Angiotensin type 2 receptor gene polymorphisms and susceptibility to preeclampsia

Background: The purpose of the study was to determine the relationship between angiotensin II type 1 receptor at position+1166 [AT1R+1166A/C; rs5186] and angi-otensin II type 2 receptor at position+1675 [AT2R+1675A/G; rs5194] gene poly-morphisms with preeclampsia in an Iranian women population

Methods: 430 women were recruited in this study including 212 preeclamptics and 218 healthy women. PCR-RFLP method was used for genotyping the polymorphisms. Chi square and Fisher exact test were used for comparing case and control groups. The p<0.05 was considered statistically significant

Results: The frequency of genotypes of the AT1R gene and AT2R gene was similar in preeclampsia and normal pregnancy. There were no significant differences in geno-type and also allele frequencies between preeclamptics and healthy women regarding the two studied polymorphisms. AT1R/AT2R genotypes combination study indicated that there was a statistically significant difference between preeclamptics and healthy women. AC/AG combination was significantly decreased, while CC/AA combination showed significant increase in patients compared with the healthy women [p<0.01]

Conclusion: The present study showed that the genetic polymorphisms within AT1R and AT2R genes may be associated with susceptibility to preeclampsia in Iranian women

T helper cell subsets and related cytokines in infertile women undergoing in vitro fertilization before and after seminal plasma exposure / 대한생식의학회지

OBJECTIVE: In vitro fertilization (IVF) is a well-known method for the treatment of infertility. The present study aimed to compare the differences between infertile women with successful and unsuccessful IVF outcomes regarding the expression of T helper (Th) cell transcription factors and a group of related cytokines before and after exposure to their husbands' seminal plasma. METHODS: This study was performed on 19 couples with unexplained infertility undergoing IVF treatment. Among the studied group, nine and 10 couples had successful and unsuccessful IVF outcomes, respectively. This study was carried out using real-time polymerase chain reaction. RESULTS: Before seminal plasma exposure, the expression levels of T-bet (p < 0.007), interferon-γ (p=0.013), and tumor necrosis factor (TNF)-α (p=0.017) were higher in the infertile women with IVF failure than in those with successful IVF outcomes, while those of GATA3 (p < 0.001), Foxp3 (p=0.001), and interleukin (IL)-35 (p < 0.003) were lower. After seminal exposure, the expression of T-bet (p=0.02), Rorc (p < 0.001), TNF-α (p=0.001), Foxp3 (p=0.02), and interferon-γ (p=0.001) increased in the unsuccessful IVF group, while the expression of Foxp3 (p=0.02), Rorc (p < 0.001), IL-23 (p=0.04), IL-17 (p=0.02), IL-6 (p < 0.001), transforming growth factor-β (p=0.01), and IL-35 (p < 0.001) increased in the successful IVF group. CONCLUSION: In summary, IVF failure was associated with imbalanced Th1/Th2/Th17/Treg responses. Moreover, our results show that seminal plasma might have a positive effect on IVF outcomes via changes in peripheral blood T cell subsets.

Genomic DNA extraction from sperm

Unlike the somatic cells, sperm DNA is very compact due to replacement of histones with protamines. Disulfide bridges formed within and between the protamines inhibit the extraction of sperm DNA through standard techniques used for the somatic cells. Furthermore, the spermatozoa themselves are protected by a membrane which is rich in disulfide bonds, making cell lysis very difficult. Following a comprehensive literature search, we developed a protocol for DNA extraction from sperm and semen fluid. The quality of extracted DNA was checked running on agarose gel, used for bisulfite conversion and PCR amplification

effect of HLA-DRB1 sharing between the couples with recurrent pregnancy loss on the pregnancy outcome after leukocyte therapy

