 (OR: 0,9). CONCLUSÕES: os dados do presente estudo sugerem que o polimorfismo PROGINS do gene dos receptores de progesterona não está relacionado ao AER.

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 percent 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 percent confidence intervals 95 percentCI. RESULTS: PROGINS genotypic frequencies were 72.3 percent T1T1 and 27.7 percent T1T2 for the RSA group and 764 percent T1T1, 22.3 percent T1T2 and 1.3 percent T2T2 for the control group. There were no differecnes 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.

CD46 polymorphism: A probable risk factor for recurrent spontaneous abortion in a northern Indian population.

Background. CD46 is a complement regulatory glycoprotein. Certain polymorphic forms of the CD46 gene have been associated with recurrent pregnancy loss in the Caucasian population. We assessed the role of CD46 polymorphism in recurrent spontaneous abortion in our setting, as this has not been done on Indian subjects till date. Methods. Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was carried out on 44 samples each from women with recurrent spontaneous abortion and normal pregnancy. Genotyping of the CD46 gene was done using 2.5% agarose gel. Statistical analysis was done using the TFPGA software. Results. The absence of CD46H*1 homozygosity was more pronounced in women with recurrent spontaneous abortion in the Indian population. Of recurrent aborters, 9% had the H*1/*1 genotype as compared to 30% of normal pregnant women. Conclusion. Although our data did not fit the Hardy– Weinberg equilibrium, this pilot study indicates that further increasing the sample size might clarify whether polymorphism in the first intron of the CD46 gene can be regarded as a risk factor for recurrent spontaneous abortion.

Polymorphisms in NOS3, ACE and PAI-1 genes and risk of spontaneous recurrent miscarriage in the Gaza Strip

This study was conducted to investigate the correlation between spontaneous recurrent miscarriage [RM] and common polymorphisms in angiotensin-converting enzyme [ACE], plasminogen activator inhibitor 1 [PAI-1] and endothelium-derived nitric oxide synthase 3 [NOS3] genes among women experiencing RM in the Gaza Strip. The presence of these genetic profiles was determined for 100 women who had had at least 3 constitutive abortions and 100 controls without any history of abortion using molecular biological techniques. The ACE D/D polymorphism was present in 49% of the study population and in 54% of the controls [p = 0.479]. Similarly, there was no significant difference detected in the distribution of polymorphisms for PAI-1, with the 4G/4G genotype present in the study group and in controls [p = 1.00]. NOS3 4a/4a was present in 4% of the study group and in none of the 100 controls [p = 0.123]. In this study, we also discovered a new variant in the NOS3 gene which was named 4c allele and was encountered in 1 patient and in 1 control subject. There was no significant association between ACEI/D, PAI-1 4G/5G and NOS3 4a/4b and the occurrence of first-trimester RM. In-depth investigation of the association of NOS3 4a/4a with RM is strongly recommended

Estudo citogenético de abortos espontâneos / Cytogenetical study on miscarriages

Os abortamentos espontâneos ocorrem por diversas causas, sendo as anomalias cromossômicas do concepto,as mais freqüentes. Assim, o estudo citogenético de seus produtos e genitores, seguido do aconselhamentogenético aos casais participantes, são condutas essenciais. O presente estudo retrospectivo teve comoobjetivo investigar os cariótipos de 574 amostras de produtos de abortamentos espontâneos, bem comosangue periférico de casais com abortamentos espontâneos recorrentes, para estimar a frequência de alteraçõescromossômicas. Os cariótipos foram previamente realizados a partir da cultura de vilosidade coriônica (abortos)e sangue periférico (casais), seguida da técnica de bandeamento G. Concluiu-se que as frequências observadasde alterações cromossômicas entre os abortos (19,69%) e os casais (7,6%) foram concordantes com a literatura,reforçando a importância da análise citogenética nesses casos.

