Biochemical Markers of Oxidative Stress in Saudi Women with Recurrent Miscarriage

This study was undertaken to investigate the antioxidant/oxidant status in recurrent miscarriage patients. Antioxidants including glutathione peroxidase (GPx), catalase (CAT), glutathione reductase (GR), reduced glutathione (GSH) and selenium (Se), as well as the oxidants hydrogen peroxide (H2O2), oxidised glutathione (GSSG) and lipid peroxidation were assayed in plasma, whole blood and placental tissue of non-pregnant women (NP), healthy pregnant women (HP), and recurrent miscarriage (RM) patients. Results indicated that all antioxidant activities and levels in plasma and whole blood of HP women were consistently moderately lower, and much more significantly lower in RM patients when both were compared to those seen in NP women (P<0.05 and P<0.001, respectively). Furthermore, whereas plasma antioxidant activities and levels were significantly lower in RM patients, those of whole blood and placental tissue were much more significantly lower when compared with HP women (P<0.001). Concurrent with these findings there were consistent increases of equal statistical significance and magnitude in the levels of all investigated oxidants assayed in all samples when compared in between subjects of the study as indicated above. Data thus illustrated a distinct shift in favor of oxidative reactions and reactive oxygen species (ROS) generation, and very significant decreases in the GSH/GSSG ratios in whole blood and placental tissue of RM patients when compared to HP and NP women (P<0.001). The above noted oxidative stress could have been a major causative factor of recurrent miscarriage.

Gene expression analysis of VEGF and its receptors and assessment of its serum level in unexplained recurrent spontaneous abortion

Unexplained recurrent spontaneous abortion [URSA] is one of the main complications of pregnancy which is usually defined as three or more consecutive pregnancy losses before the 20[th] week of gestation without a known cause. Vascular endothelial growth factor [VEGF] is a potent angiogenic factor and shown, along with its receptors [VEGFR1, 2], to play important roles in several physiologic processes including reproduction. The aim of the present study was to analyze gene expression of VEGF and VEGF receptors in endometrium of patients with a history of URSA compared with normal fertile women. In addition, serum VEGF concentration was assessed and compared between the two groups at the same time. In this case control study, endometrial and blood samples were obtained between day 19[th] and 24[th] of menstrual cycle [window of implantation] from 10 women with a history of URSA [case group] and 6 fertile women who had at least one successful pregnancy [control group]. Expression of VEGF and VEGFRs was studied by reverse transcription- polymerase chain reaction [RT-PCR] and then quantified by real time PCR. Normalization of expression levels was done by comparison with beta-actin expression level as an internal control. Relative VEGF, VEGFR1 and VEGFR2 expression quantities were compared between the two groups. Enzyme linked immunosorbent assay [ELISA] was used for serum VEGF assay. VEGF, VEGFR1 and VEGFR2 gene expression was detected in endometrial samples of both groups. The mean relative expression of VEGF gene was lower in the case group compared with control women, however, both VEGF receptors were expressed higher in endometrium of the case group. In addition, the serum level of VEGF was significantly higher in the case group compared with the controls. Alteration in gene expression of VEGF and its receptors in endometrium and changes of serum VEGF might play important roles in pathogenesis of unexplained RSA

Efecto de los anticuerpos antifosfolipídicos en la formación y degradación de la malla de fibrina en pacientes con aborto recurrente / Effect of antiphospholipid antibodies on the formation and lysis of fibrin network in patients with recurrent miscarriage

