Risk Factor For Recurrent Pregnancy Loss Research Articles

Oxidative/nitrosative stress increased the risk of recurrent pregnancy loss–Taiwan Recurrent Pregnancy Loss and Environmental Study (TREPLES)

ObjectiveOxidative stress biomarkers (OSBs) may be strongly associated with disease progression and recurrent pregnancy loss (RPL). However, the research on associations of most OSBs (e.g., 8-nitroguanine [8-NO2Gua] and 4-hydroxy-2-nonenal-mercapturic acid [HNE-MA]) with RPL is limited. Therefore, we aimed to investigate the effect of OSBs exposure on RPL risk by performing a case-control study. Material and methodsWe use our established dataset, Taiwan Recurrent Pregnancy Loss and Environmental Study (TREPLES), which included 514 Taiwanese reproductive age women (aged 20–50 years; 397 cases and 117 controls) from National Cheng Kung University Hospital. RPL is clinically defined by a history of two or more consecutive miscarriages, where a miscarriage is defined as the termination of pregnancy before 20 weeks of gestation. The urinary levels of several OSBs (e.g., 8-hydroxy-2′-deoxyguanosine [8-OHdG], 8-NO2Gua, 8-isoprostaglandin F2α [8-isoPGF2α], and HNE-MA) and malondialdehyde (MDA) were measured using isotope dilution liquid chromatography-tandem mass spectrometry and thiobarbituric acid reactive substances, respectively. ResultsThe median levels of 8-NO2Gua (6.15 vs. 3.76 ng/mL) and HNE-MA (30.12 and 21.54 ng/mL) were significantly higher in the RPL group than in the control group. By categorizing the OSBs data into tertiles, after we adjusted for age and urine creatinine levels discovered that the RPL risk associated with 8-NO2Gua and HNE-MA levels in the third tertile were approximately 2 times higher than those in the first tertile (8-NO2Gua, adjusted OR = 3.27, 95 % CI = 1.66–6.43; HNE-MA, adjusted OR = 1.96, 95 % CI = 1.05–3.64; p < 0.05). These findings suggest that the oxidative stress biomarkers of 8-NO2Gua and HNE-MA are risk factors for RPL. ConclusionOur findings indicate that specific OSBs are associated with an increased RPL risk, suggesting that reducing OSB levels can improve RPL risk. Nevertheless, more studies on preventive medicine are required to understand the exposure sources and adverse outcome pathways of OSBs associated with RPL.

O-146 Oocyte donation does not increase live birth rates in young women suffering from recurrent pregnancy loss

Abstract Study question Does oocyte donation improve reproductive outcomes in recurrent pregnancy loss (RPL) patients? Summary answer Oocyte donation increases live birth and reduces miscarriage rates in RPL women older than 35, but it does not improve reproductive outcomes in younger patients. What is known already Recurrent pregnancy loss (RPL) is defined as 2 or more pregnancy losses prior to 20-24 weeks of gestation and has an incidence of approximately 1 to 2% in couples trying to conceive. Known risk factors for RPL are embryo aneuploidy, advanced maternal age (AMA), previous miscarriages, uterine abnormalities, parental chromosomal abnormalities, antiphospholipid syndrome, endocrine factors, such as thyroid function or obesity, and thrombophilia. However, 50-70% of patients suffering from RPL do not present any known risk factor, making the management of idiopathic RPL patients particularly challenging for ART clinicians. Study design, size, duration This is a cohort retrospective study involving 2 centers. Charts from 18,273 women undergoing IVF treatment between 2011 and 2020 were reviewed. A total of 912 patients (5%) met the definition of RPL and were classified into study groups based on their age and oocyte origin: ≤35 years old receiving oocyte donation (n = 39) or using their own oocytes (n = 35), and AMA women using donor eggs (n = 716) or their own (n = 122). Participants/materials, setting, methods Demographic variables analysed included known risk factors, number of transferred embryos, day of embryo transfer (3 vs 5) and sperm origin (partner/donor). Differences in biochemical, clinical, ongoing pregnancy, miscarriage and live birth rates were assessed by Pearson’s Chi-squared or Fisher’s exact test. P-values &amp;lt;0.05 were considered significant. Main results and the role of chance RPL patients had 2.77±1.27 pregnancy losses overall (2.76±1.37 in women ≤35 and 2.77±1.26 in patients &amp;gt;35). Most RPL patients (91.9%) were of AMA. Other RPL-associated risk factors, including uterine malformations, previous pregnancy losses, chromosomal abnormalities and thrombophilia were identified in 207/912 patients (22.7%). Interestingly, these were more frequent in patients &amp;lt;35 (37.8% vs 21%, p = 0.001). Young RPL women presented higher rates of karyotype abnormalities than older patients (17.6% vs 3.8%, p &amp;lt; 1x10-04), while showing a similar incidence of uterine abnormalities (14.9% vs 12.9%, p &amp;gt; 0.63) and thrombophilia (4.1% vs 4.7%, p &amp;gt; 0.81). RPL patients &amp;gt;35 preferentially underwent oocyte donation (85.4% vs 52.7%, p &amp;lt; 1x10-04), which led to significantly higher biochemical (52.5% vs 17.1%, p &amp;lt; 1x10-04), clinical (42.4% vs. 8.5% p &amp;lt; 1x10-04), ongoing pregnancy (37.9% vs 5.1%, p &amp;lt; 1x10-04), live birth (32% vs 4%, p &amp;lt; 1x10-04) and lower miscarriage rates (10.5% vs 40%, p = 0.0186) than in autologous cycles. In contrast, RPL patients &amp;lt;35 had similar reproductive outcomes: biochemical (41% vs. 52.9), clinical (28.2% vs. 37.1%), ongoing pregnancy (25.6% vs 29.4%), live birth (25.6% vs 26.4%) and miscarriage rates (9% vs 23%), regardless of oocyte origin (donated vs own, p &amp;gt; 0.05 for all cases). Importantly, this was also true for a small subgroup of idiopathic young RPL patients (n = 13). Limitations, reasons for caution The main limitation of this study is its retrospective nature, which does not allow for full elucidation of all potential confounders. The number of young RPL patients undergoing oocyte donation may be too low to draw significant conclusions. Wider implications of the findings RPL can be resolved in AMA patients by using donor oocytes, which points to key roles of oocyte quality and aneuploidy underlying RPL aetiology. However, RPL is not ameliorated by oocyte donation in young women, suggesting an endometrial/systemic origin and highlighting the need to further study mechanisms driving this disorder. Trial registration number Not applicable

