Association of kinase insert domain-containing receptor [KDR] gene polymorphism/ haplotypes with recurrent spontaneous abortion and genetic structure

Background: Recurrent spontaneous abortion is one of the diseases that can lead to physical, psychological, and, economical problems for both individuals and society. Recently a few numbers of genetic polymorphisms in kinase insert domain-containing receptor [KDR] gene are examined that can endanger the life of the fetus in pregnant women

Objective: The risk of KDR gene polymorphisms was investigated in Iranian women with idiopathic recurrent spontaneous abortion [RSA]

Materials and Methods: A case controlled study was performed. One hundred idiopathic recurrent spontaneous abortion patients with at least two consecutive pregnancy losses before 20 weeks of gestational age with normal karyotypes were included in the study. Also, 100 healthy women with at least one natural pregnancy were studied as control group. Two functional SNPs located in KDR gene; rs1870377 [Q472H], and rs2305948 [V297I] as well as one tag SNP in the intron region [rs6838752] were genotyped by using PCR based restriction fragment length polymorphism [PCR-RFLP] technique. Haplotype frequency was determined for these three SNPs' genotypes. Analysis of genetic STRUCTURE and K means clustering were performed to study genetic variation

Results: Functional SNP [rs1870377] was highly linked to tag SNP [rs6838752] [D' value=0. 214; x2 = 16.44, p<0. 001]. K means clustering showed that k = 8 as the best fit for the optimal number of genetic subgroups in our studied materials. This result was in agreement with Neighbor Joining cluster analysis

Conclusion: In our study, the allele and genotype frequencies were not associated with RSA between patient and control individuals. Inconsistent results in different populations with different allele frequencies among RSA patients and controls may be due to ethnic variation and used sample size

Lentiviral mediating genetic engineered mesenchymal stem cells for releasing IL-27 as a gene therapy approach for autoimmune diseases

Autoimmune diseases precede a complex dysregulation of the immune system. T helper17 [Th17] and interleukin [IL]-17 have central roles in initiation of inflammation and subsequent autoimmune diseases. IL-27 significantly controls autoimmune diseases by Th17 and IL-17 suppression. In the present study we have created genetic engineered mesenchymal stem cells [MSCs] that mediate with lentiviral vectors to release IL-27 as an adequate vehicle for ex vivo gene therapy in the reduction of inflammation and autoimmune diseases. In this experimental study, we isolated adipose-derived MSCs [AD-MSCs] from lipoaspirate and subsequently characterized them by differentiation. Two subunits of IL-27 [p28 and EBI3] were cloned in a pCDH-513B-1 lentiviral vector. Expressions of p28 and EBI3 [Epstein-Barr virus induced gene 3] were determined by real time polymerase chain reaction [PCR]. MSCs were transduced by a pCDH-CMV-p28-IRESEBI3- EF-copGFP-Pur lentiviral vector and the bioassay of IL-27 was evaluated by IL-10 expression. Cell differentiation confirmed true isolation of MSCs from lipoaspirate. Restriction enzyme digestion and sequencing verified successful cloning of both p28 and EBI3 in the pCDH-513B-1 lentiviral vector. Real time PCR showed high expressions level of IL-27 and IL-10 as well as accurate activity of IL-27. The results showed transduction of functional IL-27 to AD-MSCs by means of a lentiviral vector. The lentiviral vector did not impact MSC characteristics

Compound Heterozygosity for Two Novel SLC26A4 Mutations in a Large Iranian Pedigree with Pendred Syndrome

OBJECTIVES: The aim of this study was to detect the genetic cause of deafness in a large Iranian family. Due to the importance of SLC26A4 in causing hearing loss, information about the gene mutations can be beneficial in molecular detection and management of deaf patients. METHODS: We investigated the genetic etiology in a large consanguineous family with 9 deaf patients from Fars province of Iran with no GJB2 mutations. Initially, linkage analysis was performed by four DFNB4 short tandem repeat markers. The result showed linkage to DFNB4 locus. Following that, DNA sequencing of all 21 exons, their adjacent intronic sequences and the promoter of SLC26A4 was carried out for mutation detection. RESULTS: Two novel mutations (c.863-864insT and c.881-882delAC) were identified in exon 7 of the gene, in both homozygous and compound heterozygous state in patients. CONCLUSION: Our results supported the importance of the SLC26A4 mutations in the etiology of hearing loss among the Iranian patients and therefore its mutation screening should be considered after GJB2 in the molecular diagnostics of hearing loss, especially when enlarged vestibular aqueduct or goiter is detected.

[Evaluation of the association between the C677T and A1298C polymorphisms of MTHFR gene and recurrent miscarriage]

Two factors alleged to cause thrombophilia in women with unexplained recurrent spontaneous abortion [RSA] are the MTHFR polymorphisms including C677T and A1298C. This case-control study was aimed to determine the association between RSA and two polymorphisms of MTHFR in Iranian patients. 30 patients with the background of two or more consecutive unexplained abortions and 10 women with at least two live births without a miscarriage who referred to Baqiyatallah Hospital and Avicenna Infertility Clinic were analyzed for MTHFR C677T and A1298C polymorphisms by the PCR-RFLP method. Results achieved from estimating the genotype of each polymorphism were analyzed by the SPSS program via the 2chi method. The Sperman method was also used to evaluate the correlation between the two polymorphisms. The data presented in this study have shown a significant correlation between MTHFR C677T and MTHFR A1298C polymorphisms. 17 women [56.6%] with recurrent spontaneous abortions and 5 women [50%] among the controls were heterozygote for MTFIFR C677T polymorphism. T allele frequency in the patient group was more than the control group [28.4% for patients and 25% for controls]. Frequency of the MTHFR A1298C polymorphism was 63.3% in patient and 50% in controls. For A1298C polymorphism, 43.3% of patients and 20% of controls were heterozygote. Furthermore, 20% of patients and 30% of controls were homozygote for this polymorphism. The prevalence of MTHFR C677T and MTHFR A1298C polymorphisms were slightly, but not significantly, higher in RSA patients compared to controls. These findings failed to support the relationship between thrombophilia polymorphisms and the increasing risk of RSA in evaluated Iranian women