Patterns of thrombospondin genes polymorphisms in acute myocardial infarction patients in Ismailia City

Thrombospondin [TSP] 2 and 4 are multidomain calcium-binding extracellular glycoproteins which play a role in platelet aggregation and inflammatory response. TSP-2 has chemotactic and mitogenic activities for vascular smooth muscle cells while TSP-4 mRNA is expressed by endothelial and smooth muscle cells in vascular wall, and brain endothelial cells produce the protein both in vivo and in cell culture, localization consistent with its pro-atherogenic effects. These common functions may be central to the roles of the thrombospondins in coronary artery disease and myocardial infarction [MI]. In the present study, the association of the TSP-2[3949 T-+G, rs8089] and TSP-4 [Ala387Pro 1186 G-+C, rs866389] gene variations and MI among Egyptian patients living in Ismailia city has been examined. Both rs8089 and rs 1866389 were studied in 50 acute MI patients and 50 controls using Real-Time polymerase chain reaction. The prevalence of TSP-2 and TSP-4 alleles was not different in MI patients compared to controls [P> 0.05]. Although the minor allele homozygotes [GG] of TSP-2 seems to confer reduced risk of MI [OR: 0.42 95% CI=0.095-1.89] this was not statistically significant [P> 0.05]. The distribution of different TSP-4 genotypes did not differ between MI patients and controls [P>0.05]. Total cholesterol was statistically significantly higher [P=Q.Q2] in carriers of minor allele [C] of TSP4 [GC+CC]. Although, both polymorphisms showed no statistically significant difference in MI patients regarding all other measured conventional risk factors. However, the frequency of TTGC haplotype is statistically significantly higher in MI patients [24%] than in controls [6%] [P value=0.0226]. Our data suggests that although association analysis with MI did not reach significance, an at-risk haplotype of common variants located in THBS2 and THBS4 may be part of the genetic determinants for MI in the Egyptian population living in Ismailia city

Vitamin D binding protein gene polymorphisms and risk of type 1 diabetes mellitus among Egyptians

Type 1 diabetes [T1DM] is a multifactorial autoimmune disease in which both genetic predisposition and environmental factors participate in its development. Many cellular and epidemiological studies suggest a role for vitamin D in pathogenesis and prevention of T1DM. Polymorphisms of the genes involved in the metabolism of vitamin D may predispose to T1DM. Vitamin D-binding protein [DBP] is the main systemic transporter of vitamin D and is essential for its cellular endocytosis. There are two known polymorphisms in exon 11 of the DBP gene resulting in amino acid variants: GAT->GAG substitution replaces aspartic acid by glutamic acid in codon 416; and ACG->AAG substitution in codon 420 leads to an exchange of threonine for lysine. These DBP variants lead to differences in the affinity for vitamin D. Few published studies, about the correlation between these alleles and T1DM, yielded conflicting results. Therefore, we investigated the association of these polymorphisms with T1DM in Egyptian subjects. Unrelated 59 children with T1DM and 65 healthy controls were included in this study. The sequence of DBP exon 11, which contains both examined variants, was amplified by polymerase chain reaction [PCR]. Alleles and genotypes were determined using Restriction Fragment Length Polymorphism analysis [RFLP]. At codon 416 the frequency of Glu/Asp alleles was 64.4/35.6% in T1DM patients and 55.4/44.6% in controls [P >0.05]. At codon 420 the frequency of Thr/Lys alleles were 88.1/11.9% and 87.7/12.3% [P >0.05] respectively. Distributions of genotypes at both loci, and the common haplotypes constructed by them, were also very similar in both groups [P >0.05]. It could be concluded that the studied DNA polymorphisms in the DBP gene are not associated with T1DM in Egyptian patients

Effect alpha globlin gene deletion and gamma globin gene -158 [C/T] polymorphism in beta-thalassemic patients

The beta-thalassemias [beta- thalassemias] are among the most common autosomal recessive disorders. They have a remarkably high frequency in the Mediterranean region and represent one of the most common genetic diseases in Egypt. In this study, the spectrum of beta-thalassemia mutations and genotype-to-phenotype correlations were defined in 32 beta- thalassemic patients [beta- thlassemias major and intermedia] with varying disease severity in two cities of the Suez Canal region. Ten different mutations were identified and the most frequent ones were: IVSI-6 [T-C] [37.5%], IVSI-110 [G-A] [34.4%] and both IVSI-1 [G-A], IVSII-745 [C-G] and -102 [C-G] [12.5% each]. There was a wide spectrum of phenotypic severity in all patients. We studied the Xmnl polymorphism [C/T] in gamma- globin gene position -158 of beta- thalassemia as a modulating factor of the disease severity. Presence of the polymorphism was found in two patients and this was not sufficient to explain the diversity of the phenotype encountered. Co-inheritance of alpha thalassaemia as a modulating factor was not evident in our patients. In conclusion, we have been unable to find a molecular basis for the benign clinical course in all our patients. Other genetic or acquired factors must be hypothesized which ameliorate the clinical condition

C-Reactive protein, Tumor Ncrosis Factor-alpha, interleukin-6 and cortisol in type 2 diabetes mellitus