Evaluation of platelet parameters, coagulation markers, antiphospholipid syndrome, and thyroid function in palestinian women with recurrent pregnancy loss

BackgroundMultiple etiologies contribute to recurrent pregnancy loss (RPL) including immunological, endocrine, anatomical, genetic and infection but more than 50% of cases remain unexplained. Evidences of thrombotic and inflammatory processes were observed at maternal-fetal interface and considered pathological findings in most RPL cases including unexplained cases. This study aimed to evaluate the association between RPL and several risk factors: platelet parameters, coagulation factors, antiphospholipid syndrome, and thyroid function.MethodsThis is an unmatched case-control study that included 100 RPL and 100 control women. Anthropometric and health data were collected and a gynecologist examined participants to assure fitting the inclusion criteria. Platelet parameters [including Mean Platelet Mass (MPM), Concentration (MPC) and Volume (MPV)] and ratios (MPV/Platelet, MPC/Platelet, MPM/Platelet, Platelet/Mononuclear cells), coagulation markers [Protein C (PC), Protein S (PS), Antithrombin III, D-dimer], antiphospholipid antibodies [Anti-phospholipid (APA), Anti-cardiolipin (ACA) and anti-B2-glycoprotein 1], Lupus anticoagulant, Antinuclear antibodies, and thyroid function (Thyroid stimulating hormone and anti-thyroid peroxidase) were measured.ResultsMean ages of cases and controls at marriage were 22.5 years for both, and their current ages were 29.4 and 33.0, respectively. 92% of cases and 99% of controls aged blow 30 years at marriage. 75% of cases have 3–4 miscarriages and 9% have ≥ 7 miscarriages. Our results indicated significantly lower male/female age ratio (p = .019), PC (p = .036) and PS (p = .025) in cases compared to controls. Plasma D-dimer (p = .020) and antiphospholipid antibodies [ACA (IgM and IgG), APA (IgM)] were significantly higher in cases compared to controls. No significant differences were observed between cases and controls concerning APA (IgG), anti-B2-glycoprotein 1 (IgM and IgG), Lupus anticoagulant, Antinuclear antibodies, platelet parameters, thyroid markers, family history of miscarriage, consanguineous marriage, and other health data.ConclusionsThis is the first study that investigated the association between platelet, coagulation, antiphospholipid, autoimmune and thyroid parameters, and RPL in Palestinian women. Significant associations between male/female age ratio, PC, PS, D-dimer, ACA (IgM, IgG), APA (IgM) and RPL were observed. These markers could be used in evaluating RPL. These findings confirm the heterogeneous nature of RPL and emphasize the need for further studies to find out risk factors for RPL.

Understanding recurrent pregnancy loss: recent advances on its etiology, clinical diagnosis, and management.

