Circulating microRNAs as potential non-invasive biomarkers in pediatric patients with celiac disease.

Summary: Celiac disease is an enteropathy induced by ingestion of gluten triggering an immune response in genetically predisposed individuals. MiRNAs are small non-coding RNAs that have a role as regulators of gene expression at the post transcriptional level. The aim of this study is to evaluate the possibility of using circulating miRNAs as non-invasive biomarkers in pediatric patients with celiac disease. In addition, we examine the effect of a gluten-free diet on the expression of these miRNAs in serum of CD patients. The expression pattern of miR-21 and miR-31 was estimated in serum of 25 untreated CD patients (recently diagnosed), 25 treated CD patients (on gluten-free diet) and 20 healthy controls using qRT-PCR. Our results demonstrated the significant up-regulation of microRNA-21 in the untreated celiac patients in comparison with the treated group and healthy controls. Moreover, miR-31 expression was significantly under-expressed in the untreated celiac patients in comparison with the treated group and healthy controls. Furthermore, the results showed that miR-21 expression level was significantly positively correlated with the tTG IgA auto-antibodies. In conclusion, circulating miRNA-21 and miRNA-31 could serve as potential non-invasive biomarkers for pediatric CD patients.

Potential value of circulating microRNA-126 and microRNA-210 as biomarkers for type 2 diabetes with coronary artery disease.

BACKGROUND: Macrovascular complications are the main cause of morbidity and mortality among the diabetic patients. MicroRNAs (miRNAs), a family of small non-coding RNAs, play vital roles in the regulation of blood glucose level and the concurrent cardiovascular complications of type 2 diabetes. We hypothesized that plasma miR-126 and miR-210 are linked to coronary artery disease (CAD) in these diabetes patients. METHODS: Fasting blood samples were collected from 20 healthy volunteers and 100 patients with diabetes (54 patients without CAD and 46 patients with CAD). Plasma miR-126 and miR-210 expressions were assessed by quantitative real time PCR. Specificity and sensitivity of miR-126 and miR-210 to discriminate CAD with diabetes was determined by receiver operating characteristic curve analysis. Correlations between miR-126 and miR-210 and studied characteristics in diabetes patients with and without CAD were compared. RESULTS: Plasma relative expressions of miR-126 and miR-210 were 0.38 ± 0.03 and 5.3 ± 0.56 in diabetes alone vs. 0.08 ± 0.03 and 21.44 ± 0.97 in diabetes with CAD, respectively (both p < 0.0001). Levels of miR-126 and miR-210 significantly correlated with certain glycemic and lipid indices. The miRNAs significantly discriminated between diabetes with and without CAD at cut-off values of 0.055 (sensitivity 91.3%, specificity 100%) for miR-126 and of 17.59 (sensitivity 93.5%, specificity 100%) for miR-210. CONCLUSION: Plasma miR-126 and miR-210 levels may be biomarkers for diabetes with or without CAD.

Transforming growth factor-ß1 gene polymorphism in mycosis fungoides.

BACKGROUND: Dysregulation in transforming growth factor (TGF)-ß1 signalling pathways has been linked to cancer. AIM: To study the association between single nucleotide polymorphisms (SNPs) of the TGF-ß1 gene and mycosis fungoides (MF). METHODS: Using restriction fragment length polymorphism analysis, SNPs in the TGF-ß1 gene were studied in 55 patients with MF of different stages and in 100 apparently healthy controls. RESULTS: A significant difference was found between patients and controls in distribution of the different TGF-ß1 genotypes, with mutant forms (T/C, T/T) encountered significantly more often in patients with MF (P < 0.001). The heterozygous genotype (T/C) was significantly associated with patch stage MF, whereas the homozygous genotype (T/T) was significantly associated with tumour stage (stage IIb) MF (P = 0.001), although this study included only a small number of these patients. CONCLUSIONS: Mutant TGF-ß1 genotypes are significantly associated with MF in Egyptian patients, with the homozygous genotype (T/T) having a stronger association with tumour stage (stage IIb).

Screening of dystrophin gene deletions in Egyptian patients with DMD/BMD muscular dystrophies.

Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations within the dystrophin gene. Our study has identified 100 Egyptian families collected from the Human Genetics Clinic, National Research Center, Cairo. All cases were subjected to complete clinical evaluation pedigree analysis, electromyography studies, estimation of serum creatine phosphokinase enzyme (CPK) levels and DNA analysis. Multiplex PCR using 18 pairs of specific primers were used for screening of deletion mutations within the dystrophin gene. A frequency of 55% among the families. Sixty per cent of detected deletions involved multiple exons spanning the major or the minor hot spot of the dystrophin gene. The remainder 40% which mainly involved exon 45. Comparing these findings with frequencies of other countries it was found that our figures fall within the reported range of 40%- distribution of deletions in our study and other different studies was variable and specific ethnic differences do not apparently account for specific deletions. In addition this study concluded that employment of the 18 exon analysis is a cost effective and a highly accurate (97% to launch a nationwide program.

Assay for hepatitis C virus in peripheral blood mononuclear cells enhances sensitivity of diagnosis and monitoring of HCV-associated hepatitis.

Hepatitis C virus (HCV) is a major etiological factor in chronic hepatitis affecting up to 24% of blood donors in Egypt. Since fluctuating levels of HCV RNA loads, including undetectable values, have been frequently observed in sera of chronic hepatitis patients, this study was designed to assess the sensitivity of PCR amplification for the plus- and minus-RNA strands in peripheral blood mononuclear cells (PBMC) compared to single serum PCR assay. Since the latter test detects viremia in only 79.5% of seropositive cases, the highest sensitivity for HCV diagnosis was achieved (93.20% when applying the combined triple test including PCR amplification of plus-strand in serum, together with plus-strand in PBMC and minus-strand in PBMC. The results of this study indicate that the triple test provides significant information on extrahepatic replication of HCV in a sizable sample of seropositive subjects (429 cases) and improves the assessment of HCV viremia. The cost/effectiveness and speed were upgraded by using capillary/air rapid thermal cycler. The use of the triple assay in HCV diagnosis and post-therapy monitoring is recommended.

The milder phenotype of the dystrophin gene double deletions.

OBJECTIVES: This study aimed to examine the genotypephenotype correlation in Duchenne muscular dystrophy (MD) patients with double deletion (Ddel) mutations in comparison with those having single deletions (Sdel). MATERIALS AND METHODS: The study included 250 Duchenne/Becker MD male patients from whom the 10 Ddel patients were compared with 20 Sdel subjects of same age and disease durations. The patients were subjected to neurological examination including functional disability grading scale (FDGS), molecular analysis of the dystrophin gene and immunohistochemical studies of some muscle biopsies. RESULTS: The mean FDGS value in the Ddel group was lower than that in Sdel patients. The Ddel patients had partial expression of dystrophin in their skeletal muscles, while Sdel cases showed complete absence of the protein. CONCLUSION: Patients with double deletion mutations within the dystrophin gene have a milder phenotype than patients harboring single deletions at either major or minor hot spots of the gene.

Schistosoma hematobium soluble egg antigens induce proliferation of urothelial and endothelial cells.

Bladder carcinoma accounts for 26% of reported human malignancies in Egypt, and has been strongly associated with urinary schistosomiasis. Nevertheless, the immediate role of schistosomal egg proteins in bladder carcinogenesis is unexplored. We investigated the effects of crude soluble egg antigens (SEA) of Schistosoma hematobium on urothelial cell proliferation. The proliferation of bovine endothelial Endo, human urothelial J82 and smooth muscle SMC cell lines was assessed by low-density growth assays. SEA induced proliferation of both J82 and Endo cells in a dose-dependent fashion, but not SMC. Preboiling or proteinase K treatment of SEA abolished its effect. In addition, SEA enhanced urothelial expression of B-cell translocation protein (BTG1) and human proliferating cell nuclear antigen (PCNA) mRNAs. Given the strong correlation between cell proliferation and carcinogenesis, the findings suggest that crude SEA may play some role in schistosomal bladder carcinogenesis.