En el presente trabajo se estudió el proceso de formación y disolución de la malla de fibrina y la generación de plasmina en un grupo de pacientes con aborto recurrente (AR) debido a la presencia de anticuerpos antifosfolipídicos (N= 10), mujeres con AR sin el síndrome antifosfolipídico (SAF) (N= 6) y se comparó con un grupo de mujeres sanas (N= 8). Del grupo de pacientes estudiadas con SAF, nueve fueron positivas para anticuerpos anticardiolipina (aCL), cinco para la anti-b2-glicoproteína I (anti-b2GPI), cuatro para ambos anticuerpos, una para anticuerpos antiprotrombina (aPT) y anticoagulante lúpico (AL). El proceso de formación de la fibrina y su disolución fue estudiado por turbidimetría y la generación de plasmina mediante sustrato cromogénico S2251. Las curvas de polimerización de la(s) paciente(s) con AR sin SAF y AL presentaron un incremento en la pendiente y turbidez final, comparado con las del grupo control de mujeres sanas. La velocidad de disolución del coágulo fue mayor en la paciente con AL (21 ± 0) 10-4 DDO/seg y en las AR sin SAF (19,6 ± 5,7) 10-4 DDO/seg, comparado con el grupo control (14,5 ± 2,8) 10-4 DDO/seg. La generación de plasmina estuvo incrementada solamente en las AR sin SAF (85 ± 24%) comparado con 52 ± 3% en el grupo control, p= 0,005. Los cambios observados en el proceso de polimerización y fibrinólisis de la(s) paciente(s) con AR sin SAF y AL pudieran estar relacionados con el incremento en los niveles de fibrinógeno, mientras que los de la generación de plasmina con la entidad mórbida.

The present work was intended to study the process of fibrin formation and lysis and plasmin generation in a group of patients with recurrent miscarriage (RM), due to the presence of antiphospholipid antibodies (N= 10); as well as in women with RM without the antiphospholipid syndrome (APS) (N= 6), compared with those of a group of healthy women (N= 8). In the group of patients with APS, nine were positive for antibodies against cardiolipin (aCL), five for anti-b2-glycoprotein I (anti-b2GPI), four for both antibodies, and one for antibodies against prothrombin (aPT) and lupus anticoagulant (LA). Fibrin formation and lysis was followed by turbidity and plasmin generation using chromogenic substrate S2251. The polymerization curves from RM patients without APS and the LA patient showed an increased slope and maximum turbidity compared to those of the control group. The speed of lysis was higher in the LA patient (21 ± 0) 10-4 DOD/seg and the RM patients without APS (19.6 ± 5.7) 10-4 DDO/seg, compared to that of the control group (14.5 ± 2.8) 10-4 DDO/seg. Plasmin generation increased only in RM patients without APS (85 ± 24%) against the control group (52 ± 3%), p= 0.005. The changes observed in the fibrin polymerization and lysis process of women with RM without APS and LA seem to be related to their higher fibrinogen levels, while the increased plasmin generation was related to the patients´ morbidity.

Effects of sera taken from women with recurrent spontaneous abortion on sperm motility and apoptosis

Recurrent spontaneous abortion impacts almost 1% of couples. The sera from women with unexplained recurrent spontaneous abortion [URSA] have toxic effects on embryos that grow in the uterus. Therefore, the abnormal condition of the uterus may also affect sperm qualities. The objectives of this study were to search if these sera could induce DNA denaturation in sperm nuclei and also it could reduce sperm motility. Sera of 20 women with URSA history and sera from 20 women with at least two healthy children were added to the sperms samples from 20 healthy men for 2 hours. The sperm motility was assessed after incubation with sera. The samples were stained with Tdt mediated dUTP nick end labeling [TUNEL] assay for DNA fragmentation. The samples were analyzed with flow cytometry and the percentage of the TUNEL positive sperms were calculated. The data were analyzed by t-test. The incubation of the sperm samples in sera with URSA lead to a decrease in the percentage of the motile sperm from 55% in control to 41% in the treated group, significantly [p=0.038]. The percentage of the sperm with abnormal fragmented DNA increased after incubation with URSA [26.6%] compare to the control [21.2%]; however, it was not significant. It seems that sera from URSA patients could not induce a significant increase in the percentage of the sperms with nuclei contain DNA fragmentation. However, the sera of women with URSA could affect the fertility rate by reduction of the sperm motility