Recurrent pregnancy loss (RPL) has become an important reproductive health issue worldwide. RPL affects about 2%-3% of reproductive-aged women, and makes serious threats to women's physical and mental health. However, the etiology of approximately 50% of RPL cases remains unknown (unexplained RPL), which poses a big challenge for clinical management of these patients. RPL has been widely regarded as a complex disease where its etiology has been attributed to numerous factors. Heretofore, various risk factors for RPL have been identified, such as maternal ages, genetic factors, anatomical structural abnormalities, endocrine dysfunction, prethrombotic state, immunological factors, and infection. More importantly, development and applications of next generation sequencing technology have significantly expanded opportunities to discover chromosomal aberrations and single gene variants responsible for RPL, which provides new insight into itspathogenic mechanisms. Furthermore, based upon patients' diagnostic evaluation and etiologic diagnosis, specific therapeutic recommendations have been established. This review will highlight current understanding and recent advances on RPL, with a special focus on the immunological and genetic etiologies, clinical diagnosis and therapeutic management.

O-303 Recurrent pregnancy loss is associated with unfavorable composition of endometrial and vaginal microbiota

Abstract Study question Does endometrial or vaginal microbiota composition associate with recurrent pregnancy loss (RPL)? Summary answer Reduced relative abundance of Lactobacillus crispatus in endometrium and increased relative abundance of Gardnerella vaginalis in endometrium and vagina were associated with RPL. What is known already Dysbiotic vaginal microbiota associates with sporadic miscarriage and the depletion of lactobacilli in the endometrium has been associated with poor IVF outcomes, but their association with RPL has not been studied. Study design, size, duration A nested case-control study of 47 women investigated for RPL and 39 healthy control women without a history of pregnancy loss were recruited between March 2018 and December 2020 at a university hospital. Endometrial biopsies and vaginal samples were collected from the women 6-8 days after positive ovulation test result. Participants/materials, setting, methods The endometrial and vaginal microbiota compositions, analysed using 16S rRNA gene amplicon sequencing, were compared between RPL women and controls, and between individual vaginal and endometrial samples. The mycobiota composition was analysed using internal transcribed spacer 1 amplicon sequencing for a descriptive summary. The models were adjusted for BMI, age, and parity. False discovery rate -corrected P-values (q-values) were used to define nominal statistical significance at q&amp;lt; 0.05. Main results and the role of chance Lactobacillus crispatus was less abundant in the RPL women’s endometrial samples compared to controls (mean relative abundance 17.2% vs. 45.6%, q= 0.04). Gardnerella vaginalis was more abundant in RPL women than in controls in both endometrial (12.4% vs. 5.8%, q&amp;lt; 0.001) and vaginal samples (8.7% vs. 5.7%, q&amp;lt; 0.01). The individual vaginal and endometrial microbial compositions correlated strongly (R = 0.85, P&amp;lt; 0.001). Fungi, mostly Candida albicans, were detected in 22% of the endometrial and 36% of the vaginal samples. Limitations, reasons for caution Limitations include the transcervical sampling technique with a potential risk of cervicovaginal contamination of endometrial samples, the loss of a proportion of the samples from statistical analyses due to their low read count, and the presentation of the 16S rRNA sequencing results in relative rather than absolute abundances Wider implications of the findings Our findings suggest that an unfavorable endometrial microbiota may be a novel risk factor for RPL. Further studies should elucidate the mechanisms, how microbiota may predispose to RPL, possibilities to modify microbiota by treatments, and the effects of such treatments on the success of future pregnancies in couples having RPL. Trial registration number not applicable

P–425 Recurrent pregnancy loss: what can we learn from different international guidelines?

