Factor VII R353Q (rs6046), FGA A6534G (rs6050), and FGG C10034T (rs2066865) Gene Polymorphisms and Risk of Recurrent Pregnancy Loss in Iranian Women.

Recurrent pregnancy loss is a multi factorial and heterogeneous disorder defined as two or more consecutive pregnancy losses before 20 weeks' gestation. Gene polymorphisms including factor VII R353Q (rs6046), fibrinogen alpha chain A6534G (rs6050) and fibrinogen gamma chain C10034T (rs2066865) have potential role in thrombophilia and the relation between these three polymorphisms and an increased risk of venous thrombosis have been reported. As thrombophilia is associated with a considerable proportion of pregnancy loss and the association between these gene polymorphisms and recurrent pregnancy loss remains controversial, the aim of the present study was to evaluate the relation of these polymorphisms and recurrent pregnancy loss in Iranian women. A total of 144 women with a history of two or more consecutive miscarriages as the patient group and 150 healthy women with two live births and no history of pregnancy loss as the control group were included in the study. Polymerase chain reaction and restriction fragment length polymorphism were used for genotyping. The results were validated by DNA sequencing. The SPSS, SNPStats and Finch TV were used to analyze the results. Factor VII R353Q (rs6046) gene polymorphism showed a significant difference between RPL patients and the control group according to multiple logistic regression models [codominant (OR=0.38; 95% CI=0.23-0.63, P≤0.0001), dominant (OR=0.32; 95% CI=0.20-0.52, P≤0.0001), over dominant (OR=0.46; 95% CI=0.29-0.75, P=0.0017) and log-additive (OR=0.35; 95% CI=0.23-0.53, P≤0.0001)]. Fibrinogen alpha chain A6534G (rs6050) and fibrinogen gamma chain C10034T (rs2066865) gene polymorphisms showed no correlation with recurrent pregnancy loss. Factor VII R353Q (rs6046) gene polymorphism can be considered a risk factor for recurrent pregnancy loss. Further studies in larger populations are needed to confirm the findings.

Toll-Like Receptor 4, 2, and Interleukin 1 Receptor Associated Kinase4: Possible Diagnostic Biomarkers in Myelodysplastic Syndrome Patients.

Background: Myelodysplastic syndrome (MDS) is a clonal hematologic disorder that requires the integration of morphologic, cytogenetic, hematologic, and clinical findings for a successful diagnosis. Trying to find ancillary tests such as biomarkers improve the diagnosis process. Several studies showed that a disordered immune system is associated with MDS. The chronic activated innate immune system, particularly the Toll-like receptors (TLRs) pathway could be involved in the induction of the inflammation. Materials and Methods: In the present study, we investigated the expression of TLR2, TLR4, and IRAK4 in bone marrow (BM) of MDS patients, the leukemia group, and the healthy group. For this purpose, we assessed the expression of TLR2, TLR4, and IRAK4 by real time-PCR. Results: In line with new findings, we demonstrated that the expression of TLR2, TLR4, and IRAK4 significantly increased in MDS BM compared with the healthy group. Moreover, IRAK4 expression raised significantly in MDS patients compared with other studied hematologic neoplasms. Also, the expression levels of TLR2 and TLR4 significantly increased in MDS in comparison to some studied non-MDS malignancies (P ˂ 0.05). Receiver operating characteristics (ROC) analysis and area under the curve (AUC) suggested that the expression of TLR2, TLR4, and IRAK4 (AUC = 0.702, AUC = 0.75, and AUC = 0.682, respectively) had acceptable diagnostic values to identify MDS from the other understudied leukemias. Conclusion: Overall, the expression of TLR2, TLR4, and IRAK4 could be potential biomarkers for discriminating MDS from some hematologic disorders.

Effects of glucose on the proliferation of human umbilical cord blood hematopoietic stem cells.

Hematopoietic stem cells (HSCs) can be isolated through umbilical cord blood (UCB), which can be used for HSC transplantation. Despite many advantages, the low number of UCB CD34+ cells lead to delayed engraftment. Ex-vivo CD34+HSC expansion is a potentially safe approach to increasing CD34+ cell numbers. The NLR family of pyrin domain-containing 3 (NLRP3) is an intracellular protein that plays an essential role in the innate immune response. Several blood cell types, HSCs and progenitor cells (HSPCs) express the NLRP3 inflammasome complex genes and participate in the development and proliferation of HSPCs. In this study, magnetic-activated cell sorting (MACS) beads isolated CD34+HSCs. The cell purity was evaluated by flow cytometry. CD34+ cells, under the influence of different doses of glucose, MCC950 were cultured for seven days. The qRT-PCR was used to evaluate gene expression. The results showed that in the culture medium treated with glucose concentrations, the expression of the NLRP3 inflammasome complex genes and the amount of CD34+ cells increased by more than 50%. In contrast, genes expression and the number of CD34+ cells in the culture medium treated with MCC950 decreased. UCB is a source of new therapeutic methods. This study demonstrates the relationship between glucose and the activation of the NLRP3 inflammasome. Based on these results, glucose causes the expansion of CD34+HSCs through its effect on HSCs in simultaneous culture.