Oxidative stress in recurrent pregnancy loss women

To investigate biochemical changes in lipid peroxidation, nitric oxide, and vitamin E in recurrent pregnancy loss women, and compare these with healthy pregnant, and non-pregnant women. A case control study was conducted from September 2008 to December 2009 at Al Khadimiya Teaching Hospital, Baghdad, Iraq. Ninety-six subjects were included in the study, 32 were patients with recurrent pregnancy loss [RPL], and 32 pregnant women in their third trimester, and another 32 non-pregnant women were used as controls. Blood samples were collected from each patient at the time of pregnancy loss. Serum from patients and controls was then used to estimate malondialdehyde [MDA], nitric oxide [NO], and vitamin E levels. There was a significant elevation in patient serum MDA compared with third trimester pregnant women [p=0.002] and non-pregnant women [p=0.0001]. Both serum vitamin E and NO levels in RPL patients also showed a highly significant decrease compared with third trimester pregnant, and non-pregnant women. A highly significant difference was found in the MDA/vitamin E ratio between RPL and control groups, while no significance was found between RPL and control groups' NO/vitamin E ratio. The decrease in NO production and vitamin E is a result of RPL and not a causative factor, as the RPL was without pathological cause, medication, or fibroid presence, and no significant difference was found between the NO/vitamin E ratio in RPL and controls group

Pregnancy immunology: decidual immune cells.

Human pregnancy is a complex process. Placental development depends on the function of secretory molecules produced by placental trophoblast cells as well as by maternal uterine immune cells within the decidua. These decidual immune cells are T cells, natural killer cells, macrophages and dendritic cells. The interactions between the trophoblast cells and the maternal immune cells have an impact on the outcome of the pregnancy. Knowledge about the phenotypes and functions of the maternal immune cells in normal and pathological pregnancies including recurrent spontaneous abortions, preeclampsia and hydatidiform moles may improve our understanding of the immunobiology of the normal pregnancy as a whole and may provide approaches for improving the treatment of pathological pregnancies.

Comparison of anticardiolipin antibody and antiphospholipid antibody in women with recurrent abortions

Recurrent abortion is a critical problem in which many factors play a crucial role such as anticordiolipin antibody and antiphospholipid antibody. This study was conducted to evaluate the frequency of anticardiolipin antibody and antiphospholipid antibody in pregnancy failures in women with the history of recurrent pregnancy loss. In 154 women with the history of two or more recurrent pregnancy losses, serum anticardiolipin and serum antiphospholipid were measured using ELISA method. The positive IgG anticardiolipin and IgG antiphospholipid were detected in 12. 34% [19 patients] and 6.5% [10 patients] of patients respectively. Although 16 out of 19 patients with positive IgG anticardiolipin were negative for IgG antiphospholipid and 7 out of 10 patients with positive IgG antiphosphplipid were negative IgG anticardiolipin, but there was a significant correlation between IgG anticardiolipin and IgG antiphaspholipid [r = 0.222 p=0.000]. Our data concluded that anticardiolipin antibody is found to be more important than anti phospholipid antibody in recurrent abortion

Beta2-glycoprotein I dependent anticardiolipin antibodies and lupus anticoagulant in patients with recurrent pregnancy loss.