Abstract Study question To which extent do the current international guidelines and recommendations concerning recurrent pregnancy loss (RPL) differ? Summary answer All guidelines apply definitions for RPL, however few diagnostic and therapeutic options are described. Diagnostics should be based on best evidence and current scientific knowledge. What is known already Established risk factors for RPL include anatomical, genetic, endocrine, hemostatic and immune alterations. The European Society of Reproduction and Embryology (ESHRE), American Society of Reproductive Medicine (ASRM), German/Austrian/Swiss Society of Obstetrics and Gynecology (DGGG/OEGGG/SGGG) and the Royal College of Obstetricians and Gynecologists (RCOG) published guidelines concerning diagnostic and therapeutic options in RPL. Due to the different guideline processes and date of publication actuality as well as complexity differ widely. Study design, size, duration We compared the guidelines of the ESHRE, ASRM, DGGG/OEGGG/SGGG and RCOG with regard to definition, diagnostic and therapeutic aspects. The guidelines were published between 2011 and 2018. Structured guideline processes with regular (complete) updates are only provided by the DGGG/OEGGG/SGGG. Participants/materials, setting, methods After thorough literature research (Pubmed, Embase) all existing guidelines and recommendations were analysed and compared considering the current state of knowledge. The RCOG recommendations from 2011 were updated in 2014 and 2017, the ARSM expert letter was last updated in 2012. The ESHRE guideline was published in 2017. The first version of the DGGG/OEGGG/SGGG guideline was published 2006, updated in 2013 and upgraded to a higher evidence-level in 2018 and is currently under review. Main results and the role of chance All guidelines agree that a diagnostic work-up is indicated after at least two clinical pregnancies and should exclude anatomical malformations, an antiphospholipid syndrome and thyroid dysfunction. Furthermore, lifestyle modifications are recommended by all. The general evaluation of an inherited thrombophilia is not recommended by any guideline. Exclusion of other risk factors like parental chromosomal disorders, a polycystic ovary syndrome or insulin resistance are only included in some guidelines, partly due to a lack of diagnostic criteria (luteal phase insufficiency) or due to the different year of publication of the recommendations (e.g. chronic endometritis). All guidelines recommend treating APLS by administering low-dose aspirin (75–100mg daily) in combination with unfractionated/low-molecular-weight heparin. With regard to uterine malformations whether or not a septum should be dissected is still a matter of debate: ESHRE and RCOG consider evidence insufficient, while DGGG/OEGGG/SGGG and ASRM recommend a surgical intervention. In case of chronic endometritis, the DGGG/OEGGG/SGGG recommends antibiotic therapy e.g. with doxycycline (200 mg daily for 14 days). Limitations, reasons for caution Different health economic as well as consensus aspects in the process of guideline development have a significant influence on the individual guidelines and recommendations. Wider implications of the findings: Since personalized diagnostic and therapeutic strategies in RPL patients are required, physicians have to decide when to follow the guideline and when to expand diagnostics and therapy. Therefore, the knowledge of the weaknesses of each guideline and its developmental process is helpful for treating RPL couples. Trial registration number -

GROUP B STREPTOCOCCAL INFECTION AND RPL – A CASE REPORT

Recurrent pregnancy loss (RPL) has become an important cause of pregnancy loss, with major emotional implications to the couple experiencing such an event. Increasing age of women, smoking, obesity or polycystic ovary syndrome (PCOS) and a previous history of miscarriage and various infections are also considered risk factors for RPL. A thorough clinical history and examination, maternal serum biochemistry and ultrasound ndings are important to determine the treatment options and provide valuable information for the prognosis. A woman who do not have a explanation for the RPL maybe subjected to ano-genital swab culture to identify infections, as chronic and subclinical infections can cause abortion. As bacterial vaginosis has been already established with causing recurrent pregnancy loss and preterm delivery. Here we are presenting a case of successful pregnancy outcome following treatment for GBS vaginal infection where other possible aetiologies were ruled out.

Women with Recurrent Pregnancy Loss More Often Have an Older Brother and a Previous Birth of a Boy: Is Male Microchimerism a Risk Factor?

Known etiologic factors can only be found in about 50% of patients with recurrent pregnancy loss (RPL). We hypothesized that male microchimerism is a risk factor for RPL and aimed to explore whether information on family tree and reproductive history, obtained from 383 patients with unexplained RPL, was supportive of this hypothesis. The male:female sex ratio of older siblings was 1.49 (97:65) in all RPL patients and 1.79 (52:29) in secondary RPL (sRPL) patients, which differed significantly from the expected 1.04 ratio (p = 0.027 and p = 0.019, respectively). In contrast, the sex ratio of younger siblings was close to the expected ratio. Sex ratio of the firstborn child before sRPL was 1.51 (p = 0.026). When combined, 79.1% of sRPL patients had at least one older brother, a firstborn boy, or both. This differed significantly from what we expected based on the distribution of younger siblings and a general 1.04 sex ratio of newborns (p = 0.040). We speculate whether (s)RPL patients possibly acquired male microchimerism from older brother(s) and/or previous birth of boy(s) by transplacental cell trafficking. This could potentially have a detrimental impact on their immune system, causing a harmful response against the fetus or trophoblast, resulting in RPL.

Neutrophil and Neutrophil-to-Lymphocyte Ratio as Clinically Predictive Risk Markers for Recurrent Pregnancy Loss.