Elevated CD9 expression as a potential biomarker for diagnosis of Bernard-Soulier syndrome.

Diagnosis of inherited platelet glycoprotein disorders is based on specific laboratory techniques such as aggregometry and flow cytometry. Flowcytometry is a powerful method, but equivocal results are produced in some cases. New cluster of differentiation markers could resolve the diagnostic dilemmas. Abnormal expression of CD9 in Bernard-Soulier syndrome (BSS) is recently reported. We aimed to determine the diagnostic significance of CD9 expression in a cohort of Iranian patients with inherited platelet glycoprotein defects. Twelve BSS, 21 Glanzmann thrombasthenia and 16 healthy controls were included in the present study. Flowcytometric diagnosis of BSS and Glanzmann thrombasthenia was made by analysis of CD41/61 and CD42a/42b CD markers. Moreover, phycoerythrin-labelled anti CD9 was examined in patients and healthy controls. The mean fluorescence intensity (MFI) of CD9 among the three groups was compared using suitable statistical methods and a P value of less than 0.05 considered statistically significant. Mean MFI of CD9 was 990.0 in BSS patients versus 421.2 and 317.3 in individuals with Glanzmann thrombasthenia and healthy controls, respectively (P < 0.05). Between the two-group comparison of means by the Mann-Whitney test revealed a P value of less than 0.001 for BSS group versus GT (2.4-fold) and BSS versus healthy controls (2.9-fold). CD9 molecule also expressed differently in patients with Glanzmann thrombasthenia in comparison with healthy controls (P < 0.001), although with a less magnitude (1.3-fold). According to our findings, CD9 is a potential biomarker for laboratory diagnosis of inherited glycoprotein defects, especially to elucidate the ambiguous results in BSS cases.

Determining the association of thrombophilic gene polymorphisms with recurrent pregnancy loss in Iranian women.

OBJECTIVE: Thrombophilia is known to be associated with poor pregnancy outcomes. In this study, three thrombophilic gene polymorphisms, including EPCR (Ser219Gly), F11 (rs4253417) and F7 (323 Ins10) were investigated in an Iranian population of women in order to determine the correlation between thrombophilia and recurrent pregnancy loss (RPL). METHODS: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were used to evaluate the frequency of three candidate thrombophilic risk factors for recurrent pregnancy loss. The frequencies of the polymorphisms were compared between the case (144 patients with a history of at least two miscarriages) and the control (150 healthy women with no abortion) group. RESULTS: Our results show that EPCR and FVII polymorphisms of the patient and control group have the same genotype frequency, and the difference is not statistically significant (p-value > .05). Regarding FXI polymorphism, TT genotype frequency was higher in the patient group than the control group (p-value < .05); however, CT heterozygote form was higher in the control group compared to the patient group (p-value < .05). CONCLUSION: In FXI polymorphism, T allele is possibly an RPL risk factor and C allele has a protective role. Thus, wild type FXI could be related to RPL, but EPCR and FVII polymorphism have no such correlation.

Epigenetic changes in FOXO3 and CHEK2 genes and their correlation with clinicopathological findings in myelodysplastic syndromes.

OBJECTIVES/BACKGROUND: Myelodysplastic syndromes (MDSs) are a heterogeneous disease in terms of clinical course and response to therapy. Epigenetic changes are the primary mechanism of MDS pathogenesis. FOXO3 and CHEK2 genes play significant roles in normal cellular mechanisms and are also known as tumor suppressor genes. We aimed to clarify the correlation of epigenetic changes in these genes with clinicopathologic findings in MDS. METHODS: A total of 54 newly diagnosed MDS patients referred to Shariati and Firouzgar Hospitals (Tehran, Iran) were included in the study from 2013 to 2015, comprising the following cases: 26 with refractory cytopenia with unilineage dysplasia, 10 with refractory cytopenia with multilineage dysplasia, four refractory anemia with excess blasts-1 (RAEB-1), 11 refractory anemia with excess blasts-2 (RAEB-2), and three MDS associated with isolated deletion (5q-). Risk groups were determined according to the Revised International Prognostic Scoring System (IPSS-R). The methylation status of CHEK2 and FOXO3 promoters were determined by methylation-sensitive high-resolution melting analysis of sodium bisulfite-converted DNA. Expressions of CHEK2, FOXO3, and GAPDH were measured by quantitative real-time polymerase chain reaction and fold changes were calculated using the ΔΔCT method. RESULTS: Statistical analysis revealed no promoter methylation of CHEK2 and FOXO3 in healthy control specimens. FOXO3 promoter methylation was associated with high-risk World Health Organization subgroups (p = .017), high-risk IPSS-R (p = .007), high-risk cytogenetics (p = .045), and more than 5% blasts in bone marrow (p = .001). CHEK2 promoter methylation was correlated with more than 5% blasts in bone marrow (p = .009). CONCLUSIONS: Promoter methylation of CHEK2 and especially FOXO3 is associated with adverse clinicopathological findings and disease progression in MDS.