AIM: The present study was aimed to define the incidence of antiphospholipid antibodies of different types lupus anticoagulant (LAC), venereal disease research laboratory test (VDRL) and Beta2-glycoprotein I dependent anticardiolipin antibodies Beta2 I aCL) in our cohort of population experiencing recurrent pregnancy loss (RPL) from Andhra Pradesh, South India. SETTING AND DESIGN: A referral case-control study at a tertiary centre over a period of 5 years. PARTICIPANTS: 150 couples experiencing 3 or more recurrent pregnancy losses with similar number of matched controls. MATERIAL AND METHODS: LAC activity was measured by the activated partial thromboplastin time (aPTT) according to the method of Proctor and Rapaport with relevant modifications. VDRL analysis was performed by the kit method supplied by Ranbaxy Diagnostics Limited and Beta2 Glycoprotein I dependent anticardiolipin antibodies were estimated by ELISA kit (ORGen Tech, GmbH, Germany) with human Beta2 Glycoprotein I as co-factor. STATISTICAL ANALYSIS: Statistical analysis was performed using Student's t test. RESULTS: LAC activity was found positive in 11 women (10.28%). The mean +/- SE Beta2 I aCL concentration in the study group was 14.53 (micro/ml) +/- 1.79 (range 0 to 90.4 micro/ml) which was higher than the control group with a mean +/- SE of 7.26 (micro/ml) +/- 0.40 (range 0 to 18 u/ml). The binding of the antibodies to the antigen was observed in 40.24% (n=33) of the cases compared to 6.09% (n=5) in controls. VDRL test was positive in 7(2.34%) individuals (3 couples and 1 male partner) and none among controls. CONCLUSIONS: The present study indicates the importance of antiphospholipid antibodies in women experiencing RPL and suggests the usefulness of screening for these antibodies as a mandatory routine for instituting efficient therapeutic regimens for a successful outcome of pregnancy.

A preliminary study of antiphospholipid antibodies in 50 cases of bad obstetric history.

The prevalence of the antiphospholipid antibodies in certain high risk groups like bad obstetric history is higher than previously realized. A pilot study in 50 patients of bad obstetric history, for the presence of lupus anticoagulant and anticardiolipin antibodies was carried out to establish an association between the presence of the antiphospholipid antibodies and recurrent fetal wastage. Lupus anticoagulant alone was positive in six patients (12%) and anticardiolipin antibody alone was positive in 14 patients (28%), while both lupus anticoagulant and anticardiolipin antibodies were positive in three patients (6%). Activated partial thromboplastin time (aPTT) and prothrombin time (PT) were not significantly prolonged. Platelet count was normal in all the patients. We conclude that lupus anticoagulant as based on kaolin clotting time and anticardiolipin antibodies are important tests to detect the etiology of fetal wastage.

The prevalence of antiphospholipid antibodies in cases of recurrent pregnancy loss.

Antiphospholipid antibodies (aPL) are a diverse family of autoantibodies reactive against negatively charged phospholipid-protein complexes. The clinically significant members include lupus anticoagulant (LA), anticardiolipin antibody (aCL) and reaginic antibodies causing biological false positive (BFP) venereal disease laboratory test (VDRL). Although detected in various clinical scenarios, unexplained fetal loss in women of reproductive age group is the commonest association. Fifty pregnant women of first and second trimester with a history of two or more unexplained pregnancy losses were studied for the presence of LA, aCL and reaginic antibodies. Thirty pregnant women of the same trimester without any history of fetal loss were taken as control. LA was detected in nine (18%) cases and aCL in 12 (24%) cases of the study group. The control group was negative for any autoantibody. The prevalence of aPL in the study group found to be statistically significant. Detection of aPL must be considered in women with previous pregnancies complicated by unexplained fetal wastage.

Anticorpos antifosfolípides na perda fetal recorrente / Antiphospholipids antibodies in recurrent fetal loss

OBJETIVOS: Foram pesquisados os anticorpos antifosfolípides (AFL): Anticoagulante Lúpico (LAC) eAnticorpo Anticardiolipina (ACA); os anticorpos contra antígenos nucleares: Fator anti-núcleo (FAN), Fator anti-DNA (FAD), Anticorpo Anti-Sm e Anti-SSA (Rô) e as imunoglobulinas IgG, IgM e IgA em pacientes com perda fetal recorrente. CASUISTICA E METODOLOGIA: l00 pacientes distribuídas em 4 grupos de 25: Grupo 1: Pacientes com abortamento habitual; Grupo 2: Pacientes com feto morto sem causa aparente, Grupo 3: Pacientes com feto morto associado à doença hipertensiva específica da gravidez-(DHEG) e Grupo controle: Pacientes com um ou mais filhos vivos sem perdas conceptuais. RESULTADOS:O título do LAC e o índice de ligaçao específica (ILE) do ACA foram maiores no grupo l em relaçao ao controle (p