We aimed at exploring the value of neutrophil and neutrophil-to-lymphocyte ratio (NLR) as predictive risk markers for recurrent pregnancy loss (RPL) by conducting a retrospective case-control study and a systematic review and meta-analysis. This retrospective case-control study was conducted in an academic hospital from January 2012 to June 2018 and 133 cases of women with RPL and 140 control women. In parallel, we conducted a systematic review and meta-analysis on the value of NLR and neutrophil in RPL risk assessment. Neutrophil and NLR were higher in the women with RPL than those in the controls (median [interquartile range]: 4.32 [3.31-6.18] vs. 3.76 [2.78-4.81], p = 0.001; 2.17 [1.47-3.16] vs. 1.74 [1.40-2.34], p = 0.001). After the cut-off value of the neutrophil and NLR was determined to be 6.3 × 109/L and 3.16, the prevalence of RPL became significantly increased in the high-neutrophil and the high-NLR group compared to the low-neutrophil and the low-NLR group (76.3% vs. 44.2%, p < 0.001; 66.6% vs. 44.5%, p < 0.001). Univariate logistic regression analysis indicated that high-neutrophil and high-NLR were risk factors for RPL, with ORs of 4.06 (95% CI: 1.84-8.95) and 2.49 (95% CI: 1.31-4.71), respectively. Multivariate logistic regression analysis indicated that high-neutrophil was a risk factor for RPL, with OR of 4.91 (95% CI: 1.66-14.50). Meta-analysis of 5 case-control studies (including the present study) indicated that increased neutrophil and NLR may be risk factors for RPL, with SMDs of 0.63 (95% CI: 0.45-0.80) and 0.61 (95% CI: 0.39-0.83). Based on current findings and the meta-analysis, this study indicated that neutrophil and NLR might be valuable for predicting RPL; more studies with large sample are needed to verify this conclusion. The mechanism of the association between neutrophil and RPL needs to be explored in further studies.

Antinuclear Antibody Positivity As a Risk Factor for Recurrent Pregnancy Loss: A Meta-Analysis

Background: Immunologic mechanisms have been one of the suggested causes of recurrent pregnancy loss (RPL). Antinuclear antibody positivity has been regarded as a typical feature of autoimmunity. Over past years many studies tried to clarify the association of ANA with RPL and the conclusions are controversial. The aim of this meta-analysis was to assess whether the presence of ANA directly increase the risk of RPL in women who did not have definite autoimmune diseases. Methods: We searched PubMed and Embase databases for relevant literatures, which included the data of ANA rate in RPL and control groups. Both fixed-effects and randomized-effects model were used to analyze the risk odds ratio with 95% confidence intervals (CIs) according to the heterogeneity across the selected studies. Findings: Twenty-one studies with 5038 participants including 2683 patients with RPL met the inclusion and exclusion criteria. The total positive rate of ANA was 22·4% (601/2683) in total RPL patients and 20·2% (397/1959) in unexplained RPL patients, respectively. Patients with total RPL or unexplained RPL had a significantly higher ANA positive rate than control groups [(OR =3·12, 95% CI:1·96, 4·97, I² = 77%), (OR = 3·47, 95% CI: 2·07, 5·81), I² = 74%], respectively]. Subgroup analysis demonstrated low titers of antibody (≤1: 80) were not associated with RPL patients [OR = 1·22, 95% CI (0·79, 1·90), I2 =86·1%], while higher ANA titer (≥1:160) showed a significant association between ANA and RPL patients [OR = 23·23, 95% CI (4·18,129·25, I 2 =0%)]. Higher rate of homogenous pattern in RPL group was observed [OR = 4·54, 95% CI (2·01, 10·24), I2=0%], and no significant difference in speckled pattern [OR = 1·08, 95% CI (0·69, 1·70), I 2 =0%] nor nucleolar pattern [OR = 3·52, 95% CI (0·93, 13·30), I 2 =0%] was found. Three studies detected the anti-dsDNA in two groups and the results demonstrated no significant difference between RPL group and controls [OR = 1·35, 95% CI (0·52, 3·54), I2=0%]. Interpretation: This is the first meta-analysis about the association between ANA positivity and RPL. The pooled data demonstrated increased prevalence of ANA in patients with RPL. ANA is suggested as a risk factor for RPL and should be recommended for this population. Funding Statement: The work was supported by Natural Science Foundation of China (NSFC) Grant U1605223 to Dr. Guixiu Shi, NSFC grant 81501407 to Dr. Yuan Liu; NSFC grant 81501369 to Dr. Shiju Chen; and the Leadership Program of the Technology Department of Fujian Province (Grant NO. 2015D011) to Dr. Guixiu Shi. Declaration of Interests: The authors declare no conflicts of interests. Ethics Approval Statement: The meta-analysis was performed in accordance with the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines.