Miscarriages result of several causes, but chromosomal anomalies are the most frequent. Thus, the cytogeneticstudies of its products and of the parents, followed by genetic counseling to involved couples, are crucialconducts. The present retrospective work aimed to investigate the karyotypes of 574 samples of miscarriageproducts, and the peripheral blood of couples involved with recurrent miscarriages in an attempt to estimatethe frequencies of chromosomal alterations. Karyotypes were previously made from the culture of chorionicvilli (miscarriages) and peripheral blood (couples), followed by the technique of G banding. It was concludedthat the observed frequency of abnormal karyotypes among miscarriage products (19.69%) and couples(7.6%) were both consistent with the literature, reinforcing the importance of cytogenetic study in thesecases.

Anormalidades cromossômicas em casais com história de aborto recorrente / Chromosomal abnormalities in couples with history of recurrent abortion

OBJETIVO: verificar a prevalência e as características clínicas de casais com história de abortos de repetição e anormalidade cromossômica atendidos em nosso serviço. MÉTODOS: foram avaliados retrospectivamente todos os casais encaminhados de janeiro de 1975 a junho de 2008 por história de abortos de repetição. Foram incluídos no estudo somente aqueles casais, em que a análise cromossômica feita com o cariótipo por bandas GTG foi realizada com sucesso. Foram coletados dados clínicos referentes às suas idades, bem como o número de abortamentos, natimortos, crianças polimalformadas, nativivos por casal e resultado do exame de cariótipo. Para comparação da frequência das alterações cromossômicas encontradas em nosso estudo com as da literatura, bem como entre os diferentes subgrupos de nossa amostra, foi utilizado o teste exato de Fisher (p<0,05). RESULTADOS: a amostra foi composta de 108 casais. As idades variaram de 21 a 58 anos entre os homens (média de 31,4 anos) e de 19 a 43 anos entre as mulheres (média de 29,9 anos). O número de abortos oscilou de dois a nove (média de 3,2). Alterações cromossômicas foram observadas em um dos parceiros em dez casais (9,3%) e corresponderam, respectivamente, a três casos (30%) de translocação recíproca [dois de t(5;6) e um de t(2;13)], dois (20%) de translocação Robertsoniana [um de der(13;14) e um de der(13;15)], cinco (50%) de mosaicismo (mos) [dois casos de mos 45,X/46,XX, um de mos 46,XX/47,XXX, um de mos 46,XY/47,XXY e um de mos 46,XY/47,XYY] e um (10%) de inversão cromossômica [inv(10)]. Em um dos casais, a parceira apresentava duas alterações concomitantes: t(2;13) e der(13;14). Anormalidades cromossômicas foram verificadas em 5% dos casais com história de dois abortamentos, em 10,3% com três abortos e 14,3% com quatro ou mais abortos...

PURPOSE: to asses the prevalence and clinical characteristics of couples with history of recurrent spontaneous abortion and chromosome abnormality, attended at the present service. METHODS: all the couples referred to our service due to history of recurrent spontaneous abortion, from January 1975 to June 2008, were evaluated. Only the ones whose chromosome karyotype analysis by GTG bands has been successfully made were included in the study. Clinical data on their age, as well as on the number of abortions, stillbirth, multiple malformations, livebirth per couple, and the result of the karyotype exam were collected. Fisher’s exact test (p<0.05) has been used to compare the incidence of chromosome alterations found in our study, with data in the literature. RESULTS: there were 108 couples in the sample. Their ages varied from 21 to 58 years old among the men (average of 31.4 years old), and from 19 to 43 among the women (average of 29.9 years old). In ten couples, one of the mates (9.3%) presented chromosome alterations, which corresponded respectively to three cases (30%) of reciprocal translocation [two of t(5;6) and one of t(2;13)], two (20%) of Robertsonian translocation [two of der(13;14) and one of der(13;15)], five(50%) of mosaicism (mos) [two cases of mos 45,X/46,XX, one of mos 46,XX/47,XXX, one of mos 46,XY/47,XXY and one of mos 46,XY/47,XYY] and one (10%) of chromosome inversion [inv(10)]. In one of the couples, the female presented two concomitant alterations: t(2;13) and der(13;14). Chromosome abnormalities were found in 5% of the couples with a history of two abortions, in 10.3% with three abortions, and in 14.3% with four or more abortions. CONCLUSIONS: the incidence of chromosome abnormalities seen in our study (9.3%) was similar to most of the studies carried out in the last 20 years, varying from 4.8 to 10.8%...