Recently an abundance of evidence has emerged demonstrating a close link between immunity, inflammation, obesity, insulin resistance and type 2 diabetes mellitus. Activation of innate immunity with production of inflammatory markers was suggested to provide a new model for the pathogenesis of type 2 diabetes and the metabolic syndrome. This may result in new approaches for predicting and managing of type 2 diabetes and its complications. We evaluated the state of obesity and diabetes mellitus of thirty nine male type 2 diabetic patients and nineteen age-matched male healthy subjects as control. This evaluation was performed via assessment of the body mas index [BMI], fasting and postprandial [PP] blood glucose and insulin, insulin resistance and fasting C-peptide. Then we assessed the plasma levels of the most important inflammatory markers; C-reactive protein [C-RP], tumor necrosis factor alpha [TNF-alpha], interleukin-6 [IL-6] and the total leucocytic count as well as the stress hormone cortisol. Our results showed BMI above 30 for both patients and controls which indicated obesity of the two groups. Both fasting and PP glucose were 167.9 +/- 10.3 and 289.8 +/- 16 mg/dl respectively for patients and 96.4 +/- 1.96 and 108.47 +/- 4.6 rng/dl respectively for the control whish confirmed the diagnosis of diabetes mellitus. The C-RP was significantly higher in diabetics. Although the difference did not reach statistical significance fasting and PP-insulin, insulin resistance levels were higher in the diabetic patients compared to the control. Regarding the results of the acute phase reactants and the biornarkers for inflammation, we found a significant increase in C-RP, TNF-alpha, IL-6 and cortisol in the diabetic patients compared to the control. But no change could be detected in the total leucocytic count. This association between hyperglycemia and increased inflammatory markers may indicate a relationship between them. But the question which of them preceded and led to the other is still uncertain. Further studies with different approaches may be needed to solve this puzzle

study on the role of epstein barr virus infection in the pathogenesis of nasopharyngeal carcinoma

Oral herpes virus infections are thought to be a responsible predisposing cause of nasopharyngeal cancer. Of the herpes viruses, Epstein-Barr virus [EBV] has classically been associated with nasopharyngeal carcinoma [NPC] and Burkitt's lymphoma. Recently, multiple studies have been published linking EBV with oral squamous cell carcinoma and, to a lesser extent, hypopharyngeal and laryngeal tumors. Using a sensitive method of detection, this study was conducted to analyze the presence of EBV DNA in 40 NPC cases and 35 cases with benign nasal polyps as control in serum and tissue and compared it with serological markers. Three EBV serological markers were performed by enzyme linked immunosorbant assay including EBV VCA IgM, EBV IgG and EBNA IgG. Herpes simplex virus antibodies HSV I and II IgG were examined in serum. Forty serum and tissue samples exclusive of nasopharyngeal carcinoma were examined for the presence of EBV DNA and HSV II DNA using qualitative polymerase chain reaction. Thirty-five serum and tissue samples of benign nasal polyps were submitted to all the tests as control. Serological tests for EBV: revealed that EBV VCA IgG was positive in 57.5%, EBNA IgG was positive in total of 47.5% NPC. EBV DNA was positive in serum in all EBV VCA IgG and EBNA IgG positive cases. EBV DNA by PCR in tissue was positive in 72.5% of NPC in which 70% were EBV-DNA serum positive. In the benign group 17.1% EBV-DNA tissue positive cases, only 2.9% were EBV-DNA serum positive by PCR. HSV DNA in tissue was positive in 20% of NPC and 11.4% of benign group. In NPC HSV-DNA tissue positive cases, 2.5% were HSV-DNA serum positive by PCR and 17.5% were negative. The results indicate that HSV and EBV have a role in the etiology of nasopharyngeal carcinoma. Detection of EBNA1 and HSV in the serum and corresponding tissue of nasopharyngeal carcinoma indicates the role of circulating viral DNA as an early marker for pathogenesis of nasopharyngeal carcinoma and that it could serve as good supplement to pathological diagnosis of nasopharyngeal carcinoma

Enhanced apoptosis of peripheral blood mononuclear cells from chronic hcv patients is associated with reduced caspase 3 activity

Background: Infection with hepatitis C virus [HCV] is a major cause of chronic liver disease throughout the world. Down-regulation of the immune response plays a major role in HCV persistence. Recent investigation suggest that apoptosis of peripheral blood mononuclear cells [PBMCs] contributes to such down-regulation

Objective: The current study investigates apoptotic changes in PBMCs and their relation to caspase-3 and -8 activities in patients with chronic HCV infection

Methods: Apoptosis were investigated by measuring annexin-V binding using flowcytometry and DNA fragmentation using agarose gel electrophoresis, and caspases-3 and -8 specific activities were also measured in 43 chronic HCV patients and 10 normal control subjects

Results: A significantly higher percent of annexin-V positive PBMCs was found in chronic HCV patients than controls [p<0.0001]. DNA fragmentation was detected in PBMCs from 20/43 patients [46.5%] but not from controls. There was no statistically significant difference between HCV-PCR positive and negative patients as regards the degree of PBMCs apoptosis. Caspase-3 activity was significantly lower in patients than controls [p=0.001], was significantly lower in HCV-PCR positive than controls [p=0.001] and was significantly higher in patients with PBMCs DNA fragmentation [p=0.005]. On the other hand, caspase-8 activity was comparable in both patients and control groups. However, patients with PBMCs DNA fragmentation showed statistically significant higher activity than those without [p=0.023]. There was a statistically significant direct correlation between caspase-3 and caspase-8 activities in the patients groups [r=0.56,p<0.0001]

Conclusion: Chronic HCV infection is associated with PBMCs apoptosis irrespective of the presence of viremia. However, this apoptosis is independent on activation of either caspase-3 or caspase-8