Association of Angiotensin-Converting Enzyme Insertion/Deletion Polymorphism with Recurrent Pregnancy Loss: a Meta-Analysis of 26 Case-Control Studies.

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. 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. A total of 26 case-control studies with 3,140 RPL cases and 3,370 controls were included in the meta-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. The meta-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.

The association of IL-33 and Foxp3 gene polymorphisms with recurrent pregnancy loss in Egyptian women

Deregulated immunity is one of the most important factors implicated in recurrent pregnancy loss (RPL). The possible role of interleukin-33 (IL-33) and forkhead/winged helix transcription factor (Foxp3) in RPL have not been fully investigated. We aimed to evaluate IL-33 rs1929992 and Foxp3 rs2232365 single nucleotide polymorphisms (SNPs) and their serum levels in Egyptian RPL females.Blood samples were collected from 142 RPL patients and 123 women as healthy controls. IL-33 rs1929992 SNP was determined by polymerase chain reaction restriction fragment length polymorphism and Foxp3 rs2232365 SNP was determined using allele specific polymerase chain reaction. The serum IL-33 and Foxp3 levels were measured by enzyme linked immunosorbent assay.Foxp3 rs2232365 SNP showed statistically significant association with RPL. The risk of RPL was significantly higher in women carrying Foxp3 G allele than those carrying A allele. Lower serum levels of Foxp3 and IL-33 were observed in RPL patients than controls (P < 0.001). Foxp3 serum levels were much lower in carriers of G allele than those carrying A allele in all studied groups.Foxp3 rs2232365 SNP could be considered as a risk factor for RPL. The lowered serum levels of IL-33 and Foxp3 in RPL patients suggested that they might have an important role in the pathogenesis of the disease. Therefore, we hypothesized that Foxp3 polymorphisms may be important in RPL pathogenesis.

Association of genetic variants of PD1 with recurrent pregnancy loss.

PurposeProgrammed cell death 1 (PD1) and PD ligand 1 (PDL1) are speculated to have an important role in maintaining a normal pregnancy and there are also a few reports of an association between the single‐nucleotide polymorphisms (SNPs) of PD1 or PDL1 and diseases in humans. The association was examined between the polymorphisms of PD1,PDL1,CTLA4 and recurrent pregnancy loss (RPL).MethodsThe study group consisted of 243 women with two or more pregnancy losses and 176 parous women. The frequency of three SNPs of PD1, two of PDL1, and four of CTLA4 were compared between the patients and controls. Next, the subsequent live birth rates in the patients with RPL with and without risk alleles were examined.ResultsTwo SNPs (rs36084323 and rs3481962) of PD1 were found to occur at significantly higher frequencies in the patient group than in the control group. The subsequent live birth rate of the patients with vs. without the risk alleles of rs36084323 and rs3481962 were 83.3% vs. 83.3% and 81.6% vs. 84.0%, respectively.ConclusionVariations of the PD1 gene were identified as risk factors for RPL. However, the presence or absence of the PD1 risk alleles had no predictive effect on the subsequent pregnancy outcome.

Single-nucleotide polymorphism of interleukin-27 gene: A risk factor of recurrent pregnancy loss in Iraqi women

Background: Studies have been done to investigate the association between a single-nucleotide polymorphism (SNP) of interleukin-27 (IL-27) gene and the recurrent pregnancy loss (RPL). However, different results have been found in different spots of the world. Therefore, more studies are needed to understand the variation in these results. This is the first study that shows the implication of the SNP of IL-27 gene in RPL. Objective: This study aims to investigate the association between RPL and SNP of gene (−964 A > G) in Iraqi women. Materials and Methods: From September 2013 to September 2014; 100 women, as a control group, and 100 women (with three or more consecutive pregnancy loss), as a study group, were recruited to investigate the association between the IL-27(−964 A > G) SNP and the PRL. The IL-27(−964 A > G) SNP was determined using polymerase chain reaction-restriction fragment length polymorphism technique. Genotype and allele frequencies were compared using Fisher test between the two groups. P G polymorphism was significantly different between the study and the control groups (P = 0.007). The allele frequencies of this polymorphism were A (35%), G (65%) in the RPL group versus A (61%), G (39%) in the control group. The frequencies of A and G alleles in the both groups were not significantly different. Conclusion: IL-27 (−964 A > G) polymorphism is a risk factor for RPL in a sample of Iraqi women. However, this is different from what has been found in some studies which might implicate other factors in the RPL.

Thrombophilic gene polymorphisms and recurrent pregnancy loss in Greek women.

