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

Study of single nucleotide polymorphism rs13266634 in zinc transporter solute carrier family 30, member 8 [SLC30AS] gene in type 2 diabetes patients resident in Ismailia city

Several genome-wide association studies identified a strong association of SLC30A8 with type 2 diabetes in individuals of European ancestry The effect of the association of rs 13266634 with type 2 diabetes or related glycemic traits has not been fully extended to non-European populations, and a comprehensive examination of common variants in the gene has not yet been carned out in our population. The aim of the present study was to investigate the association among the polymoiphisms of SLC30A8, and the risk of T2DM and to determine the presence and frequency of single nucleotide polymorphism [SNP] rs13266634 in SLC30A8 gene in T2D patients resident in Ismailia city. SLC30A8 SNP was genotyped using real time PCR allelic discrimination TaqMan assay. A case control study was conducted in 68 cases of type 2 diabetes [51 women and 17 men] and 29 control subjects [13 women and 16 men] from out-patients diabetic clinic of Suez Canal hospital and age and gender were matched. The SNP rs13266634 was evaluated in SLC30A8 C > T genotype. The genotypes of control subjects were 27[93%, CIT], 1 normal homozygot [3.5% C/C] and 1 mutant homozygous [3.5%, T/T]. In diabetic subjects, there were 58 subjects carriers of heterozygous [C/T. 85%], 6 normal homozygots [C/C, 9%] and 4 mutant homozygous [T/T, 6%]. There was significant difference in fasting blood glucose levels in control subjects compared to diabetic subjects' P<0.05, and between control carriers of[CT] genotype compared to genotype [CT] diabetic subjects' P<0.05, also there was significant associations between polymorphisms and the risk of type 2 diabetes in women or men, as there were significant associations in lipids profile [serum high density lipoprotein between control carrier CT genotype compared to diabetic subjects' carrier CT P<0.005, also we found significant difference in body mass index [BMI] between control and diabetic subjects' P<0.05. In addition, none of the SLC30A8 polymorphisms was significantly associated with the age and sex in the control and diabetic subjects. Also, we found higher non significant difference in triglyceride levels, total cholesterol and LDL cholesterol levels in diabetic subjects carrying the C/T genotype comparing to control subjects carrying the C/T genotype. In summary, the data in this study support substantial associations between the common SLC30A8 polymorphisms in gene and the risk of type 2 diabetes. Our results may provide evidence that SLC30A8 is a susceptible locus for type 2 diabetes in our population, and its variant can influence insulin secretion

IL-6 gene promoter polymorphism [G-174C] and type 2 diabetes in Suez canal area

Interleukin-6 [IL-6], a powerful inflammatory mediator, plays a pivotal role in the pathogenesis of insulin resistance and type 2 diabetes. The IL-6 promoter polymorphism, at position -174 [G > C], has been associated to insulin sensitivity although contrasting data have been reported. The aim of this study was to investigate the association of the IL-6 174 G > C promoter polymorphism with risk of T2DM in our populations. PCR restriction fragments length polymorphism [PCR-RFLP] was used to determine the distribution of allele and genotype frequency of this variant. We conducted a case-control study of 52 cases of type 2 diabetes [32 women and 20 men] and 32 control subjects [22 women and 10 men] from out patients diabetic clinic of Suez Canal hospital, age and gender-matched. We evaluated the IL-6 -174 G > C genotype. In control subjects, both C+ [CG + CC genotypes] and G- [GG genotype] carriers. The genotypes of control subjects were 16 [50%, G/G], 6 heterozygous [18.7% G/C] and 10 homozygous [31.3%, C/C]. In diabetic subjects, there were 26 subjects carriers of homozygoits [GG, 50%], 16 heterozygous [C/G, 30.8%] and 10 homozygous [C/C, 19.2%].There was significant difference in fasting blood glucose levels in control carriers of GG genotype compared to GG diabetic subjects' P<0.05, and between control carriers of combining [GC+CC] genotype compared to [GC+CC] diabetic subjects' P<0.05. Also there was significant associations between IL6polymorphisms and the risk of type 2 diabetes in women or men. There were significant associations in lipids profile [serum total cholesterol, s. triglyceride, s. low density lipoprotein and s. high density lipoprotein between control GG genotype compared to GG diabetic subjects' P<0.05, also we found significant difference between control carriers of combining [GC+CC] genotype and [GC+CC] diabetic subjects' P<0.05. Also we found higher non significant biochemical parameters; triglyceride levels, total cholesterol and LDL cholesterol levels in control subjects carrying the G/G genotype comparing to subjects carrying the [GC+CC] genotype In addition, none of the IL6 polymorphisms was significantly associated with the age, sex and body mass index [BMI] in the control and diabetic subjects. In summary, the data in this study support substantial associations between the common polymorphisms in IL6 gene and the risk of type 2 diabetes

Real-time PCR and flow cytometry in detection of cyclospora oocysts in fecal samples of symptomatic and asymptomatic pediatrics patients

The magnitude of Cyclospora oocysts excretion in relation to infection intensity among cyclosporiasis patients was assessed using flow cytometry and quantitative real-time PCR [RT-PCR]. Oocysts from stool samples of 25 [14.8%] gastro-intestinal symptomatic pediatrics patients [169] and of 10 [2.8%] asymptomatic gastrointestinal ones [350] were identified by modified Ziehl-Neelsen [MZN] and modified Acid Fast Trichrome [MAFT] staining methods and confirmed by its auto-fluorescent characterizations. Also, 10 infants with negative stool samples were selected as controls. The intensity of infection was calculated as number of oocysts/200 microscopic filed with immersion 400. Flow cytometry and RT-PCR assessed relation between symptoms and oocysts excretions compared to MZN and MAFT. The infection severity in symptomatic patients were identified by MZN and MAFT as mild [16%], moderate [24%] and severe [60%]. All asymptomatic patients had mild infection. Flow cytometry was done for stool samples and 100% Cyclospora oocysts were in symptomatic and asymptomatic patients. None was detected in controls, RT-PCR was done for stools with both a species-specific primer set and dual fluorescent labeled Cyclospora cayetanensis hybridization probe by unique regions of 18S rRNA gene sequence. DNA of C. cayetanensis was in 100% of symptomatic and asymptomatic patients and in 20% of controls. In repetitive examination of stools Cyclospora oocysts were neither detected by staining nor flow cytometry. Based on oocysts counts, no differences were found between flow cytometry and RT-PCR in compared to staining methods