HLA-DRB1, IRF5, and CD28 gene polymorphisms in Egyptian patients with rheumatoid arthritis: susceptibility and disease activity.

This study was established to assess the effects of IRF5 rs10488631 and CD28 rs1980422 single-nucleotide polymorphisms (SNPs) and HLA-DRB1 shared epitope (SE) allele on the prognosis and disease activity of rheumatoid arthritis (RA) patients. A total of 150 RA patients and 150 healthy controls were genotyped for the selected SNPs by real-time PCR. HLA-DRB1 SE was determined using LAB Type SSO Class II DRB1 typing. Our results suggest that HLA-DRB1, CD28, and IRF5 significantly discriminated (p < 0.001) RA patients and healthy controls (OR of single HLA-DRB1 SE allele = 2.431, CI = 1.467-4.027, OR of two SE alleles = 11.152, CI = 2.479-50.159), (OR of CD28 risk allele C = 2.794, 95% CI = 1.973-3.956) and (OR of IRF5 risk allele C = 4.925, CI = 3.26-7.439). Rheumatoid factor (RF) seropositivity was associated with HLA-DRB1 SE (p < 0.001) and IRF5 risk allele (p < 0.001). ACPA was significantly associated only with IRF5 risk allele (p < 0.001). A better response to methotrexate therapy was found in HLA-DRB1 SE non-carriers, and CD28 TT patients. This study demonstrated associations of HLA-DRB1 SE, CD28, and IRF5 with the risk of RA. HLA-DRB1 SE and CD28 rs1980422 can be used as predictors of methotrexate therapy response.

Promoter methylation and expression of intercellular adhesion molecule 1 gene in blood of autoimmune thyroiditis patients.

Growing data supported that epigenetic modifications, including altered DNA methylation have a potential role in the pathogenesis of autoimmune thyroid diseases (AITD). In the present study we aimed to investigate the methylation status of ICAM-1 gene promoter in patients with AITD. Forty patients with Graves' disease (GD), 40 patients with Hashimoto thyroiditis (HT) and 40 normal controls were included. DNA extraction from blood samples was done to analyze the ICAM-1 methylation status using methylation-specific PCR method. RNA was also extracted to determine the ICAM-1 expression values by real time PCR. We found that the differences in the frequency of ICAM-1 methylation status between GD or HT and healthy individuals were statistically significant (p = 0.04, 0.018 respectively) whereas there was no significant difference between GD and HT patients (p > 0.05). There was a correlation between decreased ICAM-1 methylation and exophthalmos (p = 0.01) and the high TSI level (p < 0.002) in GD patients. However, there was no correlation between ICAM-1 methylation and other clinicopathological features or other laboratory parameters in GD or HT. Furthermore, ICAM-1 mRNA expression showed significant up-regulation in ICAM-1 un-methylated samples compared to methylated samples in both GD and HT patients (p < 0.001 for each). Results provided the evidence of association of the hypo-methylation status of ICAM-1 gene promoter with GD and HT patients. In addition, DNA methylation may have a critical role in the ICAM-1 expression regulation of AITD patients.

Circulating long non-coding RNA GAS5 and SOX2OT as potential biomarkers for diagnosis and prognosis of non-small cell lung cancer.

Early diagnosis of non-small cell lung cancer (NSCLC) is essential for patient treatment and prognosis. Long noncoding RNA (lncRNA) have potential roles in tumor initiation and differentiation. The objective of this study was to investigate whether the circulating lncRNA, growth arrest-specific transcript 5 (GAS5) and SOX2 overlapping transcript (SOX2OT), could be used as noninvasive biomarkers for NSCLC diagnosis. Moreover, we aimed at evaluating the association between lncRNA and the clinicopathological features of NSCLC in order to predict the cancer prognosis. The results showed significant downregulation of GAS5 expression and upregulation of SOX2OT in NSCLC patients compared with controls (P < 0.001). Furthermore, the expression level of GAS5 was declined in stage IV of NSCLC, but SOX2OT expression was increased sharply in stages III and IV. The expression levels of lncRNAs were used to distinguish NSCLC patients from control with an area under curve of 0.81 (sensitivity 82.5% and specificity 80%) for GAS5 and 0.73 (sensitivity 76.3% and specificity 78.6%) for SOX2OT. The combination of GAS5 and SOX2OT showed differentiation NSCLC patients from controls with increased sensitivity (83.8) and specificity (81.4). In conclusion, the newly developed diagnostic panel involving of circulating GAS5 and SOX2OT could be perfect biomarker for diagnosis and prognosis of NSCLC.