Miscarriage is a common phenomenon complicating more than half of pregnancies. Recurrent Pregnancy Loss [RPL] is defined as three or more pregnancies lost before the twentieth week of gestation. It is believed that abnormality in maternal immune reaction to fetus and sharing of HLA antigens might be associated with RPL. To investigate the effect of HLA-DRB1 sharing between the couples with recurrent pregnancy loss on the pregnancy outcome after leukocyte therapy. Sixty primary RPL women who were immunized and followed after therapy [30 successful and 30 unsuccessful] and their husbands formed the cases of this study. In addition, one hundred healthy women were considered as the controls. HLA-DRB1 genotypes of all the cases and controls were checked by PCR-SSP method. HLA typing indicated that the prevalence of HLA-DRB1 sharing [defined as at least one allele sharing] between the couples with unsuccessful outcomes was significantly higher compared to those with successful outcomes [63.3% vs. 23.3%, p<0.004]. Moreover, HLA DRB1*07:01 allelic group was significantly more frequent in the patients with unsuccessful outcome compared to the controls [18.3% vs. 8%, p<0.04]. Our results confirmed the role of HLA sharing in RPL and revealed that HLA-DRB1 typing may be a valuable prognostic factor for the leukocyte therapy outcome

Importance of anti-GRP78 antibody in pre-eclampsia

Preeclampsia [PE] is a pregnancy specific syndrome that is associated with high maternal and fetal morbidity and mortality. Glucose regulated protein78 [GRP78] is an Endoplasmic Reticulum [ER] protein which is expressed on the cell surfaces of trophoblast cells under stress or hypoxic condition. GRP78 has a role in aggressive behavior of invasive cells and may play a role in normal placentation. To investigate the autoantibody against GRP78 in the sera of patients with PE and to assess the correlation between antibody and severity of the disease. We evaluated the anti-GRP78 antibody within the sera of fifty pre-eclamptic [12 severe and 38 mild PE] and fifty healthy pregnant women using a home-made ELISA assay. Furthermore, western blot technique was used to assess the expression of GRP78 in placenta of healthy and pre-eclamptic women in their third trimester. The presence of anti-GRP78 antibody in the serum samples from pre-eclamptic and healthy women was also assessed. GRP78 was expressed by placenta, and both healthy and preeclamptic women produced anti-GRP78 antibody. Although no significant difference was found between the pre-eclamptic and healthy women regarding the level of anti-GRP78 antibody, the difference between severe pre-eclamptic and healthy control women was statistically significant [p<0.003]. The findings of the present study indicated that measurement of anti-GRP78 antibody may provide a new marker for severe pre-eclampsia. Yet, future studies are required to confirm this notion.

Soluble cd30 in normal pregnancy pre-eclampsia and recurrent pregnancy loss

Normal pregnancy is thought to be dependent on Th2 deviation, while Recurrent Pregnancy Loss [RPL] and Pre-eclampsia [PE] appear to be biased toward the Th1 immune response. It is believed that the soluble form of CD30 [sCD30] is an index of Th2 immune response or modulator of Th1/Th2 responses. The aim of this study was determination of the sCD30 level in RPL and PE patients. The sCD30 level was measured in sera of a group of normal non-pregnant women [N=43] and compared with normal pregnancy at the first [N=42] and third [N=42] trimester. Furthermore, the level of sCD30 in the normal first and third trimester pregnancies were compared with that of RPL [N=38] and severe pre-eclamptic [N=41] patients, respectively. sCD30 levels were measured by ELISA method and student t-test was used for statistical analysis. The mean level of sCD30 at the first trimester in normal pregnancy was significantly elevated as compared with normal non-pregnant women [21.4 vs. 15.2 ng/ml, p<0.0001]. A significant difference between sCD30 concentration at the first and third trimester of normal pregnancies was also observed [21.4 vs. 14.3 ng/ml, p<0.0001]. Interestingly, the sCD30 concentration did not show any significant changes at the first trimester of normal pregnancy as compared with RPL [21.4 vs. 20.9 ng/ml] and third trimester of normal pregnancy as compared with PE [14.3 vs. 13.1 ng/ml]. The data of this study indicated that the concentration of sCD30 in serum during pregnancy period is not associated with RPL or PE and serum sCD30 is not a good correlate of Th2 immune responses in pregnancy