Recurrent pregnancy loss (RPL) is a multifactorial disorder. The aim of this study was the detection of various genetic polymorphisms and their correlation to RPL, in Greek women. The impact of 12 thrombophilic polymorphisms was evaluated, among 48 Greek women with a history of RPL, vs 27 healthy parous women. Multiplex PCR and in situ hybridization on nitrocellulose films were performed, to investigate 12 genetic polymorphisms previously reported as risk factors for RPL. Heterozygous FV Leiden, homozygous PAI-1 4G/4G, heterozygous MTHFR C677T, homozygous MTHFR A1298C, as much as the combined thrombophilic genotypes MTHFR 677T + ACE Ι/D, MTHFR 677T/1298C + ACE D/D, ACE I/D + b-fibrinogen -455G/A, FV HR2 + b-fibrinogen -455G/A showed a correlation as risk factors for RPL, whereas the rest of the investigated polymorphisms and their combinations did not render statistically significant differences between the two groups in study. The results of this study, as well as those of similar studies, concerning the detection of genetic, environmental, and physiological factors underlying RPL, will prove of critical significance in the investigation and treatment of thrombophilic predisposition, in cases of RPL.

Copy number variation profile in the placental and parental genomes of recurrent pregnancy loss families

We have previously shown an extensive load of somatic copy number variations (CNVs) in the human placental genome with the highest fraction detected in normal term pregnancies. Hereby, we hypothesized that insufficient promotion of CNVs may impair placental development and lead to recurrent pregnancy loss (RPL). RPL affects ~3% of couples aiming at childbirth and idiopathic RPL represents ~50% of cases. We analysed placental and parental CNV profiles of idiopathic RPL trios (mother-father-placenta) and duos (mother-placenta). Consistent with the hypothesis, the placental genomes of RPL cases exhibited 2-fold less CNVs compared to uncomplicated 1st trimester pregnancies (P = 0.02). This difference mainly arose from lower number of duplications. Overall, 1st trimester control placentas shared only 5.3% of identified CNV regions with RPL cases, whereas the respective fraction with term placentas was 35.1% (P = 1.1 × 10−9). Disruption of the genes NUP98 (embryonic stem cell development) and MTRR (folate metabolism) was detected exclusively in RPL placentas, potentially indicative to novel loci implicated in RPL. Interestingly, genes with higher overall expression were prone to deletions (>3-fold higher median expression compared to genes unaffected by CNVs, P = 6.69 × 10−20). Additionally, large pericentromeric and subtelomeric CNVs in parental genomes emerged as a risk factor for RPL.

Genotyping analysis for the 46 C/T polymorphism of coagulation factor XII and the involvement of factor XII activity in patients with recurrent pregnancy loss.

BackgroundEstablished causes of recurrent pregnancy loss (RPL) include antiphospholipid syndrome, uterine anomalies, parental chromosomal abnormalities, particularly translocations and abnormal embryonic karyotype. A systematic review concluded that coagulation factor XII (FXII) deficiency was associated with RPL. However, it could not be established whether the 46 C/T SNP of FXII or low activity of FXII was a risk factor for RPL, because of the small sample size.Methods and FindingsWe conducted a cross-sectional and cohort study in 279 patients with two or more unexplained consecutive pregnancy losses and 100 fertile women. The association between the lupus anticoagulant (LA) activity and FXII activity was examined. The frequency of the CC, CT and TT genotypes and the FXII activity were also compared between the patients and controls. Subsequent miscarriage rates among the CC, CT, TT genotypes and according to the FXII activity was examined. LA was associated with reduced FXII activity. The CT, but not the TT, genotype was confirmed to be a risk factor for RPL in the cross-sectional study using multivariate logistic regression analysis (OR, 2.8; 95% CI, 1.37–5.85). The plasma FXII activity in the patients was similar to that in the controls. Neither low FXII activity nor the CT genotype predicted the subsequent pregnancy outcome in the cohort study. On the other hand, and intermediate FXII activity level of 85–101% was predictive of subsequent miscarriage.ConclusionsLow FXII activity was not associated with RPL. The FXII gene was found to be one of the significant susceptibility genes for RPL, similar to the FV Leiden mutation. However, the clinical influence of the CT genotype might be relatively small, because the presence/absence of this genotype did not have any predictive value for the subsequent pregnancy outcome. This was the first study indicating the influence of FXII 46C/T on further pregnancy outcomes.