Usefulness of nucleic acid testing among negative HBs Ag blood donors in Egypt.

INTRODUCTION: Hepatitis B viral infection has been transmitted from donors with HBV infections who have negative HBs Ag. Many countries have implemented nucleic acid testing (NAT) to screen donors with non- reactive HBs Ag for detection of HBV DNA and enhance safety of blood transfusion, while it is restricted to limited blood banks in Egypt. OBJECTIVE: To evaluate the significance of NAT technology in detection of HBV DNA in the Egyptian blood donors with HBs Ag non- reactivity. METHODS: The study included 36,584 collected blood samples from volunteer blood donors at the blood bank of Zagazig University Hospitals. Each specimen was tested for HBs Ag; non- reactive sera were further tested for qualitative detection of HBV-DNA by NAT testing. All positive HBV-DNA donors were tested for anti- HBc and anti- HBs by electro-chemiluminescence immunoassay and confirmed by quantitative RT-PCR. RESULTS: Among 34,671 donors non-reactive to HBs Ag, 34,657 (99.96%) were tested negative for HBV- DNA and 14 specimens (0.04%) were positive for HBV via NAT testing. Among HBV NAT positive donors, HBs Ab reactive only in (2); HBc Ab reactive only in (3); HBs and HBc Abs reactive in (3) while HBs and HBc Ab non-reactive in (6). All tested sera 14 (100%) showed low viral load for HBV (<50 IU/ml) confirmed by RT- qPCR. CONCLUSION: Our results highlighted the significance of the HBV NAT technique to reduce the potential risk of HBV transfusion-transmission and the critical need to enforce the usage of NAT technology in all blood banks in Egypt.

Association of monocyte chemoattractant protein 1 (MCP-1) gene polymorphism with lupus nephritis in Egyptian patients.

BACKGROUND AND STUDY AIM: Monocyte chemoattractant protein-1 (MCP-1) is a member of CC chemokine that plays an important role in the recruitment of monocytes/macrophages into renal tubulointerstitium. A biallelic A/G polymorphism at position ∼2518 in the MCP-1 gene was found to regulate MCP-1 expression. MCP-1 and its A/G gene polymorphism have been implicated in the pathogenesis of some renal diseases. The aim of the study was to investigate the role of the MCP-1 gene polymorphism as early predictors of the development of glomerulonephropathy in SLE patients. We also aimed to measure the serum and urinary levels of MCP-1 in patients with SLE, to find out its relation to clinical disease activity. METHODS: 140 SLE patients (100 with nephritis and 40 without nephritis) and 80 controls were included in this study. MCP-1 gene polymorphism was analyzed by polymerase chain reaction. Serum and urine MCP-1 level were measured using high-sensitivity enzyme-linked immunosorbent assay. RESULTS: The A/A genotype was more common in controls than in SLE patients, whereas both the A/G (P<0.000) and G/G (P<0.000) genotypes were more frequent in SLE patients. Carriers of G allele of the MCP-1 ∼2518 polymorphism had more than 7 fold increased risk to develop glomerulo-nephropathy in patients with SLE. High MCP-1 circulating levels production from patients with A/G and G/G genotypes was significantly higher than in A/A genotype. In addition there were significant differences in the mean levels of serum MCP-1 (P<0.001) and urinary MCP-1 (P<0.001) between patients and controls. CONCLUSION: The present study provides a new evidence that the presence of MCP-1 A (-2518) G gene polymorphism and high circulating MCP-1 levels can play an important role in the development of SLE and nephropathy in Egyptians.