Abortamento habitual - II Fatores cromossômicos e anatômicos / Habitual abortion - II Cromossomic and anatomic factors

Embora fatores cromossômicos parentais tenham sido sugeridos como responsáveis por pequena proporção de abortamentos habituais, seu papel como causa das perdas ainda não foi convincentemente comprovado. Há evidências indiretas de que algumas mulheres teriam predisposição à não disjunção cromossômica durante a meiose, que resultaria em trissomias recorrentes nos embriões. As evidências que associam os abortamentos habituais às translocações ou inversões cromossômicas e aos mosaicismos são de baixa qualidade e controversas. Em relação ao distúrbios anatômicos, as abortadoras habituais com história típica de incompetência istmo-cervical, com mioma submucoso, sinéquias intra-uterinas ou septos não têm sido incluídas em ensaios clínicos com grupo-controle sem tratamento. Isso se deve provavelmente a um conceito empírico, fortemente estabelecido entre clínicos e pesquisadores, de que essas doenças são causas de abortamento habitual de tratamento obrigatório. Não há evidências que justifiquem a realização de metroplastias laparotômicas em pacientes com útero didelfo ou bicorno, embora algumas possam se beneficiar da cerclagem uterina

Although parental chromosomal factors have been suggested to play a role in a small proportion of habitual abortion, they have not been convincingly imputed as the cause of miscarriage. There are indirect evidences that some women are prone to chromosomal non-disjunction during meiosis yielding recurrent trisomies in embryos. The evidences associating habitual abortion to chromosomal translocations or inversions and to mosaicism are controversial and bad quality. Concerning anatomical disturbs, habitual aborters with a typical history of cervical incompetence, with submucous myomas, intra-uterine adhesions or septum have not been included in controlled trials with a no-treatment control group. This is probably due to a conception strongly established among clinicians and researchers that these are causes of habitual abortion of mandatory treatment. There is no evidence to justify laparotomic metroplasty in didelphic of bicornuate uterus, although some women could benefit from uterine cerclagem

No evidence of association of MUC-1 genetic polymorphism with embryo implantation failure

Pregnancy loss can be caused by several factors involved in human reproduction. Although up to 50 percent of cases remain unexplained, it has been postulated that the major cause of failed pregnancy is an error of embryo implantation. Transmembrane mucin-1 (MUC-1) is a glycoprotein expressed on the endometrial cell surface which acts as a barrier to implantation. The gene that codes for this molecule is composed of a polymorphic tandem repeat of 60 nucleotides. Our objective was to determine if MUC-1 genetic polymorphism is associated with implantation failure in patients with a history of recurrent abortion. The study was conducted on 10 women aged 25 to 35 years with no history of successful pregnancy and with a diagnosis of infertility. The control group consisted of 32 patients aged 25 to 35 years who had delivered at least two full-term live children and who had no history of abortions or fetal losses. MUC-1 amplicons were obtained by PCR and observed on agarose and polyacrylamide gel after electrophoresis. Statistical analysis showed no significant difference in the number of MUC-1 variable number of tandem repeats between these groups (P > 0.05). Our results suggest that there is no effect of the polymorphic MUC-1 sequence on the implantation failure. However, the data do not exclude MUC-1 relevance during embryo implantation. The process is related to several associated factors such as the mechanisms of gene expression in the uterus, specific MUC-1 post-translational modifications and appropriate interactions with other molecules during embryo implantation.