THE POTENTIAL ROLE OF EPSTEIN BARR VIRUS IN MULTIPLE SCLEROSIS MOLECULAR AND SEROLOGICAL STUDY.

OBJECTIVE: The aim: To identify the relation between EBV infection/reactivation and multiple sclerosis, compared to the normal controls. PATIENTS AND METHODS: Materials and methods: A case-control study conducted in 120 MS patients, aged between 12-42 years, and 120 apparently healthy age- and sex-matched volunteers as controls. Viral DNA was extracted from 100µl of plasma samples, and then viral DNA was detected and quantified by quantitative real time-polymerase chain reaction (q-PCR). Serum samples were used for the detection of anti-EBNA-1 IgG. RESULTS: Results: Quantitative polymerase chain reaction of EBV showed absence of EBV viremia in all MS patients and control. However, anti EBNA-1 IgG antibody was positive in 51.7% (62/120) of MS patients and 39.2% (47/120) of controls, (P=0.035). The median of anti EBNA-1 IgG level in MS patients and controls were 81.08 U/ml and 67.73 U/ml, respectively (P=0.043). Additionally, EBNA-1 antibody was significantly higher in younger age groups. Patients with the first-line and second-line treatment showed no significant differences in anti EBNA-1 IgG levels, while the median level in patients without treatment (newly diagnosed) was higher. CONCLUSION: Conclusions: EBNA-1antibody could play a significant role in development of MS, as it is significantly higher in MS patients than in controls, especially at younger age groups, at early stages of the disease and in female patients.

SNP-PCR genotyping links alterations in the GABAA receptor (GABRG3: rs208129) and RELN (rs73670) genes to autism spectrum disorder among peadiatric Iraqi Arabs.

INTRODUCTION: Autism spectrum disorder (ASD) is an increasing concern among the Iraqi Arab population. The genetic alterations that cause ASD are likely to converge at the synapse. This study investigated polymorphisms in the GABAA receptor subunit (GABRG3) and the RELN gene as putative biomarkers of ASD in a pediatric population in Iraq. METHODS: The case control study included 60 patients with a clinical diagnosis of ASD (mild, moderate, or severe) according to DSM-IV criteria and matched healthy controls (n = 60). Blood samples were collected for DNA genotyping of SNPs rs736707 and rs208129 for RELN and GABRG3 using allele specific PCR. Assessment of genotype and allele distributions in patient groups used odd ratios (OR) with 95% confidence intervals and the Chi-square test. All statistical analysis was performed used SPSS software. RESULT: The patient cohort was highly consanguineous, with increased ratio (p > 0.05) of males to females (3:1) in both ASD (mean age, 6.66 ± 3.05) and controls (mean age, 5.76 ± 2.3). Both GABRG3 rs208129 genotypes TT (OR 4.33, p = 0.0015) and TA (OR 0.259, P = 0.008), and the T and A alleles were significantly associated with ASD. The RELN rs736707 TC genotype (OR 2.626, P = 0.034) was the only significant association with ASD. CONCLUSION: GABRG3 SNP rs208129 is a leading biomarker to predict genetic vulnerability to ASD in Iraqi Arabs. Expanded SNP panels and increased sample sizes are required for future GABRG3 studies, and to reach a consensus on RELN utility. Future ASD screening programs in Iraq should include genetic metrics in addition to clinical phenotype assessments.

Interleukin-37 gene polymorphism and susceptibility to pulmonary tuberculosis among Iraqi patients.