Interaction between TP63 and MDM2 genes and the risk of recurrent pregnancy loss

ObjectiveRecent studies have investigated the role of the p53 gene family in reproductive processes. Each member of the gene family acts through different mechanisms: p53 is involved in genomic stability and regulation of blastocyst implantation; p63 acts as a regulator of the quality and maturation of oocytes; and p73 controls the meiotic spindle. Polymorphisms in the genes of the p53 family have been associated with female infertility. One polymorphism in MDM2, the main regulator of the p53 family, has also been associated with this condition. Although polymorphisms in the TP53 gene have been related to recurrent pregnancy loss (RPL), there have been no studies associating polymorphisms in p63 and p73 with RPL. Therefore, the aim of this study was to evaluate the role of polymorphisms in the TP63 (rs17506395), TP73 (rs2273953, rs1801173), and MDM2 (SNP309, rs2279744) genes as risk factors for RPL. Study designA case–control study was conducted in 153 women with RPL and 143 fertile women with at least two living children and no history of pregnancy loss. Molecular analysis was performed by TaqMan Allelic Discrimination assay. The statistical analysis was performed using SPSS software version 20.0 and the chi-square test, Student's t-test, Mann–Whitney test and logistic regression to compare the evaluated characteristics between both groups and RPL outcome. ResultsThe allelic and genotypic frequencies did not differ between the groups when analyzed separately, however, the interaction between the TP63 TT and MDM2 TT genotypes was shown to increase the risk of RPL (OR=2.19, CI 95%: 1.28–3.75, p=0.004), even when adjusted for alcohol consumption, smoking, number of pregnancies and ethnicity (OR=1.97, CI 95%: 1.27–3.58, p=0.025). ConclusionsOur results suggest that genes from the p53 family proteins, evaluated here, have an influence on the risk of RPL.

P53 signaling pathway polymorphisms associated to recurrent pregnancy loss

The p53 protein is known for performing essential functions in the maintenance of genomic stability in somatic cells and prevention of tumor formation. Studies of the p53 signaling pathway have suggested associations between some polymorphisms and infertility, post-in vitro fertilization implantation failure and recurrent abortions. The TP53 Pro72Arg polymorphism has been implicated as a risk factor for recurrent pregnancy loss (RPL); however, the association is controversial. In this study, our objective was to evaluate selected polymorphisms in genes of the p53 signalling pathway [TP53 c.215G>C (Pro72Arg), MDM2 c.14+309T>G (SNP309) and LIF c.1414T>G in the region 3' UTR] and determine their effect as risk factors for RPL. In a case-control study, we investigated 120 women with two or more pregnancy losses and 143 fertile control women reporting at least two live births and no history of pregnancy loss. When analyzed separately, the allele and genotype distributions of the polymorphisms in the two groups were not different. However, in a multivariate analysis adjusted for alcohol consumption, smoking, ethnicity, and number of pregnancies, the interaction between the genotypes TP53 Arg/Arg (rs1042522) and MDM2 TT (rs2279744) showed to be associated to RPL, increasing the risk for this condition (OR = 2.58, 95% CI: 1.31-5.07, p = 0.006). In conclusion, our study indicates that the combination of TP53 Arg/Arg (rs1042522) and MDM2 TT (rs2279744) genotypes may be a risk factor for RPL.

Independent association of the M2/ANXA5 haplotype with recurrent pregnancy loss (RPL) in PCOS patients

ObjectiveThe aim of this study was to analyze the contribution of the M2 haplotype of ANXA5 gene, previously identified as a risk factor for RPL and thrombophilia related pregnancy complications, to repeated miscarriage observed in PCOS patients. Patients/Methods100 PCOS patients, 500 fertile women and 533 random population controls were genotyped for M2/ANXA5. ResultsM2 haplotype carriers faced a 3.4 fold elevated RPL risk (odds ratio 5.3, 95% confidence interval 3–9.2) compared to female fertile controls and 2.1 (odds ratio 2.6, 95% confidence interval 1.6–4.3) compared to population controls. The relative population risks in subgroups of PCOS patients with primary and secondary RPL were 2.3 (odds ratio 2.5, 95% confidence interval 1.2–5) and 3.3 (odds ratio 3.6, 95% confidence interval 1.5–8.4) respectively. As compared to the fertile women group, the relative risks equaled 4 (odds ratio 5, 95% confidence interval 2.3–10.8) and 6 (odds ratio 7.2, 95% confidence interval 3–17.7). Estimated relative risks for M2 carriers among PCOS RPL patients matched the values previously obtained for repeated miscarriage populations. The essential phenotypes, clinically defining PCOS, associated neither with RPL in their diagnostically relevant combinations, nor with M2 carriage as RPL risk factor in the PCOS RPL subgroups. ConclusionsM2/ANXA5 seems an independent RPL risk factor in PCOS patients that progressively correlates with the number of first trimester pregnancies. From our pilot study in PCOS women it appears relevant to offer M2/ANXA5 diagnostic analysis to such patients with RPL complications, to possibly guide proper therapeutic decisions.