Anti-cardiolipin and anti-neutrophil cytoplasmic antibodies in Iranian patients with behcet's disease

The prevalence of anti-Neutrophil Cytoplasmic Antibodies [ANCAs] and anti-Cardiolipin Antibodies [anti-CL Ab] in Behcet's Disease [BD] and also their roles in vascular involvement is controversial. To assess the prevalence of ANCAs and anti-CL Ab as well as their correlations with clinical manifestations in Iranian patients with BD. In this case/control study, the sera from 88 patients with BD and 88 healthy controls were evaluated. The levels of ANCAs and anti-CL Ab were measured using indirect ELISA method. The levels of anti-CL, anti-PR3 and anti-MPO [Myeloperoxidase] IgG autoantibodies between BD patients and healthy controls were not statistically different [p=0.21, p=0.28 and p=0.74, respectively]. In addition, there were no significant deferences between BD patients with and without vascular involvement in the levels of anti-CL [1.42 +/- 1.24 GPLU/ml and 1.58 +/- 1.18 GPLU/ml, respectively; p=0.71], anti-PR3 [0.0 +/- 0.0 U/ml and 0.08 +/- 0.27 U/ml, respectively; p=0.10] and anti MPO [0.48 +/- 0.23 U/ml and 0.52 +/- 0.22 U/ml, respectively; p=0.41] IgG autoantibodies. Nevertheless, mean titer of anti-CL IgG was higher in male patients with skin rash than those without skin rash [2.2 +/- 0.88 GPLU/ml and 1.11 +/- 1.22 GPLU/ml, respectively; p=0.017]. While anti-CL, antiPR3 and anti-MPO IgG autoantibodies do not play a major role in susceptibility to BD or pathogenesis of vascular involvement in our patients, anti-CL Ab might be involved in skin lesion development in Iranian male BD patients. However, the results should be confirmed in other studies

Investigation of Fc gamma RIIA and Fc gamma RIIIA polymorphism in multiple sclerosis: a case control study

Multiple Sclerosis [MS], the most common demyelinating disease of the CNS, is immunologically mediated in genetically susceptible individuals. Receptors for the Fc fragment of IgG [Fc gamma R] might induce inflammatory responses through linking the humoral and cellular immune responses by targeting immune complexes to effector cells. Polymorphisms in some Fc gamma R genes are associated with various infectious and autoimmune diseases, probably due to their effects on different binding capacities of encoded receptors for IgG containing immune complexes. To investigate the importance of Fc gamma R polymorphisms in susceptibility to MS. One hundred and fifty MS patients and 136 age and sex matched controls were genotyped for Fc gamma RIIA and Fc gamma RIIIA gene polymorphisms using PCR-RFLP method. The allelic and genotypic frequencies of the Fc gamma RIIA and Fc gamma RIIIA did not differ significantly between the MS patients and controls. There was no association between allelic polymorphism of Fc gamma RIIIA and severity of disease based on Expanded Disability Status Scale [EDSS] score. However, significant association between inherited Fc gamma RIIA genotype and disease activity [p=0.001] or progression index was revealed [p=0.014]. EDSS values showed that Fc gamma RIIA [H/H] and [H/R] genotypes were associated with a lower EDSS score in relapsing-remitting MS and in the total MS population [P=0.001] but not [R/R] genotype. Considering the detrimental role of autoantibodies in the pathogenesis of MS, our results suggest that the inherited Fc gamma RIIA alleles could affect the severity of MS by influencing the clearance rate of immune complexes and autoantibodies. The results of the present study add the Fc gamma RIIA gene to the gene networks which determine the severity of MS in southern Iran