BACKGROUND: Control of tuberculosis (TB) depends on a balance between host's immune factors and bacterial evasion strategies. Interleukin-37 (IL-37) is among the immunomodulatory factors that have been proposed to influence susceptibility to tuberculosis. METHODS: A case-control study was conducted on 105 patients with pulmonary TB (37 active, 41 multi-drug resistant and 27 relapse) and 79 healthy controls to determine serum levels and single nucleotide polymorphisms (SNPs) of IL-37. The IL-37 level was assessed with an enzyme-linked immunosorbent kit, while DNA-sequencing was used to detect SNPs in the promoter region of IL37 gene. RESULTS: Median level of IL-37 was markedly increased in serum of TB patients compared to controls (325.0 vs. 169.1 pg/mL; p < 0.001). This increase was universally determined in subgroups of patients distributed according to gender, age groups, and clinical type of disease, while no significant differences were found between the subgroups in patients or controls. Analysis of receiver operating characteristic curve confirmed these findings and IL-37 occupied a very good area under the curve, which was 0.816 (95% CI = 0.744-0.888; p < 0.001). At a cut-off value of 185.6 pg/mL, the sensitivity and specificity of IL-37 were 81.0 and 82.3%, respectively. Of the nine detected SNPs (rs2466449 G/A, rs2466450 A/G, rs2723168 G/A, rs3811042 G/A, rs3811045 T/C, rs3811046 G/T, rs3811047 A/G, rs3811048 G/A and rs200782323 G/A), only rs3811048 showed a significant association with TB; the G allele showed a significantly decreased frequency in TB patients compared to controls (25.2 vs. 44.9%; OR = 0.41; p < 0.001). It was possible to assign five haplotypes, and three showed significant differences between patients and controls. Frequency of haplotype A-A-G-A-C-T-G-A-G (0.331 vs. 0.213; OR = 2.10; p = 0.015) was significantly increased in TB patients compared to controls. On the contrary, frequencies of haplotypes A-A-G-A-C-T-G-G-G (0.029 vs. 0.116; OR = 0.24; p = 0.01) and A-A-G-G-T-G-A-G-G (0.140 vs. 0.275; OR = 0.45; p = 0.015) were significantly decreased in patients. CONCLUSIONS: IL-37 was up-regulated in the serum of TB patients irrespective of their gender, age or clinical type of disease. SNPs in the promoter region of IL37 gene were proposed to be associated with susceptibility to TB.

Epidemiological profile of tuberculosis in Iraq during 2011-2018.

BACKGROUND: Tuberculosis (TB) is one of the most progressive infectious diseases caused by Mycobacterium tuberculosis. The pathogen is the first cause of mortality linked to a single pathogen worldwide, especially in poor and developing countries. METHODS: A cross-sectional descriptive study was conducted to estimate incidence rate (IR) of TB in Iraq during a period of eight years (2011-2018). TB data were extracted from the computer system of the National Specialized Center for Chest and Respiratory Diseases in Baghdad. RESULTS: During 2011-2018, 65,102 confirmed TB cases were reported in Iraq; 39,640 pulmonary TB (PTB) and 25,462 extra-pulmonary TB (EPTB). The average IR (case/100,000 inhabitants) of TB was 23.4 (14.2 for PTB and 9.1 for EPTB). Annual rate of TB cases showed a gradual decline over years (from 29.2 in 2011 to 18.6 in 2018). The decline in IR was more pronounced in PTB than EPTB. However PTB/EPTB ratio showed a gradual decreasing over years (from 2.04 in 2011 to 1.56 in 2018). GIS-mapping revealed that PTB and EPTB IRs show variations between the 18 governorates of Iraq. Most of the recorded PTB cases were new (average: 90.5%), followed by relapse cases (average: 7.9%). Among the reported PTB cases, percentage of males was greater than females (average: 52.1 vs. 47.9%), whereas an opposite trend was observed in EPTB (42.9 vs. 57.1%). The frequency distribution of PTB and EPTB varied between age groups, and lowest average frequency was recorded in age groups 1-4 and 5-14 year. CONCLUSIONS: TB is still a public health threat, and although a declining trend in incidence was depicted over the years 2011-2018, the disease is still out of control in Iraq, and more investments of resource are necessitated to eliminate the disease. In this context, EPTB and PTB relapse need a recognized attention.

Significance of the chemokine CXCL10 and human beta-defensin-3 as biomarkers of pulmonary tuberculosis.

The biomarker significance of IL-35, chemokines (CXCL9 and CXCL10) and human beta-defensins (hBD2 and hBD3) was determined in pulmonary tuberculosis (TB) of 105 Iraqi patients; 37 had active disease, 41 had multi-drug resistant (MDR) PTB and 27 had a relapse of TB. A control sample of 79 healthy persons was also included. Serum levels of markers were assessed using enzyme-linked immunosorbent assay kits. Kruskal-Wallis test together with Dunn-Bonferroni post hoc test revealed significance differences between patients and controls in levels of IL-35, CXCL9, CXCL10 and hBD3, while hBD2 showed no significant difference. Receiver operating characteristic analysis demonstrated that CXCL10 and hBD3 were the most significant markers in predicting TB, particularly active disease. Logistic regression analysis proposed the susceptibility role of CXCL10 in TB. Gender- and age-dependent variations were also observed. Spearman's rank correlation analysis showed different correlations between markers in each group of patients and controls. In conclusion, CXCL10 was up-regulated in serum of TB patients, while hBD3 showed down-regulated level. Both serum proteins are possible candidate biomarkers for evaluation of TB progression, particularly in active disease.